用于产生乙酸补身烷基酯化合物的方法
文献发布时间:2023-06-19 10:55:46
技术领域
本发明提供了新的方法,用于在体外或体内进行相应补身烷基(drimanyl)醇来源的乙酰化而进行乙酰基转移酶催化的乙酸补身烷基酯化合物的产生。本发明还涉及从不同的微生物和植物来源鉴定具有相应的乙酰基转移酶活性的酶。本发明还涉及提供衍生自所述新鉴定的酶的酶突变体。本发明的另一个形态涉及提供这种酶和突变体的相应编码序列、重组载体和重组宿主细胞,它们适合于产生这种乙酰基转移酶和突变体并用于进行乙酸补身烷基酯化合物的新产生方法。本发明的另一个形态涉及根据本发明获得的这种乙酸补身烷基酯作为中间体的用途,用于产生气味剂、调味剂或芳香剂或昆虫/害虫防治成分。
背景技术
萜烯存在于大多数生物(微生物、动物和植物)中。这些化合物由称为异戊二烯单元的五碳单元构成并且通过存在于它们结构中的这些单元的数目进行分类。因此,单萜、倍半萜和二萜是分别含有10、15和20个碳原子的萜烯。例如,在植物界中广泛地存在有倍半萜。许多倍半萜分子因为它们的风味和芳香特性以及它们的美容、医疗和抗菌效果而众所周知。已经鉴定出许多倍半萜烃和倍半萜类化合物。已经开发了化学合成途径,但是仍然很复杂并且不总是具有成本效益。
萜烯的生物合成产生涉及称为萜合酶的酶。在植物界中存在多种倍半萜合酶,它们均使用相同的底物(法呢基二磷酸,FPP),但是具有不同的产物构造。已经克隆了编码倍半萜合酶的基因和cDNA,并且表征了相应的重组酶。
倍半萜例如具有补身烷(drimane)结构的化合物,例如补身烷基醇,特别是折叶苔醇(albicanol)或补身醇(drimenol)的许多主要来源是天然含有倍半萜的植物或微生物。然而,这些天然来源中倍半萜的含量可能较低。即使可以得到,这样的补身烷基醇在它们的进一步加工期间也难以处理,主要是因为它们在环境温度下是固体。提供更易于处理的衍生物将是一种改进的方法,并简化了补身烷基醇的进一步加工。
Akita,H.等人在Tetrahedron:Asymmetry 11(2000).1375-1388中描述了脂肪酶催化乙酸(+)折叶苔基酯的不对称合成。这种现有技术的方法需要至少八个化学合成步骤和两个被脂肪酶催化的连续反应,以获得对映体纯的乙酸折叶苔基酯。另外,乙酰化反应在33℃下在二异丙醚和乙酸异丙烯基酯的存在下发生,该条件与生物合成途径所需的生理条件不相容。而且,在生理条件下,就我们所知,脂肪酶不能催化所需的酯交换反应。尽管通过脂肪酶形成酯是可能的,但是该活性强烈取决于所存在的水的含量。因此,在诸如本发明的体内环境的水性环境中,脂肪酶催化酯键的水解而不是乙酰化(Jaeger K.等人;FEMSMicrobiology Reviews,1994,15:1pp29-63)。
仍然需要提供产生乙酸补身烷基酯化合物的新方法,特别是可以在水性环境中例如在基于宿主细胞的方法(其通过糖底物的代谢而提供补身烷基醇前体)中用于完全生物化学合成乙酸补身烷基酯的方法。
发明内容
可以通过提供一类新的酶来解决上述问题,这些酶显示出乙酰基转移酶的活性,并通过乙酰化作用并使用乙酰基-CoA作为乙酰基供体,从相应的补身烷醇前体(如折叶苔醇或补身醇)中产生乙酸补身烷基酯,如乙酸折叶苔基酯或乙酸补身烯基(drimenyl)酯。由于它们的物理化学性质,特别是由于它们在环境温度下为液体,因此补身烷基醇的乙酰化衍生物可以用作更合适的材料。
附图说明
图1a:(+)-折叶苔醇、(-)-补身醇和双环法呢醇的结构,以及比图1c的补身烷结构更具体的补身烷部分。
图1b:反应路线,其说明了通过无环倍半萜前体FPP和折叶苔醇细胞生物学产生乙酸折叶苔基酯。
图1c:更概括的“补身烷结构”,标出了C=C-双键的潜在位置。
图2:(A/B)使用修饰的酿酒酵母菌株YST069产生的乙酸折叶苔基酯的GC-FID分析,该菌株将折叶苔醇合酶XP_007369631.1与乙酰基转移酶CrDAT(A)或乙酰基转移酶FgaAT(B)共表达。由(A)的乙酰基转移酶CrDAT产生的乙酸折叶苔基酯的MS谱图显示在(C)中,并且与来自乙酸折叶苔基酯标准品的MS谱图相同。
图3:由九种乙酰基转移酶(CrDAT,FgaAT,OAH94415.1,TcTAT,CrMAT,LiAAT-4,GAO81666.1,CfACT1-6和CfACT1-8)产生的乙酸折叶苔基酯的相对量,发现它们对于折叶苔醇有活性(如实施例2中所述)。
图4:由九种乙酰转移酶(CrDAT,FgaAT,OAH94415.1,TcTAT,CrMAT,LiAAT-4,GAO81666.1,CfACT1-6和CfACT1-8)产生的乙酸补身烯基酯的相对量,发现它们对于折叶苔醇有活性(如实施例2中所述)。
图5:表达以下每种乙酰基转移酶的酿酒酵母细胞所产生的乙酸折叶苔基酯的相对量:CrDAT,FgaAT,OAH94415.1,TcTAT,CrMAT,LiAAT-4,GAO81666.1,CfACT1-6,CfACT1-8,ERR364415-1_contig_8546和DfATC13。
图6:表达以下每种乙酰基转移酶的酿酒酵母细胞所产生的乙酸补身烯基酯的相对量:CrDAT,FgaAT,OAH94415.1,TcTAT,GAO81666.1,CfACT1-6,CfACT1-8,XP_001258079.1,ERR364415-1_contig_8546和DfATC13。
图7:通过乙酰基转移酶CrDAT产生的乙酸补身烯基酯的MS谱图显示在(A)中,其与(B)中显示的来自乙酸补身烯基酯标准品的MS谱图相同。
图8:通过乙酰基转移酶CrDAT产生的乙酸双环法呢基酯的MS谱图显示在(A)中,其与来自乙酸双环法呢基酯标准品(B)的MS谱图相同。
图9:表达以下每种乙酰基转移酶的酿酒酵母细胞所产生的乙酸双环法呢基酯的相对量:CrDAT,FgaAT,TcTAT,CrMAT,GAO81666.1,CfACT1-6,CfACT1-8,BAU61551.1,PsSalAT,XP_001217250.1,ERR364415-1_contig_8546,PYI04555.1和DfACT13。
具体实施方式
bp 碱基对
kb 千碱基
CoA 辅酶A
DNA 脱氧核糖核酸
cDNA 互补DNA
DTT 二硫苏糖醇
FPP 法呢基二磷酸
GC 气相色谱
MS 质谱仪/质谱法
MVA 甲羟戊酸
PCR 聚合酶链反应
RNA 核糖核酸
mRNA 信使核糖核酸
miRNA 微RNA
siRNA 小干扰RNA
rRNA 核糖体RNA
tRNA 转移RNA
除非另有说明,否则以下技术术语定义适用:
就本申请目的而言,“乙酰基转移酶”或“具有乙酰基转移酶活性的多肽”或“能够转移乙酰基的多肽”更概括而言是指一类酰基转移酶EC 2.3.1的酶,特别是乙酰基-CoA:醇O-乙酰基转移酶EC 2.3.1.84。它显示了用乙酰基-CoA作为乙酰基供体,乙酰化至少一种从折叶苔醇、补身醇和双环法呢醇中选出的补身烷基醇的能力。乙酸补身烷基酯可以以其任何立体异构体的形式或以其混合物的形式产生。如果相应的醇前体作为单个乙酰基受体存在,则乙酸折叶苔基酯,乙酸补身烯基酯或乙酸双环法呢基酯可能是唯一的产物,或者如果提供两种或更多种补身烷基醇的混合物并且乙酰基转移酶不是底物特异性的,则可能是两种或更多种乙酸补身烷基酯的混合物的一部分。在选择性增加的情况下,乙酰基转移酶可以主要形成一种单一的乙酸补身烷基酯。如本文所述的乙酰基转移酶可以对作为底物的不同的补身烷基醇显示相同或不同的偏好或特异性。例如,第一类型的乙酰基转移酶可主要为乙酰化折叶苔醇,第二类型的乙酰基转移酶可主要为乙酰化补身醇,而第三类型的乙酰基转移酶可主要为乙酰化双环法呢醇。在这种情况下,如果将这样的补身烷基醇的混合物用作底物,则会形成乙酸折叶苔醇酯、乙酸补身醇酯或乙酸双环法呢基酯。在底物特异性的情况下,乙酰基转移酶可以选择性地形成一种单一的乙酸补身烷基酯,即使将这种补身烷基醇的混合物用作底物。特别地,乙酰化是在保留了补身烷基醇底物的相应立体化学构型的情况下进行的。
“乙酰基供体”是指化学实体或分子,其充当待从所述供体酶促转移至受体分子的乙酰基的源,例如具有官能羟基的分子,其随后可与所述乙酰基反应,以形成相应的乙酸酯。一种特定的乙酰基供体是乙酰辅酶A(乙酰基-CoA)。
术语“补身烷倍半萜”或“补身烷”是指具有如图1a所示的补身烷样碳骨架结构的环状萜烯,或更具体而言是指图1c的更上位的结构,其中可选存在的C=C-双键的潜在位置用虚线标记。
术语“补身烷基醇”是指“补身烷倍半萜”或“补身烷”的羟基化衍生物。其例子是任何立体异构形式的折叶苔醇、补身醇和双环法呢醇。
术语“乙酸补身烷基酯”是指这样的乙酸补身烷基醇的乙酰基酯衍生物,例如乙酸折叶苔基酯,乙酸补身烯基酯和乙酸双环法呢基酯。
就本申请目的而言,“折叶苔醇”特别地涉及(+)-折叶苔醇(CAS:54632-04-1)。
就本申请目的而言,“补身醇”特别地涉及(-)-补身醇(CAS:468-68-8)。
就本申请目的而言,“双环法呢醇”特别地涉及(+)-双环法呢醇或[(4aS,8aS)-2,5,5,8a-四甲基-3,4,4a,5,6,7,8,8a-八氢-1-萘基]甲醇(IUPAC名称)
“法呢基二磷酸”是指(2E,6E)-3,7,11-三甲基十二碳-2,6,10-三烯-1-焦磷酸酯(FPP)。
就本申请目的而言,“Ambrox”涉及IUPAC名称:(-)-(3aR,5aS,9aS,9bR)-3a,6,6,9a-四甲基十二氢萘并[2,1-b]呋喃(CAS:6790-58-5)。
术语“萜合酶”或“倍半萜合酶”或“补身烷倍半萜合酶”在本文可互换使用。
术语“双功能萜合酶”或“具有双功能萜合酶活性的多肽”涉及如2018年5月31日提交的PCT/EP2018/064344中进一步定义的多肽。
术语“折叶苔基二磷酸合酶”或“具有折叶苔基二磷酸合酶活性的多肽”或“折叶苔基二磷酸合酶蛋白”或“具有产生折叶苔基二磷酸的能力”涉及一种多肽,其能够从无环萜烯焦磷酸酯,特别是法呢基二磷酸(FPP)开始,催化任何立体异构体或其混合物形式的折叶苔基二磷酸的合成。折叶苔基二磷酸可以是唯一的产品,或者可以是倍半萜混合物的一部分。所述混合物可以包含折叶苔基单磷酸和/或折叶苔醇。此类多肽描述于例如2018年5月29日提交的PCT/CN2018/088902。
如在PCT/CN2018/088902中描述的,“折叶苔基二磷酸合酶活性”是在“标准条件”下测定的。
术语“折叶苔醇合酶”或“具有折叶苔醇合酶活性的多肽”或“折叶苔醇合酶蛋白”是指一种多肽,其能够从无环萜烯焦磷酸酯,特别是法呢基二磷酸(FPP)开始,催化任何立体异构体或其混合物形式的折叶苔醇的合成。折叶苔醇可以是唯一的产物,或者可以是两种或更多种倍半萜的混合物的一部分。
术语“补身醇合酶”或“具有补身醇合酶活性的多肽”或“补身醇合酶蛋白”是指一种多肽,其能够从无环萜烯焦磷酸酯,特别是法呢基二磷酸(FPP)开始,催化任何立体异构体或其混合物形式的补身醇的合成。补身醇可以是唯一的产物,或者可以是两种或更多种倍半萜的混合物的一部分。
“折叶苔醇合酶活性”和补身醇合酶活性如例如PCT/EP2018/064344、WO2015/169871或WO 2015/176959中所述测定。
如本发明中使用的“磷酸酶”的酶具有在消耗水的情况下将正磷酸酯转化为相应的醇和正磷酸根的能力。在此包括酸性磷酸酶(具有最佳酸性反应的EC.3.1.3.2)和碱性磷酸酶(具有最佳碱性反应的EC.3.1.3.1)。
术语“生物学功能”,“功能”,“生物学活性”或“活性”是指如本文所述的萜合酶的如下能力:a)催化形成折叶苔基二磷酸和/或折叶苔醇,或化合物的混合物,其包含折叶苔基二磷酸,和/或折叶苔基单磷酸和/或折叶苔醇和/或一种或多种其他萜烯,特别是折叶苔基二磷酸;或b)催化形成补身烷基醇或两种或更多种补身烷基醇与可选地一种或多种其他萜烯的混合物。
术语“生物学功能”,“功能”,“生物学活性”或“活性”是指如本文所述的乙酰基转移酶催化形成乙酸补身烷基酯或两种或更多种乙酸补身烷基酯与可选的一种或多种其他乙酰化的化合物的混合物的能力。
术语“萜烯的混合物”或“倍半萜的混合物”是指萜烯或倍半萜的混合物,其包含折叶苔醇、补身醇和双环法呢醇中的至少一种,并且还可以包含一种或多种另外的萜烯和/或一种或多种另外的倍半萜。
“甲羟戊酸途径”也称为“异戊二烯途径”或“HMG-CoA还原酶途径”,是真核生物、古菌和某些细菌中必不可少的代谢途径。甲羟戊酸途径始于乙酰辅酶A,并产生两个五碳结构单元,称为异戊烯基焦磷酸(IPP)和二甲基烯丙基焦磷酸(DMAPP)。关键酶是乙酰乙酰基-CoA硫解酶,HMG-CoA合酶,HMG-CoA还原酶,甲羟戊酸激酶,磷酸甲羟戊酸激酶,甲羟戊酸二磷酸脱羧酶和异戊烯基焦磷酸异构酶。将甲羟戊酸途径与酶活性结合以产生萜烯前体GPP、FPP或GGPP,特别是FPP合酶,允许重组细胞生产萜烯。
如本文所用,术语“宿主细胞”或“经转化的细胞”是指一种细胞(或生物体),其被改变以携带(harbor)至少一个核酸分子,例如,编码所需蛋白质或核酸序列的重组基因,其在转录时产生执行本文所述的生物催化方法或其他重组方法所需的至少一种本发明的功能性多肽。特别地,这样的宿主细胞或经转化的细胞提供了乙酰基转移酶,其可用于从相应的补身烷基醇制备至少一种乙酸补身烷基酯。它们还可以提供其他酶,例如折叶苔基二磷酸合酶蛋白,其可用于产生折叶苔基二磷酸和/或折叶苔基单磷酸和/或折叶苔醇,或含有折叶苔基二磷酸和/或折叶苔基单磷酸和/或折叶苔醇的萜烯的相应混合物。它们还可以提供用于制备至少一种补身烷基醇的萜烯合酶。宿主细胞特别是细菌细胞、真菌细胞或植物细胞或植物。宿主细胞可以包含已整合到宿主细胞的核或细胞器基因组中的重组基因。或者,宿主可以在染色体外含有重组基因。
术语“生物(体)”是指任何非人的多细胞或单细胞生物,例如植物或微生物。特别地,微生物是细菌、酵母、藻类或真菌。
术语“植物”可互换使用以包括植物细胞,包括植物原生质体,植物组织,产生再生植物或植物部分的植物细胞组织培养物,或植物器官诸如根、茎、叶、花、花粉、胚珠、胚、果实等。任何植物均可以用来实施本文实施方案的方法。
当特定的生物体或细胞天然地产生FPP或当其不天然地产生FPP但是用本文所述的核酸转化以产生FPP时意味着“能够产生FPP”。经转化以比天然存在的生物体或细胞产生更高量的FPP的生物体或细胞也被“能够产生FPP的生物体或细胞”所涵盖。
当特定的生物体或细胞天然地产生乙酸补身烷基酯或当其不天然地产生乙酸补身烷基酯但用本文所述的核酸转化以产生乙酸补身烷基酯时意味着“能够产生乙酸补身烷基酯”。经转化以比天然存在的生物体或细胞产生更高量的乙酸补身烷基酯的生物体或细胞也被“能够产生乙酸补身烷基酯的生物体或细胞”所涵盖。
当特定的生物体或细胞天然地产生补身烷基醇或当其不天然产生补身烷基醇但经转化以产生补身烷基二磷酸,并视需要进一步用核酸转化以产生将补身烷基二磷酸转化为补身烷基醇的酶活性时意味着“能够产生补身烷基醇”。经转化以产生比天然存在的生物体或细胞更高量的补身烷基醇的生物体或细胞也被“能够产生补身烷基醇的生物体或细胞”所涵盖。
对于本文的说明书和所附权利要求,除非另有说明,否则“或”的使用意味着“和/或”。类似地,各种时态的“含”、“含有”、“包含”和“包括”是可互换的而不是限制性的。
应进一步理解,在各种实施方案的描述使用术语“包含”的情况下,本领域技术人员将理解,在一些特定情况下,可以使用“基本上由……组成”或“由……组成”的语言来替代地描述实施方案。
本文所用的术语“纯化的”、“基本上纯化的”和“分离的”是指不含其他不同化合物(本发明化合物通常以其天然状态与其缔合)的状态,因此“纯化的”、“基本上纯化的”和“分离的”物品占给定样品质量按重量计至少0.5%、1%、5%、10%或20%,或至少50%或75%。在一个实施方案中,这些术语是指本发明的化合物占给定样品质量按重量计至少95%、96%、97%、98%、99%或100%。如本文所用,当提及核酸或蛋白质时,核酸或蛋白质的术语“纯化的”、“基本上纯化的”和“分离的”也指一种纯化或浓缩状态,其不同于天然存在于例如原核或真核环境中,例如在细菌或真菌细胞中,或哺乳动物特别是人体中的状态。任何纯化程度或浓度,只要大于天然存在的纯化或浓缩程度,包括(1)从其他相关结构或化合物中的纯化,或(2)与在所述原核或真核环境中通常不相关的结构或化合物的缔合,都在“分离的”含义内。根据本领域技术人员已知的各种方法和工艺,本文所述的核酸、蛋白质或核酸或蛋白质的类别可以是分离的,或如若不然与它们通常在性质上不相关的结构或化合物相缔合。
术语“约”表示所述值的±25%的可能变化,特别是±15%、±10%、更特别是±5%、±2%或±1%。
术语“基本上”描述的值范围为约80至100%,例如85至99.9%,特别是90至99.9%,更特别是95至99.9%,或98至99.9%,尤其是99至99.9%。
“主要地”是指大于50%的范围内的比例,例如在51%至100%的范围内,特别是在75%至99.9%的范围内;尤其是85至98.5%,例如95至99%。
在本发明的上下文中,“主要产物”表示单一化合物或一组至少2种化合物,例如2、3、4、5或更多种,特别是2或3种化合物,该单一化合物或一组化合物“主要”是通过本文所述的反应制备的,并且基于由所述反应形成的产物的成分的总量,以主要比例包含在所述反应中。所述比例可以是摩尔比例,重量比例,或者优选基于色谱分析,由反应产物的相应色谱图计算的面积比例。
在本发明的上下文中,“副产物”表示单一化合物或一组至少2种化合物,例如2、3、4、5或更多种,特别是2或3种化合物,该单一化合物或一组化合物并非“主要”是通过本文所述的反应制备的。
由于酶促反应的可逆性,除非另有说明,否则本发明涉及在两个反应方向上本文所述的酶促或生物催化反应。
本文描述的多肽的“功能突变体”包括如下定义的此类多肽的“功能等同物”。
术语“立体异构体”特别包括构象异构体。
根据本发明,通常包括本文描述的化合物的所有“立体异构形式”,例如结构异构体,尤其是立体异构体及其混合物,例如旋光异构体或几何异构体,例如E和Z异构体,以及它们的组合。如果在一个分子中存在几个不对称中心,则本发明包括这些不对称中心的不同构象的所有组合,例如对映异构体对。
“立体选择性”描述了产生立体异构纯形式的化合物的特定立体异构体的能力或以本文所述的酶催化方法从多种立体异构体中特异性转化特定立体异构体的能力。更具体而言,这意味着本发明的产物相对于特定的立体异构体富集,或者离析物相对于特定的立体异构体可以贫化。这可以通过根据下式计算的纯度%ee参数进行量化:
%ee=[X
其中X
术语“有选择地转化”或“增加选择性”通常是指,在所述反应的整个过程期间(即在反应的起始和终止之间),在所述反应的某个时间点,或在所述反应的“一段”期间,特定的立体异构形式例如E-形式的不饱和烃以比相应的其他立体异构体形式例如Z-形式更高的比例或量(以摩尔为基准对比)被转化。特别地,在“一段”期间可以观察到所述选择性对应于底物初始量的1至99%,2至95%,3至90%,5至85%,10至80%,15至75%,20至70%,25至65%,30至60%,或40至50%的转化率。所述更高的比例或量可以例如以以下方式表示:
-在整个反应过程或其所述一段期间观察到的较高的异构体最大收率;
-在确定的底物转化率值百分比下,较高的异构体相对含量;和/或
-在较高的转化率值百分比下,相同的异构体相对含量;
其中的每一种优选相对于参考方法来观察,所述参考方法在其他相同条件下用已知化学或生物化学方法进行。
根据本发明,通常包括本文描述的化合物的所有“异构体形式”,例如结构异构体,尤其是立体异构体及其混合物,例如旋光异构体或几何异构体,例如E和Z异构体,以及它们的组合。如果在一个分子中存在几个不对称中心,则本发明包括这些不对称中心的不同构象的所有组合,例如对映异构体对,或立体异构体形式的任何混合形式。
根据本发明的反应的“收率”和/或“转化率”是在例如4、6、8、10、12、16、20、24、36或48小时的规定时间段内(反应在该段时间内进行)确定的。特别地,反应在精确定义的条件下进行,例如在本文定义的“标准条件”下进行。
不同的收率参数(“收率”或YP/S;“比生产率收率”;或时空收率(STY))在本领域中是众所周知的,并且如文献所述进行测定。
“收率”和“YP/S”(均以所生产的产品质量/所消耗的材料质量表示)在本文中用作同义词。
比生产率收率(specific productivity-yield)描述了每小时每L发酵液每克生物质所生产的产物的量。用WCW表示的湿细胞重量描述了生化反应中具有生物活性的微生物的数量。该值以每g WCW每小时的产品g数给出(即g/gWCW
术语“发酵产生”或“发酵”是指微生物(由所述微生物所包含或由其产生的酶活性辅助)在细胞培养物中利用添加到温育中的至少一种碳源产生化合物的能力。
术语“发酵液”应理解为是指一种液体,特别是水性溶液或水性/有机溶液,其基于发酵工艺并且未进行或进行了例如本文所述的后处理(work up)。
“酶催化”或“生物催化”方法是指所述方法在酶(包括本文所定义的酶突变体)的催化作用下进行。因此,该方法可以在分离形式的(纯化的、富集的)或粗制形式的所述酶的存在下,或者在细胞系统的存在下进行,所述细胞系统特别是包含活性形式的所述酶并具有如本文所公开的催化转化反应能力的天然或重组微生物细胞。
如果本公开涉及不同优先程度的特征、参数及其范围(包括上位的,非明确优选的特征、参数及其范围),则除非另有说明,否则这些特征、参数和范围中的两个或更多个的任意组合与它们各自的优选程度无关地涵盖在本发明的公开内容中。
a.本发明的特定实施方案
1.产生至少一种,特别是一种、两种或三种,特别是一种或两种乙酸补身烷基酯化合物的生物催化方法,
包括以下步骤:
(1)在乙酰基供体的存在下,使至少一种,特别是一种立体异构纯形式或立体异构体混合物形式的补身烷基醇与至少一种,特别是一种多肽接触,该多肽具有特别是能将乙酰基从所述乙酰基供体转移至所述至少一种,特别是一种补身烷基醇的乙酰基转移酶活性,从而获得至少一种作为主要产物的乙酸补身烷基酯,特别是一种作为主要产物的乙酸补身烷基酯;和
(2)可选地,从步骤(1)的反应产物中分离出所述至少一种,特别是一种乙酸补身烷基酯化合物。
如果形成多于一种的乙酸补身烷基酯,则可以进一步分离混合物并且可以纯化各个乙酸酯。
2.根据实施方案1所述的方法,其中,所述乙酸补身烷基酯化合物从由如下构成的群组中选出:乙酸折叶苔基酯,乙酸补身烯基酯,和乙酸双环法呢基酯,其各自为立体异构纯形式或作为其至少两种立体异构体的混合物,或它们的组合,其包括所述群组的至少两个成员。在一个特定的实施方案中,仅一种补身烷基醇用作底物,并且仅一种乙酸补身烷基酯以立体异构纯形式或作为其至少两种立体异构体的混合物,特别是立体异构纯形式的产物获得。
3.根据实施方案1或2所述的方法,其中所述补身烷基醇从由如下构成的群组中选出:折叶苔醇,特别是(+)-折叶苔醇,补身醇,特别是(-)-补身醇,和双环法呢醇,特别是(+)-双环法呢醇,其各自为立体异构纯形式或作为其至少两种立体异构体的混合物,或它们的组合,其包括所述群组的至少两个成员。在一个特定的实施方案中,仅一种特别是立体异构纯形式的补身烷基醇用作底物。
4.根据前述实施方案中任一项所述的方法,其中所述乙酰基供体是乙酰辅酶A(乙酰基-CoA)。所述供体可以被外源地添加到反应混合物中,例如在使用分离的、富集的或纯化的酶的体外过程中,或更具体而言是内源地存在的,例如在使用宿主细胞系统的体内过程中,该系统产生乙酰辅酶A系统作为代谢物并表达所需的一种或多种多肽,以进行预期的乙酰化或更复杂的多步骤过程,该过程包括所述乙酰化作为一个步骤。
5.根据前述实施方案中任一项所述的方法,其中所述乙酰基转移酶选自:
a)包含选自SEQ ID NO:9、11、13、15、17、19、21、23、25、118、121、124、127、130、133、136、143和144的氨基酸序列的多肽,和
b)具有乙酰基转移酶活性并包含与SEQ ID NO:9、11、13、15、17、19、21、23、25、118、121、124、127、130、133、136、143和144的至少一个所述氨基酸序列显示至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%,或99%序列同一性程度的氨基酸序列的多肽。
下面的列表说明了本发明的特定乙酰基转移酶产生一种或多种从由乙酸折叶苔基酯,乙酸补身烯基酯和乙酸双环法呢基酯构成的群组中选出的乙酸补身烷基酯化合物的能力:
6.根据前述实施方案中任一项所述的方法,还包括在步骤1)之前,所述至少一种,特别是一种补身烷基醇化合物的生物催化形成。
7.根据实施方案6所述的方法,其中所述补身烷基醇化合物可以内源地存在于反应混合物中,例如在使用宿主细胞系统的体内过程中,该系统产生所述补身烷基醇化合物作为代谢产物并表达所需的一种或多种多肽用于进行预期的补身烷基醇的合成或更复杂的多步骤过程,该过程包括所述补身烷基醇的合成作为一个步骤,由此从非环状倍半萜前体酶促合成所述补身烷基醇。
或者,所述补身烷基醇化合物通过化学或酶促方法产生,并被外源地添加到反应混合物中,例如在使用其形成所需的如下定义的分离的、富集的或纯化的合酶的体外过程中。
8.根据实施方案7所述的方法,其中所述非环状倍半萜前体是法呢基焦磷酸(FPP)。
9.根据实施方案7至8中任一项所述的方法,其中该补身烷基醇的所述酶促合成是通过一种或多种多肽来催化的,该多肽具有在一个或多个酶促步骤中将所述非环状倍半萜前体转化为至少一种补身烷基醇,特别是一种补身烷基醇,特别是作为主要产物的能力。
10.根据实施方案6至9中任一项所述的方法,其中所述至少一种补身烷基醇是在单个或多个,特别是两个酶促步骤中由FPP产生的。
11.根据实施方案10所述的方法,其中所述至少一种补身烷基醇是通过FPP的酶促转化,由如下物质催化产生的:
a)具有形成所述补身烷基醇的补身烷倍半萜合酶活性的多肽(单步生物合成);或者
b)形成具有至少一种磷酸补身烷基酯中间体的磷酸补身烷基酯合酶活性的多肽与具有将所述至少一种磷酸补身烷基酯(单磷酸酯和/或二磷酸酯)中间体转化为至少一种补身烷基醇的磷酸酶活性的多肽的组合(两步生物合成)。
12.根据实施方案11所述的方法,其中,
a)所述具有补身烷倍半萜合酶活性的多肽选自具有折叶苔醇合酶活性,补身醇合酶活性,双环法呢醇合酶活性或这些活性的任何组合的多肽,该多肽特别是优先显示所述活性之一,更特别地,特异性地显示所述活性之一;和
b)多肽的所述组合包括补身烷基二磷酸合酶活性,特别是折叶苔基二磷酸合酶活性和磷酸酶,例如细菌碱性磷酸酶。
13.根据实施方案12所述的方法,其中,
a)所述具有补身烷倍半萜合酶活性的多肽选自如PCT/EP2018/064344(2018年5月31日提交)中所述的补身烷合酶和如公开的WO2015/169871和WO 2015/176959中所述的补身醇合酶;
b)所述具有磷酸补身烷基酯合酶活性的多肽是2018年5月29日提交的PCT/CN2018/088902中所述的折叶苔基二磷酸合酶,包括从作为底物的法呢基二磷酸(FPP)产生折叶苔基磷酸酯衍生物如单磷酸酯,更特别是折叶苔基二磷酸的能力。
如PCT/CN2018/088902(2018年5月29日提交)中所述的折叶苔基二磷酸合酶为:
香鳞毛蕨(Dryopteris fragrans)的DfHAD,DfHAD-9(V274A),DfHAD-His_GST和DfHAD-8(K532R)
以及由其衍生的与其具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的多肽。
如PCT/EP2018/064344(2018年5月31日提交)中所述的补身烷合酶(即折叶苔醇合酶或补身醇合酶)是
以及由此衍生的与其具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的多肽。
如WO2015/169871中所述的补身醇合酶是
以及由此衍生的与其具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的多肽。
如WO 2015/176959中所述的补身醇合酶是
黑水缬草(Valeriana amurensis)的VaTPS3
以及由此衍生的与其具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的多肽。
在本说明书的末尾列出了上述合酶的各个氨基酸和核苷酸序列的SEQ ID NO。这些多肽和核酸以及由其衍生的与这些序列中的至少一个具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的多肽和核酸也是本公开的一部分。
14.根据实施方式13所述的方法,其中,所述补身烷倍半萜合酶选自:
a)具有(双功能)折叶苔醇合酶活性并且包含氨基酸序列SEQ ID NO:5的多肽,或具有折叶苔醇合酶活性并且包含与在PCT/EP2018/064344中描述的SEQ ID NO:5具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的氨基酸序列的突变体或变体多肽;
b)具有(双功能)补身醇合酶活性并且包含氨基酸序列SEQ ID NO:7的多肽,或具有补身醇合酶活性并且包含与在PCT/EP2018/064344中描述的SEQ ID NO:7具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%序列同一性的氨基酸序列的突变体或变体多肽。
15.根据前述实施方案中任一项所属的方法,其在宿主细胞培养物中在体内或在包含产生至少一种乙酸补身烷基酯所需的宿主细胞裂解物或者富集或分离的多肽的液体反应介质中在体外进行,每种均在有利于产生至少一种乙酸补身烷基酯的条件下进行。
特别地,该反应在内源形成的或外源添加的乙酰基-CoA作为乙酰基供体的存在下进行。特别地,该反应在内源形成的或外源添加的FPP的存在下进行。内源形成的FPP特别是生物转化为FPP的至少一种碳源例如糖底物的代谢的结果。细胞体内方法尤其令人感兴趣,其中FPP和乙酰基-CoA是内源形成的。
这些宿主细胞或生物体中的某些不天然地产生FPP。为了适合于实施本文所述的实施方案的方法,不天然地产生无环萜烯焦磷酸前体例如FPP的生物体或细胞被基因修饰以产生所述前体。例如,它们可以在用根据以上任何一个实施方案所述的核酸修饰之前或同时如此转化。转化生物体以使其产生无环萜烯焦磷酸前体例如FPP的方法是本领域已知的。例如,引入甲羟戊酸途径的酶活性是使生物体产生FPP的合适策略。
16.根据实施方案15所述的方法,其在能够功能性表达以下项的重组宿主细胞或重组非人宿主生物体中进行:
a)至少一种实施方案5中定义的乙酰基转移酶;可选地
b)至少一种多肽,其具有将非环状倍半萜前体FPP转化为实施方案9至14中任一项定义的至少一种补身烷基醇的能力;以及可选地
c)至少一种从如上定义的甲羟戊酸途径的酶中选择的酶。
在一个特定的实施方案中,酶a)和b)或者酶a)、b)和c)通过本发明的体内方法中应用的细胞系统功能性表达。
17.根据实施方案16所述的方法,其中所述非人宿主细胞或宿主生物体选自原核或真核微生物,或由其衍生的细胞。
18.根据实施方案17所述的方法,其中所述非人宿主细胞或宿主生物体选自细菌、真菌和植物细胞或植物。
19.根据实施方案18所述的方法,其中所述真菌细胞是酵母细胞,特别是选自酵母属(Saccharomyces)、毕赤酵母属(Pichia)或耶氏酵母属(Yarrowia),特别是选自酿酒酵母(Saccharomyces cerevisiae)、巴斯德毕赤酵母(Pichia pastoris)或解脂耶氏酵母(Yarrowia lipolytica)种。
20.根据实施方案18所述的方法,其中所述细菌细胞选自红球菌属(Rhodococcus)、假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)或埃希氏菌属(Escherichia),特别是大肠杆菌(E.coli)种。
21.根据前述实施方案中任一项所述的方法,其进一步包括作为步骤(3),使用化学或生物催化合成或两者的组合来处理步骤(1)或步骤(2)的至少一种乙酸补身烷基酯以获得衍生物。
22.根据实施方案21所述的方法,其中所述衍生物是烃,醇,二醇,三醇,缩醛,缩酮,醛,酸,醚,酰胺,酮,内酯,环氧化物,乙酸酯,糖苷,酯和/或多环化合物。
23.根据前述实施方案中任一项所述的方法,其中所述乙酸补身烷基酯包括乙酸折叶苔基酯,乙酸补身烯基酯或乙酸双环法呢基酯,特别是乙酸折叶苔基酯或乙酸补身烯基酯,作为主要的补身烷基醇产物或特别是作为单一的补身烷基醇产物。
24.根据前述实施方案中任一项所述的方法,其包括特别是通过转化向非人宿主生物体或宿主细胞提供以下物质:
a)至少一种核酸、表达构建体或载体,其包含编码至少一种具有乙酰基转移酶活性的多肽的核酸序列,该多肽能够将乙酰基从乙酰基供体转移至补身烷基醇,可选地稳定地整合到基因组中;可选地
b)至少一种核酸、表达构建体或载体,其包含编码至少一种具有补身烷基醇合酶活性的多肽的核酸序列,该多肽能够从非环状倍半萜前体产生补身烷基醇,可选地稳定地整合到基因组中;以及可选地
c)至少一种核酸、表达构建体或载体,其包含编码至少一种涉及生物合成途径的多肽的核酸序列,该多肽用于产生所述非环状倍半萜前体,可选地稳定地整合到基因组中。
在一个特定的实施方案中,非人宿主生物体或宿主细胞用a)和b)或者a)、b)和c)转化,并且更特别地包含稳定地整合到基因组中的所述核酸。该核酸a)、b)和/或c)可以位于相同或两个或更多个不同载体上。
25.一种具有乙酰基转移酶活性的多肽,其能够将乙酰基从乙酰基供体转移至补身烷基醇以产生乙酸补身烷基酯,其包含与从SEQ ID NO:9、11、13、15、17、19、21、23、25、118、121、124、127、130、133、136、143和144中选出的至少一个氨基酸序列具有40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%至少于100%序列同一性的氨基酸序列。
26.一种分离的核酸分子,
a)包含编码实施方案5的多肽的核苷酸序列;或者
b)包含与从SEQ ID NO:8、10、12、14、16、16、18、20、22、24、116、117、119、120、122、123、125、126、128、129、131、132、134和135中选出的核苷酸序列具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%,85%,90%,95%,96%,97%,98%或99%并少于100%序列同一性的核苷酸序列;或者
c)包含核苷酸序列,该核苷酸序列包含与序列a)或b)之一互补的序列;或者
d)包含在严格条件下与核苷酸序列a)、b)或c)杂交的核苷酸序列。
27.一种表达构建体,其包含至少一种实施方案26的核酸分子。
28.一种载体,其包含至少一种实施方案25的核酸分子或至少一种实施方案26的表达构建体。
29.根据实施方案28所述的载体,其中该载体是原核、病毒或真核载体。
30.根据实施方案28或29所述的载体,其中该载体是表达载体。
31.根据实施方案28至30中任一项所述的载体,其是质粒载体。
32.一种重组宿主细胞或重组非人宿主生物体,其包含:
a)至少一种实施方案26的分离的核酸分子,其可选地稳定地整合到基因组中;或者
b)至少一种实施方案27的表达构建体,其可选地稳定地整合到基因组中;或者
c)至少一种实施方案28至31中任一项的载体。
在一个特定的实施方案中,非人宿主生物体或宿主细胞用a)和b)或者a)、b)和c)转化,并且更特别地包含稳定地整合到基因组中的所述核酸。
33.根据实施方案32所述的宿主细胞或宿主生物体,其选自原核或真核微生物,或由其衍生的细胞。
34.根据实施方案33所述的宿主细胞或宿主生物体,其选自细菌、真菌和植物细胞或植物。
35.根据实施方案34所述的宿主细胞或宿主生物体,其中所述真菌细胞是酵母细胞。
36.根据实施方案35所述的宿主细胞或宿主生物体,其中所述细菌细胞选自埃希氏菌属(Escherichia),特别是大肠杆菌(E.coli)种,并且所述酵母细胞选自酵母属(Saccharomyces)、毕赤酵母属(Pichia)或耶氏酵母属(Yarrowia),特别是选自酿酒酵母(Saccharomyces cerevisiae)、巴斯德毕赤酵母(Pichia pastoris)或解脂耶氏酵母(Yarrowia lipolytica)种。
37.一种用于产生至少一种根据实施方案25的催化活性多肽的方法,包括:
a)培养实施方案32至34之一的非人宿主生物体或宿主细胞以表达或过表达根据实施方案25的至少一种多肽;和
b)可选地,从步骤a)中培养的非人宿主细胞或生物体中分离多肽。
38.根据实施方案37所述的方法,其还包括在步骤a)之前,向非人宿主生物体或细胞提供以下物质,特别是用以下物质转化非人宿主生物体或宿主细胞:根据权利要求26的至少一种核酸,或权利要求27的至少一种构建体,或权利要求28至31中任一项的至少一种载体,从而其表达或过表达根据权利要求25的多肽。
39.一种制备具有乙酰基转移酶活性的突变多肽的方法,该突变多肽能够将乙酰基从乙酰基供体转移至至少一种,特别是一种补身烷基醇,以产生至少一种,特别是一种乙酸补身烷基酯,该方法包括以下步骤:
a)选择出编码从SEQ ID NO:9、11、13、15、17、19、21、23、25、118、121、124、127、130、133、136、143和144中选出的多肽的核酸分子,
b)修饰所选择的核酸分子以获得至少一种突变核酸分子;
c)用突变核酸序列转化宿主细胞或单细胞宿主生物体以表达由该突变核酸序列编码的多肽;
d)筛选表达产物中至少一种具有乙酰基转移酶活性的突变体;和,
e)可选地,如果该多肽不具有所需突变活性,则重复步骤a)至d),直到获得具有所需突变活性的多肽;和
f)可选地,如果在步骤d)中鉴定出具有所需突变活性的多肽,则分离在步骤c)中获得的相应突变核酸。
40.前述实施方案中任一项定义的乙酰基转移酶的用途,用于制备气味剂、调味剂或芳香剂成分或昆虫/害虫防治,例如用于制备从身体护理、家庭护理或芳香剂组合物中选出的组合物。
b.根据本发明适用的多肽
在本文语境中,以下定义适用:
可以互换使用的通用术语“多肽”或“肽”是指天然的或合成的,连续的、肽方式连接的氨基酸残基的线性链或序列,其包含约10个至多于1000个残基。具有最多30个残基的短链多肽也被称为“寡肽”。
术语“蛋白(质)”是指由一种或多种多肽组成的大分子结构。其多肽的氨基酸序列代表蛋白质的“一级结构”。氨基酸序列还通过形成特殊的结构元素(例如在多肽链中形成的α-螺旋和β-折叠结构)来预先确定蛋白质的“二级结构”。多个这样的二级结构元件的排列定义了蛋白质的“三级结构”或空间排列。如果蛋白质包含多于一个的多肽链,则所述链在空间上排列形成蛋白质的“四级结构”。蛋白质正确的空间排列或“折叠”是蛋白质功能的前提。变性或展开会破坏蛋白质功能。如果这种破坏是可逆的,则可以通过重新折叠来恢复蛋白质功能。
本文所指的典型的蛋白质功能是“酶功能”,即蛋白质在底物例如化合物上充当生物催化剂,并催化所述底物向产物的转化。酶可以显示高或低程度的底物和/或产物特异性。
因此,本文中被称为具有特定“活性”的“多肽”隐含地是指正确折叠的蛋白质,其显示出所指示的活性,例如特定的酶活性。
因此,除非另有说明,否则术语“多肽”也涵盖术语“蛋白质”和“酶”。
类似地,术语“多肽片段”涵盖术语“蛋白质片段”和“酶片段”。
术语“分离的多肽”是指通过本领域已知的任何方法或这些方法(包括重组、生物化学和合成法)的组合从其天然环境中取出的氨基酸序列。
“靶肽”是指一种氨基酸序列,其将蛋白质或多肽靶向细胞内细胞器(即,线粒体或质体)或细胞外空间(分泌信号肽)。编码靶肽的核酸序列可以被融合到编码蛋白或多肽的氨基末端(例如N-末端)的核酸序列,或者可以被用来替换天然靶向多肽。
本发明还涉及本文具体描述的多肽的“功能等同物”(也称为“类似物”或“功能突变”)。
例如,“功能等同物”是指一种多肽,其在用于确定酶活性的测试中,显示与本文具体描述的并作为所述比较基础的多肽相比,至少高或低1至10%、或至少20%、或至少50%、或至少75%、或至少90%的酶活性。
根据本发明,“功能等同物”还涵盖特定的突变体,其在本文所述的氨基酸序列的至少一个序列位置中具有与具体陈述的氨基酸不同的氨基酸,但是仍然具有上述生物活性之一,例如酶活性。因此,“功能等同物”包括可通过一个或多个,例如1至20个、1至15个或5至10个氨基酸的添加、取代特别是保守取代(即,作为其结果,所讨论的氨基酸被具有相同电荷、大小、极性和/或溶解度的氨基酸取代)、缺失和/或倒置而获得的突变体,其中所述变化可以在任何序列位置上发生,只要它们导致突变体具有本发明特性的概貌。还特别地提供功能等同性,如果活性模式与在突变体和未改变的多肽之间定性地重合,即,如果例如观察到与相同的激动剂或拮抗剂或底物的相互作用,但是速率不同(即,通过EC
上述意义上的“功能等同物”也是本文所述多肽的“前体”,以及所述多肽的“功能衍生物”和“盐”。
在该情况下,“前体”是具有或不具有所期望生物活性的多肽的天然或合成前体。
表述“盐”是指根据本发明的蛋白质分子的羧基的盐以及氨基的酸加成的盐。羧基的盐可以已知的方式生产,包括无机盐,例如钠、钙、铵、铁和锌盐,以及与有机碱例如胺,如三乙醇胺、精氨酸、赖氨酸、哌啶等形成的盐。酸加成的盐,例如与无机酸例如盐酸或硫酸形成的盐,以及与有机酸例如乙酸和草酸形成的盐,也被本发明所涵盖。
根据本发明的多肽的“功能衍生物”还可以使用已知技术在功能性氨基酸侧基或它们的N末端或C末端产生。这样的衍生物包括例如:羧酸基的脂族酯,羧酸基的酰胺,它们可通过与氨或与伯或仲胺反应获得;游离氨基的N-酰基衍生物,其通过与酰基反应生成;或游离羟基的O-酰基衍生物,其通过与酰基反应生成。
“功能等同物”自然也包括可以从其他生物体获得的多肽以及天然存在的变体。例如,可以通过序列比较来确定同源序列区域的面积,并且等同的多肽可以基于本发明的具体参数来确定。
“功能等同物”还包含根据本发明的多肽的“片段”,例如单个结构域或序列基序,或N末端和/或C末端截短的形式,其可以显示或可以不显示期望的生物学功能。优选地,这样的“片段”至少定性地保持期望的生物学功能。
此外,“功能等同物”是融合蛋白,其具有本文所述的多肽序列之一或由其衍生的功能等同物,以及在功能性N-末端或C-末端缔合(即,没有融合蛋白部分的实质性相互功能受损)中具有至少一个另外的功能不同的异源序列。这些异源序列的非限制性例子是例如信号肽、组氨酸锚或酶。
根据本发明还包括的“功能等同物”是与具体公开的多肽的同源物。它们与具体公开的氨基酸序列具有至少60%,优选至少75%,特别是至少80或85%,例如90、91、92、93、94、95、96、97、98或99%的同源性(或同一性),其通过Pearson and Lipman,Proc.Natl.Acad,Sci.(USA)85(8),1988,2444-2448的算法计算。根据本发明的同源多肽的以百分比表示的同源性或同一性尤其是指基于本文具体描述的氨基酸序列之一的总长度,以氨基酸残基的百分比表示的同一性。
以百分比表示的同一性数据也可以借助于BLAST比对,算法blastp(蛋白质-蛋白质BLAST)或通过应用本文下面详述的Clustal设置来确定。
在可能的蛋白质糖基化的情况下,根据本发明的“功能等同物”包括本文所述的去糖基化或糖基化形式的多肽,以及可以通过改变糖基化模式获得的经修饰形式。
根据本发明的多肽的功能等同物或同源物可以通过诱变产生,例如通过点突变,延长或缩短蛋白质或如下文更详细描述。
根据本发明的多肽的功能等同物或同源物可以通过筛选突变体例如缩短的突变体的组合数据库来鉴定。例如,蛋白质变体的多样性数据库可以通过在核酸水平上的组合诱变,例如通过合成寡核苷酸混合物的酶促连接来产生。有许多方法可用于从简并寡核苷酸序列产生潜在同源物的数据库。简并基因序列的化学合成可以在自动DNA合成仪中进行,然后可以将合成基因连接在合适的表达载体中。简并基因组的使用使得可以提供混合物中的所有序列,其编码所需的潜在蛋白质序列集合。简并寡核苷酸的合成方法是本领域技术人员已知的。
在现有技术中,已知几种技术用于筛选通过点突变或缩短产生的组合数据库的基因产物,以及用于筛选具有选定性质的基因产物的cDNA文库。这些技术可以适用于快速筛选通过根据本发明的同源物的组合诱变产生的基因库。最常用于筛选大型基因库的基于高通量分析的技术包括在可复制的表达载体中克隆基因库,用所得载体数据库转化合适的细胞,以及在特定条件下表达组合基因,在所述条件下,所需活性的检测促进编码基因(其产物被检测)的载体的分离。递归整合诱变(REM)是一种提高数据库中功能突变体频率的技术,其可以与筛选测试结合使用,以鉴定同源物。
本文提供的实施方案提供了本文公开的多肽的直系同源物和旁系同源物,以及用于鉴定和分离此类直系同源物和旁系同源物的方法。术语“直系同源物”和“旁系同源物”的定义在下面给出,并适用于氨基酸和核酸序列。
c.根据本发明适用的编码核酸序列
在本文语境中,以下定义适用:
术语“核酸序列”、“核酸”、“核酸分子”和“多核苷酸”可互换使用,是指核苷酸的序列。核酸序列可以是任意长度的单链或双链脱氧核糖核苷酸或核糖核苷酸,并且包括基因的编码和非编码序列、外显子、内含子、有义和反义互补序列、基因组DNA、cDNA、miRNA、siRNA、mRNA、rRNA、tRNA、重组核酸序列、分离的和纯化的天然产生的DNA和/或RNA序列、合成的DNA和RNA序列、片段、引物和核酸探针。技术人员了解RNA的核酸序列与DNA序列相同,差异在于胸腺嘧啶(T)被替代为尿嘧啶(U)。术语“核苷酸序列”也应理解为包含单独的片段形式或作为较大核酸组分的多核苷酸分子或寡核苷酸分子。
“分离的核酸”或“分离的核酸序列”是指一种核酸或核酸序列,其所处的环境与天然产生的核酸或核酸序列所处的环境不同,并且可以包括基本上不含污染内源性物质的那些。
如本文使用的应用于核酸的术语“天然产生的”是指一种核酸,其在自然界的生物的细胞中发现,并且未经人类在实验室中进行有意的修饰。
多核苷酸或核酸序列的“片段”是指连续的核苷酸,其特别是本文一个实施方案的多核苷酸长度的至少15bp,至少30bp,至少40bp,至少50bp和/或至少60bp。特别地,多核苷酸的片段包含本文一个实施方案的多核苷酸的至少25个,更特别是至少50个,更特别是至少75个,更特别是至少100个,更特别是至少150个,更特别是至少200个,更特别是至少300个,更特别是至少400个,更特别是至少500个,更特别是至少600个,更特别是至少700个,更特别是至少800个,更特别是至少900个,更特别是至少1000个连续核苷酸。不受限制,本文的多核苷酸的片段可以用作PCR引物和/或探针,或用于反义基因沉默或RNAi。
如本文所用,术语“杂交”或在一定条件下杂交旨在描述杂交和洗涤的条件,在所述条件下彼此显著相同或同源的核苷酸序列保持彼此结合。该条件可以使得至少约70%、例如至少约80%、和例如至少约85%、90%或95%同一性的序列保持彼此结合。下文提供了低严格度、中等和高严格度杂交条件的定义。本领域技术人员可以通过例如Ausubel等人(1995,Current Protocols in Molecular Biology,John Wiley&Sons,sections 2,4,and6)所举例说明的那样以最少的实验来选择合适的杂交条件。另外,严格条件在Sambrook等人(1989,Molecular Cloning:A Laboratory Manual,2nd ed.,Cold Spring HarborPress,chapters 7,9,and 11)中描述。
“重组核酸序列”是通过使用实验室方法(例如分子克隆)将来自多于一个源的遗传物质组合在一起所生成的核酸序列,由此创造出或修饰出不是天然产生并且不能以其他方式在生物有机体中发现的核酸序列。
“重组DNA技术”是指用于制备重组核酸序列的分子生物学方法,例如描述于由Weigel和Glazebrook编辑的Laboratory Manuals,2002,Cold Spring Harbor Lab Press;和Sambrook等,1989Cold Spring Harbor,NY:Cold Spring Harbor Laboratory Press。
术语“基因”是指一种DNA序列,其包含可操作地连接到适当调控区域(例如启动子)的被转录为RNA分子(例如细胞中的mRNA)的区域。因此,基因可以包含几个可操作地连接的序列,诸如启动子、5’前导序列(包含例如参与翻译初始化的序列)、cDNA或基因组DNA的编码区、内含子、外显子和/或3’非翻译序列(包含例如转录终止位点)。
“多顺反子”是指可以在同一核酸分子内分别编码多于一个多肽的核酸分子,特别是mRNA。
“嵌合基因”是指通常不能在自然界的物种中发现的任何基因,特别是这样一种基因,其中核酸序列存在一个或多个部分在性质上彼此不相关联。例如,启动子在性质上与转录区的部分或全部或与另一调控区不相关联。术语“嵌合基因”应当被理解为包括表达构建体,其中启动子或转录调控序列被可操作地连接到一个或多个编码序列或反义(即有义链的反向互补链)或反向重复序列(有义和反义,由此RNA转录物在转录后形成双链RNA)。术语“嵌合基因”还包括通过组合一个或多个编码序列的部分以产生新基因而获得的基因。
“3’URT”或“3’非翻译序列”(也称为“3’未翻译区”或“3’末端”)是指在基因编码序列的下游发现的核酸序列,其包含例如转录终止位点和(在大多数但非全部的真核mRNA中)多聚腺苷酸化信号,例如AAUAAA或其变体。在转录终止后,mRNA转录物可以在多聚腺苷酸化信号的下游切去,并且可以添加poly(A)尾,其参与了mRNA向翻译位点例如细胞质的转运。
术语“引物”是指短的核酸序列,其被杂交到模板核酸序列并且被用于与该模板互补的核酸序列的聚合。
术语“可选择标记”是指在表达后能够被用来选择包括该可选择标记的一种或多种细胞的任何基因。以下描述了可选择标记的例子。本领域技术人员了解不同的抗生素、杀真菌剂、营养缺陷型或除草剂可选择标记可适用于不同的目标物种。
本发明还涉及编码如本文定义的多肽的核酸序列。
特别地,本发明还涉及编码上述多肽之一及其功能等同物的核酸序列(单链和双链DNA和RNA序列,例如cDNA、基因组DNA和mRNA),其可以通过例如使用人工核苷酸类似物来获得。
本发明既涉及分离的核酸分子,其编码根据本发明的多肽或其生物学活性区段,又涉及核酸片段,其可用作例如鉴定或扩增根据本发明的编码核酸的杂交探针或引物。
本发明还涉及与本文具体公开的序列具有一定程度的“同一性”的核酸。两个核酸之间的“同一性”是指在每种情况下在核酸的整个长度上核苷酸的同一性。
两个核苷酸序列(同样适用于肽或氨基酸序列)之间的“同一性”是当产生这两个序列的比对时,核苷酸残基(或氨基酸残基)的数目的函数,或两个序列中相同的残基数目。相同的残基被定义为两个序列中在比对的给定位置的相同的残基。本文使用的序列同一性的百分比是从最佳比对中通过将两个序列之间相同的残基数除以最短序列中的残基总数并乘以100计算得到的。最佳比对是同一性百分比最高可能性的比对。可以将空位引入到一个或两个序列中的比对的一个或多个位置中以获得最佳比对。然后将这些空位考虑为用于计算序列同一性百分比的不相同的残基。用于确定氨基酸或核酸序列同一性百分比的比对可以使用计算机程序以及例如在互联网上可公开获得的计算机程序以多种方式实现。
特别地,可使用可从National Center for Biotechnology Information(美国国家生物技术信息中心)(NCBI)于http://www.ncbi.nlm.nih.gov/BLAST/bl2seq/wblast2.cgi获得的设定为默认参数的BLAST程序(Tatiana等,FEMS Microbiol Lett.,1999,174:247-250,1999)来获得蛋白或核酸序列的最佳比对并计算序列同一性的百分比。
在另一个例子中,同一性可以通过Informax公司(美国)的Vector NTI Suite 7.1程序使用Clustal方法(Higgins DG,Sharp PM.((1989)))通过以下设置来计算:
多重比对参数:
成对比对参数:
或者,同一性可以根据Chenna et al.(2003),网页:http://www.ebi.ac.uk/Tools/clustalw/index.html#的方法和以下设置来确定:
本文提及的所有核酸序列(单链和双链DNA和RNA序列,例如cDNA和mRNA)可以以已知方式通过化学合成从核苷酸结构单元产生,例如通过双螺旋的各个重叠的互补核酸结构单元的片段缩合来实现。寡核苷酸的化学合成例如可以通过磷酰胺法(Voet,Voet,2ndedition,Wiley Press,New York,pages 896-897)以已知的方式进行。合成寡核苷酸的积累,和借助于DNA聚合酶的Klenow片段和连接反应的空位的填补,以及一般的克隆技术描述于Sambrook et al.(1989),请参阅下文。
另外,根据本发明的核酸分子可以另外包含来自编码遗传区域的3'和/或5'末端的非翻译序列。
本发明进一步涉及与具体描述的核苷酸序列或其区段互补的核酸分子。
根据本发明的核苷酸序列使得可以产生可用于鉴定和/或克隆其他细胞类型和生物体中的同源序列的探针和引物。此类探针或引物通常包含在“严格”条件下(如本文其他部分所定义)与根据本发明的核酸序列的有义链或相应的反义链的至少约12个,优选至少约25个,例如约40、50或75个连续核苷酸杂交的核苷酸序列区域。
“同源”序列包括直系同源或旁系同源序列。鉴别直系同源物或旁系同源物的方法包括现有技术中已知且在本文中描述的系统发生学方法、序列相似性和杂交方法。
“旁系同源物”或旁系同源序列来源于基因复制,其产生具有相似序列和相似功能的两种或更多种基因。旁系同源物通常聚簇在一起并且通过在相关植物物种内基因的复制而形成。使用成对Blast分析或在基因家族的系统发生分析过程中使用程序诸如CLUSTAL在类似基因的组中发现旁系同源物。在旁系同源物中,共有序列可被鉴定为其特征在于相关基因中的序列并且具有基因的类似功能。
“直系同源物”或直系同源序列是彼此相似的序列,因为它们发现于由共同的祖先传下的物种中。例如,已知具有共同祖先的植物物种含有许多具有相似序列和功能的酶。例如通过使用CLUSTAL或BLAST程序构建一个物种的基因族的系统发生树,技术人员能够鉴定直系同源序列并预测直系同源物的功能。一种用于鉴定或确认同源序列间的相似功能的方法是通过比较过表达或缺乏(在基因敲除/敲减中)相关多肽的宿主细胞或生物体(如植物或微生物)中的转录物概况。技术人员能够理解,具有相似转录物概况的基因(具有大于50%调控的共同转录物,或具有大于70%调控的共同转录物,或大于90%调控的共同转录物)会具有相似的功能。通过使宿主细胞,生物体例如植物或微生物产生萜合酶蛋白,本文所述序列的同源物、旁系同源物、直系同源物以及任何其他变体预期以类似的方式发挥作用。
术语“可选择标记”是指在表达后能够被用来选择包括该可选择标记的一种或多种细胞的任何基因。以下描述了可选择标记的例子。本领域技术人员了解不同的抗生素、杀真菌剂、营养缺陷型或除草剂可选择标记可适用于不同的目标物种。
“分离的”核酸分子与存在于核酸天然来源中的其他核酸分子分离,并且如果通过重组技术生产,则可以基本上不含其他细胞材料或培养基,或者如果通过化学合成,则可以不含化学前体或其他化学物质。
可以借助于分子生物学的标准技术和根据本发明提供的序列信息来分离根据本发明的核酸分子。例如,可以使用具体公开的完整序列之一或其片段作为杂交探针和标准杂交技术(例如,描述于Sambrook,(1989))从合适的cDNA文库中分离cDNA。
另外,包含所公开的序列之一或其片段的核酸分子可以使用基于该序列构建的寡核苷酸引物,通过聚合酶链反应来分离。以此方式扩增的核酸可以克隆到合适的载体中,并可以通过DNA测序来表征。根据本发明的寡核苷酸也可以通过标准的合成方法,例如使用自动DNA合成仪制备。
根据本发明的核酸序列或其衍生物,这些序列的同源物或部分可以例如通过常规的杂交技术或PCR技术从其他细菌中,例如通过基因组或cDNA文库分离出来。这些DNA序列在标准条件下与根据本发明的序列杂交。
“杂交”是指多核苷酸或寡核苷酸在标准条件下结合几乎互补的序列的能力,而在这些条件下非互补配对者之间不发生非特异性结合。为此,序列可以是90~100%互补的。能够彼此特异性结合的互补序列的性质被用于例如Northern印迹或Southern印迹或PCR或RT-PCR中的引物结合。
保守区的短寡核苷酸有利地用于杂交。然而,也可能使用更长的本发明核酸片段或完整序列进行杂交。这些“标准条件”取决于所使用的核酸(寡核苷酸,更长的片段或完整序列)或用于杂交的核酸类型(DNA或RNA)而有所不同。例如,DNA:DNA杂交种的解链温度比相同长度的DNA:RNA杂交种低约10℃。
例如,根据特定核酸的不同,标准条件是指温度在42至58℃,在浓度为0.1至5 xSSC(1 X SSC=0.15M NaCl,15mM柠檬酸钠,pH 7.2)的缓冲水溶液中,或另外在50%甲酰胺(例如42℃,5 x SSC,50%甲酰胺)的存在下。有利地,用于DNA:DNA杂交种的杂交条件是0.1×SSC,温度为约20℃至45℃,优选约30℃至45℃。对于DNA:RNA杂交种,杂交条件有利地为0.1×SSC,并且温度为约30℃至55℃,优选约45℃至55℃。这些所述的杂交温度是对于长度约100个核苷酸的核酸,以及在不存在甲酰胺的情况下G+C含量为50%的经计算的解链温度值的例子。DNA杂交的实验条件已在相关的遗传学教科书(例如Sambrook et al.,1989)中进行了描述,并且可以使用本领域技术人员已知的分子式来计算,例如取决于核酸的长度,杂交种的类型或G+C含量。本领域技术人员可以从以下教科书中获得有关杂交的更多信息:Ausubel et al.(eds),(1985),Brown(ed)(1991)。
“杂交”尤其可以在严格条件下进行。这样的杂交条件例如描述于Sambrook(1989),或Current Protocols in Molecular Biology,John Wiley&Sons,N.Y.(1989),6.3.1-6.3.6。
如本文所用,术语“杂交或在一定条件下杂交”旨在描述杂交和洗涤的条件,在所述条件下彼此显著相同或同源的核苷酸序列保持彼此结合。该条件可以使得至少约70%,例如至少约80%和例如至少约85%、90%或95%同一性的序列保持彼此结合。本文提供了低严格度、中等和高严格度杂交条件的定义。
本领域技术人员可以通过例如Ausubel等人(1995,Current Protocols inMolecular Biology,John Wiley&Sons,sections 2,4,and 6)所举例说明的那样以最少的实验来选择合适的杂交条件。另外,严格条件在Sambrook等人(1989,Molecular Cloning:ALaboratory Manual,2nd ed.,Cold Spring Harbor Press,chapters 7,9,and 11)中描述。
如本文所用,所限定的低严格度条件如下。含有DNA的滤膜在含有35%甲酰胺,5xSSC,50mM Tris-HCl(pH 7.5),5mM EDTA,0.1%PVP,0.1%Ficoll,1%BSA和500μg/ml变性鲑鱼精子DNA的溶液中于40℃预处理6小时。杂交在相同的溶液中进行,并进行以下修改:0.02%PVP,0.02%Ficoll,0.2%BSA,100μg/ml鲑鱼精子DNA,10%(wt/vol)硫酸葡聚糖,并使用5~20x10
如本文所用,所限定的中等严格度条件如下。含有DNA的滤膜在含有35%甲酰胺,5x SSC,50mM Tris-HCl(pH 7.5),5mM EDTA,0.1%PVP,0.1%Ficoll,1%BSA和500μg/ml变性鲑鱼精子DNA的溶液中于50℃预处理7小时。杂交在相同的溶液中进行,并进行以下修改:0.02%PVP,0.02%Ficoll,0.2%BSA,100μg/ml鲑鱼精子DNA,10%(wt/vol)硫酸葡聚糖,并使用5~20x10
如本文所用,所限定的高严格度条件如下。含DNA的滤膜在由6x SSC,50mM Tris-HCl(pH 7.5),1mM EDTA,0.02%PVP,0.02%Ficoll,0.02%BSA和500μg/ml变性鲑鱼精子DNA组成的缓冲液中于65℃下预杂交8小时至过夜。在含有100μg/ml变性鲑鱼精子DNA和5~20x10
如果上述条件不合适(例如,如用于种间杂交),则可以使用本领域众所周知的其他低、中等和高严格度条件(例如,用于种间杂交)。
用于编码本发明多肽的核酸序列的检测试剂盒可以包括对编码该多肽的核酸序列具有特异性的引物和/或探针,以及使用该引物和/或探针来检测样品中编码该多肽的核酸序列的相关方案。此种检测试剂盒可用于确定植物、生物、微生物或细胞是否已被修饰,即是否已用编码多肽的序列转化。
为了测试根据本文一个实施方案的变体DNA序列的功能,将目标序列可操作地连接到可选择的或可筛选的标记基因,并且在使用微生物或原生质体进行的瞬时表达分析中或在稳定转化的植物中测试报告基因的表达。
本发明还涉及具体公开的或可衍生的核酸序列的衍生物。
因此,根据本发明的另外的核酸序列可以衍生自本文具体公开的序列,并且可以通过一个或几个(例如1至10个)核苷酸的一个或多个,例如1至20个,特别是1至15个或5至10个添加、取代、插入或缺失而与之不同,并且还编码具有所期望特性的多肽。
本发明还包括与具体陈述的序列相比,根据特定原始或宿主生物体的密码子使用而包含所谓的沉默突变或已被改变的核酸序列。
根据本发明的特定实施方案,可以制备变体核酸以使其核苷酸序列适应特定的表达系统。例如,如果氨基酸由特定的密码子编码,则已知细菌表达系统可更有效地表达多肽。由于遗传密码的简并性,多于一个密码子可以编码相同的氨基酸序列,多个核酸序列能够编码相同的蛋白或多肽,所有这些DNA序列均被涵盖在本文一个实施方案中。在适当的情况下,编码本文所述多肽的核酸序列可以被优化以增加在宿主细胞中的表达。例如,可以使用宿主特异性的密码子合成本文一个实施方案的核酸以改善表达。
本发明还涵盖本文描述的序列的天然存在的变体,例如剪接变体或等位基因变体。
等位基因变体在所衍生的氨基酸水平上,具有在整个氨基酸范围至少60%的同源性,优选至少80%的同源性,非常特别优选至少90%的同源性(关于氨基酸水平的同源性,应参考以上对于多肽给出的详细信息)。有利地,同源性可以在序列的部分区域上更高。
本发明还涉及可通过保守核苷酸取代(即,作为其结果,所讨论的氨基酸被具有相同电荷、大小、极性和/或溶解度的氨基酸取代)获得的序列。
本发明还涉及通过序列多态性从具体公开的核酸衍生的分子。由于天然等位基因变异,这种遗传多态性可能存在于来自不同群体的细胞或来自一个群体内的细胞中。等位基因变体还可包括功能等同物。这些自然变异通常会在基因的核苷酸序列中产生1~5%的变化。所述多态性可以导致本文公开的多肽的氨基酸序列的改变。等位基因变体还可包括功能等同物。
此外,衍生物也应理解为根据本发明的核酸序列的同源物,例如动物、植物、真菌或细菌的同源物,缩短的序列,编码和非编码DNA序列的单链DNA或RNA。例如,在DNA水平上,同源物在本文具体公开的序列中给定的整个DNA区域中具有至少40%,优选至少60%,特别优选至少70%,非常特别优选至少80%的同源性。
此外,衍生物应理解为例如与启动子的融合体。尽管不损害启动子的功能或功效,添加至所述核苷酸序列的启动子可以通过至少一种核苷酸交换、至少一种插入、倒置和/或缺失来修饰。而且,启动子的功效可以通过改变它们的序列来增加,或者可以与更有效的启动子甚至是不同属的生物体的启动子完全交换。
d.功能性多肽突变体的产生
此外,本领域技术人员熟悉用于产生功能性突变体的方法,也就是说,一种核苷酸序列,其编码多肽,该多肽与本文公开的任何与氨基酸相关的SEQ ID NO具有至少40%,45%,50%,55%,60%,65%,70%,75%,80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的序列同一性;和/或由核酸分子编码,该核酸分子包含与本文公开的任何与核苷酸相关的SEQ ID NO具有至少70%序列同一性的核苷酸序列。
取决于所使用的技术,本领域技术人员可以将完全随机的或更有针对性的突变引入到基因或非编码核酸区域(例如对于调节表达很重要)中,并随后产生遗传文库。为此目的所需的分子生物学方法是技术人员已知的,例如描述于Sambrook and Russell,Molecular Cloning.3rd Edition,Cold Spring Harbor Laboratory Press 2001。
修饰基因并由此修饰由其编码的多肽的方法是本领域技术人员长期已知的,举例而言例如:
-位点特异性诱变,其中基因的单个或多个核苷酸以定向方式被替换(Trower MK(Ed.)1996;In vitro mutagenesis protocols.Humana Press,New Jersey),
-饱和诱变,其中任何氨基酸的密码子都可以在基因的任何位点交换或添加(Kegler-Ebo DM,Docktor CM,DiMaio D(1994)Nucleic Acids Res 22:1593;BarettinoD,Feigenbutz M,Valcárel R,Stunnenberg HG(1994)Nucleic Acids Res 22:541;BarikS(1995)Mol Biotechnol 3:1),
-易错聚合酶链反应,其中核苷酸序列被易错DNA聚合酶突变(Eckert KA,KunkelTA(1990)Nucleic Acids Res 18:3739);
-SeSaM法(序列饱和法),其中优选的交换被聚合酶阻止。Schenk et al.,Biospektrum,Vol.3,2006,277-279,
-突变株中的基因传代,其中例如由于DNA修复机制缺陷,核苷酸序列的突变率增加(Greener A,Callahan M,Jerpseth B(1996)An efficient random mutagenesistechnique using an E.coli mutator strain.In:Trower MK(Ed.)In vitromutagenesis protocols.Humana Press,New Jersey),或
-DNA改组,其中形成并消化一组密切相关的基因,并将这些片段用作聚合酶链反应的模板,其中通过重复的链分离和重新结合,最终生成了全长的镶嵌基因(Stemmer WPC(1994)Nature 370:389;Stemmer WPC(1994)Proc Natl Acad Sci USA91:10747)。
使用所谓的定向进化(尤其描述于Reetz MT and Jaeger K-E(1999),TopicsCurr Chem 200:31;Zhao H,Moore JC,Volkov AA,Arnold FH(1999),Methods foroptimizing industrial polypeptides by directed evolution,In:Demain AL,DaviesJE(Ed.)Manual of industrial microbiology and biotechnology.American Societyfor Microbiology),熟练的工人可以以定向的方式大规模生产功能性突变体。为此,在第一步中,首先,例如使用上文给出的方法,产生各自多肽的基因文库。基因文库以合适的方式表达,例如通过细菌或噬菌体展示系统来表达。
表达功能性突变体的宿主生物体的相关基因(其功能在很大程度上与所需的特性相对应)可以提交给另一个突变周期。突变和选择或筛选的步骤可以迭代地重复,直到本发明的功能性突变体具有足够程度的所需特性。使用该迭代过程,可以分阶段进行有限数量的突变,例如1、2、3、4或5个突变,并评估和选择它们对所研究活性的影响。然后可以以相同的方式将所选择的突变体进行进一步的突变步骤。这样,可以显著减少待研究的单个突变体的数量。
根据本发明的结果还提供了与相关多肽的结构和序列有关的重要信息,这是以靶向方式产生具有所需修饰特性的其他多肽所必需的。特别地,可以定义所谓的“热点”,即潜在地适合于通过引入靶向突变来修饰特性的序列区段。
也可以推导出有关氨基酸序列位置的信息,在该区域中可以发生可能对活性几乎没有影响的突变,并且可以将其指定为潜在的“沉默突变”。
e.表达本发明多肽的构建体
在本文语境中,以下定义适用:
“基因的表达”涵盖“异源表达”和“过表达”,并且涉及基因的转录和mRNA向蛋白质的翻译。过表达是指在转基因细胞或生物体中,以mRNA、多肽和/或酶活性水平测量的基因产物的产生超过了相似遗传背景的非转化细胞或生物体中的产生水平。
如本文所用,“表达载体”是指这样一种核酸分子,其使用分子生物学方法和重组DNA技术工程化以将外来或外源DNA递送到宿主细胞中。表达载体典型地包括正确转录核苷酸序列所需的序列。编码区通常编码目的蛋白,但是也可以编码RNA,例如反义RNA、siRNA等。
如本文所用,“表达载体”包括任何线性的或环状的重组载体,包括但不限于病毒载体、噬菌体和质粒。技术人员根据表达系统能够选择适合的载体。在一个实施方案中,表达载体包括本文实施方案的核酸,其可操作地连接到至少一个“调控序列”,其控制转录、翻译、起始和终止,例如转录启动子、操纵子或增强子,或mRNA核糖体结合位点,并且可选地包括至少一个选择标记。当调控序列功能性地涉及本文实施方案的核酸时,核苷酸序列是“可操作地连接的”。
如本文所用,“表达系统”涵盖在给定表达宿主的体内或体外共表达两种或更多种多肽所需的核酸分子的任何组合。各自的编码序列可以位于单个核酸分子或载体上,例如包含多个克隆位点的载体,或位于多顺反子核酸上,或者可以分布在两个或更多个物理上不同的载体上。
如本文所用,术语“进行扩增(amplifying)”和“扩增(amplification)”是指使用任何合适的扩增方法用于产生或检测天然表达的核酸的重组体,如下文详细描述的。例如,本发明提供用于扩增(例如,通过聚合酶链反应,PCR)天然表达的(例如,基因组DNA或mRNA)或本发明在体内、离体或体外的重组的核酸(例如cDNA)的方法和试剂(例如,特异性简并寡核苷酸引物对,寡聚dT引物)。
“调控序列”是指这样一种核酸序列,其确定本文实施方案的核酸序列的表达水平、并且能够调控可操作地连接到该调控序列的核酸序列的转录速率。调控序列包含启动子、增强子、转录因子、启动子元件等。
根据本发明,“启动子”、“具有启动子活性的核酸”或“启动子序列”应理解为是指这样一种核酸,当与要转录的核酸功能性连接时,其调节所述核酸的转录。“启动子”尤其是指一种核酸序列,其通过提供RNA聚合酶用的结合位点以及适合转录所需的其它因子,包括但不限于转录因子结合位点、抑制子和活化子蛋白结合位点,来控制编码序列的表达。术语启动子的含义还包括术语“启动子调控序列”。启动子调控序列可以包括可能影响转录、RNA加工或相关编码核酸序列的稳定性的上游和下游元件。启动子包括天然来源的和合成的序列。编码核酸序列通常位于启动子相对于以转录起始位点为起始的转录方向的下游。
在本文语境中,“功能性”或“可操作地”连接被理解为例如指具有调控序列的核酸之一的顺序排列。例如,具有启动子活性的序列,和待转录的核酸序列以及可选的其他调控元件(例如确保核酸转录的核酸序列)和例如终止子,该顺序排列的方式为使得每个调控元件都能在核酸序列转录后执行其功能。这不一定需要化学意义上的直接连接。遗传控制序列,例如增强子序列,甚至可以从更远的位置甚至从其他DNA分子上对目标序列发挥作用。优选的排列是这样的,其中待转录的核酸序列位于启动子序列的下游(即3'端),从而使两个序列共价连接在一起。启动子序列和待重组表达的核酸序列之间的距离可以小于200个碱基对,或小于100个碱基对或小于50个碱基对。
除启动子和终止子外,还可以提及以下作为其他调控元件的例子:靶向序列,增强子,聚腺苷酸化信号,选择标记,扩增信号,复制起点等。合适的调节序列描述于例如Goeddel,Gene Expression Technology:Methods in Enzymology 185,Academic Press,San Diego,CA(1990)。
术语“组成型启动子”是指不受调控的启动子,其允许其可操作地连接的核酸序列的持续转录。
如本文所用,术语“可操作地连接”是指处于功能性关系的多核苷酸元件的连接。当核酸与另一核酸序列处于功能性关系时,那么该核酸是“可操作地连接”的。例如,如果启动子或者转录调控序列能够影响编码序列的转录,那么该启动子或者转录调控序列是可操作地连接到该编码序列的。可操作地连接意味着被连接的DNA序列通常是邻接的。与启动子序列有关的核苷酸序列相对于要被转化的植物可以是同源或异源来源的。所述序列还可以是完全或部分合成的。不管来源如何,与启动子序列有关的核酸序列将根据在结合到本文实施方案的多肽后所连接的启动子性质而表达或沉默。相关核酸在所有时间或替代地在特定时间在整个生物体中或在特定组织、细胞或细胞室中可以编码需要表达或抑制的蛋白。此种核苷酸序列特别地编码将所需表型性状赋予给由其改变或转化的宿主细胞或生物体的蛋白质。更特别地,相关的核苷酸序列导致在细胞或生物体中产生如本文定义的一种或多种目的产物。特别地,核苷酸序列编码具有如本文定义的酶活性的多肽。
本文如上所述的核苷酸序列可以是“表达盒”的一部分。术语“表达盒”和“表达构建体”同义使用。(优选的重组)表达构建体包含这样的核苷酸序列,其编码根据本发明的多肽并且在调节核酸序列的遗传控制之下。
在根据本发明应用的方法中,表达盒可以是“表达载体”,特别是重组表达载体的一部分。
根据本发明,“表达单位”应理解为是指具有表达活性的核酸,其包含如本文所定义的启动子,并且在与待表达的核酸或基因功能性连接后调节表达,即所述核酸或所述基因的转录和翻译。因此在这方面也被称为“调节核酸序列”。除启动子外,还可以存在其他调节元件,例如增强子。
根据本发明,“表达盒”或“表达构建体”应理解为在功能上与要表达的核酸或要表达的基因连接的表达单元。因此,与表达单元相反,表达盒不仅包含调节转录和翻译的核酸序列,而且还包含由于转录和翻译而作为蛋白质表达的核酸序列。
在本发明的语境中,术语“表达”或“过表达”描述了微生物中一种或多种由相应DNA编码的多肽的细胞内活性的产生或增加。为此,例如可以将基因导入到生物体中,用另一个基因替代现有基因,增加基因的拷贝数,使用强启动子或使用编码具有高活性的相应多肽的基因。可选地,这些措施可以组合。
优选地,根据本发明的此类构建体包含各自编码序列5'上游的启动子和3'下游的终止子序列,以及可选地其他常见的调控元件,在每种情况下均与编码序列可操作地连接。
根据本发明的核酸构建体特别地包含编码多肽的序列,该多肽例如衍生自如本文所述的氨基酸相关的SEQ ID NO或其反向互补序列,或其衍生物和同源物,并且已经与一个或多个调节信号可操作地或功能性连接,用于有利地控制例如增加基因表达。
除了这些调控序列之外,这些序列的天然调控可能仍存在于实际的结构基因之前,并且可选地可能已经进行了遗传修饰,因此天然调控已被关闭,基因的表达得到了增强。然而,核酸构建体也可以具有更简单的构建,即,在编码序列之前没有插入额外的调节信号,并且具有调节作用的天然启动子尚未去除。相反,天然调节序列被突变,使得不再发生调节并且基因表达增加。
优选的核酸构建体有利地还包含与启动子功能性连接的一个或多个已经提及的“增强子”序列,该序列使得增强核酸序列的表达成为可能。还可以在DNA序列的3'末端插入其他有利的序列,例如其他调控元件或终止子。根据本发明的核酸的一个或多个拷贝可以存在于构建体中。在该构建体中,还可以可选地存在其他标记物,例如与营养缺陷性或抗生素抗性互补的基因,以便选择该构建体。
合适的调控序列的例子存在于启动子中,例如cos、tac、trp、tet、trp-tet、lpp、lac、lpp-lac、lacI
为了在宿主生物体中表达,将核酸构建体有利地插入到载体,例如质粒或噬菌体中,这使得基因在宿主中的最佳表达成为可能。除了质粒和噬菌体,载体还应理解为是本领域技术人员已知的所有其他载体,即例如病毒,例如SV40、CMV、杆状病毒和腺病毒、转座子、IS元件、噬粒、粘粒和线性或环状DNA或人工染色体。这些载体能够在宿主生物体中自主复制或通过染色体复制。这些载体是本发明的进一步发展。二元或cpo整合载体也是适用的。
合适的质粒是例如在大肠杆菌pLG338、pACYC184、pBR322、pUC18、pUC19、pKC30、pRep4、pHS1、pKK223-3、pDHE19.2、pHS2、pPLc236、pMBL24、pLG200、pUR290、pIN-III
在载体的进一步开发中,包含本发明的核酸构建体或本发明的核酸的载体也可以有利地以线性DNA的形式引入到微生物中并通过异源或同源重组整合到宿主生物体的基因组中。该线性DNA可以由线性化的载体例如质粒组成,或者仅由本发明的核酸构建体或核酸组成。
为了在生物体中异源基因的最佳表达,有利的是修饰核酸序列以匹配生物体中使用的特定“密码子使用”。“密码子使用”可以通过对所讨论的生物体的其他已知基因的计算机评估来容易地确定。
根据本发明的表达盒通过将合适的启动子融合到合适的编码核苷酸序列和终止子或聚腺苷酸化信号来产生。为此目的使用常规的重组和克隆技术,例如描述于T.Maniatis,E.F.Fritsch and J.Sambrook,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,Cold Spring Harbor,NY(1989)和T.J.Silhavy,M.L.Berman and L.W.Enquist,Experiments with Gene Fusions,Cold Spring HarborLaboratory,Cold Spring Harbor,NY(1984)和Ausubel,F.M.et al.,Current Protocolsin Molecular Biology,Greene Publishing Assoc.and Wiley Interscience(1987)。
为了在合适的宿主生物体中表达,将重组核酸构建体或基因构建体有利地插入到宿主特异性载体中,这使得基因在宿主中的最佳表达成为可能。载体是技术人员众所周知的,并且可以在例如“cloning vectors”(Pouwels P.H.et al.,Ed.,Elsevier,Amsterdam-New York-Oxford,1985)中找到。
本文实施方案的替代实施方案提供了一种“改变宿主细胞中的基因表达”的方法。例如,在某些语境下(例如,暴露于一定温度或培养条件下),在宿主细胞或宿主生物体中可以增强或过表达或诱导本文实施方案的多核苷酸。
本文提供的多核苷酸的表达的改变还会产生异位表达,其是在改变的以及在对照或野生型生物体中的一种不同的表达模式。表达的改变是由本文一个实施方案的多肽与外源性或内源性调节剂的接触而发生的或者是由于多肽的化学修饰导致的。该术语还指本文实施方案的多核苷酸的改变的表达模式,其被改变至低于检测水平或者完全被抑制活性。
本文提供的一个实施方案还为编码本文提供的多肽或变体多肽的分离的、重组的或合成的多核苷酸。
在一个实施方案中,多种编码多肽的核酸序列在单一宿主中共表达,特别是在不同启动子的控制下。在另一个实施方案中,多种编码多肽的核酸序列可以存在于单个转化载体上,或者可以使用分离的载体并选择包含两个嵌合基因的转化体同时进行共转化。类似地,一种或多种多肽编码基因可以与其他嵌合基因一起在单一植物、细胞、微生物或生物体中表达。
f.适用于本发明的宿主
取决于语境,术语“宿主”可以指野生型宿主或经遗传改变的重组宿主或两者。
原则上,所有的原核或真核生物都可以被认为是根据本发明的核酸或核酸构建体的宿主或重组宿主生物体。
使用根据本发明的载体,可以生产重组宿主,其例如可以用至少一种根据本发明的载体转化,并可以用于生产根据本发明的多肽。有利地,将如上所述的根据本发明的重组构建体引入到合适的宿主系统中并表达。优选地,使用本领域技术人员已知的普通克隆和转染方法,例如共沉淀、原生质体融合、电穿孔、逆转录病毒转染等,以在各自的表达系统中表达所述核酸。合适的系统描述于Current Protocols in Molecular Biology,F.Ausubelet al.,Ed.,Wiley Interscience,New York 1997,或Sambrook et al.MolecularCloning:A Laboratory Manual.2nd edition,Cold Spring Harbor Laboratory,ColdSpring Harbor Laboratory Press,Cold Spring Harbor,NY,1989。
有利地,诸如细菌、真菌或酵母的微生物被用作宿主生物体。有利地,使用革兰氏阳性或革兰氏阴性细菌,优选肠杆菌科(Enterobacteriaceae),假单胞菌科(Pseudomonadaceae),根瘤菌科(Rhizobiaceae),链霉菌科(Streptomycetaceae),链球菌科(Streptococcaceae)或诺卡氏菌科(Nocardiaceae)的细菌,特别优选埃希氏菌属(Escherichia),假单胞菌属(Pseudomonas),链霉菌属(Streptomyces),乳球菌属(Lactococcus),诺卡氏菌(Nocardia),伯克霍尔德氏菌属(Burkholderia),沙门氏菌属(Salmonella),农杆菌属(Agrobacterium),艰难梭菌(Clostridium)或红球菌属(Rhodococcus)的细菌。大肠杆菌(Escherichia coli)属和种是非常特别优选的。此外,在α-变形菌(alpha-Proteobacteria)、β-变形菌(beta-Proteobacteria)或γ-变形菌(gamma-Proteobacteria)组中发现了其他有利的细菌。有利地,诸如酵母属(Saccharomyces)或毕赤酵母(Pichia)家族的酵母也是合适的宿主。
或者,整个植物或植物细胞可以用作天然或重组宿主。作为非限制性例子,可以提及以下植物或自其衍生的细胞:烟草属(Nicotiana),特别是本氏烟草(Nicotianabenthamiana)和普通烟草(Nicotiana tabacum)(tobacco);以及拟南芥属(Arabidopsis),特别是阿拉伯芥(Arabidopsis thaliana)。
取决于宿主生物体,根据本发明的方法中使用的生物体以本领域技术人员已知的方式生长或培养。培养可以分批、半分批或连续进行。营养物可以在发酵开始时给予,也可以稍后,半连续或连续地提供。这也在下面更详细地描述。
g.根据本发明的多肽的重组生产
本发明进一步涉及重组生产根据本发明的多肽或其功能性生物学活性片段的方法,其中培养产生多肽的微生物,可选地通过施加至少一种诱导基因表达的诱导剂来诱导多肽的表达,并从培养物中分离这些多肽。如果需要,多肽也可以这种方式以工业规模生产。
根据本发明产生的微生物可以分批法或补料分批法或重复补料分批法连续或不连续培养。已知培养方法的概述可在Chmiel的教科书(Bioprozesstechnik 1.Einführungin die Bioverfahrenstechnik[Bioprocess technology 1.Introduction tobioprocess technology](Gustav Fischer Verlag,Stuttgart,1991))或在Storhas的教科书(Bioreaktoren und periphere Einrichtungen[Bioreactors and peripheralequipment](Vieweg Verlag,Braunschweig/Wiesbaden,1994))中找到。
所使用的培养基必须适当地满足各个菌株的要求。在美国细菌学学会(AmericanSociety for Bacteriology(Washington D.C.,USA,1981))的手册“Manual of Methodsfor General Bacteriology”中给出了各种微生物的培养基的描述。
可以根据本发明使用的这些培养基通常包含一种或多种碳源、氮源、无机盐、维生素和/或微量元素。
优选的碳源是糖,例如单糖、二糖或多糖。很好的碳源是例如葡萄糖,果糖,甘露糖,半乳糖,核糖,山梨糖,核酮糖,乳糖,麦芽糖,蔗糖,棉子糖,淀粉或纤维素。糖也可以通过复杂的化合物(例如糖蜜)或糖精制的其他副产品添加到培养基中。添加不同碳源的混合物也是有利的。其他可能的碳源是油和脂,例如大豆油,葵花籽油,花生油和椰子油,脂肪酸例如棕榈酸,硬脂酸或亚油酸,醇例如甘油,甲醇或乙醇,和有机酸,例如乙酸或乳酸。
氮源通常是有机或无机氮化合物或包含这些化合物的材料。氮源的例子包括氨气或铵盐,例如硫酸铵,氯化铵,磷酸铵,碳酸铵或硝酸铵,硝酸盐,尿素,氨基酸或复合氮源,例如玉米浆,大豆粉,大豆蛋白,酵母提取物,肉提取物等。氮源可以单独使用或混合使用。
可以存在于培养基中的无机盐化合物包括钙,镁,钠,钴,钼,钾,锰,锌,铜和铁的氯化物、磷或硫酸盐。
无机含硫化合物,例如硫酸盐,亚硫酸盐,连二亚硫酸盐,四硫酸盐,硫代硫酸盐,硫化物,以及有机硫化合物,例如硫醇(mercaptans)和巯类(thiols),可用作硫源。
磷酸、磷酸二氢钾或磷酸氢二钾或相应的含钠盐可用作磷源。
可以将螯合剂添加到培养基中,以将金属离子保持在溶液中。特别合适的螯合剂包括二羟基苯酚,例如儿茶酚或原儿茶酸酯,或有机酸,例如柠檬酸。
根据本发明使用的发酵培养基通常还包含其他生长因子,例如维生素或生长促进剂,其包括例如生物素,核黄素,硫胺素,叶酸,烟酸,泛酸和吡哆醇(pyridoxine)。生长因子和盐通常源自复杂培养基的成分,例如酵母提取物,糖蜜,玉米浆等。此外,可以将合适的前体添加到培养基中。化合物在培养基中的确切组成在很大程度上取决于相应的实验,并针对每种具体情况分别确定。有关培养基优化的信息可以在教科书"AppliedMicrobiol.Physiology,A Practical Approach"(Ed.P.M.Rhodes,P.F.Stanbury,IRLPress(1997)p.53-73,ISBN 0 19 963577 3)中找到。生长培养基也可以从商业供应商获得,例如Standard 1(Merck)或BHI(脑心浸液,DIFCO)等。
通过加热(在1.5bar和121℃下20分钟)或通过无菌过滤对培养基的所有成分进行灭菌。这些成分可以一起消毒,也可以根据需要单独消毒。培养基的所有成分都可以在培养开始时给予,也可以连续或分批添加。
培养物的温度通常在15℃至45℃之间,优选25℃至40℃,并且在实验过程中可以改变或保持恒定。介质的pH应在5至8.5的范围内,优选为7.0左右。生长期间的pH值可以通过添加碱性化合物(例如氢氧化钠,氢氧化钾,氨或氨水)或酸性化合物(例如磷酸或硫酸)来控制。消泡剂例如脂肪酸聚乙二醇酯可用于控制发泡。为了维持质粒的稳定性,可以向培养基中添加合适的选择性物质例如抗生素。为了维持有氧条件,将氧气或含氧气体混合物(例如环境空气)供入培养物中。培养物的温度通常在20℃至45℃的范围内。继续培养直至形成最大量的所期望产物。通常会在10到160个小时内达到此目标。
然后将发酵液进一步处理。根据需要,可以通过分离技术,例如离心、过滤、倾析或这些方法的组合,将生物质完全或部分地从发酵液中除去,或者可以完全留在其中。
如果多肽没有在培养基中分泌,那么细胞也可以被裂解,并且可以通过用于分离蛋白质的已知方法从裂解物中获得产物。可以可选地通过高频超声,高压例如在高压细胞裂解机(French press)中,通过渗透,通过去污剂、裂解酶或有机溶剂的作用,通过均化器或通过上述几种方法的组合来破坏细胞。
可以通过已知的色谱技术,例如分子筛色谱(凝胶过滤),例如Q-琼脂糖色谱,离子交换色谱和疏水色谱,以及其他常规技术,例如超滤、结晶、盐析、渗析和天然凝胶电泳来纯化多肽。合适的方法描述于例如Cooper,T.G.,Biochemische Arbeitsmethoden[Biochemical processes],Verlag Walter de Gruyter,Berlin,New York,或Scopes,R.,Protein Purification,Springer Verlag,New York,Heidelberg,Berlin。
为了分离重组蛋白,使用载体系统或寡核苷酸可能是有利的,所述载体系统或寡核苷酸通过限定的核苷酸序列延长cDNA,并因此编码改变的多肽或融合蛋白,其例如用于更容易的纯化。这种类型的合适修饰例如是充当锚的所谓“标签”,例如可以被识别为抗体抗原的被称为六-组氨酸锚或表位的修饰(例如,描述于Harlow,E.and Lane,D.,1988,Antibodies:A Laboratory Manual.Cold Spring Harbor(N.Y.)Press)。这些锚可以用于将蛋白质连接至固体载体,例如聚合物基质,其可以例如用作色谱柱中的填料,或者可以用于微量滴定板或其他载体上。
同时,这些锚也可用于识别蛋白质。为了识别蛋白质,还可以使用通常的标记物,例如荧光染料,酶标记物(与底物反应后形成可检测的反应产物),或放射性标记物,单独使用或与锚结合使用以衍生化蛋白质。
h.多肽的固定化
根据本发明的酶或多肽可以在本文描述的方法中以游离形式或固定化而使用。固定化酶是固定在惰性载体上的酶。合适的载体材料和固定在其上的酶从EP-A-1149849,EP-A-1069183和DE-OS100193773以及从其中引用的参考文献中已知。在这方面,参考这些文件的全部公开内容。合适的载体材料包括例如粘土,粘土矿物,例如高岭石,硅藻土,珍珠岩,二氧化硅,氧化铝,碳酸钠,碳酸钙,纤维素粉末,阴离子交换剂材料,合成聚合物,例如聚苯乙烯,丙烯酸树脂,酚醛树脂,聚氨酯和聚烯烃,例如聚乙烯和聚丙烯。为了制备负载的酶,通常以细分的颗粒形式,优选多孔形式使用载体材料。载体材料的粒径通常不大于5mm,特别是不大于2mm(粒径分布曲线)。类似地,当使用脱氢酶作为全细胞催化剂时,可以选择游离形式或固定形式。载体材料例如为海藻酸钙和角叉菜胶。酶和细胞也可以直接与戊二醛交联(与CLEAs交联)。相应的和其他固定化技术描述于例如J.Lalonde and A.Margolin"Immobilization of Enzymes"in K.Drauz and H.Waldmann,Enzyme Catalysis inOrganic Synthesis 2002,Vol.III,991-1032,Wiley-VCH,Weinheim中。Rehm et al.(Ed.)Biotechnology,2nd Edn,Vol 3,Chapter 17,VCH,Weinheim给出了用于进行根据本发明的方法的生物转化和生物反应器的进一步信息。
i.本发明生物催化生产方法的反应条件
本发明的反应可以在体内或体外条件下进行。
存在于本发明的方法或上文定义的多步方法的单个步骤中的至少一种多肽/酶可以天然存在于活细胞中,或于收获的细胞(即在体内条件下)中,死细胞中,透化细胞中,粗细胞提取物中,纯化提取物中,或以基本纯净或完全纯净的形式(即在体外条件下),重组产生一种或多种酶。所述至少一种酶可以以溶液形式存在或以固定在载体上的酶形式存在。一种或几种酶可以同时以可溶性和/或固定化形式存在。
根据本发明的方法可以在本领域技术人员已知的普通反应器中进行,并且可以在不同的规模范围内进行,例如从实验室规模(几毫升到几十升反应体积)到工业规模(几升到数千立方米反应体积)。如果多肽以通过无生命的、可选地透化的细胞包封的形式,以或多或少纯化的细胞提取物的形式或以纯化的形式使用,则可以使用化学反应器。化学反应器通常允许控制至少一种酶的量,至少一种底物的量,pH,温度和反应介质的循环。当活细胞中存在至少一种多肽/酶时,该过程将是发酵。在这种情况下,生物催化生产将在生物反应器(发酵罐)中进行,其中对于活细胞合适的生存条件必需的参数(例如,具有营养的培养基,温度,通气,有氧或无氧或其他气体,抗生素等)可以控制。本领域技术人员熟悉化学反应器或生物反应器,例如使用将化学或生物技术方法从实验室规模扩大到工业规模或优化工艺参数的程序,这些方法在文献中也有广泛描述(有关生物技术方法,请参见例如Crueger und Crueger,Biotechnologie–Lehrbuch der angewandten Mikrobiologie,2.Ed.,R.Oldenbourg Verlag,München,Wien,1984)。
包含至少一种酶的细胞可以通过物理或机械方式例如超声或射频脉冲,高压细胞裂解机(French press)或化学方式例如在培养基中存在的低渗介质、裂解酶和去污剂或这些方法的组合来渗透。洗涤剂的例子是洋地黄毒苷,正十二烷基麦芽糖苷,辛基糖苷,
代替活细胞,也可以将含有所需生物催化剂的非活细胞的生物质应用于本发明的生物转化反应。
如果固定了至少一种酶,则将其如上所述连接至惰性载体。
转化反应可以分批、半分批或连续进行。反应物(和可选的营养物)可以在反应开始时提供,或者可以随后半连续或连续地提供。
根据特定的反应类型,本发明的反应可以在水性、水性-有机或非水性反应介质中进行。
水性或水性-有机介质可包含合适的缓冲液,以将pH值调整为5至11,例如6至10。
在水性-有机介质中,可以使用与水可混溶、部分混溶或不混溶的有机溶剂。合适的有机溶剂的非限制性例子在下面列出。进一步的例子是一元或多元,芳族或脂族醇,特别是多元脂族醇,如甘油。
非水介质可以包含基本上不含水,即,将包含少于约1重量%或0.5重量%的水。
生物催化方法也可以在有机非水介质中进行。作为合适的有机溶剂,可以提及具有例如5至8个碳原子的脂族烃,例如戊烷,环戊烷,己烷,环己烷,庚烷,辛烷或环辛烷;芳族烃,例如苯,甲苯,二甲苯,氯苯或二氯苯,脂族无环和醚,例如二乙醚,甲基叔丁基醚,乙基叔丁基醚,二丙基醚,二异丙醚,二丁基醚;或它们的混合物。
反应物/底物的浓度可以适应于最佳反应条件,这可以取决于所应用的特定酶。例如,初始底物浓度可以为0.1至0.5M,例如10至100mM。
反应温度可以适应于最佳反应条件,这可以取决于所应用的特定酶。例如,该反应可以在0至70℃的温度下进行,例如20至50或25至40℃。反应温度的例子是约30℃,约35℃,约37℃,约40℃,约45℃,约50℃,约55℃和约60℃。
该工艺可以继续进行直到在底物和随后的产物之间达到平衡为止,但是可以更早地停止。通常的工艺时间为1分钟至25小时,特别是10分钟至6小时,例如为1小时至4小时,特别是1.5小时至3.5小时。这些参数是合适的工艺条件的非限制性例子。
如果宿主是转基因植物,则可以提供最佳的生长条件,例如最佳的光照、水和营养条件。
用于进行乙酸补身烷基酯化合物的制备的特定反应条件如下。在20至35℃和4至7的pH值下孵育的水性环境中,乙酰转移酶可以作为经纯化的多肽或全细胞系统存在。底物浓度可以在10至100mM之间变化。
k.产品分离
本发明的方法可以进一步包括回收终产物或中间产物的步骤,所述终产物或中间产物可选地为立体异构体或对映异构体的基本纯净的形式。术语“回收”包括从培养基或反应介质中提取、收获、分离或纯化化合物。化合物的回收可以根据本领域已知的任何常规分离或纯化方法进行,包括但不限于用常规树脂(例如,阴离子或阳离子交换树脂,非离子吸附树脂等)处理,用常规吸附剂(例如,活性炭,硅酸,硅胶,纤维素,氧化铝等)处理,pH值的改变,溶剂萃取(例如,使用常规溶剂,例如醇,乙酸乙酯,己烷等),蒸馏,渗析,过滤,浓缩,结晶,重结晶,pH调节,冻干等。
经分离产物的身份和纯度可以通过已知技术确定,例如高效液相色谱(HPLC),气相色谱(GC),光谱学(例如IR,UV,NMR),着色方法,TLC,NIRS,酶或微生物测定(参见例如:Patek et al.(1994)Appl.Environ.Microbiol.60:133-140;Malakhova et al.(1996)Biotekhnologiya 1127-32;und Schmidt et al.(1998)Bioprocess Engineer.19:67-70.Ullmann's Encyclopedia of Industrial Chemistry(1996)Bd.A27,VCH:Weinheim,S.89-90,S.521-540,S.540-547,S.559-566,575-581und S.581-587;Michal,G(1999)Biochemical Pathways:An Atlas of Biochemistry and Molecular Biology,JohnWiley and Sons;Fallon,A.et al.(1987)Applications of HPLC in Biochemistry in:Laboratory Techniques in Biochemistry and Molecular Biology,Bd.17.)。
可以将本文所述的任何方法生产的环状萜烯化合物转化成衍生物,例如但不限于烃,酯,酰胺,糖苷,醚,环氧化物,醛,酮,醇,二醇,缩醛或缩酮。萜烯化合物衍生物可以通过化学方法获得,例如但不限于氧化,还原,烷基化,酰化和/或重排。或者,萜烯化合物衍生物可以通过使用生化方法通过使萜烯化合物与酶接触而获得,所述酶例如但不限于氧化还原酶,单加氧酶,双加氧酶,转移酶。可以使用分离的酶,来自裂解细胞的酶在体外进行生化转化,也可以使用全细胞在体内进行生化转化。
l.乙酸补身烷基醇的发酵产生
本发明还涉及用于发酵产生乙酸补身烷基酯的方法。
根据本发明使用的发酵可以例如在搅拌的发酵罐、鼓泡塔和回路反应器中进行。有关可能的方法类型的全面概述,包括搅拌器类型和几何设计,请参见“Chmiel:Bioprozesstechnik:Einfuhrung in die Bioverfahrenstechnik,Band 1”。在本发明的方法中,可用的典型变型是本领域技术人员已知的或例如在“Chmiel,Hammes and Bailey:Biochemical Engineering”中解释的以下变型,例如分批、补料分批,重复补料分批进行或连续发酵,有或没有回收生物质。取决于生产应变,可以进行空气,氧气,二氧化碳,氢气,氮气或适当的气体混合物的喷射,以实现良好的收率(YP/S)。
要使用的培养基必须以适当的方式满足特定菌株的要求。在美国细菌学学会(American Society for Bacteriology(Washington D.C.,USA,1981))的手册“Manual ofMethods for General Bacteriology”中给出了各种微生物的培养基的描述。
可以根据本发明使用的这些培养基通常包含一种或多种碳源、氮源、无机盐、维生素和/或微量元素。
优选的碳源是糖,例如单糖、二糖或多糖。非常好的碳源是例如葡萄糖,果糖,甘露糖,半乳糖,核糖,山梨糖,核酮糖,乳糖,麦芽糖,蔗糖,棉子糖,淀粉或纤维素。糖也可以通过复杂的化合物(例如糖蜜)或糖精制的其他副产物添加到培养基中。添加各种碳源的混合物也是有利的。碳的其他可能来源是油和脂,例如大豆油,葵花籽油,花生油和椰子油,脂肪酸例如棕榈酸,硬脂酸或亚油酸,醇例如甘油,甲醇或乙醇,和有机酸,例如乙酸或乳酸。
氮源通常是有机或无机氮化合物或含有这些化合物的材料。氮源的例子包括氨气或铵盐,例如硫酸铵,氯化铵,磷酸铵,碳酸铵或硝酸铵,硝酸盐,尿素,氨基酸或复合氮源,例如玉米浆,大豆粉,大豆蛋白,酵母提取物,肉提取物等。氮源可以单独使用或混合使用。
可以存在于培养基中的无机盐化合物包括钙,镁,钠,钴,钼,钾,锰,锌,铜和铁的氯化物,磷酸盐或硫酸盐。
无机含硫化合物,例如硫酸盐,亚硫酸盐,连二亚硫酸盐,四硫酸盐,硫代硫酸盐,硫化物,以及有机硫化合物,例如硫醇(mercaptans)和巯类(thiols),可用作硫源。
磷酸、磷酸二氢钾或磷酸氢二钾或相应的含钠盐可用作磷源。
可以将螯合剂添加到培养基中,以将金属离子保持在溶液中。特别合适的螯合剂包括二羟基苯酚,例如儿茶酚或原儿茶酸酯,或有机酸,例如柠檬酸。
根据本发明使用的发酵培养基还可包含其他生长因子,例如维生素或生长促进剂,其包括例如生物素,核黄素,硫胺素,叶酸,烟酸,泛酸和吡哆醇。生长因子和盐通常来自培养基的复杂成分,例如酵母提取物,糖蜜,玉米浆等。另外,可以将合适的前体添加到培养基中。化合物在培养基中的精确组成在很大程度上取决于特定的实验,必须针对每种特定情况分别确定。有关培养基优化的信息可以在教科书"Applied Microbiol.Physiology,APractical Approach"(1997)中找到。生长培养基也可以从商业供应商那里获得,例如Standard 1(Merck)或BHI(脑心浸液,DIFCO)等。。
通过加热(在1.5bar和121℃下20分钟)或通过无菌过滤对培养基的所有成分进行灭菌。这些成分可以一起消毒,也可以根据需要单独消毒。培养基的所有成分都可以在生长开始时给予,或者可以选择连续添加或分批添加。
培养物的温度通常在15℃至45℃之间,优选25℃至40℃,并且可以在实验期间保持恒定或可以变化。介质的pH值应在5至8.5的范围内,优选7.0左右。生长期间的pH值可以通过添加碱性化合物(例如氢氧化钠,氢氧化钾,氨或氨水)或酸性化合物(例如磷酸或硫酸)来控制。消泡剂例如脂肪酸聚乙二醇酯可用于控制发泡。为了维持质粒的稳定性,可以向培养基中添加具有选择性作用的合适物质例如抗生素。为了维持有氧条件,将氧气或含氧气体混合物(例如环境空气)供入培养物中。培养物的温度通常为20℃至45℃。继续培养直至形成最大量的所期望产物。通常会在1到160个小时内达到此目标。
本发明的方法可以进一步包括回收所述乙酸补身烷基酯的步骤。
术语“回收”包括从培养基中提取、收获、分离或纯化化合物。化合物的回收可以根据本领域已知的任何常规分离或纯化方法进行,包括但不限于用常规树脂(例如,阴离子或阳离子交换树脂,非离子吸附树脂等)处理,用常规吸附剂(例如,活性炭,硅酸,硅胶,纤维素,氧化铝等)处理,pH值的改变,溶剂萃取(例如,使用常规溶剂,例如醇,乙酸乙酯,己烷等),蒸馏,渗析,过滤,浓缩,结晶,重结晶,pH调节,冻干等。
在预期的分离之前,可以除去发酵液的生物质。去除生物质的方法是本领域技术人员已知的,例如过滤、沉降和浮选。因此,可以例如通过离心机、分离器、倾析器、过滤器或在浮选设备中去除生物质。为了最大程度地回收有价值的产品,通常建议洗涤生物质,例如以渗滤的形式。方法的选择取决于发酵液中生物质的含量和生物质的性质,以及生物质与有价值产品的相互作用。
在一个实施方案中,可以将发酵液灭菌或巴氏灭菌。在另一个实施方案中,将发酵液浓缩。根据需要,该浓缩可以分批或连续进行。应该选择压力和温度范围,使得首先不会发生产品损坏,其次需要最小化设备和能源的使用。多级蒸发的压力和温度水平的熟练选择尤其可以节省能源。
以下实施例仅是说明性的,并不意味着限制本文所述的权利要求和实施方案的范围。
在考虑了本文提供的公开内容之后,对于本领域技术人员而言将立即变得显而易见的多种可能的变型方案也落入本发明的范围内。
材料:
除非另有说明,否则本文使用的所有化学和生物化学材料以及微生物或细胞均为可商购的产品。
除非另有说明,否则重组蛋白是通过标准方法克隆和表达的,所述方法例如描述于Sambrook,J.,Fritsch,E.F.and Maniatis,T.,Molecular cloning:A LaboratoryManual,2
气相色谱质谱法(GC-MS)
Agilent Intuvo 9000系列气相色谱系统配备DB-5MS UI色谱柱(10m×0.25mm×0.25μm膜厚(定制色谱柱,由Agilent Technologies Inc,Santa Clara,CA制),气相色谱通过1:1的Detector Splitter Chip(G4588-60502,Agilent Technologies Inc,SantaClara,CA)与两个检测器相连。,第一个检测器是Agilent 5977B系列质谱仪,而第二个检测器是标准的Intuvo 9000火焰离子化检测器(FID)。载气为氦气,流速为2.5ml/min。进样器以分流(1:100)模式进样,进样器温度设置为240℃。烘箱温度从150℃(保持0.1分钟)编程为以40℃/min的速度升至240℃,然后以180℃/min的温度升至325℃,并保持0.5分钟)。
乙酰基转移酶构成了遗传上多样化的一类酶,已知的代表超过8,000种(PFAM数据库:PF02458转移酶家族)。尽管被乙酰基转移酶接受为底物的分子的库很大,但据报道没有一种接受倍半萜醇作为底物。为了鉴定能够乙酰化补身烷型倍半萜醇的乙酰基转移酶,基于以下原理从数千种已知的源自植物、真菌和细菌的乙酰基转移酶中选择了五十四种(表1):
从植物乙酰基转移酶BAHD家族中获知的五个进化枝中的两种——进化枝3和5——的鉴定出的成员是用乙酰辅酶A作为酰基供体对生物碱和萜类化合物进行乙酰化(Curr Opin Plant Biol.2006,9(3):331-40)。另外,来自进化枝3和5的乙酰基转移酶使用的一些底物是大体积多环的并且带有空间位阻醇基团作为酰基受体(BMC Genomics 2011,12:236;Curr Opin Plant Biol.2006,9:331-40;Elife.2017Mar14;6:e23001;Planta.2015,242:709-19)。基于此,从植物BAHD家族进化枝3和5中选择了二十一种候选物,这些候选物包括已鉴定出的参与多环二萜紫杉醇(taxol)生物合成的乙酰基转移酶(Proc Natl Acad Sci U S A.2000,18;97(2):583–587)和参与双环赖百当烷二萜毛喉素(Forskolin)的生物合成的来自毛喉鞘蕊花(Plectranthus barbatus)的两种乙酰基转移酶(Elife 2017,14;6:e23001)。
与候选植物相似,根据其公认的或实验确定的接受乙酰辅酶A作为酰基供体的能力及其底物的结构特征,选择了七种真菌乙酰基转移酶:带有空间位阻醇基团的大体积多环化合物(FEMS Microbiol Lett.2005,251:193-201;Chembiochem.2009,10:2325-8;Biotechnol Biotechnol Equip.2014,28(5):818-826;Nat Chem.2010,2:858-64)。其中,选择了参与芳基酸酯化的补身烷型倍半萜内酯astellolide(Sci Rep.2016,6:32865)的生物合成的蛋白AstG(来自米曲霉(Aspergillus oryzae),NCBI登录号XP_023091083.1)。
另外,使用PSI Blast搜索Trends Biochem Sci.2002,27:161-4.)(标准参数和二次迭代),基于其与AstG的序列相似性,从NCBI蛋白数据库中检索出二十一种推定的真菌乙酰基转移酶。使用NCBI Blast Tree View功能在系统树上可视化显示结果,以显示AstG与500PSI-Blast衍生序列之间的同源性。通过与查询序列的同源性选择候选物,这些查询序列来自系统树的单个分支。
最后,氯霉素乙酰基转移酶是混杂酶,能够容纳比氯霉素更大体积的底物(Protein Sci.2012,21(4):520–530)。因此,从NCBI蛋白质数据库中选出五种1至3类细菌氯霉素乙酰基转移酶作为假定的氯霉素乙酰基转移酶或从文献(Biochem J.1990,272:505-10)中选择。
表1:为将补身烷倍半萜转化为补身烷基乙酰化倍半萜而选择的乙酰基转移酶
筛选每一种酶候选物,以进行体内折叶苔醇向乙酸折叶苔基酯的生物转化。为了进行筛选,在工程化的酿酒酵母菌株中将乙酰基转移酶候选物与来自污叉丝孔菌(Dichomitus squalens)的折叶苔醇合酶XP_007369631.1,NCBI登录号XP_007369631.1的编码基因共表达,在该菌株中,具有提高水平的内源性法呢基二磷酸(FPP)。
为了增加酿酒酵母细胞中内源性FPP库的水平,涉及甲羟戊酸途径的所有酵母内源基因的额外副本,从编码乙酰辅酶A C-乙酰基转移酶的ERG10到编码FPP合酶的ERG20,与Paddon et al.,Nature,2013,496:528-532中所述的相似,在半乳糖诱导型启动子的控制下,整合到酿酒酵母菌株CEN.PK2-1C(Euroscarf,Frankfurt,Germany)的基因组中。简而言之,将三个盒分别整合在LEU2、TRP1和URA3基因座中。第一个盒包含在GAL10/GAL1双向启动子控制下的ERG20基因和一个截短的HMG1(tHMG1,如Proc Natl Acad Sci USA,1997,109:E111-8所述),和同样在GAL10/GAL1启动子控制下的ERG19和ERG13基因,该盒的侧翼为两个100个核苷酸区域,分别对应于LEU2的上游和下游部分。第二个盒中,基因IDI1和tHMG1在GAL10/GAL1启动子的控制下,而基因ERG13在GAL7启动子区域的控制下,该盒的侧翼为两个100个核苷酸区域,分别对应于TRP1的上游和下游部分。第三个盒具有ERG10、ERG12、tHMG1和ERG8基因,均在GAL10/GAL1启动子的控制下,该盒的侧翼为两个100个核苷酸的区域,分别对应于URA3的上游和下游部分。三个盒中的所有基因都包含其自身终止子区域的200个核苷酸。而且,在ERG9启动子区域上游,整合了如Proc Natl Acad Sci USA,1991,88:8597-8601中所描述的,在其自身启动子的突变形式的控制下的GAL4的额外拷贝。另外,通过启动子交换修饰了ERG9的表达。使用含有带有其自身的启动子和终止子的HIS3基因的盒删除GAL7、GAL10和GAL1基因。将所得菌株与菌株CEN.PK2-1D(Euroscarf,Frankfurt,Germany)交配,获得称为YST045的二倍体菌株,其根据Solis-Escalante et al.,FEMS Yeast Res,2015,15:2诱导芽孢形成。孢子分离是通过将asci重悬于200μL、0.5M山梨糖醇和2μL的zymolyase(1000U mL
为了在YST069中表达XP_007369631.1和评估的乙酰基转移酶,如先前在Kuijperset al.,Microb Cell Fact.,2013,12:47中所述,使用酵母内源同源重组在体内构建质粒。质粒由用于酿酒酵母共转化的四个DNA片段组成。这些片段是:
a)通过用BsmBI进行酶促限制而线性化的质粒pF167(SEQ ID NO:1)。pF167以前是通过体内组装在酵母中构建的,它包含带有自身启动子和终止子的酵母标记LEU2,大肠杆菌标记AmpR,2μ酵母复制起点,大肠杆菌pUC复制起点和序列5'-GCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAG-3'(SEQ ID NO:2)和5'-AGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCAT-3'(SEQ ID NO:3)用于同源重组;
b)一片段,其由序列5'-GCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAG-3'(SEQ ID NO:2),酵母基因PGK1的终止子区域和倍半萜合酶XP_007369631.1DNA密码子(针对其在酿酒酵母中的表达进行了优化)(SEQ ID NO:4)组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025)。通过PCR重叠延伸将来自酵母的双向GAL1/GAL10启动子区域添加到该片段中(Yolov and Shabarova.,Nucleic AcidsRes.1990,18(13):3983-6);
c)一片段,其由与酵母GAL10启动子区域的头部核苷酸相对应的60bp,待评估的乙酰基转移酶DNA编码序列之一(针对其在酿酒酵母中的表达进行了密码子优化)和60bp的酵母CYC1终止子区域组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA94025);和
d)一片段,由酵母基因CYC1的终止子区域和序列5’-AGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCAT-3’(SEQ ID NO:3)组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025)。
用体内质粒组装所需的片段来转化YST069。按照Gietz and Woods,MethodsEnzymol.,2002,350:87-96中所述,用乙酸锂方案进行酵母转化。将转化混合物铺板在含有6.7g/L无氨基酸的酵母氮碱(BD Difco,New Jersey,USA),1.6g/L不含亮氨酸的滴注补充物(Sigma Aldrich,Missouri,USA),20g/L葡萄糖和20g/L琼脂的SmLeu培养基上。将板在30℃下温育3至4天。使用单个菌落,在含有如Westfall et al.,Proc Natl Acad Sci USA,2012,109:E111-118中所述的250μL培养基和50μL己二酸二异癸酯(abcr GmbH,Germany)作为有机叠加层的深孔板中产生折叶苔醇和乙酸折叶苔基酯。将深孔板在平板培养箱中在30℃下温育3天。为了提取酵母细胞产生的折叶苔醇和乙酸折叶苔基酯,用700μL含内标物的乙酸乙酯提取深孔板的每个孔。使用GC-MS分析鉴定折叶苔醇和乙酸折叶苔基酯的产生,并使用上述内标通过GC-FID进行定量。
在这些实验条件下,对于9种折叶苔醇合酶和乙酰基转移酶的组合都检出了乙酸折叶苔基酯(表2)。值得注意的是,在非优化筛选条件下,两种活性最高的乙酰基转移酶(CrDAT和FgaAT)产生的乙酸折叶苔基酯滴度超过150mg/L。GC-FID色谱图显示在图2中。此外,图2显示,酵母衍生的乙酸折叶苔基酯的MS谱图与参考乙酸折叶苔基酯的MS谱图相同。
令人感兴趣的是,我们的筛选中活性最高的乙酰基转移酶CrDAT是植物BAHD家族进化枝3的成员,它参与植物生物碱的生物合成,但是,来自同一家族和进化枝的其他测试的乙酰基转移酶PsSalAT和RsVISY,其同样参与植物生物碱的生物合成(分别为蒂巴因(Thebaine)和维诺任碱(Vinorine)),当以折叶苔醇为底物时,并没有活性。另外,来自烟曲霉(Aspergillus fumigatus)的FgaAT同样参与生物碱的生物合成(对烟曲霉文(Fumigaclavine)B具有活性),在我们的筛选中产生了大量的乙酸折叶苔基酯。相反,令人惊讶的是,推定的烟曲霉文B O-乙酰基转移酶GAO81666.1比FgaAT产生的乙酸折叶苔基酯少一个数量级。
值得注意的是,尽管AstG参与了经改性的补身烷型倍半萜Astellolide的生物合成,但乙酰基转移酶AstG或基于其与AstG相似性而选择的任何蛋白质(不包括GAO81666.1,参见上文)均未产生补身烷基乙酰化倍半萜乙酸折叶苔基酯。
在二萜生物合成中唯一鉴定出的乙酰基转移酶是那些参与紫杉醇(Taxol)和毛喉素产生的酶。从毛喉鞘蕊花(C.forskohlii)测试的八种乙酰基转移酶中,只有CfACT1-6和CfACT1-8能够乙酰化折叶苔醇。尽管基因TcTAT和TcDBAT能够在紫杉醇的生物合成中分别乙酰化结构相似的中间体(分别为乙酸紫衫-4(20),11(12)-二烯-5a-基酯和浆果赤霉素(Baccatin)III),但只有TcTAT产生乙酸折叶苔基酯。
为了增加现已证明的复杂性,以鉴定能够使用折叶苔醇作为酰基受体的乙酰基转移酶,五种受试底物通用氯霉素乙酰基转移酶中只有一种能够将折叶苔醇转化为乙酸折叶苔基酯。
表2:发现能够在酿酒酵母中将折叶苔醇转化为乙酸折叶苔酯的乙酰基转移酶
为了在酿酒酵母中产生乙酸补身烯基酯,可以将如实施例2中所示能够将折叶苔醇转化为乙酸折叶苔酯的九种乙酰基转移酶(CrDAT,FgaAT,OAH94415.1,TcTAT,CrMAT,LiAAT-4,GAO81666.1,CfACT1-6,CfACT1-8)用于评估补身醇向乙酸补身烯基酯的转化。
通过共表达每种选择的乙酰基转移酶候选物与编码来自双孢蘑菇(Agaricusbisporus)的补身醇合酶XP_006461126(NCBI登录号XP_006461126)的基因,在工程化的酿酒酵母菌株YST069中体内产生乙酸补身烯基酯,在该菌株中,具有提高水平的内源性FPP(参见实施例2)。
为了在YST069中表达XP_006461126和所选的乙酰基转移酶(CrDAT,FgaAT,OAH94415.1,TcTAT,CrMAT,LiAAT-4,GAO81666.1,CfACT1-6和CfACT1-8),如先前在Kuijpers et al.,Microb Cell Fact.,2013,12:47中所述,使用酵母内源同源重组在体内构建质粒。质粒由用于酿酒酵母共转化的四个DNA片段组成。这些片段是:
a)通过用BsmBI进行酶促限制而线性化的质粒pF167(SEQ ID NO:1)。pF167以前是通过体内组装在酵母中构建的,它包含带有自身启动子和终止子的酵母标记LEU2,大肠杆菌标记AmpR,2μ酵母复制起点,大肠杆菌pUC复制起点和序列5'-GCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAG-3'(SEQ ID NO:2)和5'-AGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCAT-3'(SEQ ID NO:3)用于同源重组;
b)一片段,其由序列5'-GCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAG-3'(SEQ ID NO:2),酵母基因PGK1的终止子区域和倍半萜合酶XP_006461126DNA密码子(针对其在酿酒酵母中的表达进行了优化)(SEQ ID NO:6)组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025)。通过PCR重叠延伸将来自酵母的双向GAL1/GAL10启动子区域添加到该片段中(Yolov and Shabarova.,Nucleic AcidsRes.1990,18(13):3983-6);
c)一片段,其由与酵母GAL10启动子区域的头部核苷酸相对应的60bp,待评估的乙酰基转移酶DNA编码序列之一(针对其在酿酒酵母中的表达进行了密码子优化)和60bp的酵母CYC1终止子区域组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025);和
d)一片段,由酵母基因CYC1的终止子区域和序列5’-AGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCAT-3’(SEQ ID NO:3)组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025)。
如用于产生折叶苔醇和乙酸折叶苔基酯的实施例2中所述进行酵母转化、筛选条件和补身醇和乙酸补身烯基酯的定量。
乙酸折叶苔基酯(来自实施例2)和乙酸补身烯基酯的相对量分别显示在图3和图4中。从测试的九种候选酶中,七种(CrDAT,FgaAT,OAH94415.1,TcTAT,GAO81666.1,CfACT1-6和CfACT1-8)产生了乙酸补身烯基酯。由于折叶苔醇和补身醇之间的结构相似性很高,因此预计会发现相似的相对转化率。
出乎意料的是,发现补身醇向其相应乙酸酯的转化发生了显著变化。特别地,发现产生最高相对量的乙酸折叶苔基酯的CrDAT是乙酸补身烯基酯的最低产生者之一。相比之下,发现FgaAT和CfACT1-8产生的乙酸酯相对量最高,而它们对折叶苔醇的活性较弱。这些令人惊讶的发现表明难以鉴定合适的乙酰基转移酶候选物以接受非生理底物。
a)实验1
为了鉴定能够乙酰化补身烷型倍半萜醇的其他乙酰基转移酶,在NCBI ProteinBlast搜索中使用了来自乙酰基转移酶CrDAT和FgaAT的氨基酸序列(分别在实施例2和3中显示),它们对折叶苔醇和补身醇有活性,寻找接近的蛋白质序列同源物。使用默认参数执行Protein Blast搜索(Tatiana et al,FEMS Microbiol Lett.,1999,174:247-250,1999)。
从Blast搜索中检索到了与CrDAT同源的植物乙酰基转移酶XP_008340165.2和与FgaAT对应的4种真菌乙酰基转移酶(KEY80391,PYI04555.1,XP_001276734.1,XP_024709055.1)。它们在表3中列出。
表3:基于与CrDAT或FgaAT的氨基酸序列同源性而选择的用于将补身烷倍半萜转化为补身烷基乙酰化倍半萜的乙酰基转移酶
b)实验2
此外,还从地钱类(liverwort)鞭苔(Bazzania trilobata)和蕨类香鳞毛蕨(Dryopteris fragrans)的转录组中检索了额外的乙酰基转移酶候选物。地钱类鞭苔是富含萜类化合物包括补身烷倍半萜的来源。此外,乙酸折叶苔基酯和咖啡酸折叶苔基酯据报道是来自鞭苔属地钱的天然产物(Asakawa et al,Phytochemistry,Volume 30,Issue 9,1991,Pages 3037-3040)。类似地,在鳞毛蕨属中报道了几种不同的天然产物,包括乙酸折叶苔基酯。(Hideyuki Ito et al.Chem.Pharm.Bull.48(8)1190-1195(2000);Froissard Det al.Nat Prod Commun.2014Jan;9(1):137-40.)
使用CLC基因组工作台(Qiagen)组装了鞭苔(Bazzania trilobata)(NCBI登录号ER364415)转录组,得到总共22083个重叠群(contigs),平均长度为1,225个碱基对。CrDAT氨基酸序列用于在鞭苔(Bazzania trilobata)转录组中搜索同源序列。对于该搜索,以默认参数使用tBlastn算法(Altschul et al.1990,J.Mol.Biol.215,403-410)。E值超过0.001的转录物被考虑在内。选择属于植物乙酰基转移酶BAHD家族的10种转录物,其与CrDAT的氨基酸序列同源性低至20%。
香鳞毛蕨(D.fragrans)的植物材料是从中国北方收集的。将香鳞毛蕨的新鲜叶子(样品ID PNLI20141074)用于转录组分析。使用来自QIAGEN的RNeasy Plant Mini Kit(目录号74904)提取香鳞毛蕨的总RNA。使用用于Illumina的
进行了乙酰基转移酶的扩展筛选,以将折叶苔醇、补身醇和双环法呢醇转化为其相应的乙酸酯衍生物。在工程化的酿酒酵母细胞中体内筛选了实施例1的表1和实施例4中描述的总共89种乙酰基转移酶。不重复如实施例2和实施例3中所述在初步数目的乙酰基转移酶上产生的筛选数据。因此,仅筛选了产生在实施例2和实施例3中未测试的折叶苔醇、补身醇或双环法呢醇的乙酰基转移酶与酿酒酵母细胞的新组合。
如上文实施例2和实施例3中所述,工程化的酿酒酵母菌株YST069用于筛选乙酸折叶苔基酯和乙酸补身烯基酯的产生。
通过共表达每种选择的乙酰基转移酶候选物与负责从Astellolide生物合成中产生双环法呢醇的酶AstC(SEQ ID NO:138)、AstI(SEQ ID NO:140)和AstK(SEQ ID NO:142)(Yasutomo Shinohara et.al.Sci Rep.2016,6:32865),在改造的酿酒酵母YST069菌株中体内产生乙酸双环法呢基酯。
为了同时表达AstC和AstI,构建了表达盒,其含有密码子和双向GAL1/GAL10启动子,该密码子针对啤酒酵母版本的基因进行了优化,该基因编码AstC(SEQ ID NO:138)和AstI(SEQ ID NO:140),将该表达盒整合到YST069的基因组中,产生称为YST216的新菌株。通过DNA合成(ATUM,Menlo Park,CA 94025)获得了AstC和AstI的密码子优化的DNA序列。通过PCR重叠延伸将来自酵母的双向GAL1/GAL10启动子区添加到这些基因中(Yolov andShabarova.,Nucleic Acids Res.1990,18(13):3983-6)。
为了在YST216中表达AstK和评估的乙酰基转移酶,如先前在Kuijpers et al.,Microb Cell Fact.,2013,12:47中所述,使用酵母内源同源重组在体内构建质粒。质粒由用于酿酒酵母共转化的四个DNA片段组成。这些片段是:
a)通过用BsmBI进行酶促限制而线性化的质粒pF167(SEQ ID NO:1)。
b)一片段,其由序列5'-GCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAG-3'(SEQ ID NO:2),酵母基因PGK1的终止子区域和AstK DNA序列密码子(针对其在酿酒酵母中的表达进行了优化)(SEQ ID NO:141)组成,该片段通过DNA合成获得(ATUM,Menlo Park,CA 94025)。通过PCR重叠延伸将来自酵母的双向GAL1/GAL10启动子区域添加到该片段中(Yolov and Shabarova.,Nucleic Acids Res.1990,18(13):3983-6);
c)一片段,其由与酵母GAL10启动子区域的头部核苷酸相对应的60bp,待评估的乙酰基转移酶DNA编码序列之一(针对其在酿酒酵母中的表达进行了密码子优化)和60bp的酵母CYC1终止子区域组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025);和
d)一片段,由酵母基因CYC1的终止子区域序列5’-AGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCAT-3’(SEQ ID NO:3)组成,该片段是通过DNA合成获得的(ATUM,Menlo Park,CA 94025)。
用体内质粒组装所需的片段转化YST216。如实施例2中所述进行酵母转化。双环法呢醇和乙酸双环法呢基酯的筛选条件和定量与实施例2中所述相同,不同之处在于使用25μL矿物油(2705-01,VWR International,LLC.)作为筛选过程中的有机覆盖物。
除了先前描述的对折叶苔醇具有活性的九种酶(CrDAT,FgaAT,OAH94415.1,TcTAT,CrMAT,LiAAT-4,GAO81666.1,CfACT1-6,CfACT1-8)(参见实施例2),在这些实验条件下,当使用来自鞭苔(Bazzania trilobata)(SEQ ID NO:124或144)的乙酰基转移酶ERR364415-1_contig_8546和来自香鳞毛蕨(Dryopteris fragrans)(SEQ ID NO:118)的DfATC13时,还检测到乙酸折叶苔基酯。但是,它们的活性低于CrDAT和FgaAT。乙酸折叶苔基酯的相对量示于图5。
除了可以将补身醇转化为乙酸补身烯基酯的七种候选酶(CrDAT,FgaAT,OAH94415.1,TcTAT,GAO81666.1,CfACT1-6和CfACT1-8)之外,还发现有三种额外的乙酰基转移酶,来自费希尔曲霉(Aspergillus fischeri)的XP_001258079.1(SEQ ID NO:127),来自鞭苔(Bazzania trilobata)的ERR364415-1_contig_8546(SEQ ID NO:124或144),以及来自香鳞毛蕨(Dryopteris fragrans)的DfATC13(SEQ ID NO:118),在扩展筛选中由补身醇产生乙酸补身烯基酯。所获得的乙酸补身烯基酯的相对量示于图6。图7显示,酵母来源的乙酸补身烯基酯的MS谱图与参考乙酸补身烯基酯的MS谱图相同。令人感兴趣的是,在这些实验条件下,ERR364415-1_contig_8546和DfATC13在所有测试的乙酰基转移酶中显示出最高的活性。
AstC、AstI和AstK与测试的89种乙酰基转移酶的13种组合检测到乙酸双环法呢基酯。图8显示,酵母衍生的乙酸双环法呢基酯的MS谱图与参考乙酸双环法呢基酯的MS谱图相同。乙酸双环法呢基酯的相对量示于图9。来自烟曲霉(Aspergillus fumigatus)的乙酰基转移酶FgaAT和来自鞭苔(Bazzania trilobata)的ERR364415-1_contig_8546显示了双环法呢醇的最高乙酰化活性。在这13种发现的乙酰基转移酶中,即如上文实施例2中所述的CrDAT,FgaAT,TcTAT,CrMAT,GAO81666.1,CfACT1-6,CfACT1-8,以及BAU61551.1(SEQ IDNO:133),PsSalAT(SEQ ID NO:136),XP_001217250.1(SEQ ID NO:130),ERR364415-1_contig_8546(SEQ ID NO:124或144),PYI04555.1(SEQ ID NO:121或143),DfACT13(SEQ IDNO:118)(它们对双环法呢醇有活性),也发现9种化合物,即CrDAT,FgaAT,TcTAT,CrMAT,GAO81666.1,CfACT1-6,CfACT1-8,ERR364415-1_contig_8546,DfACT13也对折叶苔醇和补身醇有活性。值得注意的是,参与芳酸酯化的补身烷型倍半萜内酯、astellolide(SciRep.2016,6:32865)生物合成的乙酰基转移酶AstG对测试的任何补身烷型倍半萜醇均无活性。这再次表明鉴定能够使用折叶苔醇、补身醇或双环法呢醇作为酰基受体的乙酰基转移酶的不确定性和复杂性。
交叉引用的文档内容通过引用并入于此。
本文所指的序列是:
NA=核酸 AA=氨基酸。
序列表
SEQ ID NO:1
质粒pF167
TGGTCAGCAACAACGCCGAAGAATCACTCTCGTGTTGAGAATTGCACGCCTTGACCACGACACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTTTGGCATTGGCAAAGTGCGGACTGCATAGTCACTGTGGTGCCGTACTTAGGGTACGCGTTCCTGAACGAAGCATCTGTGCTTCATTTTGTAGAACAAAAATGCAACGCGAGAGCGCTAATTTTTCAAACAAAGAATCTGAGCTGCATTTTTACAGAACAGAAATGCAACGCGAAAGCGCTATTTTACCAACGAAGAATCTGTGCTTCATTTTTGTAAAACAAAAATGCAACGCGAGAGCGCTAATTTTTCAAACAAAGAATCTGAGCTGCATTTTTACAGAACAGAAATGCAACGCGAGAGCGCTATTTTACCAACAAAGAATCTATACTTCTTTTTTGTTCTACAAAAATGCATCCCGAGAGCGCTATTTTTCTAACAAAGCATCTTAGATTACTTTTTTTCTCCTTTGTGCGCTCTATAATGCAGTCTCTTGATAACTTTTTGCACTGTAGGTCCGTTAAGGTTAGAAGAAGGCTACTTTGGTGTCTATTTTCTCTTCCATAAAAAAAGCCTGACTCCACTTCCCGCGTTTACTGATTACTAGCGAAGCTGCGGGTGCATTTTTTCAAGATAAAGGCATCCCCGATTATATTCTATACCGATGTGGATTGCGCATACTTTGTGAACAGAAAGTGATAGCGTTGATGATTCTTCATTGGTCAGAAAATTATGAACGGTTTCTTCTATTTTGTCTCTATATACTACGTATAGGAAATGTTTACATTTTCGTATTGTTTTCGATTCACTCTATGAATAGTTCTTACTACAATTTTTTTGTCTAAAGAGTAATACTAGAGATAAACATAAAAAATGTAGAGGTCGAGTTTAGATGCAAGTTCAAGGAGCGAAAGGTGGATGGGTAGGTTATATAGGGATATAGCACAGAGATATATAGCAAAGAGATACTTTTGAGCAATGTTTGTGGAAGCGGTATTCGCAATATTTTAGTAGCTCGTTACAGTCCGGTGCGTTTTTGGTTTTTTGAAAGTGCGTCTTCAGAGCGCTTTTGGTTTTCAAAAGCGCTCTGAAGTTCCTATACTTTCTAGAGAATAGGAACTTCGGAATAGGAACTTCAAAGCGTTTCCGAAAACGAGCGCTTCCGAAAATGCAACGCGAGCTGCGCACATACAGCTCACTGTTCACGTCGCACCTATATCTGCGTGTTGCCTGTATATATATATACATGAGAAGAACGGCATAGTGCGTGTTTATGCTTAAATGCGTACTTATATGCGTCTATTTATGTAGGATGAAAGGTAGTCTAGTACCTCCTGTGATATTATCCCATTCCATGCGGGGTATCGTATGCTTCCTTCAGCACTACCCTTTAGCTGTTCTATATGCTGCCACTCCTCAATTGGATTAGTCTCATCCTTCAATGCTATCATTATGTCACCCGCAGTTCTGTGTCGTAGTCATCAACATAGCACCTATCCTTTGGCATCTCGGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAGTGAGACGTTAATTAAAGTAGACCGCTCACACATGGGCGGCCGCCGTCTCAAGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCATTCGACTACGTCGTAAGGCCGTTTCTGACAGAGTAAAATTCTTGAGGGAACTTTCACCATTATGGGAAATGCTTCAAGAAGGTATTGACTTAAACTCCATCAAATGGTCAGGTCATTGAGTGTTTTTTATTTGTTGTATTTTTTTTTTTTTAGAGAAAATCCTCCAATATCAAATTAGGAATCGTAGTTTCATGATTTTCTGTTACACCTAACTTTTTGTGTGGTGCCCTCCTCCTTGTCAATATTAATGTTAAAGTGCAATTCTTTTTCCTTATCACGTTGAGCCATTAGTATCAATTTGCTTACCTGTATTCCTTTACTATCCTCCTTTTTCTCCTTCTTGATAAATGTATGTAGATTGCGTATATAGTTTCGTCTACCCTATGAACATATTCCATTTTGTAATTTCGTGTCGTTTCTATTATGAATTTCATTTATAAAGTTTATGTACAAATATCATAAAAAAAGAGAATCTTTTTAAGCAAGGATTTTCTTAACTTCTTCGGCGACAGCATCACCGACTTCGGTGGTACTGTTGGAACCACCTAAATCACCAGTTCTGATACCTGCATCCAAAACCTTTTTAACTGCATCTTCAATGGCCTTACCTTCTTCAGGCAAGTTCAATGACAATTTCAACATCATTGCAGCAGACAAGATAGTGGCGATAGGGTCAACCTTATTCTTTGGCAAATCTGGAGCAGAACCGTGGCATGGTTCGTACAAACCAAATGCGGTGTTCTTGTCTGGCAAAGAGGCCAAGGACGCAGATGGCAACAAACCCAAGGAACCTGGGATAACGGAGGCTTCATCGGAGATGATATCACCAAACATGTTGCTGGTGATTATAATACCATTTAGGTGGGTTGGGTTCTTAACTAGGATCATGGCGGCAGAATCAATCAATTGATGTTGAACCTTCAATGTAGGGAATTCGTTCTTGATGGTTTCCTCCACAGTTTTTCTCCATAATCTTGAAGAGGCCAAAAGATTAGCTTTATCCAAGGACCAAATAGGCAATGGTGGCTCATGTTGTAGGGCCATGAAAGCGGCCATTCTTGTGATTCTTTGCACTTCTGGAACGGTGTATTGTTCACTATCCCAAGCGACACCATCACCATCGTCTTCCTTTCTCTTACCAAAGTAAATACCTCCCACTAATTCTCTGACAACAACGAAGTCAGTACCTTTAGCAAATTGTGGCTTGATTGGAGATAAGTCTAAAAGAGAGTCGGATGCAAAGTTACATGGTCTTAAGTTGGCGTACAATTGAAGTTCTTTACGGATTTTTAGTAAACCTTGTTCAGGTCTAACACTACCGGTACCCCATTTAGGACCAGCCACAGCACCTAACAAAACGGCATCAACCTTCTTGGAGGCTTCCAGCGCCTCATCTGGAAGTGGGACACCTGTAGCATCGATAGCAGCACCACCAATTAAATGATTTTCGAAATCGAACTTGACATTGGAACGAACATCAGAAATAGCTTTAAGAACCTTAATGGCTTCGGCTGTGATTTCTTGACCAACGTGGTCACCTGGCAAAACGACGATCTTCTTAGGGGCAGACATAGGGGCAGACATTAGAATGGTATATCCTTGAAATATATATATATATTGCTGAAATGTAAAAGGTAAGAAAAGTTAGAAAGTAAGACGATTGCTAACCACCTATTGGAAAAAACAATAGGTCCTTAAATAATATTGTCAACTTCAAGTATTGTGATGCAAGCATTTAGTCATGAACGCTTCTCTATTCTATATGAAAAGCCGGTTCCGGCCTCTCACCTTTCCTTTTTCTCCCAATTTTTCAGTTGAAAAAGGTATATGCGTCAGGCGACCTCTGAAATTAACAAAAAATTTCCAGTCATCGAATTTGATTCTGTGCGATAGCGCCCCTGTGTGTTCTCGTTATGTTGAGGAAAAAAATAATGGTTGCTAAGAGATTCGAACTCTTGCATCTTACGATACCTGAGTATTCCCACAGTTAACTGCGGTCAAGATATTTCTTGAATCAGGCGCCTTAGACCGCTCGGCCAAACAACCAATTACTTGTTGAGAAATAGAGTATAATTATCCTATAAATATAACGTTTTTGAACACACATGAACAAGGAAGTACAGGACAATTGATTTTGAAGAGAATGTGGATTTTGATGTAATTGTTGGGATTCCATTTTTAATAAGGCAATAATATTAGGTATGTGGATATACTAGAAGTTCTCCTCGACCGTCGA
SEQ ID NO:2
用于同源重组的序列
GCACTTGCTACACTGTCAGGATAGCTTCCGTCACATGGTGGCGATCACCGTACATCTGAG
SEQ ID NO:3
用于同源重组的序列
AGGTGCAGTTCGCGTGCAATTATAACGTCGTGGCAACTGTTATCAGTCGTACCGCGCCAT
SEQ ID NO:4
XP_007369631.1污叉丝孔菌(Dichomitus squalens)折叶苔醇合酶的DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGCTTCTATCCACAGAAGATACACTACTTTGATCTTGGACTTGGGTGACGTTTTGTTCAGATGGTCTCCAAAGACTGAAACTGCTATCCCACCACAACAATTGAAGGACATCTTGTCTTCTGTTACTTGGTTCGAATACGAAAGAGGTAGATTGTCTCAAGAAGCTTGTTACGAAAGATGTGCTGAAGAATTCAAGATCGAAGCTTCTGTTATCGCTGAAGCTTTCAAGCAAGCTAGAGGTTCTTTGAGACCAAACGAAGAATTCATCGCTTTGATCAGAGACTTGAGAAGAGAAATGCACGGTGACTTGACTGTTTTGGCTTTGTCTAACATCTCTTTGCCAGACTACGAATACATCATGTCTTTGTCTTCTGACTGGACTACTGTTTTCGACAGAGTTTTCCCATCTGCTTTGGTTGGTGAAAGAAAGCCACACTTGGGTTGTTACAGAAAGGTTATCTCTGAAATGAACTTGGAACCACAAACTACTGTTTTCGTTGACGACAAGTTGGACAACGTTGCTTCTGCTAGATCTTTGGGTATGCACGGTATCGTTTTCGACAACCAAGCTAACGTTTTCAGACAATTGAGAAACATCTTCGGTGACCCAATCAGAAGAGGTCAAGAATACTTGAGAGGTCACGCTGGTAAGTTGGAATCTTCTACTGACAACGGTTTGATCTTCGAAGAAAACTTCACTCAATTGATCATCTACGAATTGACTCAAGACAGAACTTTGATCTCTTTGTCTGAATGTCCAAGAACTTGGAACTTCTTCAGAGGTGAACCATTGTTCTCTGAAACTTTCCCAGACGACGTTGACACTACTTCTGTTGCTTTGACTGTTTTGCAACCAGACAGAGCTTTGGTTAACTCTGTTTTGGACGAAATGTTGGAATACGTTGACGCTGACGGTATCATGCAAACTTACTTCGACAGATCTAGACCAAGAATGGACCCATTCGTTTGTGTTAACGTTTTGTCTTTGTTCTACGAAAACGGTAGAGGTCACGAATTGCCAAGAACTTTGGACTGGGTTTACGAAGTTTTGTTGCACAGAGCTTACCACGGTGGTTCTAGATACTACTTGTCTCCAGACTGTTTCTTGTTCTTCATGTCTAGATTGTTGAAGAGAGCTGACGACCCAGCTGTTCAAGCTAGATTGAGACCATTGTTCGTTGAAAGAGTTAACGAAAGAGTTGGTGCTGCTGGTGACTCTATGGACTTGGCTTTCAGAATCTTGGCTGCTGCTTCTGTTGGTGTTCAATGTCCAAGAGACTTGGAAAGATTGACTGCTGGTCAATGTGACGACGGTGGTTGGGACTTGTGTTGGTTCTACGTTTTCGGTTCTACTGGTGTTAAGGCTGGTAACAGAGGTTTGACTACTGCTTTGGCTGTTACTGCTATCCAAACTGCTATCGGTAGACCACCATCTCCATCTCCATCTGCTGCTTCTTCTTCTTTCAGACCATCTTCTCCATACAAGTTCTTGGGTATCTCTAGACCAGCTTCTCCAATCAGATTCGGTGACTTGTTGAGACCATGGAGAAAGATGTCTAGATCTAACTTGAAGTCTCAATAA
SEQ ID NO:5
XP_007369631.1污叉丝孔菌(Dichomitus squalens)折叶苔醇合酶的氨基酸序列
MASIHRRYTTLILDLGDVLFRWSPKTETAIPPQQLKDILSSVTWFEYERGRLSQEACYERCAEEFKIEASVIAEAFKQARGSLRPNEEFIALIRDLRREMHGDLTVLALSNISLPDYEYIMSLSSDWTTVFDRVFPSALVGERKPHLGCYRKVISEMNLEPQTTVFVDDKLDNVASARSLGMHGIVFDNQANVFRQLRNIFGDPIRRGQEYLRGHAGKLESSTDNGLIFEENFTQLIIYELTQDRTLISLSECPRTWNFFRGEPLFSETFPDDVDTTSVALTVLQPDRALVNSVLDEMLEYVDADGIMQTYFDRSRPRMDPFVCVNVLSLFYENGRGHELPRTLDWVYEVLLHRAYHGGSRYYLSPDCFLFFMSRLLKRADDPAVQARLRPLFVERVNERVGAAGDSMDLAFRILAAASVGVQCPRDLERLTAGQCDDGGWDLCWFYVFGSTGVKAGNRGLTTALAVTAIQTAIGRPPSPSPSAASSSFRPSSPYKFLGISRPASPIRFGDLLRPWRKMSRSNLKSQ
SEQ ID NO:6
XP_006461126双孢蘑菇(Agaricus bisporus)补身醇合酶DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGCTCCACCACAAAGACCATTCACTGCTATCGTTTTCGACATCGGTGACGTTTTGTTCCAATGGTCTGCTACTACTAAGACTTCTATCTCTCCAAAGACTTTGAGATCTATCTTGAACTGTCCAACTTGGTTCGACTACGAAAGAGGTAGATTGGCTGAAAACGCTTGTTACGCTGCTATCTCTCAAGAATTCAACGTTAACCCAGACGAAGTTAGAGACGCTTTCTCTCAAGCTAGAGACTCTTTGCAAGCTAACCACGACTTCATCTCTTTGATCAGAGAATTGAAGGCTCAAGCTAACGGTAGATTGAGAGTTTACGCTATGTCTAACATCTCTTTGCCAGACTGGGAAGTTTTGAGAATGAAGCCAGCTGACTGGGACATCTTCGACCACGTTTTCACTTCTGGTGCTGTTGGTGAAAGAAAGCCAAACTTGGCTTTCTACAGACACGTTATCGCTGCTACTGACTTGCAACCACACCAAACTATCTTCGTTGACGACAAGTTGGAAAACGTTTTGTCTGCTAGATCTTTGGGTTTCACTGGTATCGTTTTCGACGAACCATCTGAAGTTAAGAGAGCTTTGAGAAACTTGATCGGTGACCCAGTTCAAAGAGGTGGTGAATTCTTGGTTAGAAACGCTGGTAAGTTGGGTTCTATCACTAGAACTACTGCTAAGCACGAATCTATCCCATTGGACGAAAACTTCGCTCAATTGTTGATCTTGGAAATCACTGGTAACAGAGCTTTGGTTAACTTGGTTGAACACCCACAAACTTGGAACTTCTTCCAAGGTAAGGGTCAATTGACTACTGAAGAATTCCCATTCGACTTGGACACTACTTCTTTGGGTTTGACTATCTTGAAGAGATCTAGAGAAATCGCTGACTCTGTTATGGACGAAATGTTGGAATACGTTGACCCAGACGGTATCATCCAAACTTACTTCGACCACAGAAGACCAAGATTCGACCCAGTTGTTTGTGTTAACGCTTTGTCTTTGTTCTACGCTTACGGTAGAGGTGAACAATTGAGATCTACTTTGACTTGGGTTCACGAAGTTTTGTTGAACAGAGCTTACTTGGACGGTACTAGATACTACGAAACTGCTGAATGTTTCTTGTACTTCATGTCTAGATTGTTGGCTACTTCTGGTGACCCAGACTTGCACTCTTTGTTGAAGCCATTGTTGAAGGAAAGAGTTCAAGAAAGAATCGGTGCTGACGGTGACTCTTTGGCTTTGGCTATGAGAATCTTGGCTTGTGACTTCGTTGGTATCAGAGACGAAGTTGACTTGAGAACTTTGTTGACTTTGCAATGTGAAGACGGTGGTTGGGAAGTTGGTTGGATGTACAAGTACGGTTCTTCTGGTATCTCTATCGGTAACAGAGGTTTGGCTACTGCTTTGGCTATCAAGGCTGTTGACACTATGTTCCAACCACAAATCAGATTCTCTGAATCTCCAACTGACACTTTGGTTGAAAACGCTATCCACAAGAGAAGACCATCTTTCTCTGAAAAGTTCTTGGGTAAGAGACCAAGATCTGGTTCTTTCAGAAAGCCATTGCAATGGATCTTGCAAGGTTCTAAGTTGAGAAAGTCTGTTGAAATCGGTTCTTAA
SEQ ID NO:7
XP_006461126双孢蘑菇(Agaricus bisporus)补身醇合酶的蛋白质序列
MAPPQRPFTAIVFDIGDVLFQWSATTKTSISPKTLRSILNCPTWFDYERGRLAENACYAAISQEFNVNPDEVRDAFSQARDSLQANHDFISLIRELKAQANGRLRVYAMSNISLPDWEVLRMKPADWDIFDHVFTSGAVGERKPNLAFYRHVIAATDLQPHQTIFVDDKLENVLSARSLGFTGIVFDEPSEVKRALRNLIGDPVQRGGEFLVRNAGKLGSITRTTAKHESIPLDENFAQLLILEITGNRALVNLVEHPQTWNFFQGKGQLTTEEFPFDLDTTSLGLTILKRSREIADSVMDEMLEYVDPDGIIQTYFDHRRPRFDPVVCVNALSLFYAYGRGEQLRSTLTWVHEVLLNRAYLDGTRYYETAECFLYFMSRLLATSGDPDLHSLLKPLLKERVQERIGADGDSLALAMRILACDFVGIRDEVDLRTLLTLQCEDGGWEVGWMYKYGSSGISIGNRGLATALAIKAVDTMFQPQIRFSESPTDTLVENAIHKRRPSFSEKFLGKRPRSGSFRKPLQWILQGSKLRKSVEIGS
SEQ ID NO:8
CrDAT DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGAAAGTGGTAAAATATCTGTAGAAACGGAAACCCTATCGAAAACCCTTATCAAGCCGTCCTCGCCCACGCCACAGTCTCTTTCCCGTTACAACTTATCTTACAATGATCAGAACATTTATCAGACCTGCGTATCTGTAGGGTTCTTTTACGAAAATCCTGATGGCATCGAGATATCCACGATCCGTGAGCAGCTGCAGAACAGCCTGTCCAAAACTCTGGTCTCATACTATCCCTTCGCAGGCAAAGTAGTGAAGAACGACTACATACATTGTAACGACGATGGCATTGAGTTCGTCGAAGTCAGAATTAGATGCAGGATGAATGATATACTTAAATACGAACTACGTTCTTACGCTCGTGATTTAGTCCTGCCAAAGAGGGTTACCGTGGGCTCTGAAGATACCACCGCTATCGTGCAGCTGTCCCATTTCGACTGTGGCGGTTTGGCCGTGGCTTTTGGTATCTCCCACAAGGTGGCCGATGGTGGTACTATAGCCTCTTTCATGAAGGACTGGGCGGCCTCTGCTTGCTACTTGTCCTCAAGTCACCACGTTCCAACGCCTCTATTGGTCTCAGACAGTATTTTCCCAAGACAGGACAACATAATATGCGAACAATTTCCCACCTCAAAGAATTGCGTGGAGAAGACATTCATTTTCCCACCAGAAGCAATCGAAAAGTTAAAGAGTAAGGCCGTTGAATTCGGCATTGAGAAACCGACCAGAGTAGAGGTCCTGACTGCATTCTTATCTAGATGCGCCACCGTAGCAGGTAAGTCGGCAGCTAAGAACAACAATTGTGGTCAAAGCCTGCCCTTTCCGGTTCTACAGGCCATTAATTTGAGGCCGATTCTAGAATTGCCACAGAACTCTGTGGGTAATCTAGTTTCGATCTACTTCAGCAGGACAATTAAGGAAAACGACTACCTAAATGAAAAGGAGTACACTAAATTGGTGATAAACGAGTTGCGTAAAGAAAAGCAAAAGATCAAGAATTTGAGCAGAGAGAAGTTGACCTACGTCGCCCAGATGGAAGAGTTCGTTAAGTCACTTAAAGAGTTCGATATCTCCAACTTCCTAGATATCGATGCCTACTTAAGCGATAGTTGGTGCAGGTTTCCTTTCTACGACGTCGACTTCGGATGGGGAAAGCCTATCTGGGTCTGCCTATTCCAGCCTTACATCAAGAACTGCGTTGTTATGATGGACTATCCTTTCGGTGACGATTATGGTATCGAGGCTATTGTTTCTTTCGAGCAAGAGAAAATGTCCGCGTTCGAGAAGAACGAACAGTTACTGCAGTTTGTGTCTAACTAA
SEQ ID NO:9
CrDAT蛋白质序列
MESGKISVETETLSKTLIKPSSPTPQSLSRYNLSYNDQNIYQTCVSVGFFYENPDGIEISTIREQLQNSLSKTLVSYYPFAGKVVKNDYIHCNDDGIEFVEVRIRCRMNDILKYELRSYARDLVLPKRVTVGSEDTTAIVQLSHFDCGGLAVAFGISHKVADGGTIASFMKDWAASACYLSSSHHVPTPLLVSDSIFPRQDNIICEQFPTSKNCVEKTFIFPPEAIEKLKSKAVEFGIEKPTRVEVLTAFLSRCATVAGKSAAKNNNCGQSLPFPVLQAINLRPILELPQNSVGNLVSIYFSRTIKENDYLNEKEYTKLVINELRKEKQKIKNLSREKLTYVAQMEEFVKSLKEFDISNFLDIDAYLSDSWCRFPFYDVDFGWGKPIWVCLFQPYIKNCVVMMDYPFGDDYGIEAIVSFEQEKMSAFEKNEQLLQFVSN
SEQ ID NO:10
TcTAT DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGAGAAGACGGATTTACACGTGAACCTTATCGAAAAGGTTATGGTCGGTCCAAGCCCACCTTTGCCGAAGACTACACTACAACTATCCTCAATCGATAACCTACCTGGTGTTAGAGGCTCGATCTTTAATGCCTTGTTAATATATAATGCTTCACCGTCTCCCACGATGATCTCCGCTGATCCAGCTAAACCTATCAGAGAAGCTTTGGCCAAAATCCTGGTTTACTACCCACCTTTCGCTGGCAGGCTACGTGAGACTGAGAATGGCGATCTAGAGGTAGAGTGCACGGGAGAGGGTGCGATGTTTCTTGAAGCGATGGCCGATAATGAATTAAGTGTCTTAGGGGATTTCGACGACAGTAACCCTAGCTTCCAGCAATTGTTGTTCTCCCTTCCACTGGACACTAACTTCAAGGATCTATCCCTGTTAGTCGTGCAGGTGACAAGGTTTACATGCGGCGGGTTCGTCGTTGGGGTTTCATTTCACCACGGTGTATGCGATGGGAGAGGAGCTGCACAATTTCTGAAGGGACTAGCCGAAATGGCAAGGGGAGAAGTTAAGTTGTCGCTGGAGCCGATCTGGAATCGTGAACTAGTTAAGCTGGACGATCCCAAGTATTTGCAATTCTTTCACTTCGAATTCTTGAGAGCACCATCAATCGTCGAAAAGATAGTTCAAACCTACTTCATCATCGATTTCGAAACTATTAACTACATAAAGCAGTCAGTAATGGAAGAATGTAAAGAATTCTGCAGCTCTTTTGAAGTTGCTTCTGCAATGACGTGGATAGCTAGGACCAGGGCCTTTCAAATACCCGAGAGCGAGTACGTGAAAATCCTATTTGGTATGGACATGAGGAACTCTTTTAACCCGCCTCTGCCGTCAGGTTATTATGGAAACTCAATTGGTACGGCTTGTGCAGTGGACAATGTTCAAGACTTGTTGAGTGGTTCACTACTGAGGGCAATAATGATAATCAAGAAATCAAAGGTATCACTTAACGACAACTTTAAGAGCCGTGCTGTCGTTAAGCCTTCAGAACTTGACGTAAACATGAACCATGAGAATGTGGTGGCATTTGCTGACTGGTCAAGGCTGGGGTTCGACGAAGTCGATTTTGGCTGGGGAAACGCGGTGTCTGTTTCCCCGGTACAGCAACAGTCGGCACTTGCAATGCAGAATTACTTCCTGTTCTTGAAGCCAAGCAAGAACAAGCCCGATGGAATTAAGATTCTAATGTTCCTACCATTGTCCAAGATGAAATCCTTCAAAATTGAAATGGAAGCGATGATGAAGAAGTACGTCGCCAAGGTATAA
SEQ ID NO:11
TcTAT蛋白质序列
MEKTDLHVNLIEKVMVGPSPPLPKTTLQLSSIDNLPGVRGSIFNALLIYNASPSPTMISADPAKPIREALAKILVYYPPFAGRLRETENGDLEVECTGEGAMFLEAMADNELSVLGDFDDSNPSFQQLLFSLPLDTNFKDLSLLVVQVTRFTCGGFVVGVSFHHGVCDGRGAAQFLKGLAEMARGEVKLSLEPIWNRELVKLDDPKYLQFFHFEFLRAPSIVEKIVQTYFIIDFETINYIKQSVMEECKEFCSSFEVASAMTWIARTRAFQIPESEYVKILFGMDMRNSFNPPLPSGYYGNSIGTACAVDNVQDLLSGSLLRAIMIIKKSKVSLNDNFKSRAVVKPSELDVNMNHENVVAFADWSRLGFDEVDFGWGNAVSVSPVQQQSALAMQNYFLFLKPSKNKPDGIKILMFLPLSKMKSFKIEMEAMMKKYVAKV
SEQ ID NO:12
CrMAT DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGATTCTATTACTATGGTTGAAACCGAGACATTATCGAAGACATTGATTAAGCCATCTAGCCCAACCCCACAGAGTCTGTCACACTACAACCTGTCTTACAACGATCAGAATATATATCCAGAGTATATATTTGCAGGCTTCTTCTACAGCAACCCGGACGGACACGAGATCTCGACTATTAGAGAGCAGCTGCAGAACTCTCTTAGCAAGACATTAGTATCTTATTATCCATTCGCTGGTAAAGTGGTCAAGAACGATTACATTCACTGCAATGACGACGGAATAGAATTCGTAGACGTCAGAATCCACTGCAGGATGAACGATATTCTTAAGCCTGAATTGAGGTCTTACGCTTCCGAGCTTATTCGTCCGAATAGAAGTACAGTGGGGTCAGAGGACAGTACTGCGCTTGTCCAGTTATCTCACTTTGACTGTGGCGGGGTAGCTGTCGCATTCGGTATATCTCACAAAGTCGCAGACGCAGCGACTATTCTGTCGTTTATAAAGGATTGGGCCGCTTCTACGTGTGACTTGTCAAGTAGTCACGATGTGTCTACACCAGTACTAGTATCCGATTCCATATTCCCCCGTCAAGATAACATCATCTGTGGCCAGTTCCCTGCTTCACCGAACTGCGTGAGGAAGCGTTTCTTATTCAGCCCGGAAGCTATCGAAAGACTAAAATCGAAAGCCATTGAATTTGGGATCGAGAAGCCAACGAGGGTAGAGGTCCTGACAGCATTCTTGTGCCGTTGCGCTACCGTTGCAGGTAAATCTGCGGCCAAGAACAACAATTGTGGACAGTCACTGCCTTTCGCTGTTATTCAAGCAGTCAACCTGAGGCCCTTACTAGAACTGCCAAAGAATTCCGTCGGCAACCTTATATCAATCTACTTTTCTACAATCAAAGAAAACGACACGGTGAACATCGAACAGGAGTTCACAAAATTAGTGATCGGAGAGTTGAGGAAGGCTAAGGATAAGTTAAAGAACCTGTCGCAAGAGAAGCTGAATTACGTAGCTAGAATGCAAGATTTCGCGAATTGCCTGAAGGAATTGGACATAAGTTCATTCTTCGACATGGAAAACGTGGACATAGACGCTTATTTATTTTCGAGCTGGTGCAGGTTCCCCTTCTACGACATCGATTTCGGTCTGGGGAAGCCAATATGGGTCTGCATGTTTCAGCCTCACTTTAAGAATTGTATAATTTTAATGGATTATCCCTTTGGTGACGATTACGGCATCGAAGCCCTAATTACGTTGGAACAAGAGAAGATGCCCGCCTTCGAAAACAACGAGCTGCTGCTAAGCTTCGCCAGCAACTAA
SEQ ID NO:13
CrMAT蛋白质序列
MDSITMVETETLSKTLIKPSSPTPQSLSHYNLSYNDQNIYPEYIFAGFFYSNPDGHEISTIREQLQNSLSKTLVSYYPFAGKVVKNDYIHCNDDGIEFVDVRIHCRMNDILKPELRSYASELIRPNRSTVGSEDSTALVQLSHFDCGGVAVAFGISHKVADAATILSFIKDWAASTCDLSSSHDVSTPVLVSDSIFPRQDNIICGQFPASPNCVRKRFLFSPEAIERLKSKAIEFGIEKPTRVEVLTAFLCRCATVAGKSAAKNNNCGQSLPFAVIQAVNLRPLLELPKNSVGNLISIYFSTIKENDTVNIEQEFTKLVIGELRKAKDKLKNLSQEKLNYVARMQDFANCLKELDISSFFDMENVDIDAYLFSSWCRFPFYDIDFGLGKPIWVCMFQPHFKNCIILMDYPFGDDYGIEALITLEQEKMPAFENNELLLSFASN
SEQ ID NO:14
LiAAT-4DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGCCATGATAATCACGAAGCAAATATTAAGACCTAGTTCCCCGACGCCCCAAGCCTTTAAGAATCACAAGCTGTCCTACTTAGACCAAATACAGGCGCCTATTTACATACCTTTGTTGTTCTTCTATAAGAACGAGGAGTCAAAATACCCAGACCAGATCTCGCAAAGATTTAAGCAGAGTTTGTCCGAAATTTTGACAATATTCTACCCTTTGGCTGGTACGATGAGGCATAACTCGTTCGTGGACTGCAATGACAGGGGTGTCGAATTTGTAGAGGTCAGGGTCCATGCGAGACTAGCCCAGTTCATTCAAGATCCTAAGATGGAAGAGCTTAAGCAATTGATACCTGTGGATTGTATATCTCACACTGACGATGATTTCTTATTGCTAGTCAAGATTAGCTATTTTGACTGCGGTGAGGTCGTCGTTGGAGTCTGCATGTCGCACAAAATTGGTGACGGAATTAGTCTGGCAGCGTTTATGAACGCTTGGGCAGCAACGTGTAGGGGAGAATCGTCTAGTGAGATCATTCACCCATCTTTTGATCTTGCTTTACACTTTCCGCCTAAAGACCACTTGTCTTCAGCATCCTCATTCCGTGTGGCCATAGCCCAGGAGAACATCATGACCAAGAGGCTAGTATTTGATAGAGAAAAGTTGGAGAAGCTGCGTAAGAGAATCGCTGCCAGTTCTGATGGGGTGAGAGACCCTAGCAGAGTTGAAGCTGTATCTGTCTTTATTTGGAAAAGCTTAATTGAAGCCCACAAGGCCGAGTCACACATGACTGAGACACCAGCCGTTTCTATTGCTAGCCACGCCGTGAACTTAAGGCCTAGAACAGTCCCACAAATGGACCAAACTTTCGGTAACTGCTACGCTCCCGCTTCGGCCGTTGTCTCCTGGGATGAAGACTACGTACATCACAGTCGTTTGAGGGCGGCCCTTAGAGAAATCGACGACGACTACATTAATAAGGTCTTGAAAGCCGACAATAATTATTTAACGCAGGATCAAATTGGTGACTTGTTTAAACCAGAAAACTCAGTTCTAAGTTCGTGGTGGCGTTTCCCAGTTTACAAGGTAGACTTTGGATGGGGTAAGCCAGTTTGGGTTTCTACGACGACCATCCAATACATGAACTTGATTATATTTACTTCGACGCCCTCAGAAGACGGCATAGAGGCGTGGGTAACTACTACTCACAATTTCTTCCAAGTTCTGCAGGCAAACTATAATAAACTTGACACGTAA
SEQ ID NO:15
LiAAT-4蛋白质序列
MAMIITKQILRPSSPTPQAFKNHKLSYLDQIQAPIYIPLLFFYKNEESKYPDQISQRFKQSLSEILTIFYPLAGTMRHNSFVDCNDRGVEFVEVRVHARLAQFIQDPKMEELKQLIPVDCISHTDDDFLLLVKISYFDCGEVVVGVCMSHKIGDGISLAAFMNAWAATCRGESSSEIIHPSFDLALHFPPKDHLSSASSFRVAIAQENIMTKRLVFDREKLEKLRKRIAASSDGVRDPSRVEAVSVFIWKSLIEAHKAESHMTETPAVSIASHAVNLRPRTVPQMDQTFGNCYAPASAVVSWDEDYVHHSRLRAALREIDDDYINKVLKADNNYLTQDQIGDLFKPENSVLSSWWRFPVYKVDFGWGKPVWVSTTTIQYMNLIIFTSTPSEDGIEAWVTTTHNFFQVLQANYNKLDT
SEQ ID NO:16
FgaAT DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGAAGAAGCAGGTCACTTTTAAACCTTTCAGATTGTCACCAGTCGATCATAGTTTACCTAAGGTTTACATCTTCAAGTCTCTATATTTCAGAGGGGTAGATGACACTGGTTCTCTTAGTAGACTACAGGATGGCATTGATCGTTTGATATCTTGTCTGCCTTTCCTGTCAGGCGAGGTGGTTCCATGTGCAGACATACCAGATAAGGTCGGTGTACTTCAAGTTCAGATGCCTTGCCCTTCCTTGCAAGAAATTCCAATGCTGCTAGTCAAAAGTTATCCAAACCATACATGGCCAGCAGCTTCTACCTCGGAAAGATGGAGAAACACCGCTTTGCTAGATCAGTCGTATAGGCCATTGCCGGATTTTATACCCCCATCAAAGCCTAGACCTGTTTTGAGATTTCAGGCGAACTTCCTGGCTGATGGTTTAATGCTATGTATGGGATATAATCATTCTGTATTCGACGGAACAGGTGCGGGGAACATTCTGGAAATGTTGGCTGATTGTTGTAGAGCTAATCCAAACTCCATCTTAGCACTGCCAACAAACGGTGACATAGAGAGCGAGTTGAGAGGTTTACTGTCCAGTCCAGGTGTAGCAGTTGCAAATGCCTCTCAGGAAGCCTACGCAATTAATTGTGCACACACGGAAGTAGAACCCGAACCCAGTTCAGCTATGCTTTACTGTTGGCCTTTCTTGCTGAGTTCTGAGAAGATTGAATGTTTACAGGAAGCATGTAATAGTTTACTACCACATATCGTTCGTTTGTACTCTGGTACGCAGAGTTCGCTTATAAACCAAGATACGAACTGGCCACATATTCTTTCATCAAACGACGTGCTAACCGCTCTTTTAGCTGTTTCAATTGAAAAGGCTAGGGAAGCAACAGGTGCCCTTGGTCATATGAGCAGATCATTGGCTATGGCTGTTAACCTACGTGAGCGTTTAAAGCCTATGCCAAGACACTATTTGGGTAACCTAGTTACCACTGTGTGGGTATCACATCACCGTCCTGCCGTTAAGGACCTTGAAACAATGGTTTTACCAGTGCCAGCATGCAATAGGCACGAAATAGACAGAGACGACTTGTTGTGGATAACCCACGTGGCATTCCGTATTAGATTAGGGCTGAACGCAATAAACGAAGAACATATTAGAGGTCTGATCCACTACTTGCATTCCCAAGACGATTGGGAACAGATTGGAATACATTTCACCGATCCAATTTTCATTTCTTCCTGGCGTCACCTTAAGGTCTATGAACTAGACTTCGGACCTACTATCGGTCATGCTGAACACTTTGAGATGGATGTCGGCACCACCGATGGTGTGTGTGTTGTTATGCCTGCTAACACCAGAGCCGTCGGCAAGACTAAGAAGGCTCCTTGGGACATTAGAATCGTGTTAAACCCCGAAGTGTTACAGGCACTAATAGCTAGTGCCATCTTTGGTTGGGCTATGGTCAAGGACGCTTCGACATAA
SEQ ID NO:17
FgaAT蛋白质序列
MKKQVTFKPFRLSPVDHSLPKVYIFKSLYFRGVDDTGSLSRLQDGIDRLISCLPFLSGEVVPCADIPDKVGVLQVQMPCPSLQEIPMLLVKSYPNHTWPAASTSERWRNTALLDQSYRPLPDFIPPSKPRPVLRFQANFLADGLMLCMGYNHSVFDGTGAGNILEMLADCCRANPNSILALPTNGDIESELRGLLSSPGVAVANASQEAYAINCAHTEVEPEPSSAMLYCWPFLLSSEKIECLQEACNSLLPHIVRLYSGTQSSLINQDTNWPHILSSNDVLTALLAVSIEKAREATGALGHMSRSLAMAVNLRERLKPMPRHYLGNLVTTVWVSHHRPAVKDLETMVLPVPACNRHEIDRDDLLWITHVAFRIRLGLNAINEEHIRGLIHYLHSQDDWEQIGIHFTDPIFISSWRHLKVYELDFGPTIGHAEHFEMDVGTTDGVCVVMPANTRAVGKTKKAPWDIRIVLNPEVLQALIASAIFGWAMVKDAST
SEQ ID NO:18
GAO81666.1 DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGAAGAACATACCAGATCCTTTGAGCCATTTGACCTGGCCTGCTTAGATCATACTGTGGGGCCGGTTTTCATGAATTTCTTCTTAAGTTTCAAGCCTGCCAAGATCGAAGAGAGTCTTATCTCAATTGAAGAAGGTGTAACTAGGTTAGTCAACAGACTGCCATTCCTGGCCGGTGACGTTGTTAATTCTGAAAACGTAGACGGTCGTGTTAATGTAATGAGGATTCAGCCAAGCTCGACATTGATTAGAGAGATTCCCATGCTACACACAAAGCACCATCCTCATCATATTTTGCCGAATACACCTAGACAGATTAACCCAGGGTCCGACCAGGATCAAAGGTTTGCTCCCTTGGACGATTCGTACGTACCACCAGTGTCTTTATTACCTCTTGCTCCGGGCCCAAGGCCCGTAGTAAGATTCCAAACTAATGTTGTAATTGACGGGATTGTGCTTGCACTTGGGTTCCACCACTCAGTATTTGATGCTACCGGAGTGGGTTTGTTGATCGAAATGTTGGCCACATGTTGTTCAAGCGATTGTCCTGCCCTATCATCCCATATTGAACTGGAAGAAGAAATAAGGCTAAGGCGTTCTGTGGACAAGATAGGTAACGGCGCTACTGATTTAGCATCCCAAAGAGATGAAGTTCAGGACCCAAATGGCAGTATCACTGAGTCCCCAGCTCATGTGCCTATGGGTGACAGTAGCTGGGTACCCCCTAAACTATCTGTTTACTCCTTTAATTTATCGGCTGCCGGTTTGGCTCATTTGAAGACAGCCTGTAACAAGTTATTGCCCGCTATCCATGCGAGTCAAAATGGTTCACCACAAAGTGCGGAATCAGAAGGTGAGAAATTACAGCAAGAATTCGTTTCGACGAATGATGTTCTAACGGCTTTGTTGGCTACTTCCATTCACCAAGCGAGGTCTAGAGTTACTGAAACGGAATTAGTTCCGACAAAAGCGAAACTTGCGATGGCCGTTAATTTGAGGGAAAGGGCCAGTTCGTTACCCAAGACTTATTTGGGGAACTCGCTAACAGTTACAGAAGCTTTTGTGTATTCTTTAGCTGCTACCGACGGTTTCGATGGTGTCCCCGAGAGACATTATCATCCTGACTTGCGTAACGCCTTGCTATTAGAAATTGCTAGAGTTGCATTGCAATTGAGAAAAGGGCTGGCAGCGATAGATGACGCGTATTTTAGGCAATTTGTGTCTAGATTGAGGGCTAATCTAGACTGGTCACAGTTAGGTGCAAACCTGCCTGACACTATGGTGTCAAGCTGGAGACATTTGAAAGTATATCGTTTAGACTTCGGTGCTAGATTGGGGAGAGTTGTAGAGTTCCATCCACAAACCGCTTTGGTTGACGGTATTTGTATCATCCAACCTGAAAGGATAGCCCATGAGGATGATTCAGCTGATATGGCTCCAGAATCTGGGTGGGAAGTTTGTGTAACATTACAAAGCGATGCCATGGAGTGCTTCTTAAGAGGTGGGTTGTTCACAAGTCTTTCTCAAGGTGCAATTAGAAGAGTTTAA
SEQ ID NO:19
GAO81666.1蛋白质序列
MEEHTRSFEPFDLACLDHTVGPVFMNFFLSFKPAKIEESLISIEEGVTRLVNRLPFLAGDVVNSENVDGRVNVMRIQPSSTLIREIPMLHTKHHPHHILPNTPRQINPGSDQDQRFAPLDDSYVPPVSLLPLAPGPRPVVRFQTNVVIDGIVLALGFHHSVFDATGVGLLIEMLATCCSSDCPALSSHIELEEEIRLRRSVDKIGNGATDLASQRDEVQDPNGSITESPAHVPMGDSSWVPPKLSVYSFNLSAAGLAHLKTACNKLLPAIHASQNGSPQSAESEGEKLQQEFVSTNDVLTALLATSIHQARSRVTETELVPTKAKLAMAVNLRERASSLPKTYLGNSLTVTEAFVYSLAATDGFDGVPERHYHPDLRNALLLEIARVALQLRKGLAAIDDAYFRQFVSRLRANLDWSQLGANLPDTMVSSWRHLKVYRLDFGARLGRVVEFHPQTALVDGICIIQPERIAHEDDSADMAPESGWEVCVTLQSDAMECFLRGGLFTSLSQGAIRRV
SEQ ID NO:20
CfACT1-6 DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGAAGGTAGAACGTTTCTCAAGAAAGTTGATAAAACCCCACACGCCTACTCCCGAGAACTTAAAGAAATATAAATTGTCTCTATTGGACAAATGCCTTGGACACGACAACTTCGCGATCGTGTTGTTTTACGAATCTAAACCAAGAAATAAGAGTGAGCTTGAGGAATCGTTAGAGAAGGTCTTGGTAGACTTTTATCCCTTGGCTGGGCGTCACACGATGAACGACCATATTGTCGACTGCTCGGATGTCGGCGCTGTCTTTGTAGAAGCCGAGGCTTTAGATGTAGAGTTGACTATGGATGAGTTAGTCAAGAACATGGAAGCTCAAACTATACACCACTTGTTGCCAAATCAGTACTTCAGTGCAGATGCTCCGAACCCGCTGCTTTCCATCCAAGTTACGCACTTTCCTTCCGGTGGTTTGGCTATCGGAATCGCAGTCTCGCACGCTGTATTTGATGGTTTCTCTTTGGGCGTGTTCGTAGCAGCATGGTCAAAGGCGACCATGAACCCGGATAGGAAAATCAAAATAACACCGTCATTCGACTTACCATCACTTCTGCCCTACAAAGACGACAACTTTGGTTTGACTGCTGCTGAAATTGTCAGCCAGAGCGAGGACATCGTAGTTAAGAGATTTATCTTCGGCAAGGAAGCCATCACGAGGTTGAGAAGTAAGCTTAGTCCAAATAGGAACGGGAAGAAAATATCCAGGGTTAGGGTCGTTTGTGCAGTCATTGTAAAGGCCTTGATGGGATTGGAGCGTGCCAAACACGGTAAGACGCGTGATTTCTTAATTACTCAATCAATTAACATGAGGGAGAGAACTAAGGCGCCGCTGCAGAAACACGCCTGCGGCAACTTAGCAGTCTTGAGTTGCACGAGAAGAGTAGAGGCCGAGGAGATGATGGAGTTACAGAACCTAGTTAATTTGATCGGCGACAGTACCGAAAAGGACATCGCCGACTTTGCAGAATTATTATCACCAGATCAAGTGGGCAGAGACATTATAATCAAGATGATGAAGTCATTCATGCAGTTCTTGGACAATGACATTTACAGCGTGTGCTTTACCGATTGGTCAAAGTTTGAATTTTACGAAGCGGACTTCGGCTTCGGGAAGCCGGTTTGGATGGCAGCAGGCCCACAGAGACCCATTATCTCTACTGCAATATTGATGTCAGACAGGGAAGGCGACGGTATCGAGGCTTGGTTACATTTGAATAAGAACGACATGTTGATTTTCGAACAAGACGAGGAAATCAAGTTATTCACTACATAA
SEQ ID NO:21
CfACT1-6蛋白质序列
MKVERFSRKLIKPHTPTPENLKKYKLSLLDKCLGHDNFAIVLFYESKPRNKSELEESLEKVLVDFYPLAGRHTMNDHIVDCSDVGAVFVEAEALDVELTMDELVKNMEAQTIHHLLPNQYFSADAPNPLLSIQVTHFPSGGLAIGIAVSHAVFDGFSLGVFVAAWSKATMNPDRKIKITPSFDLPSLLPYKDDNFGLTAAEIVSQSEDIVVKRFIFGKEAITRLRSKLSPNRNGKKISRVRVVCAVIVKALMGLERAKHGKTRDFLITQSINMRERTKAPLQKHACGNLAVLSCTRRVEAEEMMELQNLVNLIGDSTEKDIADFAELLSPDQVGRDIIIKMMKSFMQFLDNDIYSVCFTDWSKFEFYEADFGFGKPVWMAAGPQRPIISTAILMSDREGDGIEAWLHLNKNDMLIFEQDEEIKLFTT
SEQ ID NO:22
CfACT1-8 DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGAAAGTCGAAAGGATTTCACGTAAATTCATCAAGCCATATACACCTACACCACAGAACCTTAAGAAGTACAAGCTATCCTTGCTGGATAAATGCATGGGACACATGGACTTCGCTGTAGTATTGTTTTACGAATCAAAGCCAAGAAACAAGAATGAGCTGGAAGAATCACTAGAGAAAGTGTTAGTCGATTTCTATCCATTGGCAGGCAGGTATACCATGAACGACCACATTGTCGATTGCAGCGATGAGGGCGCCGTTTTCGTTGAGGCAGAGGCCCCTAATGTTGAGCTTACAGTGGACCAGTTGGTTAAGAACATGGAAGCCCAGACAATCCACGACTTCTTACCAGACCAATATTTTCCTGCTGACGCACCAAATCCGTTGCTAAGTATTCAAGTAACGCACTTCCCTTGTGGTGGTTTAGCTATCGGGATTGTTGTTAGTCACGCGGTCTTTGATGGATTCTCATTGGGCGTATTCTTAGCCGCCTGGAGCAAAGCTACCATGAACCCTGAGAGGAAGATCGAAATCACCCCTTCCTTCGATTTGCCTAGTCTTCTGCCCTACAAGGATGAATCTTTCGGTTTAAATTTTAGCGAAATTGTCAAAGCTGAGAATATCGTAGTTAAACGTTTGAATTTCGGGAAAGAGGCTATTACGCGTTTGAGGTCCAAGCTGTCTCCTAACCAGAATGGTAAAACCATTTCCAGAGTGAGGGTAGTCTGCGCGGTGATAGTTAAGGCGTTGATGGGACTGGAGAGAGCAAAGACTAGAGACTTTATGATATGTCAGGGGATCAACATGAGAGAGAGGACGAAGGCGCCCCTGCAGAAGCATGCGTGTGGCAACCTAGCAGTTTCGTCTTACACTAGAAGGGTAGCGGCAGCGGAAGCAGAAGAACTGCAGTCCTTAGTGAATTTGATCGGGGACTCTATCGAAAAGTCAATCGCAGACTACGCTGATATACTTTCGAGTGATCAAGATGGGAGACACATCATTTCCACTATGATGAAGAGCTTTATGCAGTTTGCTGCACCTGATATAAAAGCCATTTCATTTACCGACTGGTCAAAGTTCGGCTTTTACCAAGTAGATTTTGGTTTTGGTAAACCAGTTTGGACCGGCGTCCGTCCAGAACGTCCAATCTTCTCAGCCGCGATATTGATGAGTAACAGGGAAGGCGATGGCATCGAGGCTTGGCTTCATTTGGACAAGAACGATATGCTAATATTTGAGCAAGACGAAGAAATTAAGTTGTTGATAACGTAA
SEQ ID NO:23
CfACT1-8蛋白质序列
MKVERISRKFIKPYTPTPQNLKKYKLSLLDKCMGHMDFAVVLFYESKPRNKNELEESLEKVLVDFYPLAGRYTMNDHIVDCSDEGAVFVEAEAPNVELTVDQLVKNMEAQTIHDFLPDQYFPADAPNPLLSIQVTHFPCGGLAIGIVVSHAVFDGFSLGVFLAAWSKATMNPERKIEITPSFDLPSLLPYKDESFGLNFSEIVKAENIVVKRLNFGKEAITRLRSKLSPNQNGKTISRVRVVCAVIVKALMGLERAKTRDFMICQGINMRERTKAPLQKHACGNLAVSSYTRRVAAAEAEELQSLVNLIGDSIEKSIADYADILSSDQDGRHIISTMMKSFMQFAAPDIKAISFTDWSKFGFYQVDFGFGKPVWTGVRPERPIFSAAILMSNREGDGIEAWLHLDKNDMLIFEQDEEIKLLITT
可选地,C-末端的“T”可缺失
SEQ ID NO:24
OAH94415.1 DNA序列,针对其在酿酒酵母中的表达经密码子优化
ATGGATACCAAGAGGGTCGGTTATACAGTCGTTGATTTGAGTCAGTGGGGCAGAAAAGAACACTTCGAAGCATTCCAGAGCTTCGCCCAGTGCACCTTTTCCCAGACTGTTCAATTAGACATAACTTCCCTGCTGAAGACTGTCAAGCAGAACGGATATAAATTCTACCCGACCTTTATCTACATAATAAGCCGTTTAGTGAACAAGCATGCCGAATTCCGTATGGCCATGAAGGATGGTGAACTGGTAATTTGGGACTCCGTAAACCCTGGTTACACAATCTTTCACGAACAGACCGAAACATTTTCATCTCTGTGGAGCTATTACCACAAGGACATCAATCAGTTTCTTAAAACGTATTCTGAAGACATCGCTCAGTACGGGGACGATCTGGCGTACTTCCCCAAGGAGTTTATTGAAAATATGTTCTTCGTGTCAGCGAACCCATGGGTGAGCTTCACAAGTTTCAACTTAAACGTGGCCAATATTAATAACTTCTTCGCACCGGTCTTTACAATCGGTAAATACTACACGCAGGGCGACAAGGTGTTGATGCCGTTAGCTATTCAGGTCCATCATGCAGTCTGCGACGGGTTTCATGTCGGCAGGTTATTGAACGAAATTCAGCAGTACTGCGATGAGGGATGCAAGTAA
SEQ ID NO:25
OAH94415.1蛋白质序列
MDTKRVGYTVVDLSQWGRKEHFEAFQSFAQCTFSQTVQLDITSLLKTVKQNGYKFYPTFIYIISRLVNKHAEFRMAMKDGELVIWDSVNPGYTIFHEQTETFSSLWSYYHKDINQFLKTYSEDIAQYGDDLAYFPKEFIENMFFVSANPWVSFTSFNLNVANINNFFAPVFTIGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRLLNEIQQYCDEGCK
SEQ ID NO:116
DfACT13天然核苷酸序列
ATGGCCTGTGGAGGCGGCGTGAGGAATATTGAGGTGAAGGCGCAGGAGCCCGTTTTGGTGCAGCCTTTATCTACAGCACAAAGCTCTGCCTATAATCTTCTTACAAAACTGGACCAGACGCTCGCACAATTGGTGGTGCAGATTGTCTTTGTCTTCGATGTGAAAAACCCCGTAACCCGCCGACCCCACGATGGCGCCGATCCTGCCAAGCTTCTGAAGGAGGCTCTGAGGAAGGTCCTTGTGCCCTTCTACCCGCTCGCTGGGCGTCTTTGCCTCTCGCCCGATGACGGCAGTCTGTTCATTGACTGCAATGCTCAGGGGGTTTCCTTTGTTGAGGCCAATGCCGACGCGGATATCTCCGAACTCGGCGACTTCTCGCAGCCCGATTTCGCGACTCTGGGCTCCCTTGTCTTCCCCTTACCTCCTATAGCTTCGGATGATGGTCCTCTTCTATCTGCGCAGGTGACCAGATTCAAGTGTGGAGGATTTGTGCTGGGCTTCGTATTCCATCATTGTTTATTTGATGGATTCGCACTTTCGGAATTCCTAAATGCGTGGGCGGAGACTGCATGCGGCGTGCCTCTTTCTACACCTCCTGTCCTCGACAGAACTTTTCCGAGGGCGCGTTCTCCCTTGCAAATCAAGTATCCCCACACCGAGTTCCTGGAAGTCGAAGATGTCTCTTTGACTCAAAATATCTCTAACGATGCCATCAACCGGTCTTTCTGCTTTACTTCGGCAAGTCTAGAGATACTCAAGAAGAAAGCCTTGGAGGATGGGGTGCTATCAAAATGCACTACTTTTGAAGCTTTATCTGGGCTGATATGGAGGGCCCGAACTAGAGCCCTGTGGAGCGATTACCCGGAGCACAAACTAAAGGTGCTCATTGTCGTCGACCCGAGAGCACGTTTTGAGCCTCGTGTGGTGCCAAAAGGGTATGTGGGCAATGCGGTGCTTTTTACATGTGCTTTCGCAAGCGCAAGGGAGCTGGAAGAAAATCCCTTGTCACATGCAGTGAAGCATGTGCAACATGCCATCGGGCGTATGACGGAGGAGTACATGTTGTCGCAAATCGACTACATGGAGCATCAGAAGGTATGGTGCCCACCGCTAGGAGCTAGTACATCTTTCATGACCAAATGGTCTAGGTTGGCCTTCAATATTCTAGACTTTGGGTGGGGCAGGCCGAAGTATGTGGGGCCGGCCACGTCGCTGTCGATGGAGACGACTACTTTTGTATCTTATGGAAAGGGCATGAGTGTGGTGTTGGCTCTTCCTCCAGAAGCAATGCGCAAATTTGAAAAAATCGTACATCCCTACCTCAATCCATGA
SEQ ID NO:117
DfACT13核苷酸序列,针对其在酿酒酵母中的表达经密码子优化.
ATGGCATGTGGCGGTGGGGTCAGGAATATCGAAGTGAAGGCTCAAGAGCCAGTCTTGGTGCAACCGTTAAGCACTGCTCAATCGTCAGCCTACAATTTGTTAACAAAGTTAGACCAGACTTTGGCGCAATTGGTTGTCCAAATAGTGTTTGTTTTCGATGTGAAGAACCCTGTGACAAGAAGGCCTCATGATGGAGCCGATCCAGCTAAGTTATTGAAAGAGGCCTTAAGGAAAGTACTAGTACCATTCTATCCCCTTGCCGGAAGACTATGTCTTTCCCCAGACGATGGTAGTCTATTCATAGACTGCAACGCTCAAGGTGTTTCATTTGTCGAAGCAAACGCTGATGCCGACATCAGTGAGTTAGGTGATTTCTCCCAACCGGATTTTGCGACCTTAGGTTCTTTAGTCTTCCCTCTTCCACCAATTGCGTCTGACGATGGACCATTGTTAAGTGCTCAAGTCACAAGATTCAAATGTGGTGGCTTCGTTCTAGGTTTCGTCTTCCATCACTGTCTATTTGATGGTTTTGCCTTATCTGAATTTCTTAATGCATGGGCTGAGACAGCGTGTGGAGTCCCGTTATCAACCCCACCTGTTTTAGATAGAACATTTCCTCGTGCTAGATCTCCCCTGCAAATAAAGTACCCACATACAGAGTTCCTAGAGGTAGAAGATGTTTCATTAACTCAGAACATCTCTAACGATGCTATCAACAGATCCTTCTGTTTTACATCAGCCTCTTTGGAAATTTTGAAGAAGAAGGCGCTAGAAGACGGGGTGTTAAGCAAATGTACGACCTTCGAAGCTCTATCTGGCTTAATATGGAGAGCACGTACCAGAGCCTTATGGAGCGACTACCCCGAGCACAAGTTAAAGGTCCTGATTGTCGTGGACCCTCGTGCCAGATTTGAACCGAGAGTGGTGCCAAAAGGCTACGTAGGGAATGCAGTCTTGTTTACTTGTGCATTTGCTTCAGCCAGAGAACTAGAAGAAAATCCATTATCTCATGCTGTTAAACACGTACAGCACGCGATCGGCCGTATGACTGAGGAATATATGCTATCACAAATTGATTACATGGAGCACCAGAAGGTTTGGTGTCCCCCACTAGGTGCGTCGACTTCTTTTATGACTAAGTGGTCAAGGCTTGCCTTTAACATCTTAGATTTTGGTTGGGGTAGACCTAAGTATGTTGGTCCGGCTACTTCATTGTCTATGGAAACAACTACATTCGTCTCCTATGGAAAGGGTATGAGCGTGGTGTTAGCCCTACCCCCAGAGGCAATGAGAAAGTTCGAAAAGATTGTACACCCTTATTTGAACCCTTAA
SEQ ID NO:118
DfACT13蛋白质序列
MACGGGVRNIEVKAQEPVLVQPLSTAQSSAYNLLTKLDQTLAQLVVQIVFVFDVKNPVTRRPHDGADPAKLLKEALRKVLVPFYPLAGRLCLSPDDGSLFIDCNAQGVSFVEANADADISELGDFSQPDFATLGSLVFPLPPIASDDGPLLSAQVTRFKCGGFVLGFVFHHCLFDGFALSEFLNAWAETACGVPLSTPPVLDRTFPRARSPLQIKYPHTEFLEVEDVSLTQNISNDAINRSFCFTSASLEILKKKALEDGVLSKCTTFEALSGLIWRARTRALWSDYPEHKLKVLIVVDPRARFEPRVVPKGYVGNAVLFTCAFASARELEENPLSHAVKHVQHAIGRMTEEYMLSQIDYMEHQKVWCPPLGASTSFMTKWSRLAFNILDFGWGRPKYVGPATSLSMETTTFVSYGKGMSVVLALPPEAMRKFEKIVHPYLNP
SEQ ID NO:119
PYI04555.1天然核苷酸序列
ATGGGTGCCAGCGTCTCTTTCCAGCCCTTTGTGCCCACTCCCCTGGACCATGCTATGCCCCCAATTTACGTGTCTCAATTTCTATGCTTTCCTACCACAACTCCGCAATCCGCTATTCAAAGTCTCCAAGTGGGAATCGAGAGATTATTCGAACGCCTGCCATTCCTGGCGGGAGAGATTCTCATCAATGAACACACCGGAGCCATCAAAGTCCAGGCTCCCAGTGCTTTGATCCGGGAAATTCCCTACATGGCCCTCCGAGCCCATCCTGATCTTTACCTTCCAGCTAAGCAATGTGCAACCACACCAATCGAGAGACAGTTGAAGACCAACAGCCTTGATGAATCCTACCATCCACTCCCGGCGGCACTTCCACTCTCCCAACCCCAGCCCGTCATCCGATTCCAAGCAAACACCCTCGCAGACGGCATTCTCTTTGCAGTCAGCTATCATCATTGCATATTCGATGGCACTGGATGCGGACAGATTCTGGAAATGTTGGCTCAGTGCTGTTCGGCCTCCGACGATAAGATCTCCCTGCCGACTGATTGCCACACCGATGTGCTCCTCCGCGAATACATCTCCAATCTAAGCCCTACTACCAACATCCCCCACGATTACACGCAAGCGTATAGCACTACGGTGCAACCGGACCCCGATGCCTCAGACCCAGACACGTCCCCCGCCATACCCTCCTCACTCTACACAGAAGCATTCACCTTTCCCTCCCAACAAATCACCACTCTCCGTGATGCATGCAACCACCTCTTGCCCAAATTACCCAGCACCAGCAACGCACATCCCCACAAACCAACACCGAATCCCCTATCATCAAATGACGTCCTCACCGCACTAATAGCCCTATGCATCACACGCGCCACCAACACCACCACCCCACCCCTCCAACCCAACAATCACAGTCTCTCAATGGCCGTCAACCTCCGAACCCGCATCCAGCCCCAAGTACCAGATCACTTTCTGGGAAACTTCGCCACGCTACTCCCAATACACTTTACCAGTCCAGTCCACACCCAACAGTCTGATCTCCTCCTCACCACAGAACCCCCCGACCCCGCCCTCATCCACCTAACCACCCTCGCCTCCCAAATCCGGTCCAGCCTATCCACAGTCAACACCGATTACATCTGCGGTCTCATGACGGATCTCCGAACCCGGCGCAATGCAGGAGAAAACAGCAGTCTTCTAATTGAAGGCATTAAGATTTCCAGTTGGAGACATCTATCCGTCTACAAGCCGGACTTTGGCCCTGGGTTGGGGAAAATAGCGGGGTTTGAGTTCCAGGCGGGGCTTATGGATAATTTGGTGGTGATTTTGCCTTGGAGAAATGGGGATTGGGATGTGCGTGTTACGTTGTTGGAGAGGGATATGCGCGGGTTTAGGGAGGATCGGTTGGTTAGGTGGGCGTTGGGGTCTGGGTAG
SEQ ID NO:120
PYI04555.1核苷酸序列,针对其在酿酒酵母中的表达经密码子优化,编码SEQ IDNO:143的C-末端扩展蛋白变体。
ATGGGTGCGTCAGTTAGTTTTCAACCTTTCGTTCCTACCCCCTTGGATCACGCCATGCCACCGATATACGTGTCACAGTTCTTGTGTTTCCCTACTACGACGCCGCAGTCAGCAATACAATCCCTACAGGTCGGTATTGAGCGTTTGTTTGAAAGACTACCTTTTCTAGCCGGGGAAATTCTGATAAATGAGCACACAGGTGCAATAAAAGTTCAAGCTCCCTCTGCTCTGATTAGAGAAATTCCGTACATGGCCTTAAGGGCTCATCCGGATTTGTATCTGCCTGCTAAGCAATGTGCAACTACTCCAATAGAGAGGCAGTTGAAAACTAACAGCCTTGATGAGTCTTATCATCCATTACCTGCGGCACTACCATTGTCTCAACCACAACCAGTCATCAGATTCCAAGCTAATACACTTGCTGACGGAATACTTTTCGCTGTCAGTTACCACCATTGTATATTTGATGGAACGGGCTGCGGACAGATCCTTGAAATGTTAGCCCAGTGTTGTTCGGCCTCAGACGATAAGATCAGCTTGCCTACGGACTGTCATACAGATGTATTGTTGAGAGAATATATTTCGAATCTATCTCCAACCACGAATATCCCGCATGACTACACTCAAGCTTATTCTACTACAGTTCAACCAGATCCTGACGCATCCGATCCTGATACTAGCCCAGCCATTCCGAGTTCGCTGTACACTGAAGCATTTACGTTTCCTTCACAACAGATTACCACACTAAGAGATGCGTGCAACCATTTATTGCCTAAATTACCATCTACTTCAAACGCACATCCCCACAAACCAACACCAAACCCGTTATCCTCAAATGACGTTTTGACGGCACTGATCGCATTGTGCATCACCAGAGCTACTAATACGACGACACCCCCATTACAACCAAACAACCATTCTCTTAGCATGGCCGTCAATCTAAGGACTCGTATTCAACCCCAAGTCCCGGACCACTTTCTTGGTAATTTTGCCACCTTGCTACCTATTCACTTCACAAGTCCAGTCCATACGCAGCAATCAGATTTATTATTGACTACTGAGCCGCCAGACCCAGCATTGATCCACCTTACAACCCTTGCATCGCAAATTAGGTCTAGTTTATCGACCGTTAACACCGATTACATATGTGGCTTGATGACAGATCTGAGGACTAGGAGAAACGCCGGTGAGAATTCCTCTTTGTTAATAGAAGGTATTAAGATTTCTTCATGGAGACATCTTAGCGTTTACAAGCCAGATTTCGGACCTGGATTAGGGAAGATTGCAGGATTTGAGTTTCAAGCTGGTTTAATGGACAACCTTGTTGTGATATTGCCTTGGAGAAACGGTGATTGGGACGTGCGTGTAACTCTTCTGGAGAGAGATATGAGGGGTTTTAGGGAAGACAGATTAGTCAGATGGGCATTAGGTAGCGGAACAGGCCCCTTTTCCTTTGTCGATATCATGTAA
SEQ ID NO:121
PYI04555.1蛋白质序列,SEQ ID NO:143的C-末端扩展蛋白变体。
MGASVSFQPFVPTPLDHAMPPIYVSQFLCFPTTTPQSAIQSLQVGIERLFERLPFLAGEILINEHTGAIKVQAPSALIREIPYMALRAHPDLYLPAKQCATTPIERQLKTNSLDESYHPLPAALPLSQPQPVIRFQANTLADGILFAVSYHHCIFDGTGCGQILEMLAQCCSASDDKISLPTDCHTDVLLREYISNLSPTTNIPHDYTQAYSTTVQPDPDASDPDTSPAIPSSLYTEAFTFPSQQITTLRDACNHLLPKLPSTSNAHPHKPTPNPLSSNDVLTALIALCITRATNTTTPPLQPNNHSLSMAVNLRTRIQPQVPDHFLGNFATLLPIHFTSPVHTQQSDLLLTTEPPDPALIHLTTLASQIRSSLSTVNTDYICGLMTDLRTRRNAGENSSLLIEGIKISSWRHLSVYKPDFGPGLGKIAGFEFQAGLMDNLVVILPWRNGDWDVRVTLLERDMRGFREDRLVRWALGSGTGPFSFVDIM
SEQ ID NO:122
ERR364415-1_contig_8546天然核苷酸序列
ATGGCCCGGGCACCACCACCTCCGCCTCCTGGTCTCAGAATGAGAGACACAGTGCTCAGCATCGTGAAGCCGATACGGAAGACACAGCATTTGGAGACGATCGACGCAACGTTCGTCGATTTGATGAGAATGGACAGCTTTATACCCGTGATTTTCGCCTACAGGCCTGCGGACAAGTCCGAGGCCGCGTACTCGCGTCTCGTGAATCGCATCAAGGAGTCGCTCCAGAAGGTTTTGGTCCCCTTTTTCGGGTTCGCGGGCCGCTGGGTTCCAAGCAGTGGCGGGAGCAGGCGGCTCTTGTGCAACGATGAGGGCGTTCCCTTCATTGAAGCGTTTGTGGACGAAGAGTTGGACTCGGTGGTGAAGGCTTCCGCCGCATTCCAGCCGGTTACGGAGCTGAATGGCTTGGGCGTCCTCGGAATGGACATGACTTCATACGATCAAAGGATGCCACCGGAAGGTGGGCAACCTTGCGTCGTTGCTCAAGTCACACGGTTCAAATGTGGGGGAGTGGTTCTGGGGGTGGCTTTCAATCACACTCACACTGACGGCCAGGGATTCTACACCTTCATGCGAGCATGGTCCGACTTCTCTCGAACCAACGGAACGGCAATCAAGGTGGACCACAACCGGGCCCTGCCAGAACTGGCTTCCCTCTCACAGTTCTTCATCAAACAGCACGACCGAATAGGAGGCAAAACTTCTACCGATCGAGTCAACGATCATTGTTCTAAAGTTCCGGAACGGCTGGCTTTGAAAGCTTTCGAGGTTCGTGCGTCTAAGATCAAAGCCGCAAAGCTAGCAGCCGAAGATGGAGGGGTTGGGTATGTCAGCACGGTAGATTGCATTGTGGCTCACTTATGGAAAACTCTTGCCAGATTGCCGCCCGTCGTGTTGGATGGGAGGGAGATTACGGTCTTCTCGCCTGTGGAGGGGAGGAACAGATTCTTGGACCCGCCAAGACCCAATATGTGTGGAAATTGTTTTGCAGCAATGGTGACCCCCAAAATCCCAACCCAGGAGTTGCTGGAGATGCCTCTCGCTGCAATTGCAGGCAAGCAACGGGAGAAATTATCCACAACCCGAAGGGAGGAATGGTTTGGACAGCAAAGCTTTAGGGAGCTGGCCTCCGCGATGAACACCAGCAAATCTGCTCTACTTATTGTGACCTCGTGGTTCAACTTTCCCATGTATGAGATCGACTTTGGAGCTGGCAAACCATTTTTTGCATCCACTACGAACATGATTTCTCCTATCAACGGCGTGTGTTGTGGAGTCATTGCACCCCCAACTCCTGGGAGCTGCTCCTCCATTGCCACTCTGTACATTTTGTGCCTTCCCGCGGTACTAGAGGCTCTTGAAAATGTTCCAGATTTCCTATCCTTCTTCGTTCCTCACCCAAATCACAAAGATAACTCGCAATAG
SEQ ID NO:123
ERR364415-1_contig_8546核苷酸序列,针对其在酿酒酵母中的表达经密码子优化,编码SEQ ID NO:144的C-末端扩展蛋白变体。
ATGGCCAGGGCTCCACCCCCACCACCACCGGGCTTGCGTATGAGAGATACGGTCCTATCAATCGTCAAACCTATAAGAAAGACTCAACACCTAGAGACAATTGATGCAACATTCGTAGATTTAATGAGAATGGACTCATTTATCCCTGTGATCTTCGCTTATAGACCTGCAGATAAGAGCGAGGCTGCGTACTCTAGATTAGTTAATAGGATAAAAGAAAGCTTACAGAAAGTACTAGTACCATTCTTTGGATTTGCTGGAAGGTGGGTGCCTTCTTCCGGTGGCTCAAGGCGTCTGCTATGCAATGACGAAGGTGTCCCTTTTATTGAGGCGTTTGTAGACGAAGAACTGGACTCTGTCGTTAAGGCTTCAGCTGCCTTCCAACCTGTAACTGAACTTAATGGTCTAGGTGTGTTGGGTATGGATATGACTAGTTATGATCAAAGAATGCCCCCTGAAGGCGGTCAACCGTGCGTCGTAGCTCAGGTAACGAGATTTAAATGCGGCGGTGTGGTATTGGGTGTAGCATTCAACCATACCCATACTGACGGGCAGGGCTTTTATACTTTTATGCGTGCATGGTCGGATTTCAGTAGAACGAATGGTACCGCAATTAAAGTCGATCACAACAGGGCACTACCCGAATTAGCATCTTTGAGTCAGTTCTTTATTAAGCAGCATGATAGAATTGGTGGGAAAACTTCCACCGATAGAGTCAATGACCACTGTAGCAAAGTCCCTGAAAGACTAGCACTAAAGGCTTTTGAAGTTAGGGCGTCCAAAATCAAGGCAGCAAAACTAGCCGCAGAAGATGGTGGTGTAGGCTACGTCTCGACGGTGGATTGTATTGTTGCTCATCTATGGAAGACACTAGCTCGTTTGCCACCAGTTGTGTTAGACGGTCGTGAGATCACTGTGTTTAGCCCAGTAGAAGGCAGAAATAGGTTTCTTGATCCGCCCCGTCCGAACATGTGTGGTAATTGTTTCGCTGCAATGGTAACCCCTAAAATCCCAACACAAGAATTGTTAGAGATGCCATTAGCCGCCATTGCCGGCAAGCAGAGAGAGAAACTATCTACCACGAGACGTGAAGAGTGGTTCGGACAGCAATCATTCAGGGAGTTGGCTTCAGCTATGAATACTTCTAAATCAGCTTTGTTAATCGTGACATCTTGGTTTAACTTCCCGATGTATGAAATCGATTTTGGTGCCGGTAAGCCGTTCTTCGCTAGTACGACTAATATGATTTCTCCTATAAATGGAGTTTGTTGTGGTGTCATAGCCCCGCCCACCCCCGGTTCCTGTTCATCCATAGCGACATTATACATTTTATGTTTACCAGCCGTGTTAGAAGCTCTTGAAAATGTCCCAGATTTCCTTTCGTTCTTCGTACCGCATCCAAACCATAAAGACAACAGCCAAACAGGCCCCTTTTCCTTTGTCGATATCATGTAA
SEQ ID NO:124
ERR364415-1_contig_8546蛋白质序列,SEQ ID NO:144的C-末端扩展蛋白变体。
MARAPPPPPPGLRMRDTVLSIVKPIRKTQHLETIDATFVDLMRMDSFIPVIFAYRPADKSEAAYSRLVNRIKESLQKVLVPFFGFAGRWVPSSGGSRRLLCNDEGVPFIEAFVDEELDSVVKASAAFQPVTELNGLGVLGMDMTSYDQRMPPEGGQPCVVAQVTRFKCGGVVLGVAFNHTHTDGQGFYTFMRAWSDFSRTNGTAIKVDHNRALPELASLSQFFIKQHDRIGGKTSTDRVNDHCSKVPERLALKAFEVRASKIKAAKLAAEDGGVGYVSTVDCIVAHLWKTLARLPPVVLDGREITVFSPVEGRNRFLDPPRPNMCGNCFAAMVTPKIPTQELLEMPLAAIAGKQREKLSTTRREEWFGQQSFRELASAMNTSKSALLIVTSWFNFPMYEIDFGAGKPFFASTTNMISPINGVCCGVIAPPTPGSCSSIATLYILCLPAVLEALENVPDFLSFFVPHPNHKDNSQTGPFSFVDIM
SEQ ID NO:125
XP_001258079.1天然核苷酸序列
ATGACCGTGACCATCAGTTTCGAGCCATATGTGGGCTCCTCTGTCGATGCTCTAAGCATCCCTCTCTATCTTCGATGTCAACTCGTCTTCAAACTTTCTAAGCCACTTGCTGCGGTGCCTCTGCTTGAGTCTGGAGTTAATCGTCTTGTACAAGCGTTACCCTTCCTCTCGGGCGAGTTCACGGCCGTGCCAGCATCCGACGGTGGGAAAGAAATTCTTCTCGTTCGCCCTGTGCTCAACTTCGAGCTCAGCCGTATACTCAAGATCAAGTACCATGAAACATCCCTACGACATGTATGCAAACAGATGAACAGGCCAAGCAGCCAGGGTGGTGACCTTCCGCATGAGCCATACATGCCCTACCCACGACTTCCAGATCCTTCACGCCCTCAACCCATCGTCGGGTTCCAAGTCAACGTTCACACGGATGGCATCATTCTCTCCGTTGCTACGCATCACTGTTCCTTTGACGCAACAGGGATGGGATCAATCGTCCAAAACCTCGCGGCTTGTTGCCGTTCTCCTCCGAGCGACGAGCCTGACTTGACCACGTCGCCAGCCCAGGAAGCAGAAGCAAGAAAAGTCCTCTCGCAAGTCCGCGAGACGCCTTTTGATCCAAAGATGTTCCCGGAGTACAGACCCTTGGACAGTATGCTGTCCTATTACAAAGGCGTCCAGTCAGCGCTCCAGGGTCGTCAAACCACTATCGTCAATCGATGTTTCACAATCGCCGCCGACAAGATCAACGCGCTCAAAAGGCGCTGCAATCAACTGATACCGGAAATGGTGAAGAAGTATGGGCTTTCAACTGAGGATGCCATTGGGAGCGCCTGGGTCTCGAGTAATGATGTTGTTGCTGCCCTCTTGTGGACATGTATCAATCGAGCGCGATATCCCGAAATCCGCGAGCGCAGCGTTCACCAGCTCCCACCAGACCTCCTACATGCGACATCTAGCCTGGGTGTGCCAGTGAACGTTCGCTCTCGACTGTCGCCGCCCTTACCCAAATCGACTTTAGGTAACGCCGTGTGTCTTCTCCGGGAGAAGGTCCCGCTGCAATTTTTCGCTTTGCCTAGTCATGCCAACATGGAGGCCACTTCTAGCGTTTGCGCAGACCATTCCGGAGACGACGAATGGGCCTTGTCCTTCTGTCGAGTCGCCTACGGACTCAGAGCGAAGCTGAACGCAATTGATGACGACTATATCCGCGACTATATCTCCTACGTGCAAAAGTCTCCGTGCCATCTGTCAGTGACACTGGATACAGAGAACCTGTACCTCAGTAACTGGCGCGAGATCGGTGTGTATGATGCTGATTTTGGAGGCATGCTGGGCAAGCCGCTACGGATGAGAGCTCCGGATGGATACACCGATGGCCTGATTTTTGTGATGGCGCAGCGGAGCGAAGATAAGTCTGCACCGTGGGAGTTTAATATCTCGCTGGAGGCATCGACAATGAAGCGTATTGTGCATGATCCCCTCTGGTGCAAGTATGTTGAGCTGGATGCGTTCTGGCATGGAGAAGAATGA
SEQ ID NO:126
XP_001258079.1核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGACCGTCACGATTTCATTCGAACCATATGTGGGATCTAGCGTGGACGCGCTGTCCATACCCTTGTATCTAAGGTGCCAACTGGTGTTCAAATTATCCAAGCCTCTTGCAGCCGTGCCCCTGCTAGAATCTGGGGTAAACCGTCTAGTACAAGCATTACCATTCTTGTCTGGTGAATTTACTGCTGTCCCTGCAAGCGATGGTGGGAAGGAAATCTTACTTGTTAGACCAGTCTTAAATTTCGAACTAAGTCGTATACTGAAGATCAAATACCACGAAACATCCTTAAGACACGTATGCAAGCAGATGAACAGACCATCCTCGCAAGGTGGTGACTTGCCACACGAACCGTATATGCCATACCCCAGGTTACCAGATCCATCTAGGCCTCAACCAATCGTGGGTTTTCAAGTCAATGTCCACACTGACGGTATAATCCTGAGTGTAGCAACTCACCATTGCAGTTTTGACGCCACGGGGATGGGAAGTATTGTACAGAACTTGGCCGCATGCTGTAGATCTCCGCCATCGGACGAGCCAGATTTAACTACAAGCCCTGCTCAAGAAGCAGAAGCTAGGAAGGTCCTGAGCCAAGTTAGAGAAACACCATTCGACCCAAAGATGTTTCCCGAATATAGGCCCTTAGACTCTATGTTATCTTATTACAAAGGTGTCCAGTCTGCTTTGCAGGGTCGTCAAACTACTATCGTTAACAGATGTTTCACTATCGCTGCTGATAAGATAAACGCCTTAAAGAGGAGATGTAACCAACTTATTCCGGAAATGGTAAAGAAATACGGATTGAGTACAGAAGATGCTATCGGTTCTGCATGGGTCTCTTCTAATGACGTGGTTGCCGCACTTTTGTGGACCTGTATTAATCGTGCTAGATACCCAGAGATTAGAGAAAGAAGTGTACATCAGCTTCCACCAGACTTGTTGCATGCTACATCTTCATTGGGTGTACCTGTTAATGTTAGATCTAGACTTTCACCACCCTTACCCAAATCCACTCTTGGGAATGCGGTCTGCCTGCTAAGGGAGAAGGTACCTTTGCAATTCTTCGCTCTTCCGAGTCACGCAAACATGGAAGCAACCAGTTCAGTTTGCGCTGACCATTCAGGGGATGATGAATGGGCGTTGAGCTTTTGTAGGGTAGCTTATGGTCTGCGTGCAAAATTGAACGCGATCGATGATGATTACATAAGGGATTACATTAGTTATGTACAGAAGTCGCCCTGTCATCTTTCAGTTACACTAGATACTGAGAACTTATACCTGTCTAATTGGAGAGAAATAGGCGTCTACGATGCAGACTTCGGTGGTATGCTAGGGAAACCATTGCGTATGAGAGCTCCGGATGGCTACACTGACGGTTTGATTTTCGTTATGGCCCAAAGATCTGAAGACAAGTCAGCTCCGTGGGAATTCAACATATCCCTTGAGGCTTCTACAATGAAAAGGATTGTACATGATCCGCTGTGGTGTAAATATGTTGAATTGGATGCCTTTTGGCATGGGGAAGAGTAA
SEQ ID NO:127
XP_001258079.1蛋白质序列
MTVTISFEPYVGSSVDALSIPLYLRCQLVFKLSKPLAAVPLLESGVNRLVQALPFLSGEFTAVPASDGGKEILLVRPVLNFELSRILKIKYHETSLRHVCKQMNRPSSQGGDLPHEPYMPYPRLPDPSRPQPIVGFQVNVHTDGIILSVATHHCSFDATGMGSIVQNLAACCRSPPSDEPDLTTSPAQEAEARKVLSQVRETPFDPKMFPEYRPLDSMLSYYKGVQSALQGRQTTIVNRCFTIAADKINALKRRCNQLIPEMVKKYGLSTEDAIGSAWVSSNDVVAALLWTCINRARYPEIRERSVHQLPPDLLHATSSLGVPVNVRSRLSPPLPKSTLGNAVCLLREKVPLQFFALPSHANMEATSSVCADHSGDDEWALSFCRVAYGLRAKLNAIDDDYIRDYISYVQKSPCHLSVTLDTENLYLSNWREIGVYDADFGGMLGKPLRMRAPDGYTDGLIFVMAQRSEDKSAPWEFNISLEASTMKRIVHDPLWCKYVELDAFWHGEE
SEQ ID NO:128
XP_001217250.1天然核苷酸序列
ATGGCAACCTTCGACCATATCGAGGATGTCATCGGCCAGCTACCTATGCTGAAGAGCTACACCCATATCTTGCTGTGCTTTCCCCTCGCTGAGAGCCAACTCAATGAAGCCATCGAGAGCCTTGAATCTGCTGTACGTCAGGTTATAAAAACCTTTTCGTTCTTGGCTGGCAAAGTAGTCAACGAAGGCAAGGGACCCAACAGCTCGGGTACTTTCAGGGTCGCTCCCTGTGAGACGTGGGAGTCCCCAGATCATCAATTTGTGCGAGTCGTTGATCGCTCTTTCATGCTGGCCTCCTACGATGAGATCCGCGGAGCACAGGCACCTGCTTCCATGCTCCCAGGAAGTCAACTCGGGTATCGGGTGGCTTTTCCAGCGCATTATCACGAGACAGAGGACGATCCCGCGCCGGTCCTGGACATTCAGTGCAATTTGATACGGGGCGGACTGCTACTTGACATCGCTGCCCAACACAACATTATCGATGCGAGCGGTATCTTTCAAATTGCCAGCTTGATCGCTCTTTCCATGCGAGGCGAGTCAATTCCTGAGGATGTCATCAAGGAAGGGAACCGTGATCGACGCAATATCATTCCACTACTAGAGGCAGATGAGCCTCTTCTTGATCACAGCGAGCTCAAAGCCAGCAGCGCGGTGCAAAATCCGCCGCCCGTCAACTTCCTCCAGGGGTATAAATGGCAAATCTTCAAGCTCTCTGCCGAGGTGTTAACTCGAATTACCGCTGAAGGACGACGACAGCCACAGGAGTTTGTCCCCTCCGTCACATTTGTCTCGGCAAACGATTGCTTGACAGCCTTTCTGTGGCAACGGGTGATAGCCATGCGCCTGAAGCGGCTCCATACGCCCGAGGCCGTATCCAAACTGAGCCGTGCTGTTGATCTTCGGCGGGCGATGGGCATTACCCCAGCATACATGGGCCATATGATTCGTGTCGCAAATACTAGTCTCACTTTTCAAGAAATTGTGGCATGCTCTTTATCCAGGCTTGCATCTCTGCTCCGCAAGAGCATCATCGATGTCAGCCAGCCGTATGCGATTCGGAGCTACGTGACCTTTATTGCAAATGAGACGGATAAATCAAAGATTGCGTATGCGGGTGCTTTCAATCCTTGCACCGACATGTCGTGCTCGTCCATTGCACACATCACTGCTCCTGAATTCGGTCGTTTGGGAGCGCCTGACTTTATAAGGAGGCCTACCTACGGGCCTCTGCCGTGCTGCACCTATGTCGCTCCCGATAAGAACGATGGAGCTTTGGATCAAAACCAGGCATGGTCAGACGTTGTGAAGCGCATCGGTTGA
SEQ ID NO:129
XP_001217250.1核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGGCCACCTTCGACCACATTGAGGACGTGATAGGACAATTGCCCATGCTAAAGAGCTACACACATATTTTACTTTGCTTTCCGTTGGCAGAATCTCAACTGAACGAGGCGATTGAATCTTTGGAGTCAGCAGTCAGACAAGTAATTAAGACGTTTTCATTCCTTGCAGGTAAGGTTGTTAATGAAGGCAAAGGTCCGAATTCCTCTGGTACGTTCCGTGTTGCTCCGTGTGAAACTTGGGAATCTCCTGATCATCAATTTGTTCGTGTTGTAGATAGGAGCTTTATGTTGGCCTCGTATGATGAGATCCGTGGTGCTCAAGCCCCCGCTTCCATGTTACCTGGGTCACAACTGGGGTATAGGGTAGCATTCCCAGCCCATTATCATGAAACTGAAGACGATCCAGCTCCAGTTCTAGATATCCAGTGTAATTTGATAAGAGGTGGTCTGCTTCTAGATATAGCGGCCCAACACAACATAATTGATGCTTCCGGGATATTTCAAATCGCTAGTCTGATCGCCTTAAGCATGAGAGGAGAAAGTATCCCTGAAGATGTTATCAAAGAAGGAAACAGAGATAGAAGGAATATCATTCCGTTATTGGAAGCCGATGAGCCTTTATTAGACCATAGTGAATTGAAGGCATCCAGCGCCGTTCAGAACCCGCCACCAGTTAATTTCTTGCAAGGTTATAAATGGCAGATTTTCAAACTGTCCGCTGAGGTATTGACCCGTATTACTGCCGAAGGACGTAGACAACCACAAGAATTTGTGCCCTCAGTGACCTTTGTGTCCGCAAACGACTGCTTGACTGCTTTCTTGTGGCAGCGTGTGATAGCTATGAGGCTAAAGAGATTGCATACCCCCGAGGCCGTTTCCAAGCTATCTAGAGCCGTGGATTTAAGAAGGGCCATGGGTATAACCCCTGCATACATGGGCCATATGATAAGAGTCGCAAACACATCCTTAACCTTTCAAGAAATTGTCGCATGTAGTTTATCACGTCTTGCGAGCTTACTTAGAAAATCTATCATTGATGTGTCCCAACCATATGCGATCAGATCATACGTCACCTTCATAGCAAACGAAACAGACAAATCTAAGATAGCCTATGCCGGGGCATTTAACCCGTGTACTGATATGTCATGTTCATCCATAGCTCATATTACGGCACCGGAGTTTGGTAGGTTAGGGGCGCCTGACTTCATTAGGAGACCCACTTATGGCCCACTACCTTGCTGTACTTACGTAGCTCCTGACAAGAATGACGGTGCATTGGATCAGAATCAAGCATGGTCAGATGTCGTAAAGAGGATAGGTTAA
SEQ ID NO:130
XP_001217250.1蛋白质序列
MATFDHIEDVIGQLPMLKSYTHILLCFPLAESQLNEAIESLESAVRQVIKTFSFLAGKVVNEGKGPNSSGTFRVAPCETWESPDHQFVRVVDRSFMLASYDEIRGAQAPASMLPGSQLGYRVAFPAHYHETEDDPAPVLDIQCNLIRGGLLLDIAAQHNIIDASGIFQIASLIALSMRGESIPEDVIKEGNRDRRNIIPLLEADEPLLDHSELKASSAVQNPPPVNFLQGYKWQIFKLSAEVLTRITAEGRRQPQEFVPSVTFVSANDCLTAFLWQRVIAMRLKRLHTPEAVSKLSRAVDLRRAMGITPAYMGHMIRVANTSLTFQEIVACSLSRLASLLRKSIIDVSQPYAIRSYVTFIANETDKSKIAYAGAFNPCTDMSCSSIAHITAPEFGRLGAPDFIRRPTYGPLPCCTYVAPDKNDGALDQNQAWSDVVKRIG
SEQ ID NO:131
BAU61551.1天然核苷酸序列
ATGTCCAAGCCCTTATTCGAAGCGTATCCTCTCACAGGGCTTGATCATACGATTCCTCCATGTTATGTTCGCTTCCTTCTAACTTTCCCCGTGCCGGATGTGGCATTGGCTGTCAATCAGCTGCAAAAGGGAGCTGAAAATTTAATCGAGAAACTTCCTTTCCTGGCCGGATATTTGGCTTCATGCGAGACCCCCGGCGTACGCCCAGGGCAGCTCGAGATTCGACCTCCCGCTGGGGAAAGGAGACCTGTCTGCCTCGTCGCACATCACTCGAACTCCTATCTCGCAGATTCCAGTGCGACGTCGACGACGGAACAGCTGGGCACCGCCAACGAGAACTATCTCCCTGTCCCGTTCTTCCCGGAGCTAGACAAGCCGGTGCCCATCTTCCGGGTTAAGGTGAATGCCATGACGGACGGCATCATTCTGGGATTTGCGTTCCACCATAGCGTGATCGATGCCACCGGGATGGGCACCATTGTCCGGGACTTTGCCAGATGCTGCCGTGGCCCTGATGGCGGTCCCCTGGAAATCAGTCTGGAGTCTCAGCAGGACTCTAGAGAGAAGCTGAGACACTCCGGAGGACCTCCCGATCCGCGGTTTGACCACAATGGGGAGTACCCTCTCGTGGCGTCTCTGCCCGCCGACCTCGAAGCCATGAAGCAGGTCTTGATCCAGACGGCCCGTCTCATGTCAACGCAGTATTTCCGCATCCCTGCCAGCCTAGTTAACACGCTAAAGGAATCCTGCAATCGGATGCTTCGGGAATCACCAGCGCTCAGGGACGAAGGGGAGAATCCATGGATTTCGAGCAACGATCTGGTGGTGTCGCTGTTGTGGCTGTGTCTGAATCGCGTTCGGTATCCTGAAGATAATACCAACGTCATTCCTCCTTCCGATTCCTCGGTCTGCATGGCCGTGAATATCCGAGGGCGTTTGCAGTCGCCCATTGATCCAGGATACGTTGGCAACGCCATCGTCCTTCTCCGGGAGAGCGTTGGCATGAATGCTTTTCTGCATAAACCGGGCGACGATGATCCCCTGGGCGCCCAATGTTACGAAACAGCGAAACGGCTAGGCCGAGAAGCGTGGGAAGCAGCCCTGGTGCGCATCGCCCTGGCCATCCGCCGCAAGCTCAACACCATAAACGCGAGTTACGTGCGCAGTGTTATATCCTATCTGGAGGACGTGCCCGACCTGTCCACTGTGGCGTTTGGCCAGACGGACTACCACATCAGCAGCTGGCGGGATATTGGCGTCTACGAGGCTGATTTTGGTGGCCACATGGGCCATCCCAGCGAAATGCGAGTACCAGATGGGATGGTCGATGGCATGTTTTACATCTTACCTCGAAGGCAGGGAACACACCCTTGCTGGGAGATCCATGTTACTATCCACCAGGACACAATGAAGCGACTCATTGCAGACCCTGTGTGGGCACGATATACAGTGAGAAAGCCTTCATCACTCTGCCGGGATGAATGA
SEQ ID NO:132
BAU61551.1核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGTCAAAGCCTCTGTTTGAAGCATACCCCCTAACTGGTCTAGATCATACTATACCCCCTTGTTACGTCAGGTTCTTATTAACTTTCCCAGTTCCAGATGTAGCCCTTGCTGTTAACCAATTACAGAAAGGCGCCGAAAATTTAATAGAGAAATTGCCTTTTCTTGCAGGGTACCTAGCTAGCTGTGAAACACCTGGCGTAAGACCGGGGCAGTTAGAAATCAGGCCACCAGCAGGTGAAAGAAGACCAGTGTGTTTGGTCGCACATCACAGTAACTCGTACCTTGCGGATTCTTCTGCAACAAGTACGACGGAGCAATTGGGAACAGCTAACGAAAATTACCTTCCCGTACCATTCTTTCCAGAGTTGGACAAACCTGTTCCAATATTCCGTGTCAAAGTTAACGCTATGACAGATGGGATTATCTTGGGGTTCGCTTTCCACCATAGTGTAATAGATGCTACGGGAATGGGTACAATAGTTAGGGACTTCGCCAGGTGCTGCAGAGGTCCTGATGGTGGGCCCTTAGAGATAAGTCTAGAAAGCCAACAAGATTCGAGAGAAAAGCTGAGGCACTCAGGCGGTCCGCCAGATCCCAGATTCGATCATAACGGAGAATACCCATTGGTGGCCTCACTGCCAGCGGACTTAGAAGCTATGAAACAAGTTTTAATCCAAACAGCGAGGCTGATGAGTACACAATACTTTAGAATACCTGCTAGCCTTGTGAACACTTTAAAAGAGTCATGTAATAGAATGCTTCGTGAATCCCCTGCACTGAGGGATGAAGGTGAAAACCCGTGGATTAGTTCTAACGATTTAGTAGTGAGTCTACTGTGGCTTTGTTTGAACAGGGTGAGGTACCCCGAAGACAATACAAATGTGATTCCACCCTCTGACAGTTCTGTTTGCATGGCTGTAAATATAAGAGGGAGATTACAGTCGCCGATCGATCCAGGTTATGTTGGTAATGCTATTGTATTATTAAGAGAATCTGTTGGAATGAATGCCTTTCTACATAAACCTGGTGATGACGACCCGCTTGGTGCCCAATGTTACGAGACAGCTAAAAGACTTGGAAGAGAAGCATGGGAAGCAGCATTAGTCAGGATTGCTTTGGCAATTAGGCGTAAGTTGAACACTATTAATGCTTCCTATGTCAGATCAGTTATTAGCTACTTAGAGGATGTGCCCGATCTATCAACCGTTGCCTTCGGGCAAACTGATTATCATATCTCCAGTTGGAGAGACATTGGAGTTTACGAGGCTGACTTTGGTGGTCATATGGGCCATCCATCTGAGATGAGAGTCCCTGACGGGATGGTCGATGGTATGTTTTACATACTACCTAGAAGACAAGGTACTCACCCATGTTGGGAAATTCATGTGACTATACACCAGGATACCATGAAAAGACTGATCGCTGATCCGGTTTGGGCAAGATATACCGTTAGAAAGCCTAGTTCTTTGTGCAGGGACGAGTAA
SEQ ID NO:133
BAU61551.1蛋白质序列
MSKPLFEAYPLTGLDHTIPPCYVRFLLTFPVPDVALAVNQLQKGAENLIEKLPFLAGYLASCETPGVRPGQLEIRPPAGERRPVCLVAHHSNSYLADSSATSTTEQLGTANENYLPVPFFPELDKPVPIFRVKVNAMTDGIILGFAFHHSVIDATGMGTIVRDFARCCRGPDGGPLEISLESQQDSREKLRHSGGPPDPRFDHNGEYPLVASLPADLEAMKQVLIQTARLMSTQYFRIPASLVNTLKESCNRMLRESPALRDEGENPWISSNDLVVSLLWLCLNRVRYPEDNTNVIPPSDSSVCMAVNIRGRLQSPIDPGYVGNAIVLLRESVGMNAFLHKPGDDDPLGAQCYETAKRLGREAWEAALVRIALAIRRKLNTINASYVRSVISYLEDVPDLSTVAFGQTDYHISSWRDIGVYEADFGGHMGHPSEMRVPDGMVDGMFYILPRRQGTHPCWEIHVTIHQDTMKRLIADPVWARYTVRKPSSLCRDE
SEQ ID NO:134
PsSalAT天然核苷酸序列
ATGGCAACAATGTATAGTGCTGCTGTTGAAGTGATCTCTAAGGAAACCATTAAACCCACAACTCCAACCCCATCTCAACTTAAAAACTTCAATCTGTCACTTCTCGATCAATGTTTTCCTTTATATTATTATGTTCCAATCATTCTTTTCTACCCAGCCACCGCCGCTAATAGTACCGGTAGCAGTAACCATCATGATGATCTTGACTTGCTTAAGAGTTCTCTTTCCAAAACACTAGTTCACTTTTATCCAATGGCTGGTAGGATGATAGACAATATTCTGGTCGACTGTCATGACCAAGGGATTAACTTTTACAAAGTTAAAATTAGAGGTAAAATGTGTGAGTTCATGTCGCAACCGGATGTGCCACTAAGCCAGCTTCTTCCCTCTGAAGTTGTTTCCGCGAGTGTCCCTAAGGAAGCACTGGTGATCGTTCAAGTGAACATGTTTGACTGTGGTGGAACAGCCATTTGTTCGAGTGTATCACATAAGATTGCCGATGCAGCTACAATGAGTACGTTCATTCGTAGTTGGGCAAGCACCACTAAAACATCTCGTAGTGGGGGTTCAACTGCTGCCGTTACAGATCAGAAATTGATTCCTTCTTTCGACTCGGCATCTCTATTCCCACCTAGTGAACGATTGACATCTCCATCAGGGATGTCAGAGATACCATTTTCCAGTACCCCAGAGGATACAGAAGATGATAAAACTGTCAGCAAGAGATTTGTGTTCGATTTTGCAAAGATAACATCTGTACGTGAAAAGTTGCAAGTATTGATGCATGATAACTACAAAAGCCGCAGGCAAACAAGGGTTGAGGTGGTTACTTCTCTAATATGGAAGTCCGTGATGAAATCCACTCCAGCCGGTTTTTTACCAGTGGTACATCATGCCGTGAACCTTAGAAAGAAAATGGACCCACCATTACAAGATGTTTCATTCGGAAATCTATCTGTAACTGTTTCGGCGTTCTTACCAGCAACAACAACGACAACAACAAATGCGGTCAACAAGACAATCAATAGTACGAGTAGTGAATCACAAGTGGTACTTCATGAGTTACATGATTTTATAGCTCAGATGAGGAGTGAAATAGATAAGGTCAAGGGTGATAAAGGTAGCTTGGAGAAAGTCATTCAAAATTTTGCTTCTGGTCATGATGCTTCAATAAAGAAAATCAATGATGTTGAAGTGATAAACTTTTGGATAAGTAGCTGGTGCAGGATGGGATTATACGAGATTGATTTTGGTTGGGGAAAGCCAATTTGGGTAACAGTTGATCCAAATATCAAGCCGAACAAGAATTGTTTTTTCATGAATGATACGAAATGTGGTGAAGGAATAGAAGTTTGGGCGAGCTTTCTTGAGGATGATATGGCTAAGTTCGAGCTTCACCTAAGTGAAATCCTTGAATTGATTTGA
SEQ ID NO:135
PsSalAT核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGGCAACAATGTACTCAGCTGCAGTTGAGGTTATATCTAAGGAAACGATAAAACCAACCACTCCAACCCCAAGCCAATTGAAGAATTTCAATTTATCTTTATTAGACCAGTGCTTTCCCTTGTACTACTATGTCCCCATCATCTTGTTCTACCCTGCGACTGCTGCAAACTCCACTGGTTCCTCGAACCACCATGATGATCTAGATCTTCTGAAGAGCTCCCTTAGCAAGACACTTGTTCACTTCTACCCTATGGCCGGTAGGATGATCGATAACATATTGGTTGACTGCCACGATCAGGGTATCAATTTCTATAAAGTTAAAATCAGGGGCAAGATGTGTGAATTCATGTCTCAGCCTGATGTGCCACTGTCGCAGCTGCTACCTAGTGAAGTGGTATCCGCATCTGTCCCAAAAGAGGCCTTGGTCATAGTCCAAGTTAATATGTTCGATTGCGGTGGGACGGCCATCTGCTCGTCGGTCAGTCATAAGATCGCAGACGCCGCAACCATGTCAACATTTATTAGATCTTGGGCGAGTACCACCAAAACTTCAAGGTCTGGCGGGTCAACCGCCGCTGTTACTGACCAGAAGTTGATTCCTAGCTTTGATTCGGCAAGCTTATTCCCACCTTCCGAAAGGTTGACTTCACCAAGCGGGATGTCTGAGATACCATTTTCTTCAACCCCTGAAGATACCGAGGACGACAAAACAGTTAGCAAAAGATTCGTGTTTGACTTTGCAAAGATAACATCTGTTAGAGAAAAGCTTCAGGTATTAATGCACGACAACTACAAAAGCAGGCGTCAGACCAGGGTTGAGGTCGTAACGAGCCTGATCTGGAAGAGTGTCATGAAGTCAACACCAGCTGGGTTCCTTCCCGTCGTGCATCATGCGGTAAATTTGAGGAAGAAGATGGACCCACCATTGCAGGATGTCTCCTTCGGCAACCTGAGTGTTACTGTCTCAGCATTCTTGCCCGCCACAACTACCACCACCACAAATGCCGTTAATAAGACAATAAATTCAACTTCATCGGAGAGTCAGGTGGTGCTACATGAATTGCACGATTTTATCGCACAAATGAGAAGTGAGATAGACAAAGTTAAAGGCGATAAGGGTAGCCTAGAAAAGGTGATTCAGAACTTTGCCTCTGGTCATGACGCTTCAATAAAGAAAATAAATGACGTAGAGGTCATAAATTTCTGGATATCTTCATGGTGCAGAATGGGCTTGTATGAGATCGACTTCGGCTGGGGCAAGCCGATTTGGGTTACAGTTGACCCTAATATCAAGCCAAATAAGAATTGCTTCTTTATGAACGACACAAAATGCGGAGAAGGAATAGAGGTATGGGCAAGCTTCCTGGAAGACGACATGGCCAAGTTTGAATTGCACTTATCGGAGATATTGGAATTGATCTAA
SEQ ID NO:136
PsSalAT蛋白质序列
MATMYSAAVEVISKETIKPTTPTPSQLKNFNLSLLDQCFPLYYYVPIILFYPATAANSTGSSNHHDDLDLLKSSLSKTLVHFYPMAGRMIDNILVDCHDQGINFYKVKIRGKMCEFMSQPDVPLSQLLPSEVVSASVPKEALVIVQVNMFDCGGTAICSSVSHKIADAATMSTFIRSWASTTKTSRSGGSTAAVTDQKLIPSFDSASLFPPSERLTSPSGMSEIPFSSTPEDTEDDKTVSKRFVFDFAKITSVREKLQVLMHDNYKSRRQTRVEVVTSLIWKSVMKSTPAGFLPVVHHAVNLRKKMDPPLQDVSFGNLSVTVSAFLPATTTTTTNAVNKTINSTSSESQVVLHELHDFIAQMRSEIDKVKGDKGSLEKVIQNFASGHDASIKKINDVEVINFWISSWCRMGLYEIDFGWGKPIWVTVDPNIKPNKNCFFMNDTKCGEGIEVWASFLEDDMAKFELHLSEILELI
SEQ ID NO:137
AstC核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGACTAAGATCAACCCATACAAGGGTATCTTGGTTGAATTGAAGGACATCGTTTTCACTTCTTCTTCTGACCAAATCAAGTTGCCAATCAACACTTTCAAGTCTATCTTGTGTTGTGGTGCTACTGCTCAATACCAATGTGGTAAGATCAACAGAGCTCAATACTACTCTAGATTGGCTAGAGACTTCGCTTTGTCTTTGGCTGACGTTACTGCTTTGTTCGACACTGTTCAAGCTACTATCAGACCAGAAGAATCTTTCTTGGCTTTCTTGGCTGAATTGAAGTCTAGATTCGGTGAACAATTGAAGTTGTACGCTGTTGCTAACATGTCTAGAGAAGACTACGCTATGTTGAAGTCTTTGCCAATCGACTGGTCTTTGTTCGACGGTGTTTTCTTGTCTGCTGACTTGGGTATGAGAAAGCCAGAATTGAGATTCTTCAGACACGTTTTGGAATCTATCTCTATGAAGCCAGAAGACACTATCTTGGTTGACAACGACACTGACAACATCTTGTGTGCTTTGTCTATGGGTTTGAAGGGTATCTTGTTCGGTTCTACTTCTGTTCCACAAGCTTTGACTAACTTGTTGGAATACGACCACATCTCTAGAGCTGAACAATTCTTGAGATCTCACGCTAAGTCTTTGCACTCTGTTACTCACACTGGTGTTACTATCAGAGAAAACTTCGCTCAATTGTTGATCTTGGAAGCTACTGGTGACATCGACTTGGTTGAATTGGAATACCACCCAACTACTTGGAACTACTTCATCGGTACTCCAGTTTTGACTCAAACTGAATTCCCACACGACTTGGACACTACTTCTTTGGCTACTACTGTTTTGGACAGACCAAAGGACATCGCTAACGAAATCATGGACGAAATGTTGAAGTACAGATCTGACGACGACTTGATGTTGACTTTCTTCACTGACTTCAAGAACAGAGTTGACCCAGTTGTTTGTTGTAACGTTTTGTCTTTGTTCTACAAGTACGGTAGAGGTCACGAATTGCACCACACTTTGGCTTGGGTTAGACAAGTTTTGATCAGAAGAGCTTACATCAACGGTACTGCTTTCTACCCAATGCCAGAAGCTTTCTTGTACTTCTTCTTCAGATTCTTGCAACACATCACTCACTTGCCACAATTGTACGACGGTTTGAAGGTTTTGTTGAAGGAAAGATTGCAAGAAAGAGTTGGTGTTCCAGTTGACCCAATCTCTTTGTCTATGAGATTGATCGCTTGTAACGGTGTTGGTATCCACGACAGAATGGGTTTGAACGCTTTGTTGTCTATGCAAAACCCAGACGGTTCTTGGGACTTGGGTACTATGTACCACTACGCTTCTAAGAGATTGCCAATCGGTAACCAAGGTGTTTCTACTGCTATGGCTATCAAGGCTATCAAGCAATGTCAAGCTAACCAATGTGCTGGTATCTAA
SEQ ID NO:138
AstC蛋白质序列
MTKINPYKGILVELKDIVFTSSSDQIKLPINTFKSILCCGATAQYQCGKINRAQYYSRLARDFALSLADVTALFDTVQATIRPEESFLAFLAELKSRFGEQLKLYAVANMSREDYAMLKSLPIDWSLFDGVFLSADLGMRKPELRFFRHVLESISMKPEDTILVDNDTDNILCALSMGLKGILFGSTSVPQALTNLLEYDHISRAEQFLRSHAKSLHSVTHTGVTIRENFAQLLILEATGDIDLVELEYHPTTWNYFIGTPVLTQTEFPHDLDTTSLATTVLDRPKDIANEIMDEMLKYRSDDDLMLTFFTDFKNRVDPVVCCNVLSLFYKYGRGHELHHTLAWVRQVLIRRAYINGTAFYPMPEAFLYFFFRFLQHITHLPQLYDGLKVLLKERLQERVGVPVDPISLSMRLIACNGVGIHDRMGLNALLSMQNPDGSWDLGTMYHYASKRLPIGNQGVSTAMAIKAIKQCQANQCAGI
SEQ ID NO:139
AstI核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGACTAGACAATCTCACTACCAAGCTATCATCTTGGACTTGGGTAACGTTGTTTTCGAATGGGACACTTCTCAAAACCCACCAACTGCTGCTCCAAACCAAATCTCTTTGTTGAGAACTTCTATGAAGTCTCCAGTTTACCACTCTTACGAAAGAGGTCAATTGTCTACTGAAGAATGTCACAGATTGTTGGGTGAATCTTTGCACGTTGACCCAGGTCAAATCAAGGAAGCTTTCGACTTGGCTAGACAATCTTTGAGATCTAACCCAGCTTTGTTGGACTTCATCAGACAATTGAAGCAAACTAGAGGTGTTGCTGTTTACGCTATGTCTAACATCCCACAAGCTGAAATCGAATACTTGAAGGAATCTAGAGCTGGTGACATGGAAGTTTTCGACGAAGTTTTCGCTTCTGGTTACGTTGGTTCTAGAAAGCCAGAAACTGAATTCTACAGAAGAGTTATGGGTGAAATCGGTTTGAAGGCTGAAAGAGTTGTTTTCGTTGACGACAAGGAAGAAAACGTTGACGTTGCTAGAGGTTTGGGTTTGTACGGTGTTTGTTTCGGTGGTGTTGAAGAATTGAGAGGTCACTTGTTGGGTATCTAA
SEQ ID NO:140
AstI蛋白质序列
MTRQSHYQAIILDLGNVVFEWDTSQNPPTAAPNQISLLRTSMKSPVYHSYERGQLSTEECHRLLGESLHVDPGQIKEAFDLARQSLRSNPALLDFIRQLKQTRGVAVYAMSNIPQAEIEYLKESRAGDMEVFDEVFASGYVGSRKPETEFYRRVMGEIGLKAERVVFVDDKEENVDVARGLGLYGVCFGGVEELRGHLLGI
SEQ ID NO:141
AstK核苷酸序列,针对其在酿酒酵母中的表达经密码子优化。
ATGTGTACTACTTTCAAGGCTGCTATCTTCGACATGGGTGGTGTTTTGTTCACTTGGAACCCAATCGTTGACACTCAAGTTTCTTTGAAGGACTTGGGTACTATCATCAACTCTGAAACTTGGGAACAATTCGAAAGAGGTAAGATCGAACCAGACGACTGTTACCACCAATTGGGTTCTCAAATCGGTTTGCCAGGTTCTGAAATCGCTGCTACTTTCAGACAAACTACTGGTTGTTTGAGACCAGACGCTAGAATGACTTCTTTGTTGAGAGAATTGAAGGGTCAAGGTGTTGCTGTTTACATGATGACTAACATCCCAGCTCCAGACTTCCACCAATTGAGAGAAATGCACTACGAATGGGACTTGTTCGACGGTATCTTCGCTTCTGCTTTGGAAGGTATGAGAAAGCCAGACTTGGAATTCTACGAACACGTTTTGAAGCAAATCGACACTTCTGCTGCTGAAACTATCTTCGTTGACGACAAGTTGGAAAACGTTATCGCTGCTCAAGCTGTTGGTATGGTTGGTTTGCACTTGACTGACTCTTTGGCTACTTGTATGGAATTGAGACAATTGGTTGGTTGTTAA
SEQ ID NO:142
AstK蛋白质序列
MCTTFKAAIFDMGGVLFTWNPIVDTQVSLKDLGTIINSETWEQFERGKIEPDDCYHQLGSQIGLPGSEIAATFRQTTGCLRPDARMTSLLRELKGQGVAVYMMTNIPAPDFHQLREMHYEWDLFDGIFASALEGMRKPDLEFYEHVLKQIDTSAAETIFVDDKLENVIAAQAVGMVGLHLTDSLATCMELRQLVGC
SEQ ID NO:143
PYI04555.1蛋白质序列,未经C-末端扩展。
MGASVSFQPFVPTPLDHAMPPIYVSQFLCFPTTTPQSAIQSLQVGIERLFERLPFLAGEILINEHTGAIKVQAPSALIREIPYMALRAHPDLYLPAKQCATTPIERQLKTNSLDESYHPLPAALPLSQPQPVIRFQANTLADGILFAVSYHHCIFDGTGCGQILEMLAQCCSASDDKISLPTDCHTDVLLREYISNLSPTTNIPHDYTQAYSTTVQPDPDASDPDTSPAIPSSLYTEAFTFPSQQITTLRDACNHLLPKLPSTSNAHPHKPTPNPLSSNDVLTALIALCITRATNTTTPPLQPNNHSLSMAVNLRTRIQPQVPDHFLGNFATLLPIHFTSPVHTQQSDLLLTTEPPDPALIHLTTLASQIRSSLSTVNTDYICGLMTDLRTRRNAGENSSLLIEGIKISSWRHLSVYKPDFGPGLGKIAGFEFQAGLMDNLVVILPWRNGDWDVRVTLLERDMRGFREDRLVRWALGSG
SEQ ID NO:144
ERR364415-1_contig_8546蛋白质序列,未经C-末端扩展。
MARAPPPPPPGLRMRDTVLSIVKPIRKTQHLETIDATFVDLMRMDSFIPVIFAYRPADKSEAAYSRLVNRIKESLQKVLVPFFGFAGRWVPSSGGSRRLLCNDEGVPFIEAFVDEELDSVVKASAAFQPVTELNGLGVLGMDMTSYDQRMPPEGGQPCVVAQVTRFKCGGVVLGVAFNHTHTDGQGFYTFMRAWSDFSRTNGTAIKVDHNRALPELASLSQFFIKQHDRIGGKTSTDRVNDHCSKVPERLALKAFEVRASKIKAAKLAAEDGGVGYVSTVDCIVAHLWKTLARLPPVVLDGREITVFSPVEGRNRFLDPPRPNMCGNCFAAMVTPKIPTQELLEMPLAAIAGKQREKLSTTRREEWFGQQSFRELASAMNTSKSALLIVTSWFNFPMYEIDFGAGKPFFASTTNMISPINGVCCGVIAPPTPGSCSSIATLYILCLPAVLEALENVPDFLSFFVPHPNHKDNSQ
适用于本发明的其他序列在下面列出。“先前的”SEQ ID NO是指在相应专利文件中使用的那些。
1.PCT/CN2018/088902(2018年5月29日提交)中所述的折叶苔基二磷酸合酶为:
香鳞毛蕨(Dryopteris fragrans)的DfHAD、DfHAD-9(V274A)和DfHAD-8(K532R)
NA=核酸
AA=氨基酸
2.PCT/EP2018/064344(2018年5月31日提交)中所述的补身烷合酶(即折叶苔醇合酶或补身醇合酶)是
AA=氨基酸
3.WO2015/169871中所述的补身醇合酶是
4.WO 2015/176959中所述的补身醇合酶是
黑水缬草(Valeriana amurensis)的VaTPS3(SEQ ID NO:6)
序列表
<110> 弗门尼舍有限公司
<120> 用于产生乙酸补身烷基酯化合物的方法
<130> 11110/WO
<160> 144
<170> PatentIn version 3.5
<210> 1
<211> 5689
<212> DNA
<213> 质粒 pF167
<400> 1
tggtcagcaa caacgccgaa gaatcactct cgtgttgaga attgcacgcc ttgaccacga 60
cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 120
attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 180
accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 240
ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 300
gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 360
agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 420
ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 480
ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 540
gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 600
ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 660
tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 720
tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 780
cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 840
tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 900
gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 960
tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 1020
ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 1080
attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 1140
cgcgcacatt tttggcattg gcaaagtgcg gactgcatag tcactgtggt gccgtactta 1200
gggtacgcgt tcctgaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg 1260
agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac 1320
gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca 1380
acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat 1440
gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa 1500
atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt 1560
gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga 1620
agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc 1680
gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt 1740
atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga 1800
ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta 1860
taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca 1920
atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta 1980
gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga 2040
tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag 2100
tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt 2160
ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact tcggaatagg 2220
aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata 2280
cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag 2340
aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag 2400
gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg 2460
cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc 2520
tcatccttca atgctatcat tatgtcaccc gcagttctgt gtcgtagtca tcaacatagc 2580
acctatcctt tggcatctcg gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 2640
gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 2700
cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 2760
ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 2820
tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 2880
gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 2940
tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 3000
ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 3060
ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 3120
ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 3180
accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 3240
tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagcactt gctacactgt 3300
caggatagct tccgtcacat ggtggcgatc accgtacatc tgagtgagac gttaattaaa 3360
gtagaccgct cacacatggg cggccgccgt ctcaaggtgc agttcgcgtg caattataac 3420
gtcgtggcaa ctgttatcag tcgtaccgcg ccattcgact acgtcgtaag gccgtttctg 3480
acagagtaaa attcttgagg gaactttcac cattatggga aatgcttcaa gaaggtattg 3540
acttaaactc catcaaatgg tcaggtcatt gagtgttttt tatttgttgt attttttttt 3600
ttttagagaa aatcctccaa tatcaaatta ggaatcgtag tttcatgatt ttctgttaca 3660
cctaactttt tgtgtggtgc cctcctcctt gtcaatatta atgttaaagt gcaattcttt 3720
ttccttatca cgttgagcca ttagtatcaa tttgcttacc tgtattcctt tactatcctc 3780
ctttttctcc ttcttgataa atgtatgtag attgcgtata tagtttcgtc taccctatga 3840
acatattcca ttttgtaatt tcgtgtcgtt tctattatga atttcattta taaagtttat 3900
gtacaaatat cataaaaaaa gagaatcttt ttaagcaagg attttcttaa cttcttcggc 3960
gacagcatca ccgacttcgg tggtactgtt ggaaccacct aaatcaccag ttctgatacc 4020
tgcatccaaa acctttttaa ctgcatcttc aatggcctta ccttcttcag gcaagttcaa 4080
tgacaatttc aacatcattg cagcagacaa gatagtggcg atagggtcaa ccttattctt 4140
tggcaaatct ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg tgttcttgtc 4200
tggcaaagag gccaaggacg cagatggcaa caaacccaag gaacctggga taacggaggc 4260
ttcatcggag atgatatcac caaacatgtt gctggtgatt ataataccat ttaggtgggt 4320
tgggttctta actaggatca tggcggcaga atcaatcaat tgatgttgaa ccttcaatgt 4380
agggaattcg ttcttgatgg tttcctccac agtttttctc cataatcttg aagaggccaa 4440
aagattagct ttatccaagg accaaatagg caatggtggc tcatgttgta gggccatgaa 4500
agcggccatt cttgtgattc tttgcacttc tggaacggtg tattgttcac tatcccaagc 4560
gacaccatca ccatcgtctt cctttctctt accaaagtaa atacctccca ctaattctct 4620
gacaacaacg aagtcagtac ctttagcaaa ttgtggcttg attggagata agtctaaaag 4680
agagtcggat gcaaagttac atggtcttaa gttggcgtac aattgaagtt ctttacggat 4740
ttttagtaaa ccttgttcag gtctaacact accggtaccc catttaggac cagccacagc 4800
acctaacaaa acggcatcaa ccttcttgga ggcttccagc gcctcatctg gaagtgggac 4860
acctgtagca tcgatagcag caccaccaat taaatgattt tcgaaatcga acttgacatt 4920
ggaacgaaca tcagaaatag ctttaagaac cttaatggct tcggctgtga tttcttgacc 4980
aacgtggtca cctggcaaaa cgacgatctt cttaggggca gacatagggg cagacattag 5040
aatggtatat ccttgaaata tatatatata ttgctgaaat gtaaaaggta agaaaagtta 5100
gaaagtaaga cgattgctaa ccacctattg gaaaaaacaa taggtcctta aataatattg 5160
tcaacttcaa gtattgtgat gcaagcattt agtcatgaac gcttctctat tctatatgaa 5220
aagccggttc cggcctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat 5280
atgcgtcagg cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg 5340
cgatagcgcc cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat 5400
tcgaactctt gcatcttacg atacctgagt attcccacag ttaactgcgg tcaagatatt 5460
tcttgaatca ggcgccttag accgctcggc caaacaacca attacttgtt gagaaataga 5520
gtataattat cctataaata taacgttttt gaacacacat gaacaaggaa gtacaggaca 5580
attgattttg aagagaatgt ggattttgat gtaattgttg ggattccatt tttaataagg 5640
caataatatt aggtatgtgg atatactaga agttctcctc gaccgtcga 5689
<210> 2
<211> 60
<212> DNA
<213> 用于同源重组的序列
<400> 2
gcacttgcta cactgtcagg atagcttccg tcacatggtg gcgatcaccg tacatctgag 60
<210> 3
<211> 60
<212> DNA
<213> 用于同源重组的序列
<400> 3
aggtgcagtt cgcgtgcaat tataacgtcg tggcaactgt tatcagtcgt accgcgccat 60
<210> 4
<211> 1584
<212> DNA
<213> XP_007369631.1 污叉丝孔菌(Dichomitus squalens)折叶苔醇合酶,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1584)
<400> 4
atg gct tct atc cac aga aga tac act act ttg atc ttg gac ttg ggt 48
Met Ala Ser Ile His Arg Arg Tyr Thr Thr Leu Ile Leu Asp Leu Gly
1 5 10 15
gac gtt ttg ttc aga tgg tct cca aag act gaa act gct atc cca cca 96
Asp Val Leu Phe Arg Trp Ser Pro Lys Thr Glu Thr Ala Ile Pro Pro
20 25 30
caa caa ttg aag gac atc ttg tct tct gtt act tgg ttc gaa tac gaa 144
Gln Gln Leu Lys Asp Ile Leu Ser Ser Val Thr Trp Phe Glu Tyr Glu
35 40 45
aga ggt aga ttg tct caa gaa gct tgt tac gaa aga tgt gct gaa gaa 192
Arg Gly Arg Leu Ser Gln Glu Ala Cys Tyr Glu Arg Cys Ala Glu Glu
50 55 60
ttc aag atc gaa gct tct gtt atc gct gaa gct ttc aag caa gct aga 240
Phe Lys Ile Glu Ala Ser Val Ile Ala Glu Ala Phe Lys Gln Ala Arg
65 70 75 80
ggt tct ttg aga cca aac gaa gaa ttc atc gct ttg atc aga gac ttg 288
Gly Ser Leu Arg Pro Asn Glu Glu Phe Ile Ala Leu Ile Arg Asp Leu
85 90 95
aga aga gaa atg cac ggt gac ttg act gtt ttg gct ttg tct aac atc 336
Arg Arg Glu Met His Gly Asp Leu Thr Val Leu Ala Leu Ser Asn Ile
100 105 110
tct ttg cca gac tac gaa tac atc atg tct ttg tct tct gac tgg act 384
Ser Leu Pro Asp Tyr Glu Tyr Ile Met Ser Leu Ser Ser Asp Trp Thr
115 120 125
act gtt ttc gac aga gtt ttc cca tct gct ttg gtt ggt gaa aga aag 432
Thr Val Phe Asp Arg Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys
130 135 140
cca cac ttg ggt tgt tac aga aag gtt atc tct gaa atg aac ttg gaa 480
Pro His Leu Gly Cys Tyr Arg Lys Val Ile Ser Glu Met Asn Leu Glu
145 150 155 160
cca caa act act gtt ttc gtt gac gac aag ttg gac aac gtt gct tct 528
Pro Gln Thr Thr Val Phe Val Asp Asp Lys Leu Asp Asn Val Ala Ser
165 170 175
gct aga tct ttg ggt atg cac ggt atc gtt ttc gac aac caa gct aac 576
Ala Arg Ser Leu Gly Met His Gly Ile Val Phe Asp Asn Gln Ala Asn
180 185 190
gtt ttc aga caa ttg aga aac atc ttc ggt gac cca atc aga aga ggt 624
Val Phe Arg Gln Leu Arg Asn Ile Phe Gly Asp Pro Ile Arg Arg Gly
195 200 205
caa gaa tac ttg aga ggt cac gct ggt aag ttg gaa tct tct act gac 672
Gln Glu Tyr Leu Arg Gly His Ala Gly Lys Leu Glu Ser Ser Thr Asp
210 215 220
aac ggt ttg atc ttc gaa gaa aac ttc act caa ttg atc atc tac gaa 720
Asn Gly Leu Ile Phe Glu Glu Asn Phe Thr Gln Leu Ile Ile Tyr Glu
225 230 235 240
ttg act caa gac aga act ttg atc tct ttg tct gaa tgt cca aga act 768
Leu Thr Gln Asp Arg Thr Leu Ile Ser Leu Ser Glu Cys Pro Arg Thr
245 250 255
tgg aac ttc ttc aga ggt gaa cca ttg ttc tct gaa act ttc cca gac 816
Trp Asn Phe Phe Arg Gly Glu Pro Leu Phe Ser Glu Thr Phe Pro Asp
260 265 270
gac gtt gac act act tct gtt gct ttg act gtt ttg caa cca gac aga 864
Asp Val Asp Thr Thr Ser Val Ala Leu Thr Val Leu Gln Pro Asp Arg
275 280 285
gct ttg gtt aac tct gtt ttg gac gaa atg ttg gaa tac gtt gac gct 912
Ala Leu Val Asn Ser Val Leu Asp Glu Met Leu Glu Tyr Val Asp Ala
290 295 300
gac ggt atc atg caa act tac ttc gac aga tct aga cca aga atg gac 960
Asp Gly Ile Met Gln Thr Tyr Phe Asp Arg Ser Arg Pro Arg Met Asp
305 310 315 320
cca ttc gtt tgt gtt aac gtt ttg tct ttg ttc tac gaa aac ggt aga 1008
Pro Phe Val Cys Val Asn Val Leu Ser Leu Phe Tyr Glu Asn Gly Arg
325 330 335
ggt cac gaa ttg cca aga act ttg gac tgg gtt tac gaa gtt ttg ttg 1056
Gly His Glu Leu Pro Arg Thr Leu Asp Trp Val Tyr Glu Val Leu Leu
340 345 350
cac aga gct tac cac ggt ggt tct aga tac tac ttg tct cca gac tgt 1104
His Arg Ala Tyr His Gly Gly Ser Arg Tyr Tyr Leu Ser Pro Asp Cys
355 360 365
ttc ttg ttc ttc atg tct aga ttg ttg aag aga gct gac gac cca gct 1152
Phe Leu Phe Phe Met Ser Arg Leu Leu Lys Arg Ala Asp Asp Pro Ala
370 375 380
gtt caa gct aga ttg aga cca ttg ttc gtt gaa aga gtt aac gaa aga 1200
Val Gln Ala Arg Leu Arg Pro Leu Phe Val Glu Arg Val Asn Glu Arg
385 390 395 400
gtt ggt gct gct ggt gac tct atg gac ttg gct ttc aga atc ttg gct 1248
Val Gly Ala Ala Gly Asp Ser Met Asp Leu Ala Phe Arg Ile Leu Ala
405 410 415
gct gct tct gtt ggt gtt caa tgt cca aga gac ttg gaa aga ttg act 1296
Ala Ala Ser Val Gly Val Gln Cys Pro Arg Asp Leu Glu Arg Leu Thr
420 425 430
gct ggt caa tgt gac gac ggt ggt tgg gac ttg tgt tgg ttc tac gtt 1344
Ala Gly Gln Cys Asp Asp Gly Gly Trp Asp Leu Cys Trp Phe Tyr Val
435 440 445
ttc ggt tct act ggt gtt aag gct ggt aac aga ggt ttg act act gct 1392
Phe Gly Ser Thr Gly Val Lys Ala Gly Asn Arg Gly Leu Thr Thr Ala
450 455 460
ttg gct gtt act gct atc caa act gct atc ggt aga cca cca tct cca 1440
Leu Ala Val Thr Ala Ile Gln Thr Ala Ile Gly Arg Pro Pro Ser Pro
465 470 475 480
tct cca tct gct gct tct tct tct ttc aga cca tct tct cca tac aag 1488
Ser Pro Ser Ala Ala Ser Ser Ser Phe Arg Pro Ser Ser Pro Tyr Lys
485 490 495
ttc ttg ggt atc tct aga cca gct tct cca atc aga ttc ggt gac ttg 1536
Phe Leu Gly Ile Ser Arg Pro Ala Ser Pro Ile Arg Phe Gly Asp Leu
500 505 510
ttg aga cca tgg aga aag atg tct aga tct aac ttg aag tct caa taa 1584
Leu Arg Pro Trp Arg Lys Met Ser Arg Ser Asn Leu Lys Ser Gln
515 520 525
<210> 5
<211> 527
<212> PRT
<213> XP_007369631.1 污叉丝孔菌(Dichomitus squalens)折叶苔醇合酶,针对其在酿酒酵母中的表达经密码子优化
<400> 5
Met Ala Ser Ile His Arg Arg Tyr Thr Thr Leu Ile Leu Asp Leu Gly
1 5 10 15
Asp Val Leu Phe Arg Trp Ser Pro Lys Thr Glu Thr Ala Ile Pro Pro
20 25 30
Gln Gln Leu Lys Asp Ile Leu Ser Ser Val Thr Trp Phe Glu Tyr Glu
35 40 45
Arg Gly Arg Leu Ser Gln Glu Ala Cys Tyr Glu Arg Cys Ala Glu Glu
50 55 60
Phe Lys Ile Glu Ala Ser Val Ile Ala Glu Ala Phe Lys Gln Ala Arg
65 70 75 80
Gly Ser Leu Arg Pro Asn Glu Glu Phe Ile Ala Leu Ile Arg Asp Leu
85 90 95
Arg Arg Glu Met His Gly Asp Leu Thr Val Leu Ala Leu Ser Asn Ile
100 105 110
Ser Leu Pro Asp Tyr Glu Tyr Ile Met Ser Leu Ser Ser Asp Trp Thr
115 120 125
Thr Val Phe Asp Arg Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys
130 135 140
Pro His Leu Gly Cys Tyr Arg Lys Val Ile Ser Glu Met Asn Leu Glu
145 150 155 160
Pro Gln Thr Thr Val Phe Val Asp Asp Lys Leu Asp Asn Val Ala Ser
165 170 175
Ala Arg Ser Leu Gly Met His Gly Ile Val Phe Asp Asn Gln Ala Asn
180 185 190
Val Phe Arg Gln Leu Arg Asn Ile Phe Gly Asp Pro Ile Arg Arg Gly
195 200 205
Gln Glu Tyr Leu Arg Gly His Ala Gly Lys Leu Glu Ser Ser Thr Asp
210 215 220
Asn Gly Leu Ile Phe Glu Glu Asn Phe Thr Gln Leu Ile Ile Tyr Glu
225 230 235 240
Leu Thr Gln Asp Arg Thr Leu Ile Ser Leu Ser Glu Cys Pro Arg Thr
245 250 255
Trp Asn Phe Phe Arg Gly Glu Pro Leu Phe Ser Glu Thr Phe Pro Asp
260 265 270
Asp Val Asp Thr Thr Ser Val Ala Leu Thr Val Leu Gln Pro Asp Arg
275 280 285
Ala Leu Val Asn Ser Val Leu Asp Glu Met Leu Glu Tyr Val Asp Ala
290 295 300
Asp Gly Ile Met Gln Thr Tyr Phe Asp Arg Ser Arg Pro Arg Met Asp
305 310 315 320
Pro Phe Val Cys Val Asn Val Leu Ser Leu Phe Tyr Glu Asn Gly Arg
325 330 335
Gly His Glu Leu Pro Arg Thr Leu Asp Trp Val Tyr Glu Val Leu Leu
340 345 350
His Arg Ala Tyr His Gly Gly Ser Arg Tyr Tyr Leu Ser Pro Asp Cys
355 360 365
Phe Leu Phe Phe Met Ser Arg Leu Leu Lys Arg Ala Asp Asp Pro Ala
370 375 380
Val Gln Ala Arg Leu Arg Pro Leu Phe Val Glu Arg Val Asn Glu Arg
385 390 395 400
Val Gly Ala Ala Gly Asp Ser Met Asp Leu Ala Phe Arg Ile Leu Ala
405 410 415
Ala Ala Ser Val Gly Val Gln Cys Pro Arg Asp Leu Glu Arg Leu Thr
420 425 430
Ala Gly Gln Cys Asp Asp Gly Gly Trp Asp Leu Cys Trp Phe Tyr Val
435 440 445
Phe Gly Ser Thr Gly Val Lys Ala Gly Asn Arg Gly Leu Thr Thr Ala
450 455 460
Leu Ala Val Thr Ala Ile Gln Thr Ala Ile Gly Arg Pro Pro Ser Pro
465 470 475 480
Ser Pro Ser Ala Ala Ser Ser Ser Phe Arg Pro Ser Ser Pro Tyr Lys
485 490 495
Phe Leu Gly Ile Ser Arg Pro Ala Ser Pro Ile Arg Phe Gly Asp Leu
500 505 510
Leu Arg Pro Trp Arg Lys Met Ser Arg Ser Asn Leu Lys Ser Gln
515 520 525
<210> 6
<211> 1623
<212> DNA
<213> XP_006461126 双孢蘑菇(Agaricus bisporus)补身醇合酶序列,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1623)
<400> 6
atg gct cca cca caa aga cca ttc act gct atc gtt ttc gac atc ggt 48
Met Ala Pro Pro Gln Arg Pro Phe Thr Ala Ile Val Phe Asp Ile Gly
1 5 10 15
gac gtt ttg ttc caa tgg tct gct act act aag act tct atc tct cca 96
Asp Val Leu Phe Gln Trp Ser Ala Thr Thr Lys Thr Ser Ile Ser Pro
20 25 30
aag act ttg aga tct atc ttg aac tgt cca act tgg ttc gac tac gaa 144
Lys Thr Leu Arg Ser Ile Leu Asn Cys Pro Thr Trp Phe Asp Tyr Glu
35 40 45
aga ggt aga ttg gct gaa aac gct tgt tac gct gct atc tct caa gaa 192
Arg Gly Arg Leu Ala Glu Asn Ala Cys Tyr Ala Ala Ile Ser Gln Glu
50 55 60
ttc aac gtt aac cca gac gaa gtt aga gac gct ttc tct caa gct aga 240
Phe Asn Val Asn Pro Asp Glu Val Arg Asp Ala Phe Ser Gln Ala Arg
65 70 75 80
gac tct ttg caa gct aac cac gac ttc atc tct ttg atc aga gaa ttg 288
Asp Ser Leu Gln Ala Asn His Asp Phe Ile Ser Leu Ile Arg Glu Leu
85 90 95
aag gct caa gct aac ggt aga ttg aga gtt tac gct atg tct aac atc 336
Lys Ala Gln Ala Asn Gly Arg Leu Arg Val Tyr Ala Met Ser Asn Ile
100 105 110
tct ttg cca gac tgg gaa gtt ttg aga atg aag cca gct gac tgg gac 384
Ser Leu Pro Asp Trp Glu Val Leu Arg Met Lys Pro Ala Asp Trp Asp
115 120 125
atc ttc gac cac gtt ttc act tct ggt gct gtt ggt gaa aga aag cca 432
Ile Phe Asp His Val Phe Thr Ser Gly Ala Val Gly Glu Arg Lys Pro
130 135 140
aac ttg gct ttc tac aga cac gtt atc gct gct act gac ttg caa cca 480
Asn Leu Ala Phe Tyr Arg His Val Ile Ala Ala Thr Asp Leu Gln Pro
145 150 155 160
cac caa act atc ttc gtt gac gac aag ttg gaa aac gtt ttg tct gct 528
His Gln Thr Ile Phe Val Asp Asp Lys Leu Glu Asn Val Leu Ser Ala
165 170 175
aga tct ttg ggt ttc act ggt atc gtt ttc gac gaa cca tct gaa gtt 576
Arg Ser Leu Gly Phe Thr Gly Ile Val Phe Asp Glu Pro Ser Glu Val
180 185 190
aag aga gct ttg aga aac ttg atc ggt gac cca gtt caa aga ggt ggt 624
Lys Arg Ala Leu Arg Asn Leu Ile Gly Asp Pro Val Gln Arg Gly Gly
195 200 205
gaa ttc ttg gtt aga aac gct ggt aag ttg ggt tct atc act aga act 672
Glu Phe Leu Val Arg Asn Ala Gly Lys Leu Gly Ser Ile Thr Arg Thr
210 215 220
act gct aag cac gaa tct atc cca ttg gac gaa aac ttc gct caa ttg 720
Thr Ala Lys His Glu Ser Ile Pro Leu Asp Glu Asn Phe Ala Gln Leu
225 230 235 240
ttg atc ttg gaa atc act ggt aac aga gct ttg gtt aac ttg gtt gaa 768
Leu Ile Leu Glu Ile Thr Gly Asn Arg Ala Leu Val Asn Leu Val Glu
245 250 255
cac cca caa act tgg aac ttc ttc caa ggt aag ggt caa ttg act act 816
His Pro Gln Thr Trp Asn Phe Phe Gln Gly Lys Gly Gln Leu Thr Thr
260 265 270
gaa gaa ttc cca ttc gac ttg gac act act tct ttg ggt ttg act atc 864
Glu Glu Phe Pro Phe Asp Leu Asp Thr Thr Ser Leu Gly Leu Thr Ile
275 280 285
ttg aag aga tct aga gaa atc gct gac tct gtt atg gac gaa atg ttg 912
Leu Lys Arg Ser Arg Glu Ile Ala Asp Ser Val Met Asp Glu Met Leu
290 295 300
gaa tac gtt gac cca gac ggt atc atc caa act tac ttc gac cac aga 960
Glu Tyr Val Asp Pro Asp Gly Ile Ile Gln Thr Tyr Phe Asp His Arg
305 310 315 320
aga cca aga ttc gac cca gtt gtt tgt gtt aac gct ttg tct ttg ttc 1008
Arg Pro Arg Phe Asp Pro Val Val Cys Val Asn Ala Leu Ser Leu Phe
325 330 335
tac gct tac ggt aga ggt gaa caa ttg aga tct act ttg act tgg gtt 1056
Tyr Ala Tyr Gly Arg Gly Glu Gln Leu Arg Ser Thr Leu Thr Trp Val
340 345 350
cac gaa gtt ttg ttg aac aga gct tac ttg gac ggt act aga tac tac 1104
His Glu Val Leu Leu Asn Arg Ala Tyr Leu Asp Gly Thr Arg Tyr Tyr
355 360 365
gaa act gct gaa tgt ttc ttg tac ttc atg tct aga ttg ttg gct act 1152
Glu Thr Ala Glu Cys Phe Leu Tyr Phe Met Ser Arg Leu Leu Ala Thr
370 375 380
tct ggt gac cca gac ttg cac tct ttg ttg aag cca ttg ttg aag gaa 1200
Ser Gly Asp Pro Asp Leu His Ser Leu Leu Lys Pro Leu Leu Lys Glu
385 390 395 400
aga gtt caa gaa aga atc ggt gct gac ggt gac tct ttg gct ttg gct 1248
Arg Val Gln Glu Arg Ile Gly Ala Asp Gly Asp Ser Leu Ala Leu Ala
405 410 415
atg aga atc ttg gct tgt gac ttc gtt ggt atc aga gac gaa gtt gac 1296
Met Arg Ile Leu Ala Cys Asp Phe Val Gly Ile Arg Asp Glu Val Asp
420 425 430
ttg aga act ttg ttg act ttg caa tgt gaa gac ggt ggt tgg gaa gtt 1344
Leu Arg Thr Leu Leu Thr Leu Gln Cys Glu Asp Gly Gly Trp Glu Val
435 440 445
ggt tgg atg tac aag tac ggt tct tct ggt atc tct atc ggt aac aga 1392
Gly Trp Met Tyr Lys Tyr Gly Ser Ser Gly Ile Ser Ile Gly Asn Arg
450 455 460
ggt ttg gct act gct ttg gct atc aag gct gtt gac act atg ttc caa 1440
Gly Leu Ala Thr Ala Leu Ala Ile Lys Ala Val Asp Thr Met Phe Gln
465 470 475 480
cca caa atc aga ttc tct gaa tct cca act gac act ttg gtt gaa aac 1488
Pro Gln Ile Arg Phe Ser Glu Ser Pro Thr Asp Thr Leu Val Glu Asn
485 490 495
gct atc cac aag aga aga cca tct ttc tct gaa aag ttc ttg ggt aag 1536
Ala Ile His Lys Arg Arg Pro Ser Phe Ser Glu Lys Phe Leu Gly Lys
500 505 510
aga cca aga tct ggt tct ttc aga aag cca ttg caa tgg atc ttg caa 1584
Arg Pro Arg Ser Gly Ser Phe Arg Lys Pro Leu Gln Trp Ile Leu Gln
515 520 525
ggt tct aag ttg aga aag tct gtt gaa atc ggt tct taa 1623
Gly Ser Lys Leu Arg Lys Ser Val Glu Ile Gly Ser
530 535 540
<210> 7
<211> 540
<212> PRT
<213> XP_006461126 双孢蘑菇(Agaricus bisporus)补身醇合酶序列,针对其在酿酒酵母中的表达经密码子优化
<400> 7
Met Ala Pro Pro Gln Arg Pro Phe Thr Ala Ile Val Phe Asp Ile Gly
1 5 10 15
Asp Val Leu Phe Gln Trp Ser Ala Thr Thr Lys Thr Ser Ile Ser Pro
20 25 30
Lys Thr Leu Arg Ser Ile Leu Asn Cys Pro Thr Trp Phe Asp Tyr Glu
35 40 45
Arg Gly Arg Leu Ala Glu Asn Ala Cys Tyr Ala Ala Ile Ser Gln Glu
50 55 60
Phe Asn Val Asn Pro Asp Glu Val Arg Asp Ala Phe Ser Gln Ala Arg
65 70 75 80
Asp Ser Leu Gln Ala Asn His Asp Phe Ile Ser Leu Ile Arg Glu Leu
85 90 95
Lys Ala Gln Ala Asn Gly Arg Leu Arg Val Tyr Ala Met Ser Asn Ile
100 105 110
Ser Leu Pro Asp Trp Glu Val Leu Arg Met Lys Pro Ala Asp Trp Asp
115 120 125
Ile Phe Asp His Val Phe Thr Ser Gly Ala Val Gly Glu Arg Lys Pro
130 135 140
Asn Leu Ala Phe Tyr Arg His Val Ile Ala Ala Thr Asp Leu Gln Pro
145 150 155 160
His Gln Thr Ile Phe Val Asp Asp Lys Leu Glu Asn Val Leu Ser Ala
165 170 175
Arg Ser Leu Gly Phe Thr Gly Ile Val Phe Asp Glu Pro Ser Glu Val
180 185 190
Lys Arg Ala Leu Arg Asn Leu Ile Gly Asp Pro Val Gln Arg Gly Gly
195 200 205
Glu Phe Leu Val Arg Asn Ala Gly Lys Leu Gly Ser Ile Thr Arg Thr
210 215 220
Thr Ala Lys His Glu Ser Ile Pro Leu Asp Glu Asn Phe Ala Gln Leu
225 230 235 240
Leu Ile Leu Glu Ile Thr Gly Asn Arg Ala Leu Val Asn Leu Val Glu
245 250 255
His Pro Gln Thr Trp Asn Phe Phe Gln Gly Lys Gly Gln Leu Thr Thr
260 265 270
Glu Glu Phe Pro Phe Asp Leu Asp Thr Thr Ser Leu Gly Leu Thr Ile
275 280 285
Leu Lys Arg Ser Arg Glu Ile Ala Asp Ser Val Met Asp Glu Met Leu
290 295 300
Glu Tyr Val Asp Pro Asp Gly Ile Ile Gln Thr Tyr Phe Asp His Arg
305 310 315 320
Arg Pro Arg Phe Asp Pro Val Val Cys Val Asn Ala Leu Ser Leu Phe
325 330 335
Tyr Ala Tyr Gly Arg Gly Glu Gln Leu Arg Ser Thr Leu Thr Trp Val
340 345 350
His Glu Val Leu Leu Asn Arg Ala Tyr Leu Asp Gly Thr Arg Tyr Tyr
355 360 365
Glu Thr Ala Glu Cys Phe Leu Tyr Phe Met Ser Arg Leu Leu Ala Thr
370 375 380
Ser Gly Asp Pro Asp Leu His Ser Leu Leu Lys Pro Leu Leu Lys Glu
385 390 395 400
Arg Val Gln Glu Arg Ile Gly Ala Asp Gly Asp Ser Leu Ala Leu Ala
405 410 415
Met Arg Ile Leu Ala Cys Asp Phe Val Gly Ile Arg Asp Glu Val Asp
420 425 430
Leu Arg Thr Leu Leu Thr Leu Gln Cys Glu Asp Gly Gly Trp Glu Val
435 440 445
Gly Trp Met Tyr Lys Tyr Gly Ser Ser Gly Ile Ser Ile Gly Asn Arg
450 455 460
Gly Leu Ala Thr Ala Leu Ala Ile Lys Ala Val Asp Thr Met Phe Gln
465 470 475 480
Pro Gln Ile Arg Phe Ser Glu Ser Pro Thr Asp Thr Leu Val Glu Asn
485 490 495
Ala Ile His Lys Arg Arg Pro Ser Phe Ser Glu Lys Phe Leu Gly Lys
500 505 510
Arg Pro Arg Ser Gly Ser Phe Arg Lys Pro Leu Gln Trp Ile Leu Gln
515 520 525
Gly Ser Lys Leu Arg Lys Ser Val Glu Ile Gly Ser
530 535 540
<210> 8
<211> 1320
<212> DNA
<213> CrDAT,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1320)
<400> 8
atg gaa agt ggt aaa ata tct gta gaa acg gaa acc cta tcg aaa acc 48
Met Glu Ser Gly Lys Ile Ser Val Glu Thr Glu Thr Leu Ser Lys Thr
1 5 10 15
ctt atc aag ccg tcc tcg ccc acg cca cag tct ctt tcc cgt tac aac 96
Leu Ile Lys Pro Ser Ser Pro Thr Pro Gln Ser Leu Ser Arg Tyr Asn
20 25 30
tta tct tac aat gat cag aac att tat cag acc tgc gta tct gta ggg 144
Leu Ser Tyr Asn Asp Gln Asn Ile Tyr Gln Thr Cys Val Ser Val Gly
35 40 45
ttc ttt tac gaa aat cct gat ggc atc gag ata tcc acg atc cgt gag 192
Phe Phe Tyr Glu Asn Pro Asp Gly Ile Glu Ile Ser Thr Ile Arg Glu
50 55 60
cag ctg cag aac agc ctg tcc aaa act ctg gtc tca tac tat ccc ttc 240
Gln Leu Gln Asn Ser Leu Ser Lys Thr Leu Val Ser Tyr Tyr Pro Phe
65 70 75 80
gca ggc aaa gta gtg aag aac gac tac ata cat tgt aac gac gat ggc 288
Ala Gly Lys Val Val Lys Asn Asp Tyr Ile His Cys Asn Asp Asp Gly
85 90 95
att gag ttc gtc gaa gtc aga att aga tgc agg atg aat gat ata ctt 336
Ile Glu Phe Val Glu Val Arg Ile Arg Cys Arg Met Asn Asp Ile Leu
100 105 110
aaa tac gaa cta cgt tct tac gct cgt gat tta gtc ctg cca aag agg 384
Lys Tyr Glu Leu Arg Ser Tyr Ala Arg Asp Leu Val Leu Pro Lys Arg
115 120 125
gtt acc gtg ggc tct gaa gat acc acc gct atc gtg cag ctg tcc cat 432
Val Thr Val Gly Ser Glu Asp Thr Thr Ala Ile Val Gln Leu Ser His
130 135 140
ttc gac tgt ggc ggt ttg gcc gtg gct ttt ggt atc tcc cac aag gtg 480
Phe Asp Cys Gly Gly Leu Ala Val Ala Phe Gly Ile Ser His Lys Val
145 150 155 160
gcc gat ggt ggt act ata gcc tct ttc atg aag gac tgg gcg gcc tct 528
Ala Asp Gly Gly Thr Ile Ala Ser Phe Met Lys Asp Trp Ala Ala Ser
165 170 175
gct tgc tac ttg tcc tca agt cac cac gtt cca acg cct cta ttg gtc 576
Ala Cys Tyr Leu Ser Ser Ser His His Val Pro Thr Pro Leu Leu Val
180 185 190
tca gac agt att ttc cca aga cag gac aac ata ata tgc gaa caa ttt 624
Ser Asp Ser Ile Phe Pro Arg Gln Asp Asn Ile Ile Cys Glu Gln Phe
195 200 205
ccc acc tca aag aat tgc gtg gag aag aca ttc att ttc cca cca gaa 672
Pro Thr Ser Lys Asn Cys Val Glu Lys Thr Phe Ile Phe Pro Pro Glu
210 215 220
gca atc gaa aag tta aag agt aag gcc gtt gaa ttc ggc att gag aaa 720
Ala Ile Glu Lys Leu Lys Ser Lys Ala Val Glu Phe Gly Ile Glu Lys
225 230 235 240
ccg acc aga gta gag gtc ctg act gca ttc tta tct aga tgc gcc acc 768
Pro Thr Arg Val Glu Val Leu Thr Ala Phe Leu Ser Arg Cys Ala Thr
245 250 255
gta gca ggt aag tcg gca gct aag aac aac aat tgt ggt caa agc ctg 816
Val Ala Gly Lys Ser Ala Ala Lys Asn Asn Asn Cys Gly Gln Ser Leu
260 265 270
ccc ttt ccg gtt cta cag gcc att aat ttg agg ccg att cta gaa ttg 864
Pro Phe Pro Val Leu Gln Ala Ile Asn Leu Arg Pro Ile Leu Glu Leu
275 280 285
cca cag aac tct gtg ggt aat cta gtt tcg atc tac ttc agc agg aca 912
Pro Gln Asn Ser Val Gly Asn Leu Val Ser Ile Tyr Phe Ser Arg Thr
290 295 300
att aag gaa aac gac tac cta aat gaa aag gag tac act aaa ttg gtg 960
Ile Lys Glu Asn Asp Tyr Leu Asn Glu Lys Glu Tyr Thr Lys Leu Val
305 310 315 320
ata aac gag ttg cgt aaa gaa aag caa aag atc aag aat ttg agc aga 1008
Ile Asn Glu Leu Arg Lys Glu Lys Gln Lys Ile Lys Asn Leu Ser Arg
325 330 335
gag aag ttg acc tac gtc gcc cag atg gaa gag ttc gtt aag tca ctt 1056
Glu Lys Leu Thr Tyr Val Ala Gln Met Glu Glu Phe Val Lys Ser Leu
340 345 350
aaa gag ttc gat atc tcc aac ttc cta gat atc gat gcc tac tta agc 1104
Lys Glu Phe Asp Ile Ser Asn Phe Leu Asp Ile Asp Ala Tyr Leu Ser
355 360 365
gat agt tgg tgc agg ttt cct ttc tac gac gtc gac ttc gga tgg gga 1152
Asp Ser Trp Cys Arg Phe Pro Phe Tyr Asp Val Asp Phe Gly Trp Gly
370 375 380
aag cct atc tgg gtc tgc cta ttc cag cct tac atc aag aac tgc gtt 1200
Lys Pro Ile Trp Val Cys Leu Phe Gln Pro Tyr Ile Lys Asn Cys Val
385 390 395 400
gtt atg atg gac tat cct ttc ggt gac gat tat ggt atc gag gct att 1248
Val Met Met Asp Tyr Pro Phe Gly Asp Asp Tyr Gly Ile Glu Ala Ile
405 410 415
gtt tct ttc gag caa gag aaa atg tcc gcg ttc gag aag aac gaa cag 1296
Val Ser Phe Glu Gln Glu Lys Met Ser Ala Phe Glu Lys Asn Glu Gln
420 425 430
tta ctg cag ttt gtg tct aac taa 1320
Leu Leu Gln Phe Val Ser Asn
435
<210> 9
<211> 439
<212> PRT
<213> CrDAT,针对其在酿酒酵母中的表达经密码子优化
<400> 9
Met Glu Ser Gly Lys Ile Ser Val Glu Thr Glu Thr Leu Ser Lys Thr
1 5 10 15
Leu Ile Lys Pro Ser Ser Pro Thr Pro Gln Ser Leu Ser Arg Tyr Asn
20 25 30
Leu Ser Tyr Asn Asp Gln Asn Ile Tyr Gln Thr Cys Val Ser Val Gly
35 40 45
Phe Phe Tyr Glu Asn Pro Asp Gly Ile Glu Ile Ser Thr Ile Arg Glu
50 55 60
Gln Leu Gln Asn Ser Leu Ser Lys Thr Leu Val Ser Tyr Tyr Pro Phe
65 70 75 80
Ala Gly Lys Val Val Lys Asn Asp Tyr Ile His Cys Asn Asp Asp Gly
85 90 95
Ile Glu Phe Val Glu Val Arg Ile Arg Cys Arg Met Asn Asp Ile Leu
100 105 110
Lys Tyr Glu Leu Arg Ser Tyr Ala Arg Asp Leu Val Leu Pro Lys Arg
115 120 125
Val Thr Val Gly Ser Glu Asp Thr Thr Ala Ile Val Gln Leu Ser His
130 135 140
Phe Asp Cys Gly Gly Leu Ala Val Ala Phe Gly Ile Ser His Lys Val
145 150 155 160
Ala Asp Gly Gly Thr Ile Ala Ser Phe Met Lys Asp Trp Ala Ala Ser
165 170 175
Ala Cys Tyr Leu Ser Ser Ser His His Val Pro Thr Pro Leu Leu Val
180 185 190
Ser Asp Ser Ile Phe Pro Arg Gln Asp Asn Ile Ile Cys Glu Gln Phe
195 200 205
Pro Thr Ser Lys Asn Cys Val Glu Lys Thr Phe Ile Phe Pro Pro Glu
210 215 220
Ala Ile Glu Lys Leu Lys Ser Lys Ala Val Glu Phe Gly Ile Glu Lys
225 230 235 240
Pro Thr Arg Val Glu Val Leu Thr Ala Phe Leu Ser Arg Cys Ala Thr
245 250 255
Val Ala Gly Lys Ser Ala Ala Lys Asn Asn Asn Cys Gly Gln Ser Leu
260 265 270
Pro Phe Pro Val Leu Gln Ala Ile Asn Leu Arg Pro Ile Leu Glu Leu
275 280 285
Pro Gln Asn Ser Val Gly Asn Leu Val Ser Ile Tyr Phe Ser Arg Thr
290 295 300
Ile Lys Glu Asn Asp Tyr Leu Asn Glu Lys Glu Tyr Thr Lys Leu Val
305 310 315 320
Ile Asn Glu Leu Arg Lys Glu Lys Gln Lys Ile Lys Asn Leu Ser Arg
325 330 335
Glu Lys Leu Thr Tyr Val Ala Gln Met Glu Glu Phe Val Lys Ser Leu
340 345 350
Lys Glu Phe Asp Ile Ser Asn Phe Leu Asp Ile Asp Ala Tyr Leu Ser
355 360 365
Asp Ser Trp Cys Arg Phe Pro Phe Tyr Asp Val Asp Phe Gly Trp Gly
370 375 380
Lys Pro Ile Trp Val Cys Leu Phe Gln Pro Tyr Ile Lys Asn Cys Val
385 390 395 400
Val Met Met Asp Tyr Pro Phe Gly Asp Asp Tyr Gly Ile Glu Ala Ile
405 410 415
Val Ser Phe Glu Gln Glu Lys Met Ser Ala Phe Glu Lys Asn Glu Gln
420 425 430
Leu Leu Gln Phe Val Ser Asn
435
<210> 10
<211> 1320
<212> DNA
<213> TcTAT,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1320)
<400> 10
atg gag aag acg gat tta cac gtg aac ctt atc gaa aag gtt atg gtc 48
Met Glu Lys Thr Asp Leu His Val Asn Leu Ile Glu Lys Val Met Val
1 5 10 15
ggt cca agc cca cct ttg ccg aag act aca cta caa cta tcc tca atc 96
Gly Pro Ser Pro Pro Leu Pro Lys Thr Thr Leu Gln Leu Ser Ser Ile
20 25 30
gat aac cta cct ggt gtt aga ggc tcg atc ttt aat gcc ttg tta ata 144
Asp Asn Leu Pro Gly Val Arg Gly Ser Ile Phe Asn Ala Leu Leu Ile
35 40 45
tat aat gct tca ccg tct ccc acg atg atc tcc gct gat cca gct aaa 192
Tyr Asn Ala Ser Pro Ser Pro Thr Met Ile Ser Ala Asp Pro Ala Lys
50 55 60
cct atc aga gaa gct ttg gcc aaa atc ctg gtt tac tac cca cct ttc 240
Pro Ile Arg Glu Ala Leu Ala Lys Ile Leu Val Tyr Tyr Pro Pro Phe
65 70 75 80
gct ggc agg cta cgt gag act gag aat ggc gat cta gag gta gag tgc 288
Ala Gly Arg Leu Arg Glu Thr Glu Asn Gly Asp Leu Glu Val Glu Cys
85 90 95
acg gga gag ggt gcg atg ttt ctt gaa gcg atg gcc gat aat gaa tta 336
Thr Gly Glu Gly Ala Met Phe Leu Glu Ala Met Ala Asp Asn Glu Leu
100 105 110
agt gtc tta ggg gat ttc gac gac agt aac cct agc ttc cag caa ttg 384
Ser Val Leu Gly Asp Phe Asp Asp Ser Asn Pro Ser Phe Gln Gln Leu
115 120 125
ttg ttc tcc ctt cca ctg gac act aac ttc aag gat cta tcc ctg tta 432
Leu Phe Ser Leu Pro Leu Asp Thr Asn Phe Lys Asp Leu Ser Leu Leu
130 135 140
gtc gtg cag gtg aca agg ttt aca tgc ggc ggg ttc gtc gtt ggg gtt 480
Val Val Gln Val Thr Arg Phe Thr Cys Gly Gly Phe Val Val Gly Val
145 150 155 160
tca ttt cac cac ggt gta tgc gat ggg aga gga gct gca caa ttt ctg 528
Ser Phe His His Gly Val Cys Asp Gly Arg Gly Ala Ala Gln Phe Leu
165 170 175
aag gga cta gcc gaa atg gca agg gga gaa gtt aag ttg tcg ctg gag 576
Lys Gly Leu Ala Glu Met Ala Arg Gly Glu Val Lys Leu Ser Leu Glu
180 185 190
ccg atc tgg aat cgt gaa cta gtt aag ctg gac gat ccc aag tat ttg 624
Pro Ile Trp Asn Arg Glu Leu Val Lys Leu Asp Asp Pro Lys Tyr Leu
195 200 205
caa ttc ttt cac ttc gaa ttc ttg aga gca cca tca atc gtc gaa aag 672
Gln Phe Phe His Phe Glu Phe Leu Arg Ala Pro Ser Ile Val Glu Lys
210 215 220
ata gtt caa acc tac ttc atc atc gat ttc gaa act att aac tac ata 720
Ile Val Gln Thr Tyr Phe Ile Ile Asp Phe Glu Thr Ile Asn Tyr Ile
225 230 235 240
aag cag tca gta atg gaa gaa tgt aaa gaa ttc tgc agc tct ttt gaa 768
Lys Gln Ser Val Met Glu Glu Cys Lys Glu Phe Cys Ser Ser Phe Glu
245 250 255
gtt gct tct gca atg acg tgg ata gct agg acc agg gcc ttt caa ata 816
Val Ala Ser Ala Met Thr Trp Ile Ala Arg Thr Arg Ala Phe Gln Ile
260 265 270
ccc gag agc gag tac gtg aaa atc cta ttt ggt atg gac atg agg aac 864
Pro Glu Ser Glu Tyr Val Lys Ile Leu Phe Gly Met Asp Met Arg Asn
275 280 285
tct ttt aac ccg cct ctg ccg tca ggt tat tat gga aac tca att ggt 912
Ser Phe Asn Pro Pro Leu Pro Ser Gly Tyr Tyr Gly Asn Ser Ile Gly
290 295 300
acg gct tgt gca gtg gac aat gtt caa gac ttg ttg agt ggt tca cta 960
Thr Ala Cys Ala Val Asp Asn Val Gln Asp Leu Leu Ser Gly Ser Leu
305 310 315 320
ctg agg gca ata atg ata atc aag aaa tca aag gta tca ctt aac gac 1008
Leu Arg Ala Ile Met Ile Ile Lys Lys Ser Lys Val Ser Leu Asn Asp
325 330 335
aac ttt aag agc cgt gct gtc gtt aag cct tca gaa ctt gac gta aac 1056
Asn Phe Lys Ser Arg Ala Val Val Lys Pro Ser Glu Leu Asp Val Asn
340 345 350
atg aac cat gag aat gtg gtg gca ttt gct gac tgg tca agg ctg ggg 1104
Met Asn His Glu Asn Val Val Ala Phe Ala Asp Trp Ser Arg Leu Gly
355 360 365
ttc gac gaa gtc gat ttt ggc tgg gga aac gcg gtg tct gtt tcc ccg 1152
Phe Asp Glu Val Asp Phe Gly Trp Gly Asn Ala Val Ser Val Ser Pro
370 375 380
gta cag caa cag tcg gca ctt gca atg cag aat tac ttc ctg ttc ttg 1200
Val Gln Gln Gln Ser Ala Leu Ala Met Gln Asn Tyr Phe Leu Phe Leu
385 390 395 400
aag cca agc aag aac aag ccc gat gga att aag att cta atg ttc cta 1248
Lys Pro Ser Lys Asn Lys Pro Asp Gly Ile Lys Ile Leu Met Phe Leu
405 410 415
cca ttg tcc aag atg aaa tcc ttc aaa att gaa atg gaa gcg atg atg 1296
Pro Leu Ser Lys Met Lys Ser Phe Lys Ile Glu Met Glu Ala Met Met
420 425 430
aag aag tac gtc gcc aag gta taa 1320
Lys Lys Tyr Val Ala Lys Val
435
<210> 11
<211> 439
<212> PRT
<213> TcTAT,针对其在酿酒酵母中的表达经密码子优化
<400> 11
Met Glu Lys Thr Asp Leu His Val Asn Leu Ile Glu Lys Val Met Val
1 5 10 15
Gly Pro Ser Pro Pro Leu Pro Lys Thr Thr Leu Gln Leu Ser Ser Ile
20 25 30
Asp Asn Leu Pro Gly Val Arg Gly Ser Ile Phe Asn Ala Leu Leu Ile
35 40 45
Tyr Asn Ala Ser Pro Ser Pro Thr Met Ile Ser Ala Asp Pro Ala Lys
50 55 60
Pro Ile Arg Glu Ala Leu Ala Lys Ile Leu Val Tyr Tyr Pro Pro Phe
65 70 75 80
Ala Gly Arg Leu Arg Glu Thr Glu Asn Gly Asp Leu Glu Val Glu Cys
85 90 95
Thr Gly Glu Gly Ala Met Phe Leu Glu Ala Met Ala Asp Asn Glu Leu
100 105 110
Ser Val Leu Gly Asp Phe Asp Asp Ser Asn Pro Ser Phe Gln Gln Leu
115 120 125
Leu Phe Ser Leu Pro Leu Asp Thr Asn Phe Lys Asp Leu Ser Leu Leu
130 135 140
Val Val Gln Val Thr Arg Phe Thr Cys Gly Gly Phe Val Val Gly Val
145 150 155 160
Ser Phe His His Gly Val Cys Asp Gly Arg Gly Ala Ala Gln Phe Leu
165 170 175
Lys Gly Leu Ala Glu Met Ala Arg Gly Glu Val Lys Leu Ser Leu Glu
180 185 190
Pro Ile Trp Asn Arg Glu Leu Val Lys Leu Asp Asp Pro Lys Tyr Leu
195 200 205
Gln Phe Phe His Phe Glu Phe Leu Arg Ala Pro Ser Ile Val Glu Lys
210 215 220
Ile Val Gln Thr Tyr Phe Ile Ile Asp Phe Glu Thr Ile Asn Tyr Ile
225 230 235 240
Lys Gln Ser Val Met Glu Glu Cys Lys Glu Phe Cys Ser Ser Phe Glu
245 250 255
Val Ala Ser Ala Met Thr Trp Ile Ala Arg Thr Arg Ala Phe Gln Ile
260 265 270
Pro Glu Ser Glu Tyr Val Lys Ile Leu Phe Gly Met Asp Met Arg Asn
275 280 285
Ser Phe Asn Pro Pro Leu Pro Ser Gly Tyr Tyr Gly Asn Ser Ile Gly
290 295 300
Thr Ala Cys Ala Val Asp Asn Val Gln Asp Leu Leu Ser Gly Ser Leu
305 310 315 320
Leu Arg Ala Ile Met Ile Ile Lys Lys Ser Lys Val Ser Leu Asn Asp
325 330 335
Asn Phe Lys Ser Arg Ala Val Val Lys Pro Ser Glu Leu Asp Val Asn
340 345 350
Met Asn His Glu Asn Val Val Ala Phe Ala Asp Trp Ser Arg Leu Gly
355 360 365
Phe Asp Glu Val Asp Phe Gly Trp Gly Asn Ala Val Ser Val Ser Pro
370 375 380
Val Gln Gln Gln Ser Ala Leu Ala Met Gln Asn Tyr Phe Leu Phe Leu
385 390 395 400
Lys Pro Ser Lys Asn Lys Pro Asp Gly Ile Lys Ile Leu Met Phe Leu
405 410 415
Pro Leu Ser Lys Met Lys Ser Phe Lys Ile Glu Met Glu Ala Met Met
420 425 430
Lys Lys Tyr Val Ala Lys Val
435
<210> 12
<211> 1332
<212> DNA
<213> CrMAT,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1332)
<400> 12
atg gat tct att act atg gtt gaa acc gag aca tta tcg aag aca ttg 48
Met Asp Ser Ile Thr Met Val Glu Thr Glu Thr Leu Ser Lys Thr Leu
1 5 10 15
att aag cca tct agc cca acc cca cag agt ctg tca cac tac aac ctg 96
Ile Lys Pro Ser Ser Pro Thr Pro Gln Ser Leu Ser His Tyr Asn Leu
20 25 30
tct tac aac gat cag aat ata tat cca gag tat ata ttt gca ggc ttc 144
Ser Tyr Asn Asp Gln Asn Ile Tyr Pro Glu Tyr Ile Phe Ala Gly Phe
35 40 45
ttc tac agc aac ccg gac gga cac gag atc tcg act att aga gag cag 192
Phe Tyr Ser Asn Pro Asp Gly His Glu Ile Ser Thr Ile Arg Glu Gln
50 55 60
ctg cag aac tct ctt agc aag aca tta gta tct tat tat cca ttc gct 240
Leu Gln Asn Ser Leu Ser Lys Thr Leu Val Ser Tyr Tyr Pro Phe Ala
65 70 75 80
ggt aaa gtg gtc aag aac gat tac att cac tgc aat gac gac gga ata 288
Gly Lys Val Val Lys Asn Asp Tyr Ile His Cys Asn Asp Asp Gly Ile
85 90 95
gaa ttc gta gac gtc aga atc cac tgc agg atg aac gat att ctt aag 336
Glu Phe Val Asp Val Arg Ile His Cys Arg Met Asn Asp Ile Leu Lys
100 105 110
cct gaa ttg agg tct tac gct tcc gag ctt att cgt ccg aat aga agt 384
Pro Glu Leu Arg Ser Tyr Ala Ser Glu Leu Ile Arg Pro Asn Arg Ser
115 120 125
aca gtg ggg tca gag gac agt act gcg ctt gtc cag tta tct cac ttt 432
Thr Val Gly Ser Glu Asp Ser Thr Ala Leu Val Gln Leu Ser His Phe
130 135 140
gac tgt ggc ggg gta gct gtc gca ttc ggt ata tct cac aaa gtc gca 480
Asp Cys Gly Gly Val Ala Val Ala Phe Gly Ile Ser His Lys Val Ala
145 150 155 160
gac gca gcg act att ctg tcg ttt ata aag gat tgg gcc gct tct acg 528
Asp Ala Ala Thr Ile Leu Ser Phe Ile Lys Asp Trp Ala Ala Ser Thr
165 170 175
tgt gac ttg tca agt agt cac gat gtg tct aca cca gta cta gta tcc 576
Cys Asp Leu Ser Ser Ser His Asp Val Ser Thr Pro Val Leu Val Ser
180 185 190
gat tcc ata ttc ccc cgt caa gat aac atc atc tgt ggc cag ttc cct 624
Asp Ser Ile Phe Pro Arg Gln Asp Asn Ile Ile Cys Gly Gln Phe Pro
195 200 205
gct tca ccg aac tgc gtg agg aag cgt ttc tta ttc agc ccg gaa gct 672
Ala Ser Pro Asn Cys Val Arg Lys Arg Phe Leu Phe Ser Pro Glu Ala
210 215 220
atc gaa aga cta aaa tcg aaa gcc att gaa ttt ggg atc gag aag cca 720
Ile Glu Arg Leu Lys Ser Lys Ala Ile Glu Phe Gly Ile Glu Lys Pro
225 230 235 240
acg agg gta gag gtc ctg aca gca ttc ttg tgc cgt tgc gct acc gtt 768
Thr Arg Val Glu Val Leu Thr Ala Phe Leu Cys Arg Cys Ala Thr Val
245 250 255
gca ggt aaa tct gcg gcc aag aac aac aat tgt gga cag tca ctg cct 816
Ala Gly Lys Ser Ala Ala Lys Asn Asn Asn Cys Gly Gln Ser Leu Pro
260 265 270
ttc gct gtt att caa gca gtc aac ctg agg ccc tta cta gaa ctg cca 864
Phe Ala Val Ile Gln Ala Val Asn Leu Arg Pro Leu Leu Glu Leu Pro
275 280 285
aag aat tcc gtc ggc aac ctt ata tca atc tac ttt tct aca atc aaa 912
Lys Asn Ser Val Gly Asn Leu Ile Ser Ile Tyr Phe Ser Thr Ile Lys
290 295 300
gaa aac gac acg gtg aac atc gaa cag gag ttc aca aaa tta gtg atc 960
Glu Asn Asp Thr Val Asn Ile Glu Gln Glu Phe Thr Lys Leu Val Ile
305 310 315 320
gga gag ttg agg aag gct aag gat aag tta aag aac ctg tcg caa gag 1008
Gly Glu Leu Arg Lys Ala Lys Asp Lys Leu Lys Asn Leu Ser Gln Glu
325 330 335
aag ctg aat tac gta gct aga atg caa gat ttc gcg aat tgc ctg aag 1056
Lys Leu Asn Tyr Val Ala Arg Met Gln Asp Phe Ala Asn Cys Leu Lys
340 345 350
gaa ttg gac ata agt tca ttc ttc gac atg gaa aac gtg gac ata gac 1104
Glu Leu Asp Ile Ser Ser Phe Phe Asp Met Glu Asn Val Asp Ile Asp
355 360 365
gct tat tta ttt tcg agc tgg tgc agg ttc ccc ttc tac gac atc gat 1152
Ala Tyr Leu Phe Ser Ser Trp Cys Arg Phe Pro Phe Tyr Asp Ile Asp
370 375 380
ttc ggt ctg ggg aag cca ata tgg gtc tgc atg ttt cag cct cac ttt 1200
Phe Gly Leu Gly Lys Pro Ile Trp Val Cys Met Phe Gln Pro His Phe
385 390 395 400
aag aat tgt ata att tta atg gat tat ccc ttt ggt gac gat tac ggc 1248
Lys Asn Cys Ile Ile Leu Met Asp Tyr Pro Phe Gly Asp Asp Tyr Gly
405 410 415
atc gaa gcc cta att acg ttg gaa caa gag aag atg ccc gcc ttc gaa 1296
Ile Glu Ala Leu Ile Thr Leu Glu Gln Glu Lys Met Pro Ala Phe Glu
420 425 430
aac aac gag ctg ctg cta agc ttc gcc agc aac taa 1332
Asn Asn Glu Leu Leu Leu Ser Phe Ala Ser Asn
435 440
<210> 13
<211> 443
<212> PRT
<213> CrMAT,针对其在酿酒酵母中的表达经密码子优化
<400> 13
Met Asp Ser Ile Thr Met Val Glu Thr Glu Thr Leu Ser Lys Thr Leu
1 5 10 15
Ile Lys Pro Ser Ser Pro Thr Pro Gln Ser Leu Ser His Tyr Asn Leu
20 25 30
Ser Tyr Asn Asp Gln Asn Ile Tyr Pro Glu Tyr Ile Phe Ala Gly Phe
35 40 45
Phe Tyr Ser Asn Pro Asp Gly His Glu Ile Ser Thr Ile Arg Glu Gln
50 55 60
Leu Gln Asn Ser Leu Ser Lys Thr Leu Val Ser Tyr Tyr Pro Phe Ala
65 70 75 80
Gly Lys Val Val Lys Asn Asp Tyr Ile His Cys Asn Asp Asp Gly Ile
85 90 95
Glu Phe Val Asp Val Arg Ile His Cys Arg Met Asn Asp Ile Leu Lys
100 105 110
Pro Glu Leu Arg Ser Tyr Ala Ser Glu Leu Ile Arg Pro Asn Arg Ser
115 120 125
Thr Val Gly Ser Glu Asp Ser Thr Ala Leu Val Gln Leu Ser His Phe
130 135 140
Asp Cys Gly Gly Val Ala Val Ala Phe Gly Ile Ser His Lys Val Ala
145 150 155 160
Asp Ala Ala Thr Ile Leu Ser Phe Ile Lys Asp Trp Ala Ala Ser Thr
165 170 175
Cys Asp Leu Ser Ser Ser His Asp Val Ser Thr Pro Val Leu Val Ser
180 185 190
Asp Ser Ile Phe Pro Arg Gln Asp Asn Ile Ile Cys Gly Gln Phe Pro
195 200 205
Ala Ser Pro Asn Cys Val Arg Lys Arg Phe Leu Phe Ser Pro Glu Ala
210 215 220
Ile Glu Arg Leu Lys Ser Lys Ala Ile Glu Phe Gly Ile Glu Lys Pro
225 230 235 240
Thr Arg Val Glu Val Leu Thr Ala Phe Leu Cys Arg Cys Ala Thr Val
245 250 255
Ala Gly Lys Ser Ala Ala Lys Asn Asn Asn Cys Gly Gln Ser Leu Pro
260 265 270
Phe Ala Val Ile Gln Ala Val Asn Leu Arg Pro Leu Leu Glu Leu Pro
275 280 285
Lys Asn Ser Val Gly Asn Leu Ile Ser Ile Tyr Phe Ser Thr Ile Lys
290 295 300
Glu Asn Asp Thr Val Asn Ile Glu Gln Glu Phe Thr Lys Leu Val Ile
305 310 315 320
Gly Glu Leu Arg Lys Ala Lys Asp Lys Leu Lys Asn Leu Ser Gln Glu
325 330 335
Lys Leu Asn Tyr Val Ala Arg Met Gln Asp Phe Ala Asn Cys Leu Lys
340 345 350
Glu Leu Asp Ile Ser Ser Phe Phe Asp Met Glu Asn Val Asp Ile Asp
355 360 365
Ala Tyr Leu Phe Ser Ser Trp Cys Arg Phe Pro Phe Tyr Asp Ile Asp
370 375 380
Phe Gly Leu Gly Lys Pro Ile Trp Val Cys Met Phe Gln Pro His Phe
385 390 395 400
Lys Asn Cys Ile Ile Leu Met Asp Tyr Pro Phe Gly Asp Asp Tyr Gly
405 410 415
Ile Glu Ala Leu Ile Thr Leu Glu Gln Glu Lys Met Pro Ala Phe Glu
420 425 430
Asn Asn Glu Leu Leu Leu Ser Phe Ala Ser Asn
435 440
<210> 14
<211> 1254
<212> DNA
<213> LiAAT-4 DNA序列,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1254)
<400> 14
atg gcc atg ata atc acg aag caa ata tta aga cct agt tcc ccg acg 48
Met Ala Met Ile Ile Thr Lys Gln Ile Leu Arg Pro Ser Ser Pro Thr
1 5 10 15
ccc caa gcc ttt aag aat cac aag ctg tcc tac tta gac caa ata cag 96
Pro Gln Ala Phe Lys Asn His Lys Leu Ser Tyr Leu Asp Gln Ile Gln
20 25 30
gcg cct att tac ata cct ttg ttg ttc ttc tat aag aac gag gag tca 144
Ala Pro Ile Tyr Ile Pro Leu Leu Phe Phe Tyr Lys Asn Glu Glu Ser
35 40 45
aaa tac cca gac cag atc tcg caa aga ttt aag cag agt ttg tcc gaa 192
Lys Tyr Pro Asp Gln Ile Ser Gln Arg Phe Lys Gln Ser Leu Ser Glu
50 55 60
att ttg aca ata ttc tac cct ttg gct ggt acg atg agg cat aac tcg 240
Ile Leu Thr Ile Phe Tyr Pro Leu Ala Gly Thr Met Arg His Asn Ser
65 70 75 80
ttc gtg gac tgc aat gac agg ggt gtc gaa ttt gta gag gtc agg gtc 288
Phe Val Asp Cys Asn Asp Arg Gly Val Glu Phe Val Glu Val Arg Val
85 90 95
cat gcg aga cta gcc cag ttc att caa gat cct aag atg gaa gag ctt 336
His Ala Arg Leu Ala Gln Phe Ile Gln Asp Pro Lys Met Glu Glu Leu
100 105 110
aag caa ttg ata cct gtg gat tgt ata tct cac act gac gat gat ttc 384
Lys Gln Leu Ile Pro Val Asp Cys Ile Ser His Thr Asp Asp Asp Phe
115 120 125
tta ttg cta gtc aag att agc tat ttt gac tgc ggt gag gtc gtc gtt 432
Leu Leu Leu Val Lys Ile Ser Tyr Phe Asp Cys Gly Glu Val Val Val
130 135 140
gga gtc tgc atg tcg cac aaa att ggt gac gga att agt ctg gca gcg 480
Gly Val Cys Met Ser His Lys Ile Gly Asp Gly Ile Ser Leu Ala Ala
145 150 155 160
ttt atg aac gct tgg gca gca acg tgt agg gga gaa tcg tct agt gag 528
Phe Met Asn Ala Trp Ala Ala Thr Cys Arg Gly Glu Ser Ser Ser Glu
165 170 175
atc att cac cca tct ttt gat ctt gct tta cac ttt ccg cct aaa gac 576
Ile Ile His Pro Ser Phe Asp Leu Ala Leu His Phe Pro Pro Lys Asp
180 185 190
cac ttg tct tca gca tcc tca ttc cgt gtg gcc ata gcc cag gag aac 624
His Leu Ser Ser Ala Ser Ser Phe Arg Val Ala Ile Ala Gln Glu Asn
195 200 205
atc atg acc aag agg cta gta ttt gat aga gaa aag ttg gag aag ctg 672
Ile Met Thr Lys Arg Leu Val Phe Asp Arg Glu Lys Leu Glu Lys Leu
210 215 220
cgt aag aga atc gct gcc agt tct gat ggg gtg aga gac cct agc aga 720
Arg Lys Arg Ile Ala Ala Ser Ser Asp Gly Val Arg Asp Pro Ser Arg
225 230 235 240
gtt gaa gct gta tct gtc ttt att tgg aaa agc tta att gaa gcc cac 768
Val Glu Ala Val Ser Val Phe Ile Trp Lys Ser Leu Ile Glu Ala His
245 250 255
aag gcc gag tca cac atg act gag aca cca gcc gtt tct att gct agc 816
Lys Ala Glu Ser His Met Thr Glu Thr Pro Ala Val Ser Ile Ala Ser
260 265 270
cac gcc gtg aac tta agg cct aga aca gtc cca caa atg gac caa act 864
His Ala Val Asn Leu Arg Pro Arg Thr Val Pro Gln Met Asp Gln Thr
275 280 285
ttc ggt aac tgc tac gct ccc gct tcg gcc gtt gtc tcc tgg gat gaa 912
Phe Gly Asn Cys Tyr Ala Pro Ala Ser Ala Val Val Ser Trp Asp Glu
290 295 300
gac tac gta cat cac agt cgt ttg agg gcg gcc ctt aga gaa atc gac 960
Asp Tyr Val His His Ser Arg Leu Arg Ala Ala Leu Arg Glu Ile Asp
305 310 315 320
gac gac tac att aat aag gtc ttg aaa gcc gac aat aat tat tta acg 1008
Asp Asp Tyr Ile Asn Lys Val Leu Lys Ala Asp Asn Asn Tyr Leu Thr
325 330 335
cag gat caa att ggt gac ttg ttt aaa cca gaa aac tca gtt cta agt 1056
Gln Asp Gln Ile Gly Asp Leu Phe Lys Pro Glu Asn Ser Val Leu Ser
340 345 350
tcg tgg tgg cgt ttc cca gtt tac aag gta gac ttt gga tgg ggt aag 1104
Ser Trp Trp Arg Phe Pro Val Tyr Lys Val Asp Phe Gly Trp Gly Lys
355 360 365
cca gtt tgg gtt tct acg acg acc atc caa tac atg aac ttg att ata 1152
Pro Val Trp Val Ser Thr Thr Thr Ile Gln Tyr Met Asn Leu Ile Ile
370 375 380
ttt act tcg acg ccc tca gaa gac ggc ata gag gcg tgg gta act act 1200
Phe Thr Ser Thr Pro Ser Glu Asp Gly Ile Glu Ala Trp Val Thr Thr
385 390 395 400
act cac aat ttc ttc caa gtt ctg cag gca aac tat aat aaa ctt gac 1248
Thr His Asn Phe Phe Gln Val Leu Gln Ala Asn Tyr Asn Lys Leu Asp
405 410 415
acg taa 1254
Thr
<210> 15
<211> 417
<212> PRT
<213> LiAAT-4 DNA序列,针对其在酿酒酵母中的表达经密码子优化
<400> 15
Met Ala Met Ile Ile Thr Lys Gln Ile Leu Arg Pro Ser Ser Pro Thr
1 5 10 15
Pro Gln Ala Phe Lys Asn His Lys Leu Ser Tyr Leu Asp Gln Ile Gln
20 25 30
Ala Pro Ile Tyr Ile Pro Leu Leu Phe Phe Tyr Lys Asn Glu Glu Ser
35 40 45
Lys Tyr Pro Asp Gln Ile Ser Gln Arg Phe Lys Gln Ser Leu Ser Glu
50 55 60
Ile Leu Thr Ile Phe Tyr Pro Leu Ala Gly Thr Met Arg His Asn Ser
65 70 75 80
Phe Val Asp Cys Asn Asp Arg Gly Val Glu Phe Val Glu Val Arg Val
85 90 95
His Ala Arg Leu Ala Gln Phe Ile Gln Asp Pro Lys Met Glu Glu Leu
100 105 110
Lys Gln Leu Ile Pro Val Asp Cys Ile Ser His Thr Asp Asp Asp Phe
115 120 125
Leu Leu Leu Val Lys Ile Ser Tyr Phe Asp Cys Gly Glu Val Val Val
130 135 140
Gly Val Cys Met Ser His Lys Ile Gly Asp Gly Ile Ser Leu Ala Ala
145 150 155 160
Phe Met Asn Ala Trp Ala Ala Thr Cys Arg Gly Glu Ser Ser Ser Glu
165 170 175
Ile Ile His Pro Ser Phe Asp Leu Ala Leu His Phe Pro Pro Lys Asp
180 185 190
His Leu Ser Ser Ala Ser Ser Phe Arg Val Ala Ile Ala Gln Glu Asn
195 200 205
Ile Met Thr Lys Arg Leu Val Phe Asp Arg Glu Lys Leu Glu Lys Leu
210 215 220
Arg Lys Arg Ile Ala Ala Ser Ser Asp Gly Val Arg Asp Pro Ser Arg
225 230 235 240
Val Glu Ala Val Ser Val Phe Ile Trp Lys Ser Leu Ile Glu Ala His
245 250 255
Lys Ala Glu Ser His Met Thr Glu Thr Pro Ala Val Ser Ile Ala Ser
260 265 270
His Ala Val Asn Leu Arg Pro Arg Thr Val Pro Gln Met Asp Gln Thr
275 280 285
Phe Gly Asn Cys Tyr Ala Pro Ala Ser Ala Val Val Ser Trp Asp Glu
290 295 300
Asp Tyr Val His His Ser Arg Leu Arg Ala Ala Leu Arg Glu Ile Asp
305 310 315 320
Asp Asp Tyr Ile Asn Lys Val Leu Lys Ala Asp Asn Asn Tyr Leu Thr
325 330 335
Gln Asp Gln Ile Gly Asp Leu Phe Lys Pro Glu Asn Ser Val Leu Ser
340 345 350
Ser Trp Trp Arg Phe Pro Val Tyr Lys Val Asp Phe Gly Trp Gly Lys
355 360 365
Pro Val Trp Val Ser Thr Thr Thr Ile Gln Tyr Met Asn Leu Ile Ile
370 375 380
Phe Thr Ser Thr Pro Ser Glu Asp Gly Ile Glu Ala Trp Val Thr Thr
385 390 395 400
Thr His Asn Phe Phe Gln Val Leu Gln Ala Asn Tyr Asn Lys Leu Asp
405 410 415
Thr
<210> 16
<211> 1485
<212> DNA
<213> FgaAT,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1485)
<400> 16
atg aag aag cag gtc act ttt aaa cct ttc aga ttg tca cca gtc gat 48
Met Lys Lys Gln Val Thr Phe Lys Pro Phe Arg Leu Ser Pro Val Asp
1 5 10 15
cat agt tta cct aag gtt tac atc ttc aag tct cta tat ttc aga ggg 96
His Ser Leu Pro Lys Val Tyr Ile Phe Lys Ser Leu Tyr Phe Arg Gly
20 25 30
gta gat gac act ggt tct ctt agt aga cta cag gat ggc att gat cgt 144
Val Asp Asp Thr Gly Ser Leu Ser Arg Leu Gln Asp Gly Ile Asp Arg
35 40 45
ttg ata tct tgt ctg cct ttc ctg tca ggc gag gtg gtt cca tgt gca 192
Leu Ile Ser Cys Leu Pro Phe Leu Ser Gly Glu Val Val Pro Cys Ala
50 55 60
gac ata cca gat aag gtc ggt gta ctt caa gtt cag atg cct tgc cct 240
Asp Ile Pro Asp Lys Val Gly Val Leu Gln Val Gln Met Pro Cys Pro
65 70 75 80
tcc ttg caa gaa att cca atg ctg cta gtc aaa agt tat cca aac cat 288
Ser Leu Gln Glu Ile Pro Met Leu Leu Val Lys Ser Tyr Pro Asn His
85 90 95
aca tgg cca gca gct tct acc tcg gaa aga tgg aga aac acc gct ttg 336
Thr Trp Pro Ala Ala Ser Thr Ser Glu Arg Trp Arg Asn Thr Ala Leu
100 105 110
cta gat cag tcg tat agg cca ttg ccg gat ttt ata ccc cca tca aag 384
Leu Asp Gln Ser Tyr Arg Pro Leu Pro Asp Phe Ile Pro Pro Ser Lys
115 120 125
cct aga cct gtt ttg aga ttt cag gcg aac ttc ctg gct gat ggt tta 432
Pro Arg Pro Val Leu Arg Phe Gln Ala Asn Phe Leu Ala Asp Gly Leu
130 135 140
atg cta tgt atg gga tat aat cat tct gta ttc gac gga aca ggt gcg 480
Met Leu Cys Met Gly Tyr Asn His Ser Val Phe Asp Gly Thr Gly Ala
145 150 155 160
ggg aac att ctg gaa atg ttg gct gat tgt tgt aga gct aat cca aac 528
Gly Asn Ile Leu Glu Met Leu Ala Asp Cys Cys Arg Ala Asn Pro Asn
165 170 175
tcc atc tta gca ctg cca aca aac ggt gac ata gag agc gag ttg aga 576
Ser Ile Leu Ala Leu Pro Thr Asn Gly Asp Ile Glu Ser Glu Leu Arg
180 185 190
ggt tta ctg tcc agt cca ggt gta gca gtt gca aat gcc tct cag gaa 624
Gly Leu Leu Ser Ser Pro Gly Val Ala Val Ala Asn Ala Ser Gln Glu
195 200 205
gcc tac gca att aat tgt gca cac acg gaa gta gaa ccc gaa ccc agt 672
Ala Tyr Ala Ile Asn Cys Ala His Thr Glu Val Glu Pro Glu Pro Ser
210 215 220
tca gct atg ctt tac tgt tgg cct ttc ttg ctg agt tct gag aag att 720
Ser Ala Met Leu Tyr Cys Trp Pro Phe Leu Leu Ser Ser Glu Lys Ile
225 230 235 240
gaa tgt tta cag gaa gca tgt aat agt tta cta cca cat atc gtt cgt 768
Glu Cys Leu Gln Glu Ala Cys Asn Ser Leu Leu Pro His Ile Val Arg
245 250 255
ttg tac tct ggt acg cag agt tcg ctt ata aac caa gat acg aac tgg 816
Leu Tyr Ser Gly Thr Gln Ser Ser Leu Ile Asn Gln Asp Thr Asn Trp
260 265 270
cca cat att ctt tca tca aac gac gtg cta acc gct ctt tta gct gtt 864
Pro His Ile Leu Ser Ser Asn Asp Val Leu Thr Ala Leu Leu Ala Val
275 280 285
tca att gaa aag gct agg gaa gca aca ggt gcc ctt ggt cat atg agc 912
Ser Ile Glu Lys Ala Arg Glu Ala Thr Gly Ala Leu Gly His Met Ser
290 295 300
aga tca ttg gct atg gct gtt aac cta cgt gag cgt tta aag cct atg 960
Arg Ser Leu Ala Met Ala Val Asn Leu Arg Glu Arg Leu Lys Pro Met
305 310 315 320
cca aga cac tat ttg ggt aac cta gtt acc act gtg tgg gta tca cat 1008
Pro Arg His Tyr Leu Gly Asn Leu Val Thr Thr Val Trp Val Ser His
325 330 335
cac cgt cct gcc gtt aag gac ctt gaa aca atg gtt tta cca gtg cca 1056
His Arg Pro Ala Val Lys Asp Leu Glu Thr Met Val Leu Pro Val Pro
340 345 350
gca tgc aat agg cac gaa ata gac aga gac gac ttg ttg tgg ata acc 1104
Ala Cys Asn Arg His Glu Ile Asp Arg Asp Asp Leu Leu Trp Ile Thr
355 360 365
cac gtg gca ttc cgt att aga tta ggg ctg aac gca ata aac gaa gaa 1152
His Val Ala Phe Arg Ile Arg Leu Gly Leu Asn Ala Ile Asn Glu Glu
370 375 380
cat att aga ggt ctg atc cac tac ttg cat tcc caa gac gat tgg gaa 1200
His Ile Arg Gly Leu Ile His Tyr Leu His Ser Gln Asp Asp Trp Glu
385 390 395 400
cag att gga ata cat ttc acc gat cca att ttc att tct tcc tgg cgt 1248
Gln Ile Gly Ile His Phe Thr Asp Pro Ile Phe Ile Ser Ser Trp Arg
405 410 415
cac ctt aag gtc tat gaa cta gac ttc gga cct act atc ggt cat gct 1296
His Leu Lys Val Tyr Glu Leu Asp Phe Gly Pro Thr Ile Gly His Ala
420 425 430
gaa cac ttt gag atg gat gtc ggc acc acc gat ggt gtg tgt gtt gtt 1344
Glu His Phe Glu Met Asp Val Gly Thr Thr Asp Gly Val Cys Val Val
435 440 445
atg cct gct aac acc aga gcc gtc ggc aag act aag aag gct cct tgg 1392
Met Pro Ala Asn Thr Arg Ala Val Gly Lys Thr Lys Lys Ala Pro Trp
450 455 460
gac att aga atc gtg tta aac ccc gaa gtg tta cag gca cta ata gct 1440
Asp Ile Arg Ile Val Leu Asn Pro Glu Val Leu Gln Ala Leu Ile Ala
465 470 475 480
agt gcc atc ttt ggt tgg gct atg gtc aag gac gct tcg aca taa 1485
Ser Ala Ile Phe Gly Trp Ala Met Val Lys Asp Ala Ser Thr
485 490
<210> 17
<211> 494
<212> PRT
<213> FgaAT,针对其在酿酒酵母中的表达经密码子优化
<400> 17
Met Lys Lys Gln Val Thr Phe Lys Pro Phe Arg Leu Ser Pro Val Asp
1 5 10 15
His Ser Leu Pro Lys Val Tyr Ile Phe Lys Ser Leu Tyr Phe Arg Gly
20 25 30
Val Asp Asp Thr Gly Ser Leu Ser Arg Leu Gln Asp Gly Ile Asp Arg
35 40 45
Leu Ile Ser Cys Leu Pro Phe Leu Ser Gly Glu Val Val Pro Cys Ala
50 55 60
Asp Ile Pro Asp Lys Val Gly Val Leu Gln Val Gln Met Pro Cys Pro
65 70 75 80
Ser Leu Gln Glu Ile Pro Met Leu Leu Val Lys Ser Tyr Pro Asn His
85 90 95
Thr Trp Pro Ala Ala Ser Thr Ser Glu Arg Trp Arg Asn Thr Ala Leu
100 105 110
Leu Asp Gln Ser Tyr Arg Pro Leu Pro Asp Phe Ile Pro Pro Ser Lys
115 120 125
Pro Arg Pro Val Leu Arg Phe Gln Ala Asn Phe Leu Ala Asp Gly Leu
130 135 140
Met Leu Cys Met Gly Tyr Asn His Ser Val Phe Asp Gly Thr Gly Ala
145 150 155 160
Gly Asn Ile Leu Glu Met Leu Ala Asp Cys Cys Arg Ala Asn Pro Asn
165 170 175
Ser Ile Leu Ala Leu Pro Thr Asn Gly Asp Ile Glu Ser Glu Leu Arg
180 185 190
Gly Leu Leu Ser Ser Pro Gly Val Ala Val Ala Asn Ala Ser Gln Glu
195 200 205
Ala Tyr Ala Ile Asn Cys Ala His Thr Glu Val Glu Pro Glu Pro Ser
210 215 220
Ser Ala Met Leu Tyr Cys Trp Pro Phe Leu Leu Ser Ser Glu Lys Ile
225 230 235 240
Glu Cys Leu Gln Glu Ala Cys Asn Ser Leu Leu Pro His Ile Val Arg
245 250 255
Leu Tyr Ser Gly Thr Gln Ser Ser Leu Ile Asn Gln Asp Thr Asn Trp
260 265 270
Pro His Ile Leu Ser Ser Asn Asp Val Leu Thr Ala Leu Leu Ala Val
275 280 285
Ser Ile Glu Lys Ala Arg Glu Ala Thr Gly Ala Leu Gly His Met Ser
290 295 300
Arg Ser Leu Ala Met Ala Val Asn Leu Arg Glu Arg Leu Lys Pro Met
305 310 315 320
Pro Arg His Tyr Leu Gly Asn Leu Val Thr Thr Val Trp Val Ser His
325 330 335
His Arg Pro Ala Val Lys Asp Leu Glu Thr Met Val Leu Pro Val Pro
340 345 350
Ala Cys Asn Arg His Glu Ile Asp Arg Asp Asp Leu Leu Trp Ile Thr
355 360 365
His Val Ala Phe Arg Ile Arg Leu Gly Leu Asn Ala Ile Asn Glu Glu
370 375 380
His Ile Arg Gly Leu Ile His Tyr Leu His Ser Gln Asp Asp Trp Glu
385 390 395 400
Gln Ile Gly Ile His Phe Thr Asp Pro Ile Phe Ile Ser Ser Trp Arg
405 410 415
His Leu Lys Val Tyr Glu Leu Asp Phe Gly Pro Thr Ile Gly His Ala
420 425 430
Glu His Phe Glu Met Asp Val Gly Thr Thr Asp Gly Val Cys Val Val
435 440 445
Met Pro Ala Asn Thr Arg Ala Val Gly Lys Thr Lys Lys Ala Pro Trp
450 455 460
Asp Ile Arg Ile Val Leu Asn Pro Glu Val Leu Gln Ala Leu Ile Ala
465 470 475 480
Ser Ala Ile Phe Gly Trp Ala Met Val Lys Asp Ala Ser Thr
485 490
<210> 18
<211> 1548
<212> DNA
<213> GAO81666.1,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1548)
<400> 18
atg gaa gaa cat acc aga tcc ttt gag cca ttt gac ctg gcc tgc tta 48
Met Glu Glu His Thr Arg Ser Phe Glu Pro Phe Asp Leu Ala Cys Leu
1 5 10 15
gat cat act gtg ggg ccg gtt ttc atg aat ttc ttc tta agt ttc aag 96
Asp His Thr Val Gly Pro Val Phe Met Asn Phe Phe Leu Ser Phe Lys
20 25 30
cct gcc aag atc gaa gag agt ctt atc tca att gaa gaa ggt gta act 144
Pro Ala Lys Ile Glu Glu Ser Leu Ile Ser Ile Glu Glu Gly Val Thr
35 40 45
agg tta gtc aac aga ctg cca ttc ctg gcc ggt gac gtt gtt aat tct 192
Arg Leu Val Asn Arg Leu Pro Phe Leu Ala Gly Asp Val Val Asn Ser
50 55 60
gaa aac gta gac ggt cgt gtt aat gta atg agg att cag cca agc tcg 240
Glu Asn Val Asp Gly Arg Val Asn Val Met Arg Ile Gln Pro Ser Ser
65 70 75 80
aca ttg att aga gag att ccc atg cta cac aca aag cac cat cct cat 288
Thr Leu Ile Arg Glu Ile Pro Met Leu His Thr Lys His His Pro His
85 90 95
cat att ttg ccg aat aca cct aga cag att aac cca ggg tcc gac cag 336
His Ile Leu Pro Asn Thr Pro Arg Gln Ile Asn Pro Gly Ser Asp Gln
100 105 110
gat caa agg ttt gct ccc ttg gac gat tcg tac gta cca cca gtg tct 384
Asp Gln Arg Phe Ala Pro Leu Asp Asp Ser Tyr Val Pro Pro Val Ser
115 120 125
tta tta cct ctt gct ccg ggc cca agg ccc gta gta aga ttc caa act 432
Leu Leu Pro Leu Ala Pro Gly Pro Arg Pro Val Val Arg Phe Gln Thr
130 135 140
aat gtt gta att gac ggg att gtg ctt gca ctt ggg ttc cac cac tca 480
Asn Val Val Ile Asp Gly Ile Val Leu Ala Leu Gly Phe His His Ser
145 150 155 160
gta ttt gat gct acc gga gtg ggt ttg ttg atc gaa atg ttg gcc aca 528
Val Phe Asp Ala Thr Gly Val Gly Leu Leu Ile Glu Met Leu Ala Thr
165 170 175
tgt tgt tca agc gat tgt cct gcc cta tca tcc cat att gaa ctg gaa 576
Cys Cys Ser Ser Asp Cys Pro Ala Leu Ser Ser His Ile Glu Leu Glu
180 185 190
gaa gaa ata agg cta agg cgt tct gtg gac aag ata ggt aac ggc gct 624
Glu Glu Ile Arg Leu Arg Arg Ser Val Asp Lys Ile Gly Asn Gly Ala
195 200 205
act gat tta gca tcc caa aga gat gaa gtt cag gac cca aat ggc agt 672
Thr Asp Leu Ala Ser Gln Arg Asp Glu Val Gln Asp Pro Asn Gly Ser
210 215 220
atc act gag tcc cca gct cat gtg cct atg ggt gac agt agc tgg gta 720
Ile Thr Glu Ser Pro Ala His Val Pro Met Gly Asp Ser Ser Trp Val
225 230 235 240
ccc cct aaa cta tct gtt tac tcc ttt aat tta tcg gct gcc ggt ttg 768
Pro Pro Lys Leu Ser Val Tyr Ser Phe Asn Leu Ser Ala Ala Gly Leu
245 250 255
gct cat ttg aag aca gcc tgt aac aag tta ttg ccc gct atc cat gcg 816
Ala His Leu Lys Thr Ala Cys Asn Lys Leu Leu Pro Ala Ile His Ala
260 265 270
agt caa aat ggt tca cca caa agt gcg gaa tca gaa ggt gag aaa tta 864
Ser Gln Asn Gly Ser Pro Gln Ser Ala Glu Ser Glu Gly Glu Lys Leu
275 280 285
cag caa gaa ttc gtt tcg acg aat gat gtt cta acg gct ttg ttg gct 912
Gln Gln Glu Phe Val Ser Thr Asn Asp Val Leu Thr Ala Leu Leu Ala
290 295 300
act tcc att cac caa gcg agg tct aga gtt act gaa acg gaa tta gtt 960
Thr Ser Ile His Gln Ala Arg Ser Arg Val Thr Glu Thr Glu Leu Val
305 310 315 320
ccg aca aaa gcg aaa ctt gcg atg gcc gtt aat ttg agg gaa agg gcc 1008
Pro Thr Lys Ala Lys Leu Ala Met Ala Val Asn Leu Arg Glu Arg Ala
325 330 335
agt tcg tta ccc aag act tat ttg ggg aac tcg cta aca gtt aca gaa 1056
Ser Ser Leu Pro Lys Thr Tyr Leu Gly Asn Ser Leu Thr Val Thr Glu
340 345 350
gct ttt gtg tat tct tta gct gct acc gac ggt ttc gat ggt gtc ccc 1104
Ala Phe Val Tyr Ser Leu Ala Ala Thr Asp Gly Phe Asp Gly Val Pro
355 360 365
gag aga cat tat cat cct gac ttg cgt aac gcc ttg cta tta gaa att 1152
Glu Arg His Tyr His Pro Asp Leu Arg Asn Ala Leu Leu Leu Glu Ile
370 375 380
gct aga gtt gca ttg caa ttg aga aaa ggg ctg gca gcg ata gat gac 1200
Ala Arg Val Ala Leu Gln Leu Arg Lys Gly Leu Ala Ala Ile Asp Asp
385 390 395 400
gcg tat ttt agg caa ttt gtg tct aga ttg agg gct aat cta gac tgg 1248
Ala Tyr Phe Arg Gln Phe Val Ser Arg Leu Arg Ala Asn Leu Asp Trp
405 410 415
tca cag tta ggt gca aac ctg cct gac act atg gtg tca agc tgg aga 1296
Ser Gln Leu Gly Ala Asn Leu Pro Asp Thr Met Val Ser Ser Trp Arg
420 425 430
cat ttg aaa gta tat cgt tta gac ttc ggt gct aga ttg ggg aga gtt 1344
His Leu Lys Val Tyr Arg Leu Asp Phe Gly Ala Arg Leu Gly Arg Val
435 440 445
gta gag ttc cat cca caa acc gct ttg gtt gac ggt att tgt atc atc 1392
Val Glu Phe His Pro Gln Thr Ala Leu Val Asp Gly Ile Cys Ile Ile
450 455 460
caa cct gaa agg ata gcc cat gag gat gat tca gct gat atg gct cca 1440
Gln Pro Glu Arg Ile Ala His Glu Asp Asp Ser Ala Asp Met Ala Pro
465 470 475 480
gaa tct ggg tgg gaa gtt tgt gta aca tta caa agc gat gcc atg gag 1488
Glu Ser Gly Trp Glu Val Cys Val Thr Leu Gln Ser Asp Ala Met Glu
485 490 495
tgc ttc tta aga ggt ggg ttg ttc aca agt ctt tct caa ggt gca att 1536
Cys Phe Leu Arg Gly Gly Leu Phe Thr Ser Leu Ser Gln Gly Ala Ile
500 505 510
aga aga gtt taa 1548
Arg Arg Val
515
<210> 19
<211> 515
<212> PRT
<213> GAO81666.1,针对其在酿酒酵母中的表达经密码子优化
<400> 19
Met Glu Glu His Thr Arg Ser Phe Glu Pro Phe Asp Leu Ala Cys Leu
1 5 10 15
Asp His Thr Val Gly Pro Val Phe Met Asn Phe Phe Leu Ser Phe Lys
20 25 30
Pro Ala Lys Ile Glu Glu Ser Leu Ile Ser Ile Glu Glu Gly Val Thr
35 40 45
Arg Leu Val Asn Arg Leu Pro Phe Leu Ala Gly Asp Val Val Asn Ser
50 55 60
Glu Asn Val Asp Gly Arg Val Asn Val Met Arg Ile Gln Pro Ser Ser
65 70 75 80
Thr Leu Ile Arg Glu Ile Pro Met Leu His Thr Lys His His Pro His
85 90 95
His Ile Leu Pro Asn Thr Pro Arg Gln Ile Asn Pro Gly Ser Asp Gln
100 105 110
Asp Gln Arg Phe Ala Pro Leu Asp Asp Ser Tyr Val Pro Pro Val Ser
115 120 125
Leu Leu Pro Leu Ala Pro Gly Pro Arg Pro Val Val Arg Phe Gln Thr
130 135 140
Asn Val Val Ile Asp Gly Ile Val Leu Ala Leu Gly Phe His His Ser
145 150 155 160
Val Phe Asp Ala Thr Gly Val Gly Leu Leu Ile Glu Met Leu Ala Thr
165 170 175
Cys Cys Ser Ser Asp Cys Pro Ala Leu Ser Ser His Ile Glu Leu Glu
180 185 190
Glu Glu Ile Arg Leu Arg Arg Ser Val Asp Lys Ile Gly Asn Gly Ala
195 200 205
Thr Asp Leu Ala Ser Gln Arg Asp Glu Val Gln Asp Pro Asn Gly Ser
210 215 220
Ile Thr Glu Ser Pro Ala His Val Pro Met Gly Asp Ser Ser Trp Val
225 230 235 240
Pro Pro Lys Leu Ser Val Tyr Ser Phe Asn Leu Ser Ala Ala Gly Leu
245 250 255
Ala His Leu Lys Thr Ala Cys Asn Lys Leu Leu Pro Ala Ile His Ala
260 265 270
Ser Gln Asn Gly Ser Pro Gln Ser Ala Glu Ser Glu Gly Glu Lys Leu
275 280 285
Gln Gln Glu Phe Val Ser Thr Asn Asp Val Leu Thr Ala Leu Leu Ala
290 295 300
Thr Ser Ile His Gln Ala Arg Ser Arg Val Thr Glu Thr Glu Leu Val
305 310 315 320
Pro Thr Lys Ala Lys Leu Ala Met Ala Val Asn Leu Arg Glu Arg Ala
325 330 335
Ser Ser Leu Pro Lys Thr Tyr Leu Gly Asn Ser Leu Thr Val Thr Glu
340 345 350
Ala Phe Val Tyr Ser Leu Ala Ala Thr Asp Gly Phe Asp Gly Val Pro
355 360 365
Glu Arg His Tyr His Pro Asp Leu Arg Asn Ala Leu Leu Leu Glu Ile
370 375 380
Ala Arg Val Ala Leu Gln Leu Arg Lys Gly Leu Ala Ala Ile Asp Asp
385 390 395 400
Ala Tyr Phe Arg Gln Phe Val Ser Arg Leu Arg Ala Asn Leu Asp Trp
405 410 415
Ser Gln Leu Gly Ala Asn Leu Pro Asp Thr Met Val Ser Ser Trp Arg
420 425 430
His Leu Lys Val Tyr Arg Leu Asp Phe Gly Ala Arg Leu Gly Arg Val
435 440 445
Val Glu Phe His Pro Gln Thr Ala Leu Val Asp Gly Ile Cys Ile Ile
450 455 460
Gln Pro Glu Arg Ile Ala His Glu Asp Asp Ser Ala Asp Met Ala Pro
465 470 475 480
Glu Ser Gly Trp Glu Val Cys Val Thr Leu Gln Ser Asp Ala Met Glu
485 490 495
Cys Phe Leu Arg Gly Gly Leu Phe Thr Ser Leu Ser Gln Gly Ala Ile
500 505 510
Arg Arg Val
515
<210> 20
<211> 1284
<212> DNA
<213> CfACT1-6 DNA序列,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1284)
<400> 20
atg aag gta gaa cgt ttc tca aga aag ttg ata aaa ccc cac acg cct 48
Met Lys Val Glu Arg Phe Ser Arg Lys Leu Ile Lys Pro His Thr Pro
1 5 10 15
act ccc gag aac tta aag aaa tat aaa ttg tct cta ttg gac aaa tgc 96
Thr Pro Glu Asn Leu Lys Lys Tyr Lys Leu Ser Leu Leu Asp Lys Cys
20 25 30
ctt gga cac gac aac ttc gcg atc gtg ttg ttt tac gaa tct aaa cca 144
Leu Gly His Asp Asn Phe Ala Ile Val Leu Phe Tyr Glu Ser Lys Pro
35 40 45
aga aat aag agt gag ctt gag gaa tcg tta gag aag gtc ttg gta gac 192
Arg Asn Lys Ser Glu Leu Glu Glu Ser Leu Glu Lys Val Leu Val Asp
50 55 60
ttt tat ccc ttg gct ggg cgt cac acg atg aac gac cat att gtc gac 240
Phe Tyr Pro Leu Ala Gly Arg His Thr Met Asn Asp His Ile Val Asp
65 70 75 80
tgc tcg gat gtc ggc gct gtc ttt gta gaa gcc gag gct tta gat gta 288
Cys Ser Asp Val Gly Ala Val Phe Val Glu Ala Glu Ala Leu Asp Val
85 90 95
gag ttg act atg gat gag tta gtc aag aac atg gaa gct caa act ata 336
Glu Leu Thr Met Asp Glu Leu Val Lys Asn Met Glu Ala Gln Thr Ile
100 105 110
cac cac ttg ttg cca aat cag tac ttc agt gca gat gct ccg aac ccg 384
His His Leu Leu Pro Asn Gln Tyr Phe Ser Ala Asp Ala Pro Asn Pro
115 120 125
ctg ctt tcc atc caa gtt acg cac ttt cct tcc ggt ggt ttg gct atc 432
Leu Leu Ser Ile Gln Val Thr His Phe Pro Ser Gly Gly Leu Ala Ile
130 135 140
gga atc gca gtc tcg cac gct gta ttt gat ggt ttc tct ttg ggc gtg 480
Gly Ile Ala Val Ser His Ala Val Phe Asp Gly Phe Ser Leu Gly Val
145 150 155 160
ttc gta gca gca tgg tca aag gcg acc atg aac ccg gat agg aaa atc 528
Phe Val Ala Ala Trp Ser Lys Ala Thr Met Asn Pro Asp Arg Lys Ile
165 170 175
aaa ata aca ccg tca ttc gac tta cca tca ctt ctg ccc tac aaa gac 576
Lys Ile Thr Pro Ser Phe Asp Leu Pro Ser Leu Leu Pro Tyr Lys Asp
180 185 190
gac aac ttt ggt ttg act gct gct gaa att gtc agc cag agc gag gac 624
Asp Asn Phe Gly Leu Thr Ala Ala Glu Ile Val Ser Gln Ser Glu Asp
195 200 205
atc gta gtt aag aga ttt atc ttc ggc aag gaa gcc atc acg agg ttg 672
Ile Val Val Lys Arg Phe Ile Phe Gly Lys Glu Ala Ile Thr Arg Leu
210 215 220
aga agt aag ctt agt cca aat agg aac ggg aag aaa ata tcc agg gtt 720
Arg Ser Lys Leu Ser Pro Asn Arg Asn Gly Lys Lys Ile Ser Arg Val
225 230 235 240
agg gtc gtt tgt gca gtc att gta aag gcc ttg atg gga ttg gag cgt 768
Arg Val Val Cys Ala Val Ile Val Lys Ala Leu Met Gly Leu Glu Arg
245 250 255
gcc aaa cac ggt aag acg cgt gat ttc tta att act caa tca att aac 816
Ala Lys His Gly Lys Thr Arg Asp Phe Leu Ile Thr Gln Ser Ile Asn
260 265 270
atg agg gag aga act aag gcg ccg ctg cag aaa cac gcc tgc ggc aac 864
Met Arg Glu Arg Thr Lys Ala Pro Leu Gln Lys His Ala Cys Gly Asn
275 280 285
tta gca gtc ttg agt tgc acg aga aga gta gag gcc gag gag atg atg 912
Leu Ala Val Leu Ser Cys Thr Arg Arg Val Glu Ala Glu Glu Met Met
290 295 300
gag tta cag aac cta gtt aat ttg atc ggc gac agt acc gaa aag gac 960
Glu Leu Gln Asn Leu Val Asn Leu Ile Gly Asp Ser Thr Glu Lys Asp
305 310 315 320
atc gcc gac ttt gca gaa tta tta tca cca gat caa gtg ggc aga gac 1008
Ile Ala Asp Phe Ala Glu Leu Leu Ser Pro Asp Gln Val Gly Arg Asp
325 330 335
att ata atc aag atg atg aag tca ttc atg cag ttc ttg gac aat gac 1056
Ile Ile Ile Lys Met Met Lys Ser Phe Met Gln Phe Leu Asp Asn Asp
340 345 350
att tac agc gtg tgc ttt acc gat tgg tca aag ttt gaa ttt tac gaa 1104
Ile Tyr Ser Val Cys Phe Thr Asp Trp Ser Lys Phe Glu Phe Tyr Glu
355 360 365
gcg gac ttc ggc ttc ggg aag ccg gtt tgg atg gca gca ggc cca cag 1152
Ala Asp Phe Gly Phe Gly Lys Pro Val Trp Met Ala Ala Gly Pro Gln
370 375 380
aga ccc att atc tct act gca ata ttg atg tca gac agg gaa ggc gac 1200
Arg Pro Ile Ile Ser Thr Ala Ile Leu Met Ser Asp Arg Glu Gly Asp
385 390 395 400
ggt atc gag gct tgg tta cat ttg aat aag aac gac atg ttg att ttc 1248
Gly Ile Glu Ala Trp Leu His Leu Asn Lys Asn Asp Met Leu Ile Phe
405 410 415
gaa caa gac gag gaa atc aag tta ttc act aca taa 1284
Glu Gln Asp Glu Glu Ile Lys Leu Phe Thr Thr
420 425
<210> 21
<211> 427
<212> PRT
<213> CfACT1-6 DNA序列,针对其在酿酒酵母中的表达经密码子优化
<400> 21
Met Lys Val Glu Arg Phe Ser Arg Lys Leu Ile Lys Pro His Thr Pro
1 5 10 15
Thr Pro Glu Asn Leu Lys Lys Tyr Lys Leu Ser Leu Leu Asp Lys Cys
20 25 30
Leu Gly His Asp Asn Phe Ala Ile Val Leu Phe Tyr Glu Ser Lys Pro
35 40 45
Arg Asn Lys Ser Glu Leu Glu Glu Ser Leu Glu Lys Val Leu Val Asp
50 55 60
Phe Tyr Pro Leu Ala Gly Arg His Thr Met Asn Asp His Ile Val Asp
65 70 75 80
Cys Ser Asp Val Gly Ala Val Phe Val Glu Ala Glu Ala Leu Asp Val
85 90 95
Glu Leu Thr Met Asp Glu Leu Val Lys Asn Met Glu Ala Gln Thr Ile
100 105 110
His His Leu Leu Pro Asn Gln Tyr Phe Ser Ala Asp Ala Pro Asn Pro
115 120 125
Leu Leu Ser Ile Gln Val Thr His Phe Pro Ser Gly Gly Leu Ala Ile
130 135 140
Gly Ile Ala Val Ser His Ala Val Phe Asp Gly Phe Ser Leu Gly Val
145 150 155 160
Phe Val Ala Ala Trp Ser Lys Ala Thr Met Asn Pro Asp Arg Lys Ile
165 170 175
Lys Ile Thr Pro Ser Phe Asp Leu Pro Ser Leu Leu Pro Tyr Lys Asp
180 185 190
Asp Asn Phe Gly Leu Thr Ala Ala Glu Ile Val Ser Gln Ser Glu Asp
195 200 205
Ile Val Val Lys Arg Phe Ile Phe Gly Lys Glu Ala Ile Thr Arg Leu
210 215 220
Arg Ser Lys Leu Ser Pro Asn Arg Asn Gly Lys Lys Ile Ser Arg Val
225 230 235 240
Arg Val Val Cys Ala Val Ile Val Lys Ala Leu Met Gly Leu Glu Arg
245 250 255
Ala Lys His Gly Lys Thr Arg Asp Phe Leu Ile Thr Gln Ser Ile Asn
260 265 270
Met Arg Glu Arg Thr Lys Ala Pro Leu Gln Lys His Ala Cys Gly Asn
275 280 285
Leu Ala Val Leu Ser Cys Thr Arg Arg Val Glu Ala Glu Glu Met Met
290 295 300
Glu Leu Gln Asn Leu Val Asn Leu Ile Gly Asp Ser Thr Glu Lys Asp
305 310 315 320
Ile Ala Asp Phe Ala Glu Leu Leu Ser Pro Asp Gln Val Gly Arg Asp
325 330 335
Ile Ile Ile Lys Met Met Lys Ser Phe Met Gln Phe Leu Asp Asn Asp
340 345 350
Ile Tyr Ser Val Cys Phe Thr Asp Trp Ser Lys Phe Glu Phe Tyr Glu
355 360 365
Ala Asp Phe Gly Phe Gly Lys Pro Val Trp Met Ala Ala Gly Pro Gln
370 375 380
Arg Pro Ile Ile Ser Thr Ala Ile Leu Met Ser Asp Arg Glu Gly Asp
385 390 395 400
Gly Ile Glu Ala Trp Leu His Leu Asn Lys Asn Asp Met Leu Ile Phe
405 410 415
Glu Gln Asp Glu Glu Ile Lys Leu Phe Thr Thr
420 425
<210> 22
<211> 1272
<212> DNA
<213> CfACT1-8,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1272)
<400> 22
atg aaa gtc gaa agg att tca cgt aaa ttc atc aag cca tat aca cct 48
Met Lys Val Glu Arg Ile Ser Arg Lys Phe Ile Lys Pro Tyr Thr Pro
1 5 10 15
aca cca cag aac ctt aag aag tac aag cta tcc ttg ctg gat aaa tgc 96
Thr Pro Gln Asn Leu Lys Lys Tyr Lys Leu Ser Leu Leu Asp Lys Cys
20 25 30
atg gga cac atg gac ttc gct gta gta ttg ttt tac gaa tca aag cca 144
Met Gly His Met Asp Phe Ala Val Val Leu Phe Tyr Glu Ser Lys Pro
35 40 45
aga aac aag aat gag ctg gaa gaa tca cta gag aaa gtg tta gtc gat 192
Arg Asn Lys Asn Glu Leu Glu Glu Ser Leu Glu Lys Val Leu Val Asp
50 55 60
ttc tat cca ttg gca ggc agg tat acc atg aac gac cac att gtc gat 240
Phe Tyr Pro Leu Ala Gly Arg Tyr Thr Met Asn Asp His Ile Val Asp
65 70 75 80
tgc agc gat gag ggc gcc gtt ttc gtt gag gca gag gcc cct aat gtt 288
Cys Ser Asp Glu Gly Ala Val Phe Val Glu Ala Glu Ala Pro Asn Val
85 90 95
gag ctt aca gtg gac cag ttg gtt aag aac atg gaa gcc cag aca atc 336
Glu Leu Thr Val Asp Gln Leu Val Lys Asn Met Glu Ala Gln Thr Ile
100 105 110
cac gac ttc tta cca gac caa tat ttt cct gct gac gca cca aat ccg 384
His Asp Phe Leu Pro Asp Gln Tyr Phe Pro Ala Asp Ala Pro Asn Pro
115 120 125
ttg cta agt att caa gta acg cac ttc cct tgt ggt ggt tta gct atc 432
Leu Leu Ser Ile Gln Val Thr His Phe Pro Cys Gly Gly Leu Ala Ile
130 135 140
ggg att gtt gtt agt cac gcg gtc ttt gat gga ttc tca ttg ggc gta 480
Gly Ile Val Val Ser His Ala Val Phe Asp Gly Phe Ser Leu Gly Val
145 150 155 160
ttc tta gcc gcc tgg agc aaa gct acc atg aac cct gag agg aag atc 528
Phe Leu Ala Ala Trp Ser Lys Ala Thr Met Asn Pro Glu Arg Lys Ile
165 170 175
gaa atc acc cct tcc ttc gat ttg cct agt ctt ctg ccc tac aag gat 576
Glu Ile Thr Pro Ser Phe Asp Leu Pro Ser Leu Leu Pro Tyr Lys Asp
180 185 190
gaa tct ttc ggt tta aat ttt agc gaa att gtc aaa gct gag aat atc 624
Glu Ser Phe Gly Leu Asn Phe Ser Glu Ile Val Lys Ala Glu Asn Ile
195 200 205
gta gtt aaa cgt ttg aat ttc ggg aaa gag gct att acg cgt ttg agg 672
Val Val Lys Arg Leu Asn Phe Gly Lys Glu Ala Ile Thr Arg Leu Arg
210 215 220
tcc aag ctg tct cct aac cag aat ggt aaa acc att tcc aga gtg agg 720
Ser Lys Leu Ser Pro Asn Gln Asn Gly Lys Thr Ile Ser Arg Val Arg
225 230 235 240
gta gtc tgc gcg gtg ata gtt aag gcg ttg atg gga ctg gag aga gca 768
Val Val Cys Ala Val Ile Val Lys Ala Leu Met Gly Leu Glu Arg Ala
245 250 255
aag act aga gac ttt atg ata tgt cag ggg atc aac atg aga gag agg 816
Lys Thr Arg Asp Phe Met Ile Cys Gln Gly Ile Asn Met Arg Glu Arg
260 265 270
acg aag gcg ccc ctg cag aag cat gcg tgt ggc aac cta gca gtt tcg 864
Thr Lys Ala Pro Leu Gln Lys His Ala Cys Gly Asn Leu Ala Val Ser
275 280 285
tct tac act aga agg gta gcg gca gcg gaa gca gaa gaa ctg cag tcc 912
Ser Tyr Thr Arg Arg Val Ala Ala Ala Glu Ala Glu Glu Leu Gln Ser
290 295 300
tta gtg aat ttg atc ggg gac tct atc gaa aag tca atc gca gac tac 960
Leu Val Asn Leu Ile Gly Asp Ser Ile Glu Lys Ser Ile Ala Asp Tyr
305 310 315 320
gct gat ata ctt tcg agt gat caa gat ggg aga cac atc att tcc act 1008
Ala Asp Ile Leu Ser Ser Asp Gln Asp Gly Arg His Ile Ile Ser Thr
325 330 335
atg atg aag agc ttt atg cag ttt gct gca cct gat ata aaa gcc att 1056
Met Met Lys Ser Phe Met Gln Phe Ala Ala Pro Asp Ile Lys Ala Ile
340 345 350
tca ttt acc gac tgg tca aag ttc ggc ttt tac caa gta gat ttt ggt 1104
Ser Phe Thr Asp Trp Ser Lys Phe Gly Phe Tyr Gln Val Asp Phe Gly
355 360 365
ttt ggt aaa cca gtt tgg acc ggc gtc cgt cca gaa cgt cca atc ttc 1152
Phe Gly Lys Pro Val Trp Thr Gly Val Arg Pro Glu Arg Pro Ile Phe
370 375 380
tca gcc gcg ata ttg atg agt aac agg gaa ggc gat ggc atc gag gct 1200
Ser Ala Ala Ile Leu Met Ser Asn Arg Glu Gly Asp Gly Ile Glu Ala
385 390 395 400
tgg ctt cat ttg gac aag aac gat atg cta ata ttt gag caa gac gaa 1248
Trp Leu His Leu Asp Lys Asn Asp Met Leu Ile Phe Glu Gln Asp Glu
405 410 415
gaa att aag ttg ttg ata acg taa 1272
Glu Ile Lys Leu Leu Ile Thr
420
<210> 23
<211> 423
<212> PRT
<213> CfACT1-8,针对其在酿酒酵母中的表达经密码子优化
<400> 23
Met Lys Val Glu Arg Ile Ser Arg Lys Phe Ile Lys Pro Tyr Thr Pro
1 5 10 15
Thr Pro Gln Asn Leu Lys Lys Tyr Lys Leu Ser Leu Leu Asp Lys Cys
20 25 30
Met Gly His Met Asp Phe Ala Val Val Leu Phe Tyr Glu Ser Lys Pro
35 40 45
Arg Asn Lys Asn Glu Leu Glu Glu Ser Leu Glu Lys Val Leu Val Asp
50 55 60
Phe Tyr Pro Leu Ala Gly Arg Tyr Thr Met Asn Asp His Ile Val Asp
65 70 75 80
Cys Ser Asp Glu Gly Ala Val Phe Val Glu Ala Glu Ala Pro Asn Val
85 90 95
Glu Leu Thr Val Asp Gln Leu Val Lys Asn Met Glu Ala Gln Thr Ile
100 105 110
His Asp Phe Leu Pro Asp Gln Tyr Phe Pro Ala Asp Ala Pro Asn Pro
115 120 125
Leu Leu Ser Ile Gln Val Thr His Phe Pro Cys Gly Gly Leu Ala Ile
130 135 140
Gly Ile Val Val Ser His Ala Val Phe Asp Gly Phe Ser Leu Gly Val
145 150 155 160
Phe Leu Ala Ala Trp Ser Lys Ala Thr Met Asn Pro Glu Arg Lys Ile
165 170 175
Glu Ile Thr Pro Ser Phe Asp Leu Pro Ser Leu Leu Pro Tyr Lys Asp
180 185 190
Glu Ser Phe Gly Leu Asn Phe Ser Glu Ile Val Lys Ala Glu Asn Ile
195 200 205
Val Val Lys Arg Leu Asn Phe Gly Lys Glu Ala Ile Thr Arg Leu Arg
210 215 220
Ser Lys Leu Ser Pro Asn Gln Asn Gly Lys Thr Ile Ser Arg Val Arg
225 230 235 240
Val Val Cys Ala Val Ile Val Lys Ala Leu Met Gly Leu Glu Arg Ala
245 250 255
Lys Thr Arg Asp Phe Met Ile Cys Gln Gly Ile Asn Met Arg Glu Arg
260 265 270
Thr Lys Ala Pro Leu Gln Lys His Ala Cys Gly Asn Leu Ala Val Ser
275 280 285
Ser Tyr Thr Arg Arg Val Ala Ala Ala Glu Ala Glu Glu Leu Gln Ser
290 295 300
Leu Val Asn Leu Ile Gly Asp Ser Ile Glu Lys Ser Ile Ala Asp Tyr
305 310 315 320
Ala Asp Ile Leu Ser Ser Asp Gln Asp Gly Arg His Ile Ile Ser Thr
325 330 335
Met Met Lys Ser Phe Met Gln Phe Ala Ala Pro Asp Ile Lys Ala Ile
340 345 350
Ser Phe Thr Asp Trp Ser Lys Phe Gly Phe Tyr Gln Val Asp Phe Gly
355 360 365
Phe Gly Lys Pro Val Trp Thr Gly Val Arg Pro Glu Arg Pro Ile Phe
370 375 380
Ser Ala Ala Ile Leu Met Ser Asn Arg Glu Gly Asp Gly Ile Glu Ala
385 390 395 400
Trp Leu His Leu Asp Lys Asn Asp Met Leu Ile Phe Glu Gln Asp Glu
405 410 415
Glu Ile Lys Leu Leu Ile Thr
420
<210> 24
<211> 654
<212> DNA
<213> OAH94415.1,针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(654)
<400> 24
atg gat acc aag agg gtc ggt tat aca gtc gtt gat ttg agt cag tgg 48
Met Asp Thr Lys Arg Val Gly Tyr Thr Val Val Asp Leu Ser Gln Trp
1 5 10 15
ggc aga aaa gaa cac ttc gaa gca ttc cag agc ttc gcc cag tgc acc 96
Gly Arg Lys Glu His Phe Glu Ala Phe Gln Ser Phe Ala Gln Cys Thr
20 25 30
ttt tcc cag act gtt caa tta gac ata act tcc ctg ctg aag act gtc 144
Phe Ser Gln Thr Val Gln Leu Asp Ile Thr Ser Leu Leu Lys Thr Val
35 40 45
aag cag aac gga tat aaa ttc tac ccg acc ttt atc tac ata ata agc 192
Lys Gln Asn Gly Tyr Lys Phe Tyr Pro Thr Phe Ile Tyr Ile Ile Ser
50 55 60
cgt tta gtg aac aag cat gcc gaa ttc cgt atg gcc atg aag gat ggt 240
Arg Leu Val Asn Lys His Ala Glu Phe Arg Met Ala Met Lys Asp Gly
65 70 75 80
gaa ctg gta att tgg gac tcc gta aac cct ggt tac aca atc ttt cac 288
Glu Leu Val Ile Trp Asp Ser Val Asn Pro Gly Tyr Thr Ile Phe His
85 90 95
gaa cag acc gaa aca ttt tca tct ctg tgg agc tat tac cac aag gac 336
Glu Gln Thr Glu Thr Phe Ser Ser Leu Trp Ser Tyr Tyr His Lys Asp
100 105 110
atc aat cag ttt ctt aaa acg tat tct gaa gac atc gct cag tac ggg 384
Ile Asn Gln Phe Leu Lys Thr Tyr Ser Glu Asp Ile Ala Gln Tyr Gly
115 120 125
gac gat ctg gcg tac ttc ccc aag gag ttt att gaa aat atg ttc ttc 432
Asp Asp Leu Ala Tyr Phe Pro Lys Glu Phe Ile Glu Asn Met Phe Phe
130 135 140
gtg tca gcg aac cca tgg gtg agc ttc aca agt ttc aac tta aac gtg 480
Val Ser Ala Asn Pro Trp Val Ser Phe Thr Ser Phe Asn Leu Asn Val
145 150 155 160
gcc aat att aat aac ttc ttc gca ccg gtc ttt aca atc ggt aaa tac 528
Ala Asn Ile Asn Asn Phe Phe Ala Pro Val Phe Thr Ile Gly Lys Tyr
165 170 175
tac acg cag ggc gac aag gtg ttg atg ccg tta gct att cag gtc cat 576
Tyr Thr Gln Gly Asp Lys Val Leu Met Pro Leu Ala Ile Gln Val His
180 185 190
cat gca gtc tgc gac ggg ttt cat gtc ggc agg tta ttg aac gaa att 624
His Ala Val Cys Asp Gly Phe His Val Gly Arg Leu Leu Asn Glu Ile
195 200 205
cag cag tac tgc gat gag gga tgc aag taa 654
Gln Gln Tyr Cys Asp Glu Gly Cys Lys
210 215
<210> 25
<211> 217
<212> PRT
<213> OAH94415.1,针对其在酿酒酵母中的表达经密码子优化
<400> 25
Met Asp Thr Lys Arg Val Gly Tyr Thr Val Val Asp Leu Ser Gln Trp
1 5 10 15
Gly Arg Lys Glu His Phe Glu Ala Phe Gln Ser Phe Ala Gln Cys Thr
20 25 30
Phe Ser Gln Thr Val Gln Leu Asp Ile Thr Ser Leu Leu Lys Thr Val
35 40 45
Lys Gln Asn Gly Tyr Lys Phe Tyr Pro Thr Phe Ile Tyr Ile Ile Ser
50 55 60
Arg Leu Val Asn Lys His Ala Glu Phe Arg Met Ala Met Lys Asp Gly
65 70 75 80
Glu Leu Val Ile Trp Asp Ser Val Asn Pro Gly Tyr Thr Ile Phe His
85 90 95
Glu Gln Thr Glu Thr Phe Ser Ser Leu Trp Ser Tyr Tyr His Lys Asp
100 105 110
Ile Asn Gln Phe Leu Lys Thr Tyr Ser Glu Asp Ile Ala Gln Tyr Gly
115 120 125
Asp Asp Leu Ala Tyr Phe Pro Lys Glu Phe Ile Glu Asn Met Phe Phe
130 135 140
Val Ser Ala Asn Pro Trp Val Ser Phe Thr Ser Phe Asn Leu Asn Val
145 150 155 160
Ala Asn Ile Asn Asn Phe Phe Ala Pro Val Phe Thr Ile Gly Lys Tyr
165 170 175
Tyr Thr Gln Gly Asp Lys Val Leu Met Pro Leu Ala Ile Gln Val His
180 185 190
His Ala Val Cys Asp Gly Phe His Val Gly Arg Leu Leu Asn Glu Ile
195 200 205
Gln Gln Tyr Cys Asp Glu Gly Cys Lys
210 215
<210> 26
<211> 1755
<212> DNA
<213> DfHAD_香鳞毛蕨(Dryopteris fragrans)
<220>
<221> CDS
<222> (1)..(1755)
<400> 26
atg gag ttc tct gcc tct gct cct cct cct agg cta gcc agt gtc ata 48
Met Glu Phe Ser Ala Ser Ala Pro Pro Pro Arg Leu Ala Ser Val Ile
1 5 10 15
ata ttg gag cct ctc ggc ttc ctc ctc aca cca cac tac tcc tct cag 96
Ile Leu Glu Pro Leu Gly Phe Leu Leu Thr Pro His Tyr Ser Ser Gln
20 25 30
ctt ccc aaa aag ctg ctc cgt cgc ctg ttg tgc act aga atc tgg cac 144
Leu Pro Lys Lys Leu Leu Arg Arg Leu Leu Cys Thr Arg Ile Trp His
35 40 45
agg tat cag cga ggc cgc ctt cgc ctg cgt gac gct gct atg ctg ctc 192
Arg Tyr Gln Arg Gly Arg Leu Arg Leu Arg Asp Ala Ala Met Leu Leu
50 55 60
gcc cag ctc cca ttc cta gct gtg tct gat cac ccc tgg gct ctg gac 240
Ala Gln Leu Pro Phe Leu Ala Val Ser Asp His Pro Trp Ala Leu Asp
65 70 75 80
aat ctc gca agc ctg ctc cgc ccc aca gct gtg cgt gcg gtg cca tgg 288
Asn Leu Ala Ser Leu Leu Arg Pro Thr Ala Val Arg Ala Val Pro Trp
85 90 95
atg ctg ctg ctg ctc gac ttc cta cga gac gag ctc cat ctg aag gta 336
Met Leu Leu Leu Leu Asp Phe Leu Arg Asp Glu Leu His Leu Lys Val
100 105 110
gtc tgc gcg acc aac tcc tcc cca gaa gag ctg caa gag ctg cgc cac 384
Val Cys Ala Thr Asn Ser Ser Pro Glu Glu Leu Gln Glu Leu Arg His
115 120 125
cag ttt ccg gcc ctc ttt gcc aag gtc gat gcc acc gtt tct tca ggc 432
Gln Phe Pro Ala Leu Phe Ala Lys Val Asp Ala Thr Val Ser Ser Gly
130 135 140
gag gag ggc gtg ggc aag ccg tcc gtg cgc ttc ctg cag gct gcg ttg 480
Glu Glu Gly Val Gly Lys Pro Ser Val Arg Phe Leu Gln Ala Ala Leu
145 150 155 160
gac aaa gcc ggt gtc cac gcg cag caa acc ttg tat ctt gac tct ttt 528
Asp Lys Ala Gly Val His Ala Gln Gln Thr Leu Tyr Leu Asp Ser Phe
165 170 175
gac agc ttg gag acc atc atg gct gca cgc tct ctt ggc atg cat gca 576
Asp Ser Leu Glu Thr Ile Met Ala Ala Arg Ser Leu Gly Met His Ala
180 185 190
cta tct gta gag cca tgc cac att gat gag ctc acc gcc agg gcc tct 624
Leu Ser Val Glu Pro Cys His Ile Asp Glu Leu Thr Ala Arg Ala Ser
195 200 205
tcc ggc cag cta aga gat gca cag ctt ata agg cgt att gtg tgc gcc 672
Ser Gly Gln Leu Arg Asp Ala Gln Leu Ile Arg Arg Ile Val Cys Ala
210 215 220
atg cac ggg cca gca gta tct gca gtt gtg tcg ggc agt atc aca tcg 720
Met His Gly Pro Ala Val Ser Ala Val Val Ser Gly Ser Ile Thr Ser
225 230 235 240
tcc ggc cca cag aca gca aag atc gag gaa ttg cca aca gct gct gat 768
Ser Gly Pro Gln Thr Ala Lys Ile Glu Glu Leu Pro Thr Ala Ala Asp
245 250 255
agt cat ctc cgc agc gca gct ctc act tct gct cag cag ttt ttc ctc 816
Ser His Leu Arg Ser Ala Ala Leu Thr Ser Ala Gln Gln Phe Phe Leu
260 265 270
aaa gtt att gct cca cat cgt cct gag aag cca ttc gtc cag ctt cca 864
Lys Val Ile Ala Pro His Arg Pro Glu Lys Pro Phe Val Gln Leu Pro
275 280 285
tct ctc acc tcg gag ggc atc cga ata tac gac acc ttt gca cag ttt 912
Ser Leu Thr Ser Glu Gly Ile Arg Ile Tyr Asp Thr Phe Ala Gln Phe
290 295 300
gtc ata gcc gac ctg ctc gac gac acc cgc ttc cta ccc atg caa tct 960
Val Ile Ala Asp Leu Leu Asp Asp Thr Arg Phe Leu Pro Met Gln Ser
305 310 315 320
cct cct ccc aat ggg ctc atc acc ttt gtt aac cca agc gcg tac ctt 1008
Pro Pro Pro Asn Gly Leu Ile Thr Phe Val Asn Pro Ser Ala Tyr Leu
325 330 335
gct gat gat ata aag aat ggc aac agc cat att gtc ccg ggt gtg caa 1056
Ala Asp Asp Ile Lys Asn Gly Asn Ser His Ile Val Pro Gly Val Gln
340 345 350
ttt tac gca tcc gat gcg tgc act ctc atc gac atc cca cat gac cta 1104
Phe Tyr Ala Ser Asp Ala Cys Thr Leu Ile Asp Ile Pro His Asp Leu
355 360 365
gac acc acc tcc gtt ggc ttg tca gta ctg cac aag ttt gga aag gtg 1152
Asp Thr Thr Ser Val Gly Leu Ser Val Leu His Lys Phe Gly Lys Val
370 375 380
gac aag gac aca ctc aac aaa gtg cta gac aga atg ctc gag caa gtg 1200
Asp Lys Asp Thr Leu Asn Lys Val Leu Asp Arg Met Leu Glu Gln Val
385 390 395 400
agt gaa gac gac ggc att ctg cag gtg tat ttt gat gtg gag cgt ccg 1248
Ser Glu Asp Asp Gly Ile Leu Gln Val Tyr Phe Asp Val Glu Arg Pro
405 410 415
cgc atc gat cca gtt gtg gtg gca aac acg gtg ttt ctg ttc cac ttg 1296
Arg Ile Asp Pro Val Val Val Ala Asn Thr Val Phe Leu Phe His Leu
420 425 430
gga aag aga ggg cat gag gtg gcg agg agt gag aag ttt gtg gag agt 1344
Gly Lys Arg Gly His Glu Val Ala Arg Ser Glu Lys Phe Val Glu Ser
435 440 445
gtg ctg ctg cag agg gca tac gaa gaa ggg acg ttg tat tac aac ctg 1392
Val Leu Leu Gln Arg Ala Tyr Glu Glu Gly Thr Leu Tyr Tyr Asn Leu
450 455 460
ggg gaa gca ttt ttg gtg agt gtg gcg agg ctg gtg cac gag ttt aag 1440
Gly Glu Ala Phe Leu Val Ser Val Ala Arg Leu Val His Glu Phe Lys
465 470 475 480
gag cac ttt aca agg agc ggc atg agg agg gca ctg gag gag agg cta 1488
Glu His Phe Thr Arg Ser Gly Met Arg Arg Ala Leu Glu Glu Arg Leu
485 490 495
aga gag cgg gca agg gcg ggc atg caa gag agg gat gat gcg ctg gcg 1536
Arg Glu Arg Ala Arg Ala Gly Met Gln Glu Arg Asp Asp Ala Leu Ala
500 505 510
cta gcc atg cgc att cgt gca tgc gct ttg tgt ggc ctg gcc gga gag 1584
Leu Ala Met Arg Ile Arg Ala Cys Ala Leu Cys Gly Leu Ala Gly Glu
515 520 525
ggc ctc aca aaa gca gca gag cag gag ctt ttg cgc ctg cag tgc aag 1632
Gly Leu Thr Lys Ala Ala Glu Gln Glu Leu Leu Arg Leu Gln Cys Lys
530 535 540
tcc aag ggc tgt tgg ggg tgc cac cct ttc tat cgc aat ggc agt aat 1680
Ser Lys Gly Cys Trp Gly Cys His Pro Phe Tyr Arg Asn Gly Ser Asn
545 550 555 560
gtg ctc agc tgg atc ggc agt gag gcc ctt acc act gct tac gct att 1728
Val Leu Ser Trp Ile Gly Ser Glu Ala Leu Thr Thr Ala Tyr Ala Ile
565 570 575
gct gcg cta cag ccc att gat att taa 1755
Ala Ala Leu Gln Pro Ile Asp Ile
580
<210> 27
<211> 584
<212> PRT
<213> DfHAD_香鳞毛蕨(Dryopteris fragrans)
<400> 27
Met Glu Phe Ser Ala Ser Ala Pro Pro Pro Arg Leu Ala Ser Val Ile
1 5 10 15
Ile Leu Glu Pro Leu Gly Phe Leu Leu Thr Pro His Tyr Ser Ser Gln
20 25 30
Leu Pro Lys Lys Leu Leu Arg Arg Leu Leu Cys Thr Arg Ile Trp His
35 40 45
Arg Tyr Gln Arg Gly Arg Leu Arg Leu Arg Asp Ala Ala Met Leu Leu
50 55 60
Ala Gln Leu Pro Phe Leu Ala Val Ser Asp His Pro Trp Ala Leu Asp
65 70 75 80
Asn Leu Ala Ser Leu Leu Arg Pro Thr Ala Val Arg Ala Val Pro Trp
85 90 95
Met Leu Leu Leu Leu Asp Phe Leu Arg Asp Glu Leu His Leu Lys Val
100 105 110
Val Cys Ala Thr Asn Ser Ser Pro Glu Glu Leu Gln Glu Leu Arg His
115 120 125
Gln Phe Pro Ala Leu Phe Ala Lys Val Asp Ala Thr Val Ser Ser Gly
130 135 140
Glu Glu Gly Val Gly Lys Pro Ser Val Arg Phe Leu Gln Ala Ala Leu
145 150 155 160
Asp Lys Ala Gly Val His Ala Gln Gln Thr Leu Tyr Leu Asp Ser Phe
165 170 175
Asp Ser Leu Glu Thr Ile Met Ala Ala Arg Ser Leu Gly Met His Ala
180 185 190
Leu Ser Val Glu Pro Cys His Ile Asp Glu Leu Thr Ala Arg Ala Ser
195 200 205
Ser Gly Gln Leu Arg Asp Ala Gln Leu Ile Arg Arg Ile Val Cys Ala
210 215 220
Met His Gly Pro Ala Val Ser Ala Val Val Ser Gly Ser Ile Thr Ser
225 230 235 240
Ser Gly Pro Gln Thr Ala Lys Ile Glu Glu Leu Pro Thr Ala Ala Asp
245 250 255
Ser His Leu Arg Ser Ala Ala Leu Thr Ser Ala Gln Gln Phe Phe Leu
260 265 270
Lys Val Ile Ala Pro His Arg Pro Glu Lys Pro Phe Val Gln Leu Pro
275 280 285
Ser Leu Thr Ser Glu Gly Ile Arg Ile Tyr Asp Thr Phe Ala Gln Phe
290 295 300
Val Ile Ala Asp Leu Leu Asp Asp Thr Arg Phe Leu Pro Met Gln Ser
305 310 315 320
Pro Pro Pro Asn Gly Leu Ile Thr Phe Val Asn Pro Ser Ala Tyr Leu
325 330 335
Ala Asp Asp Ile Lys Asn Gly Asn Ser His Ile Val Pro Gly Val Gln
340 345 350
Phe Tyr Ala Ser Asp Ala Cys Thr Leu Ile Asp Ile Pro His Asp Leu
355 360 365
Asp Thr Thr Ser Val Gly Leu Ser Val Leu His Lys Phe Gly Lys Val
370 375 380
Asp Lys Asp Thr Leu Asn Lys Val Leu Asp Arg Met Leu Glu Gln Val
385 390 395 400
Ser Glu Asp Asp Gly Ile Leu Gln Val Tyr Phe Asp Val Glu Arg Pro
405 410 415
Arg Ile Asp Pro Val Val Val Ala Asn Thr Val Phe Leu Phe His Leu
420 425 430
Gly Lys Arg Gly His Glu Val Ala Arg Ser Glu Lys Phe Val Glu Ser
435 440 445
Val Leu Leu Gln Arg Ala Tyr Glu Glu Gly Thr Leu Tyr Tyr Asn Leu
450 455 460
Gly Glu Ala Phe Leu Val Ser Val Ala Arg Leu Val His Glu Phe Lys
465 470 475 480
Glu His Phe Thr Arg Ser Gly Met Arg Arg Ala Leu Glu Glu Arg Leu
485 490 495
Arg Glu Arg Ala Arg Ala Gly Met Gln Glu Arg Asp Asp Ala Leu Ala
500 505 510
Leu Ala Met Arg Ile Arg Ala Cys Ala Leu Cys Gly Leu Ala Gly Glu
515 520 525
Gly Leu Thr Lys Ala Ala Glu Gln Glu Leu Leu Arg Leu Gln Cys Lys
530 535 540
Ser Lys Gly Cys Trp Gly Cys His Pro Phe Tyr Arg Asn Gly Ser Asn
545 550 555 560
Val Leu Ser Trp Ile Gly Ser Glu Ala Leu Thr Thr Ala Tyr Ala Ile
565 570 575
Ala Ala Leu Gln Pro Ile Asp Ile
580
<210> 28
<211> 1755
<212> DNA
<213> DfHAD-8(K532R)_香鳞毛蕨(Dryopteris fragrans)
<220>
<221> CDS
<222> (1)..(1755)
<400> 28
atg gag ttc tct gcc tct gct cct cct cct agg cta gcc agt gtc ata 48
Met Glu Phe Ser Ala Ser Ala Pro Pro Pro Arg Leu Ala Ser Val Ile
1 5 10 15
ata ttg gag cct ctc ggc ttc ctc ctc aca cca cac tac tcc tct cag 96
Ile Leu Glu Pro Leu Gly Phe Leu Leu Thr Pro His Tyr Ser Ser Gln
20 25 30
ctt ccc aaa aag ctg ctc cgt cgc ctg ttg tgc act aga atc tgg cac 144
Leu Pro Lys Lys Leu Leu Arg Arg Leu Leu Cys Thr Arg Ile Trp His
35 40 45
agg tat cag cga ggc cgc ctt cgc ctg cgt gac gct gct atg ctg ctc 192
Arg Tyr Gln Arg Gly Arg Leu Arg Leu Arg Asp Ala Ala Met Leu Leu
50 55 60
gcc cag ctc cca ttc cta gct gtg tct gat cac ccc tgg gct ctg gac 240
Ala Gln Leu Pro Phe Leu Ala Val Ser Asp His Pro Trp Ala Leu Asp
65 70 75 80
aat ctc gca agc ctg ctc cgc ccc aca gct gtg cgt gcg gtg cca tgg 288
Asn Leu Ala Ser Leu Leu Arg Pro Thr Ala Val Arg Ala Val Pro Trp
85 90 95
atg ctg ctg ctg ctc gac ttc cta cga gac gag ctc cat ctg aag gta 336
Met Leu Leu Leu Leu Asp Phe Leu Arg Asp Glu Leu His Leu Lys Val
100 105 110
gtc tgc gcg acc aac tcc tcc cca gaa gag ctg caa gag ctg cgc cac 384
Val Cys Ala Thr Asn Ser Ser Pro Glu Glu Leu Gln Glu Leu Arg His
115 120 125
cag ttt ccg gcc ctc ttt gcc aag gtc gat gcc acc gtt tct tca ggc 432
Gln Phe Pro Ala Leu Phe Ala Lys Val Asp Ala Thr Val Ser Ser Gly
130 135 140
gag gag ggc gtg ggc aag ccg tcc gtg cgc ttc ctg cag gct gcg ttg 480
Glu Glu Gly Val Gly Lys Pro Ser Val Arg Phe Leu Gln Ala Ala Leu
145 150 155 160
gac aaa gcc ggt gtc cac gcg cag caa acc ttg tat ctt gac tct ttt 528
Asp Lys Ala Gly Val His Ala Gln Gln Thr Leu Tyr Leu Asp Ser Phe
165 170 175
gac agc ttg gag acc atc atg gct gca cgc tct ctt ggc atg cat gca 576
Asp Ser Leu Glu Thr Ile Met Ala Ala Arg Ser Leu Gly Met His Ala
180 185 190
cta tct gta gag cca tgc cac att gat gag ctc acc gcc agg gcc tct 624
Leu Ser Val Glu Pro Cys His Ile Asp Glu Leu Thr Ala Arg Ala Ser
195 200 205
tcc ggc cag cta aga gat gca cag ctt ata agg cgt att gtg tgc gcc 672
Ser Gly Gln Leu Arg Asp Ala Gln Leu Ile Arg Arg Ile Val Cys Ala
210 215 220
atg cac ggg cca gca gta tct gca gtt gtg tcg ggc agt atc aca tcg 720
Met His Gly Pro Ala Val Ser Ala Val Val Ser Gly Ser Ile Thr Ser
225 230 235 240
tcc ggc cca cag aca gca aag atc gag gaa ttg cca aca gct gct gat 768
Ser Gly Pro Gln Thr Ala Lys Ile Glu Glu Leu Pro Thr Ala Ala Asp
245 250 255
agt cat ctc cgc agc gca gct ctc act tct gct cag cag ttt ttc ctc 816
Ser His Leu Arg Ser Ala Ala Leu Thr Ser Ala Gln Gln Phe Phe Leu
260 265 270
aaa gtt att gct cca cat cgt cct gag aag cca ttc gtc cag ctt cca 864
Lys Val Ile Ala Pro His Arg Pro Glu Lys Pro Phe Val Gln Leu Pro
275 280 285
tct ctc acc tcg gag ggc atc cga ata tac gac acc ttt gca cag ttt 912
Ser Leu Thr Ser Glu Gly Ile Arg Ile Tyr Asp Thr Phe Ala Gln Phe
290 295 300
gtc ata gcc gac ctg ctc gac gac acc cgc ttc cta ccc atg caa tct 960
Val Ile Ala Asp Leu Leu Asp Asp Thr Arg Phe Leu Pro Met Gln Ser
305 310 315 320
cct cct ccc aat ggg ctc atc acc ttt gtt aac cca agc gcg tac ctt 1008
Pro Pro Pro Asn Gly Leu Ile Thr Phe Val Asn Pro Ser Ala Tyr Leu
325 330 335
gct gat gat ata aag aat ggc aac agc cat att gtc ccg ggt gtg caa 1056
Ala Asp Asp Ile Lys Asn Gly Asn Ser His Ile Val Pro Gly Val Gln
340 345 350
ttt tac gca tcc gat gcg tgc act ctc atc gac atc cca cat gac cta 1104
Phe Tyr Ala Ser Asp Ala Cys Thr Leu Ile Asp Ile Pro His Asp Leu
355 360 365
gac acc acc tcc gtt ggc ttg tca gta ctg cac aag ttt gga aag gtg 1152
Asp Thr Thr Ser Val Gly Leu Ser Val Leu His Lys Phe Gly Lys Val
370 375 380
gac aag gac aca ctc aac aaa gtg cta gac aga atg ctc gag caa gtg 1200
Asp Lys Asp Thr Leu Asn Lys Val Leu Asp Arg Met Leu Glu Gln Val
385 390 395 400
agt gaa gac gac ggc att ctg cag gtg tat ttt gat gtg gag cgt ccg 1248
Ser Glu Asp Asp Gly Ile Leu Gln Val Tyr Phe Asp Val Glu Arg Pro
405 410 415
cgc atc gat cca gtt gtg gtg gca aac acg gtg ttt ctg ttc cac ttg 1296
Arg Ile Asp Pro Val Val Val Ala Asn Thr Val Phe Leu Phe His Leu
420 425 430
gga aag aga ggg cat gag gtg gcg agg agt gag aag ttt gtg gag agt 1344
Gly Lys Arg Gly His Glu Val Ala Arg Ser Glu Lys Phe Val Glu Ser
435 440 445
gtg ctg ctg cag agg gca tac gaa gaa ggg acg ttg tat tac aac ctg 1392
Val Leu Leu Gln Arg Ala Tyr Glu Glu Gly Thr Leu Tyr Tyr Asn Leu
450 455 460
ggg gaa gca ttt ttg gtg agt gtg gcg agg ctg gtg cac gag ttt aag 1440
Gly Glu Ala Phe Leu Val Ser Val Ala Arg Leu Val His Glu Phe Lys
465 470 475 480
gag cac ttt aca agg agc ggc atg agg agg gca ctg gag gag agg cta 1488
Glu His Phe Thr Arg Ser Gly Met Arg Arg Ala Leu Glu Glu Arg Leu
485 490 495
aga gag cgg gca agg gcg ggc atg caa gag agg gat gat gcg ctg gcg 1536
Arg Glu Arg Ala Arg Ala Gly Met Gln Glu Arg Asp Asp Ala Leu Ala
500 505 510
ctg gcc atg cgc att cgt gca tgc gct ttg tgt ggc ctg gcc gga gag 1584
Leu Ala Met Arg Ile Arg Ala Cys Ala Leu Cys Gly Leu Ala Gly Glu
515 520 525
ggc ctc aca aga gca gca gag cag gag cta ctg cgc ctg cag tgc aag 1632
Gly Leu Thr Arg Ala Ala Glu Gln Glu Leu Leu Arg Leu Gln Cys Lys
530 535 540
tcc aag ggc tgt tgg ggg tgc cac cct ttc tat cgc aat ggc agt aat 1680
Ser Lys Gly Cys Trp Gly Cys His Pro Phe Tyr Arg Asn Gly Ser Asn
545 550 555 560
gtg ctc agc tgg atc ggc agt gag gcc ctt acc act gct tac gct att 1728
Val Leu Ser Trp Ile Gly Ser Glu Ala Leu Thr Thr Ala Tyr Ala Ile
565 570 575
gct gcg cta cag ccc att gat att taa 1755
Ala Ala Leu Gln Pro Ile Asp Ile
580
<210> 29
<211> 584
<212> PRT
<213> DfHAD-8(K532R)_香鳞毛蕨(Dryopteris fragrans)
<400> 29
Met Glu Phe Ser Ala Ser Ala Pro Pro Pro Arg Leu Ala Ser Val Ile
1 5 10 15
Ile Leu Glu Pro Leu Gly Phe Leu Leu Thr Pro His Tyr Ser Ser Gln
20 25 30
Leu Pro Lys Lys Leu Leu Arg Arg Leu Leu Cys Thr Arg Ile Trp His
35 40 45
Arg Tyr Gln Arg Gly Arg Leu Arg Leu Arg Asp Ala Ala Met Leu Leu
50 55 60
Ala Gln Leu Pro Phe Leu Ala Val Ser Asp His Pro Trp Ala Leu Asp
65 70 75 80
Asn Leu Ala Ser Leu Leu Arg Pro Thr Ala Val Arg Ala Val Pro Trp
85 90 95
Met Leu Leu Leu Leu Asp Phe Leu Arg Asp Glu Leu His Leu Lys Val
100 105 110
Val Cys Ala Thr Asn Ser Ser Pro Glu Glu Leu Gln Glu Leu Arg His
115 120 125
Gln Phe Pro Ala Leu Phe Ala Lys Val Asp Ala Thr Val Ser Ser Gly
130 135 140
Glu Glu Gly Val Gly Lys Pro Ser Val Arg Phe Leu Gln Ala Ala Leu
145 150 155 160
Asp Lys Ala Gly Val His Ala Gln Gln Thr Leu Tyr Leu Asp Ser Phe
165 170 175
Asp Ser Leu Glu Thr Ile Met Ala Ala Arg Ser Leu Gly Met His Ala
180 185 190
Leu Ser Val Glu Pro Cys His Ile Asp Glu Leu Thr Ala Arg Ala Ser
195 200 205
Ser Gly Gln Leu Arg Asp Ala Gln Leu Ile Arg Arg Ile Val Cys Ala
210 215 220
Met His Gly Pro Ala Val Ser Ala Val Val Ser Gly Ser Ile Thr Ser
225 230 235 240
Ser Gly Pro Gln Thr Ala Lys Ile Glu Glu Leu Pro Thr Ala Ala Asp
245 250 255
Ser His Leu Arg Ser Ala Ala Leu Thr Ser Ala Gln Gln Phe Phe Leu
260 265 270
Lys Val Ile Ala Pro His Arg Pro Glu Lys Pro Phe Val Gln Leu Pro
275 280 285
Ser Leu Thr Ser Glu Gly Ile Arg Ile Tyr Asp Thr Phe Ala Gln Phe
290 295 300
Val Ile Ala Asp Leu Leu Asp Asp Thr Arg Phe Leu Pro Met Gln Ser
305 310 315 320
Pro Pro Pro Asn Gly Leu Ile Thr Phe Val Asn Pro Ser Ala Tyr Leu
325 330 335
Ala Asp Asp Ile Lys Asn Gly Asn Ser His Ile Val Pro Gly Val Gln
340 345 350
Phe Tyr Ala Ser Asp Ala Cys Thr Leu Ile Asp Ile Pro His Asp Leu
355 360 365
Asp Thr Thr Ser Val Gly Leu Ser Val Leu His Lys Phe Gly Lys Val
370 375 380
Asp Lys Asp Thr Leu Asn Lys Val Leu Asp Arg Met Leu Glu Gln Val
385 390 395 400
Ser Glu Asp Asp Gly Ile Leu Gln Val Tyr Phe Asp Val Glu Arg Pro
405 410 415
Arg Ile Asp Pro Val Val Val Ala Asn Thr Val Phe Leu Phe His Leu
420 425 430
Gly Lys Arg Gly His Glu Val Ala Arg Ser Glu Lys Phe Val Glu Ser
435 440 445
Val Leu Leu Gln Arg Ala Tyr Glu Glu Gly Thr Leu Tyr Tyr Asn Leu
450 455 460
Gly Glu Ala Phe Leu Val Ser Val Ala Arg Leu Val His Glu Phe Lys
465 470 475 480
Glu His Phe Thr Arg Ser Gly Met Arg Arg Ala Leu Glu Glu Arg Leu
485 490 495
Arg Glu Arg Ala Arg Ala Gly Met Gln Glu Arg Asp Asp Ala Leu Ala
500 505 510
Leu Ala Met Arg Ile Arg Ala Cys Ala Leu Cys Gly Leu Ala Gly Glu
515 520 525
Gly Leu Thr Arg Ala Ala Glu Gln Glu Leu Leu Arg Leu Gln Cys Lys
530 535 540
Ser Lys Gly Cys Trp Gly Cys His Pro Phe Tyr Arg Asn Gly Ser Asn
545 550 555 560
Val Leu Ser Trp Ile Gly Ser Glu Ala Leu Thr Thr Ala Tyr Ala Ile
565 570 575
Ala Ala Leu Gln Pro Ile Asp Ile
580
<210> 30
<211> 1755
<212> DNA
<213> DfHAD-9(V274A)_香鳞毛蕨(Dryopteris fragrans)
<220>
<221> CDS
<222> (1)..(1755)
<400> 30
atg gag ttc tct gcc tct gct cct cct cct agg cta gcc agt gtc ata 48
Met Glu Phe Ser Ala Ser Ala Pro Pro Pro Arg Leu Ala Ser Val Ile
1 5 10 15
ata ttg gag cct ctc ggc ttc ctc ctc aca cca cac tac tcc tct cag 96
Ile Leu Glu Pro Leu Gly Phe Leu Leu Thr Pro His Tyr Ser Ser Gln
20 25 30
ctt ccc aaa aag ctg ctc cgt cgc ctg ttg tgc act aga atc tgg cac 144
Leu Pro Lys Lys Leu Leu Arg Arg Leu Leu Cys Thr Arg Ile Trp His
35 40 45
agg tat cag cga ggc cgc ctt cgc ctg cgt gac gct gct atg ctg ctc 192
Arg Tyr Gln Arg Gly Arg Leu Arg Leu Arg Asp Ala Ala Met Leu Leu
50 55 60
gcc cag ctc cca ttc cta gct gtg tct gat cac ccc tgg gct ctg gac 240
Ala Gln Leu Pro Phe Leu Ala Val Ser Asp His Pro Trp Ala Leu Asp
65 70 75 80
aat ctc gca agc ctg ctc cgc ccc aca gct gtg cgt gcg gtg cca tgg 288
Asn Leu Ala Ser Leu Leu Arg Pro Thr Ala Val Arg Ala Val Pro Trp
85 90 95
atg ctg ctg ctg ctc gac ttc cta cga gac gag ctc cat ctg aag gta 336
Met Leu Leu Leu Leu Asp Phe Leu Arg Asp Glu Leu His Leu Lys Val
100 105 110
gtc tgc gcg acc aac tcc tcc cca gaa gag ctg caa gag ctg cgc cac 384
Val Cys Ala Thr Asn Ser Ser Pro Glu Glu Leu Gln Glu Leu Arg His
115 120 125
cag ttt ccg gcc ctc ttt gcc aag gtc gat gcc acc gtt tct tca ggc 432
Gln Phe Pro Ala Leu Phe Ala Lys Val Asp Ala Thr Val Ser Ser Gly
130 135 140
gag gag ggc gtg ggc aag ccg tcc gtg cgc ttc ctg cag gct gcg ttg 480
Glu Glu Gly Val Gly Lys Pro Ser Val Arg Phe Leu Gln Ala Ala Leu
145 150 155 160
gac aaa gcc ggt gtc cac gcg cag caa acc ttg tat ctt gac tct ttt 528
Asp Lys Ala Gly Val His Ala Gln Gln Thr Leu Tyr Leu Asp Ser Phe
165 170 175
gac agc ttg gag acc atc atg gct gca cgc tct ctt ggc atg cat gca 576
Asp Ser Leu Glu Thr Ile Met Ala Ala Arg Ser Leu Gly Met His Ala
180 185 190
cta tct gta gag cca tgc cac att gat gag ctc acc gcc agg gcc tct 624
Leu Ser Val Glu Pro Cys His Ile Asp Glu Leu Thr Ala Arg Ala Ser
195 200 205
tcc ggc cag cta aga gat gca cag ctt ata agg cgt att gtg tgc gcc 672
Ser Gly Gln Leu Arg Asp Ala Gln Leu Ile Arg Arg Ile Val Cys Ala
210 215 220
atg cac ggg cca gca gta tct gca gtt gtg tcg ggc agt atc aca tcg 720
Met His Gly Pro Ala Val Ser Ala Val Val Ser Gly Ser Ile Thr Ser
225 230 235 240
tcc ggc cca cag aca gca aag atc gag gaa ttg cca aca gct gct gat 768
Ser Gly Pro Gln Thr Ala Lys Ile Glu Glu Leu Pro Thr Ala Ala Asp
245 250 255
agt cat ctc cgc agc gca gct ctc act tct gct cag cag ttt ttc ctc 816
Ser His Leu Arg Ser Ala Ala Leu Thr Ser Ala Gln Gln Phe Phe Leu
260 265 270
aaa gct att gct cca cat cgt cct gag aag cca ttc gtc cag ctt cca 864
Lys Ala Ile Ala Pro His Arg Pro Glu Lys Pro Phe Val Gln Leu Pro
275 280 285
tct ctc acc tcg gag ggc atc cga ata tac gac acc ttt gca cag ttt 912
Ser Leu Thr Ser Glu Gly Ile Arg Ile Tyr Asp Thr Phe Ala Gln Phe
290 295 300
gtc ata gcc gac ctg ctc gac gac acc cgc ttc cta ccc atg caa tct 960
Val Ile Ala Asp Leu Leu Asp Asp Thr Arg Phe Leu Pro Met Gln Ser
305 310 315 320
cct cct ccc aat ggg ctc atc acc ttt gtt aac cca agc gcg tac ctt 1008
Pro Pro Pro Asn Gly Leu Ile Thr Phe Val Asn Pro Ser Ala Tyr Leu
325 330 335
gct gat gat ata aag aat ggc aac agc cat att gtc ccg ggt gtg caa 1056
Ala Asp Asp Ile Lys Asn Gly Asn Ser His Ile Val Pro Gly Val Gln
340 345 350
ttt tac gca tct gat gcg tgc act ctc atc gac atc cca cat gac cta 1104
Phe Tyr Ala Ser Asp Ala Cys Thr Leu Ile Asp Ile Pro His Asp Leu
355 360 365
gac acc acc tcc gtt ggc ttg tca gta ctg cac aag ttt gga aag gtg 1152
Asp Thr Thr Ser Val Gly Leu Ser Val Leu His Lys Phe Gly Lys Val
370 375 380
gac aag gac aca ctc aac aaa gtg cta gac aga atg ctg gag caa gtg 1200
Asp Lys Asp Thr Leu Asn Lys Val Leu Asp Arg Met Leu Glu Gln Val
385 390 395 400
agt gaa gac gac ggc att ctc cag gtg tat ttt gat gtg gag cgt ccg 1248
Ser Glu Asp Asp Gly Ile Leu Gln Val Tyr Phe Asp Val Glu Arg Pro
405 410 415
cgc atc gat cca gtt gtg gtg gca aac acg gtg ttt ctg ttc cac ttg 1296
Arg Ile Asp Pro Val Val Val Ala Asn Thr Val Phe Leu Phe His Leu
420 425 430
gga aag aga ggg cat gag gtg gcg agg agt gag aag ttt gtg gag agt 1344
Gly Lys Arg Gly His Glu Val Ala Arg Ser Glu Lys Phe Val Glu Ser
435 440 445
gtg ctg ctg cag agg gca tac gaa gaa ggg acg ttg tat tac aac ctg 1392
Val Leu Leu Gln Arg Ala Tyr Glu Glu Gly Thr Leu Tyr Tyr Asn Leu
450 455 460
ggg gaa gca ttt ttg gtg agt gtg gcg agg ctg gtg cac gag ttt aag 1440
Gly Glu Ala Phe Leu Val Ser Val Ala Arg Leu Val His Glu Phe Lys
465 470 475 480
gag cac ttt aca agg agc ggc atg agg agg gca ctg gag gag agg cta 1488
Glu His Phe Thr Arg Ser Gly Met Arg Arg Ala Leu Glu Glu Arg Leu
485 490 495
aga gag cgg gca agg gcg ggc atg caa gag agg gat gat gcg ctg gcg 1536
Arg Glu Arg Ala Arg Ala Gly Met Gln Glu Arg Asp Asp Ala Leu Ala
500 505 510
ctg gcc atg cgc att cgt gca tgc gct ttg tgt ggc ctg gcc gga gag 1584
Leu Ala Met Arg Ile Arg Ala Cys Ala Leu Cys Gly Leu Ala Gly Glu
515 520 525
ggc ctc aca aaa gca gca gag cag gag cta ctg cgc ctg cag tgc aag 1632
Gly Leu Thr Lys Ala Ala Glu Gln Glu Leu Leu Arg Leu Gln Cys Lys
530 535 540
tcc aag ggc tgt tgg ggg tgc cac cct ttc tat cgc aat ggc agt aat 1680
Ser Lys Gly Cys Trp Gly Cys His Pro Phe Tyr Arg Asn Gly Ser Asn
545 550 555 560
gtg ctc agc tgg atc ggc agt gag gcc ctt acc act gct tac gct att 1728
Val Leu Ser Trp Ile Gly Ser Glu Ala Leu Thr Thr Ala Tyr Ala Ile
565 570 575
gct gcg cta cag ccc att gat att taa 1755
Ala Ala Leu Gln Pro Ile Asp Ile
580
<210> 31
<211> 584
<212> PRT
<213> DfHAD-9(V274A)_香鳞毛蕨(Dryopteris fragrans)
<400> 31
Met Glu Phe Ser Ala Ser Ala Pro Pro Pro Arg Leu Ala Ser Val Ile
1 5 10 15
Ile Leu Glu Pro Leu Gly Phe Leu Leu Thr Pro His Tyr Ser Ser Gln
20 25 30
Leu Pro Lys Lys Leu Leu Arg Arg Leu Leu Cys Thr Arg Ile Trp His
35 40 45
Arg Tyr Gln Arg Gly Arg Leu Arg Leu Arg Asp Ala Ala Met Leu Leu
50 55 60
Ala Gln Leu Pro Phe Leu Ala Val Ser Asp His Pro Trp Ala Leu Asp
65 70 75 80
Asn Leu Ala Ser Leu Leu Arg Pro Thr Ala Val Arg Ala Val Pro Trp
85 90 95
Met Leu Leu Leu Leu Asp Phe Leu Arg Asp Glu Leu His Leu Lys Val
100 105 110
Val Cys Ala Thr Asn Ser Ser Pro Glu Glu Leu Gln Glu Leu Arg His
115 120 125
Gln Phe Pro Ala Leu Phe Ala Lys Val Asp Ala Thr Val Ser Ser Gly
130 135 140
Glu Glu Gly Val Gly Lys Pro Ser Val Arg Phe Leu Gln Ala Ala Leu
145 150 155 160
Asp Lys Ala Gly Val His Ala Gln Gln Thr Leu Tyr Leu Asp Ser Phe
165 170 175
Asp Ser Leu Glu Thr Ile Met Ala Ala Arg Ser Leu Gly Met His Ala
180 185 190
Leu Ser Val Glu Pro Cys His Ile Asp Glu Leu Thr Ala Arg Ala Ser
195 200 205
Ser Gly Gln Leu Arg Asp Ala Gln Leu Ile Arg Arg Ile Val Cys Ala
210 215 220
Met His Gly Pro Ala Val Ser Ala Val Val Ser Gly Ser Ile Thr Ser
225 230 235 240
Ser Gly Pro Gln Thr Ala Lys Ile Glu Glu Leu Pro Thr Ala Ala Asp
245 250 255
Ser His Leu Arg Ser Ala Ala Leu Thr Ser Ala Gln Gln Phe Phe Leu
260 265 270
Lys Ala Ile Ala Pro His Arg Pro Glu Lys Pro Phe Val Gln Leu Pro
275 280 285
Ser Leu Thr Ser Glu Gly Ile Arg Ile Tyr Asp Thr Phe Ala Gln Phe
290 295 300
Val Ile Ala Asp Leu Leu Asp Asp Thr Arg Phe Leu Pro Met Gln Ser
305 310 315 320
Pro Pro Pro Asn Gly Leu Ile Thr Phe Val Asn Pro Ser Ala Tyr Leu
325 330 335
Ala Asp Asp Ile Lys Asn Gly Asn Ser His Ile Val Pro Gly Val Gln
340 345 350
Phe Tyr Ala Ser Asp Ala Cys Thr Leu Ile Asp Ile Pro His Asp Leu
355 360 365
Asp Thr Thr Ser Val Gly Leu Ser Val Leu His Lys Phe Gly Lys Val
370 375 380
Asp Lys Asp Thr Leu Asn Lys Val Leu Asp Arg Met Leu Glu Gln Val
385 390 395 400
Ser Glu Asp Asp Gly Ile Leu Gln Val Tyr Phe Asp Val Glu Arg Pro
405 410 415
Arg Ile Asp Pro Val Val Val Ala Asn Thr Val Phe Leu Phe His Leu
420 425 430
Gly Lys Arg Gly His Glu Val Ala Arg Ser Glu Lys Phe Val Glu Ser
435 440 445
Val Leu Leu Gln Arg Ala Tyr Glu Glu Gly Thr Leu Tyr Tyr Asn Leu
450 455 460
Gly Glu Ala Phe Leu Val Ser Val Ala Arg Leu Val His Glu Phe Lys
465 470 475 480
Glu His Phe Thr Arg Ser Gly Met Arg Arg Ala Leu Glu Glu Arg Leu
485 490 495
Arg Glu Arg Ala Arg Ala Gly Met Gln Glu Arg Asp Asp Ala Leu Ala
500 505 510
Leu Ala Met Arg Ile Arg Ala Cys Ala Leu Cys Gly Leu Ala Gly Glu
515 520 525
Gly Leu Thr Lys Ala Ala Glu Gln Glu Leu Leu Arg Leu Gln Cys Lys
530 535 540
Ser Lys Gly Cys Trp Gly Cys His Pro Phe Tyr Arg Asn Gly Ser Asn
545 550 555 560
Val Leu Ser Trp Ile Gly Ser Glu Ala Leu Thr Thr Ala Tyr Ala Ile
565 570 575
Ala Ala Leu Gln Pro Ile Asp Ile
580
<210> 32
<211> 1755
<212> DNA
<213> 通过大肠杆菌Genscript遗传密码子频率对DfHAD进行密码子优化的序列
<400> 32
atggagttca gcgcgagcgc tccgccgccg cgtctggcga gcgtgatcat tctggaaccg 60
ctgggttttc tgctgacccc gcactacagc agccagctgc cgaagaaact gctgcgtcgt 120
ctgctgtgca cccgtatctg gcaccgttat cagcgtggcc gtctgcgtct gcgtgacgcg 180
gcgatgctgc tggcgcaact gccgttcctg gcggttagcg accacccgtg ggcgctggat 240
aacctggcga gcctgctgcg tccgaccgcg gttcgtgcgg tgccgtggat gctgctgctg 300
ctggactttc tgcgtgatga gctgcacctg aaagtggttt gcgcgaccaa cagcagcccg 360
gaggaactgc aggaactgcg tcaccaattc ccggcgctgt ttgcgaaggt tgacgcgacc 420
gtgagcagcg gcgaggaagg tgttggcaaa ccgagcgtgc gtttcctgca agcggcgctg 480
gataaggcgg gcgtgcacgc gcagcaaacc ctgtacctgg acagctttga tagcctggag 540
accatcatgg cggcgcgtag cctgggtatg cacgcgctga gcgttgagcc gtgccacatt 600
gacgaactga ccgcgcgtgc gagcagcggt cagctgcgtg atgcgcaact gatccgtcgt 660
attgtttgcg cgatgcacgg tccggctgtg agcgcggtgg ttagcggtag catcaccagc 720
agcggtccgc agaccgcgaa aattgaggaa ctgccgaccg cggcggacag ccacctgcgt 780
agcgcggcgc tgaccagcgc gcagcaattc tttctgaaag tgattgcgcc gcaccgtccg 840
gagaagccgt tcgttcaact gccgagcctg accagcgaag gtatccgtat ttatgacacc 900
ttcgcgcagt ttgtgatcgc ggatctgctg gacgataccc gtttcctgcc gatgcaaagc 960
ccgccgccga acggcctgat tacctttgtt aacccgagcg cgtacctggc ggacgatatc 1020
aaaaacggta acagccacat tgttccgggc gtgcagttct atgcgagcga cgcgtgcacc 1080
ctgatcgata ttccgcacga cctggatacc accagcgttg gtctgagcgt gctgcacaag 1140
tttggcaaag ttgacaagga taccctgaac aaggtgctgg atcgtatgct ggagcaagtt 1200
agcgaagacg atggtatcct gcaagtttac tttgacgtgg agcgtccgcg tattgatccg 1260
gtggttgtgg cgaacaccgt gttcctgttt cacctgggta aacgtggcca cgaagttgcg 1320
cgtagcgaga agttcgttga aagcgtgctg ctgcagcgtg cgtacgagga aggcaccctg 1380
tactataacc tgggcgaagc gtttctggtt agcgtggcgc gtctggtgca cgagttcaaa 1440
gaacacttta cccgtagcgg tatgcgtcgt gcgctggagg aacgtctgcg tgagcgtgcg 1500
cgtgcgggta tgcaagaacg tgacgatgcg ctggcgctgg cgatgcgtat ccgtgcgtgc 1560
gcgctgtgcg gtctggcggg cgagggtctg accaaggcgg cggagcagga actgctgcgt 1620
ctgcaatgca agagcaaagg ttgctggggc tgccacccgt tctaccgtaa cggtagcaac 1680
gttctgagct ggatcggcag cgaagcgctg accaccgcgt atgcgattgc ggcgctgcag 1740
ccgatcgaca tttaa 1755
<210> 33
<211> 1755
<212> DNA
<213> 通过烟草Genscript遗传密码子频率对DfHAD进行密码子优化的序列
<400> 33
atggaatttt ctgcttcagc tccacctcca agacttgctt cagttattat tcttgagcct 60
ttgggatttc ttttgactcc acattactct tcacaattgc ctaagaaact tttgagaagg 120
cttttgtgta caagaatttg gcataggtac caaaggggta ggcttagatt gagggatgct 180
gctatgcttt tggctcaact tccatttttg gctgtttcag atcatccttg ggctcttgat 240
aatttggctt ctcttttgag accaactgct gttagggctg ttccttggat gcttttgctt 300
ttggattttc ttagagatga acttcatttg aaggttgttt gcgctactaa ttcttcacca 360
gaagagcttc aagagttgag gcatcaattt cctgctttgt ttgctaaggt tgatgctaca 420
gtttcttcag gagaagaggg agttggtaaa ccatctgtta gatttcttca agctgctttg 480
gataaggctg gtgttcatgc tcaacaaact ctttatttgg attctttcga ttcacttgaa 540
acaattatgg ctgctaggtc attgggaatg catgctcttt ctgttgaacc atgtcatatt 600
gatgagttga ctgctagagc ttcttcagga caattgaggg atgctcaact tattagaagg 660
attgtttgcg ctatgcatgg tcctgctgtt tcagctgttg tttctggatc aattacttct 720
tcaggtccac aaacagctaa aattgaagag cttcctactg ctgctgattc tcatttgaga 780
tcagctgctc ttacatctgc tcaacaattt ttccttaaag ttattgctcc acatagacct 840
gaaaagccat ttgttcaact tccttctttg acttcagagg gaatcaggat ctatgataca 900
ttcgctcaat tcgttatcgc tgatcttttg gatgatacta ggtttttgcc aatgcaatca 960
cctccaccta atggtcttat cacattcgtt aacccttctg cttatttggc tgatgatatt 1020
aaaaatggta actcacatat tgttccaggt gttcaatttt acgcttctga tgcttgtact 1080
ttgattgata ttcctcatga tcttgatact acatctgttg gactttcagt tttgcataag 1140
ttcggtaaag ttgataagga tacacttaat aaggttttgg atagaatgct tgaacaagtt 1200
tcagaggatg atggaatcct tcaagtttac ttcgatgttg aaagacctag gattgatcca 1260
gttgttgttg ctaacactgt ttttcttttc catttgggaa aaagaggtca tgaggttgct 1320
agatcagaaa agtttgttga gtctgttctt ttgcaaagag cttacgaaga gggaactttg 1380
tattacaatc ttggtgaagc ttttcttgtt tctgttgcta gacttgttca tgagtttaag 1440
gagcatttta caaggtctgg aatgagaagg gctttggaag agagacttag ggaaagagct 1500
agggctggta tgcaagagag agatgatgct cttgctttgg ctatgagaat tagggcttgt 1560
gctctttgcg gtttggctgg agaaggtctt acaaaggctg ctgaacaaga gcttttgaga 1620
ttgcaatgca agtctaaagg atgttggggt tgccatccat tctacaggaa tggttctaac 1680
gttttgtcat ggattggttc tgaggctctt actacagctt acgctattgc tgctcttcaa 1740
cctattgata tttga 1755
<210> 34
<211> 1755
<212> DNA
<213> 通过大肠杆菌Genscript遗传密码子频率对DfHAD-8(K532R)进行密码子优化的序列
<400> 34
atggagttca gcgcgagcgc tccgccgccg cgtctggcga gcgtgatcat tctggaaccg 60
ctgggttttc tgctgacccc gcactacagc agccagctgc cgaagaaact gctgcgtcgt 120
ctgctgtgca cccgtatctg gcaccgttat cagcgtggcc gtctgcgtct gcgtgacgcg 180
gcgatgctgc tggcgcaact gccgttcctg gcggttagcg accacccgtg ggcgctggat 240
aacctggcga gcctgctgcg tccgaccgcg gttcgtgcgg tgccgtggat gctgctgctg 300
ctggactttc tgcgtgatga gctgcacctg aaagtggttt gcgcgaccaa cagcagcccg 360
gaggaactgc aggaactgcg tcaccaattc ccggcgctgt ttgcgaaggt tgacgcgacc 420
gtgagcagcg gcgaggaagg tgttggcaaa ccgagcgtgc gtttcctgca agcggcgctg 480
gataaggcgg gcgtgcacgc gcagcaaacc ctgtacctgg acagctttga tagcctggag 540
accatcatgg cggcgcgtag cctgggtatg cacgcgctga gcgttgagcc gtgccacatt 600
gacgaactga ccgcgcgtgc gagcagcggt cagctgcgtg atgcgcaact gatccgtcgt 660
attgtttgcg cgatgcacgg tccggctgtg agcgcggtgg ttagcggtag catcaccagc 720
agcggtccgc agaccgcgaa aattgaggaa ctgccgaccg cggcggacag ccacctgcgt 780
agcgcggcgc tgaccagcgc gcagcaattc tttctgaaag tgattgcgcc gcaccgtccg 840
gagaagccgt tcgttcaact gccgagcctg accagcgaag gtatccgtat ttatgacacc 900
ttcgcgcagt ttgtgatcgc ggatctgctg gacgataccc gtttcctgcc gatgcaaagc 960
ccgccgccga acggcctgat tacctttgtt aacccgagcg cgtacctggc ggacgatatc 1020
aaaaacggta acagccacat tgttccgggc gtgcagttct atgcgagcga cgcgtgcacc 1080
ctgatcgata ttccgcacga cctggatacc accagcgttg gtctgagcgt gctgcacaag 1140
tttggcaaag ttgacaagga taccctgaac aaggtgctgg atcgtatgct ggagcaagtt 1200
agcgaagacg atggtatcct gcaagtttac tttgacgtgg agcgtccgcg tattgatccg 1260
gtggttgtgg cgaacaccgt gttcctgttt cacctgggta aacgtggcca cgaagttgcg 1320
cgtagcgaga agttcgttga aagcgtgctg ctgcagcgtg cgtacgagga aggcaccctg 1380
tactataacc tgggcgaagc gtttctggtt agcgtggcgc gtctggtgca cgagttcaaa 1440
gaacacttta cccgtagcgg tatgcgtcgt gcgctggagg aacgtctgcg tgagcgtgcg 1500
cgtgcgggta tgcaagaacg tgacgatgcg ctggcgctgg cgatgcgtat ccgtgcgtgc 1560
gcgctgtgcg gtctggcggg cgagggtctg acccgggcgg cggagcagga actgctgcgt 1620
ctgcaatgca agagcaaagg ttgctggggc tgccacccgt tctaccgtaa cggtagcaac 1680
gttctgagct ggatcggcag cgaagcgctg accaccgcgt atgcgattgc ggcgctgcag 1740
ccgatcgaca tttaa 1755
<210> 35
<211> 1755
<212> DNA
<213> 通过大肠杆菌Genscript遗传密码子频率对DfHAD-9(V274A)进行密码子优化的序列
<400> 35
atggagttca gcgcgagcgc tccgccgccg cgtctggcga gcgtgatcat tctggaaccg 60
ctgggttttc tgctgacccc gcactacagc agccagctgc cgaagaaact gctgcgtcgt 120
ctgctgtgca cccgtatctg gcaccgttat cagcgtggcc gtctgcgtct gcgtgacgcg 180
gcgatgctgc tggcgcaact gccgttcctg gcggttagcg accacccgtg ggcgctggat 240
aacctggcga gcctgctgcg tccgaccgcg gttcgtgcgg tgccgtggat gctgctgctg 300
ctggactttc tgcgtgatga gctgcacctg aaagtggttt gcgcgaccaa cagcagcccg 360
gaggaactgc aggaactgcg tcaccaattc ccggcgctgt ttgcgaaggt tgacgcgacc 420
gtgagcagcg gcgaggaagg tgttggcaaa ccgagcgtgc gtttcctgca agcggcgctg 480
gataaggcgg gcgtgcacgc gcagcaaacc ctgtacctgg acagctttga tagcctggag 540
accatcatgg cggcgcgtag cctgggtatg cacgcgctga gcgttgagcc gtgccacatt 600
gacgaactga ccgcgcgtgc gagcagcggt cagctgcgtg atgcgcaact gatccgtcgt 660
attgtttgcg cgatgcacgg tccggctgtg agcgcggtgg ttagcggtag catcaccagc 720
agcggtccgc agaccgcgaa aattgaggaa ctgccgaccg cggcggacag ccacctgcgt 780
agcgcggcgc tgaccagcgc gcagcaattc tttctgaaag cgattgcgcc gcaccgtccg 840
gagaagccgt tcgttcaact gccgagcctg accagcgaag gtatccgtat ttatgacacc 900
ttcgcgcagt ttgtgatcgc ggatctgctg gacgataccc gtttcctgcc gatgcaaagc 960
ccgccgccga acggcctgat tacctttgtt aacccgagcg cgtacctggc ggacgatatc 1020
aaaaacggta acagccacat tgttccgggc gtgcagttct atgcgagcga cgcgtgcacc 1080
ctgatcgata ttccgcacga cctggatacc accagcgttg gtctgagcgt gctgcacaag 1140
tttggcaaag ttgacaagga taccctgaac aaggtgctgg atcgtatgct ggagcaagtt 1200
agcgaagacg atggtatcct gcaagtttac tttgacgtgg agcgtccgcg tattgatccg 1260
gtggttgtgg cgaacaccgt gttcctgttt cacctgggta aacgtggcca cgaagttgcg 1320
cgtagcgaga agttcgttga aagcgtgctg ctgcagcgtg cgtacgagga aggcaccctg 1380
tactataacc tgggcgaagc gtttctggtt agcgtggcgc gtctggtgca cgagttcaaa 1440
gaacacttta cccgtagcgg tatgcgtcgt gcgctggagg aacgtctgcg tgagcgtgcg 1500
cgtgcgggta tgcaagaacg tgacgatgcg ctggcgctgg cgatgcgtat ccgtgcgtgc 1560
gcgctgtgcg gtctggcggg cgagggtctg accaaggcgg cggagcagga actgctgcgt 1620
ctgcaatgca agagcaaagg ttgctggggc tgccacccgt tctaccgtaa cggtagcaac 1680
gttctgagct ggatcggcag cgaagcgctg accaccgcgt atgcgattgc ggcgctgcag 1740
ccgatcgaca tttaa 1755
<210> 36
<211> 2496
<212> DNA
<213> 通过大肠杆菌Genscript遗传密码子频率对DfHAD-6His-GST进行密码子优化的序列
<220>
<221> CDS
<222> (1)..(2496)
<400> 36
atg tct ggt tct cat cat cat cat cat cat agc agc ggt atg tcc cct 48
Met Ser Gly Ser His His His His His His Ser Ser Gly Met Ser Pro
1 5 10 15
ata cta ggt tat tgg aaa att aag ggc ctt gtg caa ccc act cga ctt 96
Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu Val Gln Pro Thr Arg Leu
20 25 30
ctt ttg gaa tat ctt gaa gaa aaa tat gaa gag cat ttg tat gag cgc 144
Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg
35 40 45
gat gaa ggt gat aaa tgg cga aac aaa aag ttt gaa ttg ggt ttg gag 192
Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu Gly Leu Glu
50 55 60
ttt ccc aat ctt cct tat tat att gat ggt gat gtt aaa tta aca cag 240
Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly Asp Val Lys Leu Thr Gln
65 70 75 80
tct atg gcc atc ata cgt tat ata gct gac aag cac aac atg ttg ggt 288
Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp Lys His Asn Met Leu Gly
85 90 95
ggt tgt cca aaa gag cgt gca gag att tca atg ctt gaa gga gcg gtt 336
Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser Met Leu Glu Gly Ala Val
100 105 110
ttg gat att aga tac ggt gtt tcg aga att gca tat agt aaa gac ttt 384
Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr Ser Lys Asp Phe
115 120 125
gaa act ctc aaa gtt gat ttt ctt agc aag cta cct gaa atg ctg aaa 432
Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu Met Leu Lys
130 135 140
atg ttc gaa gat cgt tta tgt cat aaa aca tat tta aat ggt gat cat 480
Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn Gly Asp His
145 150 155 160
gta acc cat cct gac ttc atg ttg tat gac gct ctt gat gtt gtt tta 528
Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp Val Val Leu
165 170 175
tac atg gac cca atg tgc ctg gat gcg ttc cca aaa tta gtt tgt ttt 576
Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu Val Cys Phe
180 185 190
aaa aaa cgt att gaa gct atc cca caa att gat aag tac ttg aaa tcc 624
Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile Asp Lys Tyr Leu Lys Ser
195 200 205
agc aag tat ata gca tgg cct ttg cag ggc tgg caa gcc acg ttt ggt 672
Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly Trp Gln Ala Thr Phe Gly
210 215 220
ggt ggc gac cat cct cca aaa tcg gat ctg ggc cac aca ggc cat aga 720
Gly Gly Asp His Pro Pro Lys Ser Asp Leu Gly His Thr Gly His Arg
225 230 235 240
tct gac gac gac gac aag cat atg gag ttc agc gcg agc gct ccg ccg 768
Ser Asp Asp Asp Asp Lys His Met Glu Phe Ser Ala Ser Ala Pro Pro
245 250 255
ccg cgt ctg gcg agc gtg atc att ctg gaa ccg ctg ggt ttt ctg ctg 816
Pro Arg Leu Ala Ser Val Ile Ile Leu Glu Pro Leu Gly Phe Leu Leu
260 265 270
acc ccg cac tac agc agc cag ctg ccg aag aaa ctg ctg cgt cgt ctg 864
Thr Pro His Tyr Ser Ser Gln Leu Pro Lys Lys Leu Leu Arg Arg Leu
275 280 285
ctg tgc acc cgt atc tgg cac cgt tat cag cgt ggc cgt ctg cgt ctg 912
Leu Cys Thr Arg Ile Trp His Arg Tyr Gln Arg Gly Arg Leu Arg Leu
290 295 300
cgt gac gcg gcg atg ctg ctg gcg caa ctg ccg ttc ctg gcg gtt agc 960
Arg Asp Ala Ala Met Leu Leu Ala Gln Leu Pro Phe Leu Ala Val Ser
305 310 315 320
gac cac ccg tgg gcg ctg gat aac ctg gcg agc ctg ctg cgt ccg acc 1008
Asp His Pro Trp Ala Leu Asp Asn Leu Ala Ser Leu Leu Arg Pro Thr
325 330 335
gcg gtt cgt gcg gtg ccg tgg atg ctg ctg ctg ctg gac ttt ctg cgt 1056
Ala Val Arg Ala Val Pro Trp Met Leu Leu Leu Leu Asp Phe Leu Arg
340 345 350
gat gag ctg cac ctg aaa gtg gtt tgc gcg acc aac agc agc ccg gag 1104
Asp Glu Leu His Leu Lys Val Val Cys Ala Thr Asn Ser Ser Pro Glu
355 360 365
gaa ctg cag gaa ctg cgt cac caa ttc ccg gcg ctg ttt gcg aag gtt 1152
Glu Leu Gln Glu Leu Arg His Gln Phe Pro Ala Leu Phe Ala Lys Val
370 375 380
gac gcg acc gtg agc agc ggc gag gaa ggt gtt ggc aaa ccg agc gtg 1200
Asp Ala Thr Val Ser Ser Gly Glu Glu Gly Val Gly Lys Pro Ser Val
385 390 395 400
cgt ttc ctg caa gcg gcg ctg gat aag gcg ggc gtg cac gcg cag caa 1248
Arg Phe Leu Gln Ala Ala Leu Asp Lys Ala Gly Val His Ala Gln Gln
405 410 415
acc ctg tac ctg gac agc ttt gat agc ctg gag acc atc atg gcg gcg 1296
Thr Leu Tyr Leu Asp Ser Phe Asp Ser Leu Glu Thr Ile Met Ala Ala
420 425 430
cgt agc ctg ggt atg cac gcg ctg agc gtt gag ccg tgc cac att gac 1344
Arg Ser Leu Gly Met His Ala Leu Ser Val Glu Pro Cys His Ile Asp
435 440 445
gaa ctg acc gcg cgt gcg agc agc ggt cag ctg cgt gat gcg caa ctg 1392
Glu Leu Thr Ala Arg Ala Ser Ser Gly Gln Leu Arg Asp Ala Gln Leu
450 455 460
atc cgt cgt att gtt tgc gcg atg cac ggt ccg gct gtg agc gcg gtg 1440
Ile Arg Arg Ile Val Cys Ala Met His Gly Pro Ala Val Ser Ala Val
465 470 475 480
gtt agc ggt agc atc acc agc agc ggt ccg cag acc gcg aaa att gag 1488
Val Ser Gly Ser Ile Thr Ser Ser Gly Pro Gln Thr Ala Lys Ile Glu
485 490 495
gaa ctg ccg acc gcg gcg gac agc cac ctg cgt agc gcg gcg ctg acc 1536
Glu Leu Pro Thr Ala Ala Asp Ser His Leu Arg Ser Ala Ala Leu Thr
500 505 510
agc gcg cag caa ttc ttt ctg aaa gtg att gcg ccg cac cgt ccg gag 1584
Ser Ala Gln Gln Phe Phe Leu Lys Val Ile Ala Pro His Arg Pro Glu
515 520 525
aag ccg ttc gtt caa ctg ccg agc ctg acc agc gaa ggt atc cgt att 1632
Lys Pro Phe Val Gln Leu Pro Ser Leu Thr Ser Glu Gly Ile Arg Ile
530 535 540
tat gac acc ttc gcg cag ttt gtg atc gcg gat ctg ctg gac gat acc 1680
Tyr Asp Thr Phe Ala Gln Phe Val Ile Ala Asp Leu Leu Asp Asp Thr
545 550 555 560
cgt ttc ctg ccg atg caa agc ccg ccg ccg aac ggc ctg att acc ttt 1728
Arg Phe Leu Pro Met Gln Ser Pro Pro Pro Asn Gly Leu Ile Thr Phe
565 570 575
gtt aac ccg agc gcg tac ctg gcg gac gat atc aaa aac ggt aac agc 1776
Val Asn Pro Ser Ala Tyr Leu Ala Asp Asp Ile Lys Asn Gly Asn Ser
580 585 590
cac att gtt ccg ggc gtg cag ttc tat gcg agc gac gcg tgc acc ctg 1824
His Ile Val Pro Gly Val Gln Phe Tyr Ala Ser Asp Ala Cys Thr Leu
595 600 605
atc gat att ccg cac gac ctg gat acc acc agc gtt ggt ctg agc gtg 1872
Ile Asp Ile Pro His Asp Leu Asp Thr Thr Ser Val Gly Leu Ser Val
610 615 620
ctg cac aag ttt ggc aaa gtt gac aag gat acc ctg aac aag gtg ctg 1920
Leu His Lys Phe Gly Lys Val Asp Lys Asp Thr Leu Asn Lys Val Leu
625 630 635 640
gat cgt atg ctg gag caa gtt agc gaa gac gat ggt atc ctg caa gtt 1968
Asp Arg Met Leu Glu Gln Val Ser Glu Asp Asp Gly Ile Leu Gln Val
645 650 655
tac ttt gac gtg gag cgt ccg cgt att gat ccg gtg gtt gtg gcg aac 2016
Tyr Phe Asp Val Glu Arg Pro Arg Ile Asp Pro Val Val Val Ala Asn
660 665 670
acc gtg ttc ctg ttt cac ctg ggt aaa cgt ggc cac gaa gtt gcg cgt 2064
Thr Val Phe Leu Phe His Leu Gly Lys Arg Gly His Glu Val Ala Arg
675 680 685
agc gag aag ttc gtt gaa agc gtg ctg ctg cag cgt gcg tac gag gaa 2112
Ser Glu Lys Phe Val Glu Ser Val Leu Leu Gln Arg Ala Tyr Glu Glu
690 695 700
ggc acc ctg tac tat aac ctg ggc gaa gcg ttt ctg gtt agc gtg gcg 2160
Gly Thr Leu Tyr Tyr Asn Leu Gly Glu Ala Phe Leu Val Ser Val Ala
705 710 715 720
cgt ctg gtg cac gag ttc aaa gaa cac ttt acc cgt agc ggt atg cgt 2208
Arg Leu Val His Glu Phe Lys Glu His Phe Thr Arg Ser Gly Met Arg
725 730 735
cgt gcg ctg gag gaa cgt ctg cgt gag cgt gcg cgt gcg ggt atg caa 2256
Arg Ala Leu Glu Glu Arg Leu Arg Glu Arg Ala Arg Ala Gly Met Gln
740 745 750
gaa cgt gac gat gcg ctg gcg ctg gcg atg cgt atc cgt gcg tgc gcg 2304
Glu Arg Asp Asp Ala Leu Ala Leu Ala Met Arg Ile Arg Ala Cys Ala
755 760 765
ctg tgc ggt ctg gcg ggc gag ggt ctg acc aag gcg gcg gag cag gaa 2352
Leu Cys Gly Leu Ala Gly Glu Gly Leu Thr Lys Ala Ala Glu Gln Glu
770 775 780
ctg ctg cgt ctg caa tgc aag agc aaa ggt tgc tgg ggc tgc cac ccg 2400
Leu Leu Arg Leu Gln Cys Lys Ser Lys Gly Cys Trp Gly Cys His Pro
785 790 795 800
ttc tac cgt aac ggt agc aac gtt ctg agc tgg atc ggc agc gaa gcg 2448
Phe Tyr Arg Asn Gly Ser Asn Val Leu Ser Trp Ile Gly Ser Glu Ala
805 810 815
ctg acc acc gcg tat gcg att gcg gcg ctg cag ccg atc gac att taa 2496
Leu Thr Thr Ala Tyr Ala Ile Ala Ala Leu Gln Pro Ile Asp Ile
820 825 830
<210> 37
<211> 831
<212> PRT
<213> 通过大肠杆菌Genscript遗传密码子频率对DfHAD-6His-GST进行密码子优化的序列
<400> 37
Met Ser Gly Ser His His His His His His Ser Ser Gly Met Ser Pro
1 5 10 15
Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu Val Gln Pro Thr Arg Leu
20 25 30
Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg
35 40 45
Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu Gly Leu Glu
50 55 60
Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly Asp Val Lys Leu Thr Gln
65 70 75 80
Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp Lys His Asn Met Leu Gly
85 90 95
Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser Met Leu Glu Gly Ala Val
100 105 110
Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr Ser Lys Asp Phe
115 120 125
Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu Met Leu Lys
130 135 140
Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn Gly Asp His
145 150 155 160
Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp Val Val Leu
165 170 175
Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu Val Cys Phe
180 185 190
Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile Asp Lys Tyr Leu Lys Ser
195 200 205
Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly Trp Gln Ala Thr Phe Gly
210 215 220
Gly Gly Asp His Pro Pro Lys Ser Asp Leu Gly His Thr Gly His Arg
225 230 235 240
Ser Asp Asp Asp Asp Lys His Met Glu Phe Ser Ala Ser Ala Pro Pro
245 250 255
Pro Arg Leu Ala Ser Val Ile Ile Leu Glu Pro Leu Gly Phe Leu Leu
260 265 270
Thr Pro His Tyr Ser Ser Gln Leu Pro Lys Lys Leu Leu Arg Arg Leu
275 280 285
Leu Cys Thr Arg Ile Trp His Arg Tyr Gln Arg Gly Arg Leu Arg Leu
290 295 300
Arg Asp Ala Ala Met Leu Leu Ala Gln Leu Pro Phe Leu Ala Val Ser
305 310 315 320
Asp His Pro Trp Ala Leu Asp Asn Leu Ala Ser Leu Leu Arg Pro Thr
325 330 335
Ala Val Arg Ala Val Pro Trp Met Leu Leu Leu Leu Asp Phe Leu Arg
340 345 350
Asp Glu Leu His Leu Lys Val Val Cys Ala Thr Asn Ser Ser Pro Glu
355 360 365
Glu Leu Gln Glu Leu Arg His Gln Phe Pro Ala Leu Phe Ala Lys Val
370 375 380
Asp Ala Thr Val Ser Ser Gly Glu Glu Gly Val Gly Lys Pro Ser Val
385 390 395 400
Arg Phe Leu Gln Ala Ala Leu Asp Lys Ala Gly Val His Ala Gln Gln
405 410 415
Thr Leu Tyr Leu Asp Ser Phe Asp Ser Leu Glu Thr Ile Met Ala Ala
420 425 430
Arg Ser Leu Gly Met His Ala Leu Ser Val Glu Pro Cys His Ile Asp
435 440 445
Glu Leu Thr Ala Arg Ala Ser Ser Gly Gln Leu Arg Asp Ala Gln Leu
450 455 460
Ile Arg Arg Ile Val Cys Ala Met His Gly Pro Ala Val Ser Ala Val
465 470 475 480
Val Ser Gly Ser Ile Thr Ser Ser Gly Pro Gln Thr Ala Lys Ile Glu
485 490 495
Glu Leu Pro Thr Ala Ala Asp Ser His Leu Arg Ser Ala Ala Leu Thr
500 505 510
Ser Ala Gln Gln Phe Phe Leu Lys Val Ile Ala Pro His Arg Pro Glu
515 520 525
Lys Pro Phe Val Gln Leu Pro Ser Leu Thr Ser Glu Gly Ile Arg Ile
530 535 540
Tyr Asp Thr Phe Ala Gln Phe Val Ile Ala Asp Leu Leu Asp Asp Thr
545 550 555 560
Arg Phe Leu Pro Met Gln Ser Pro Pro Pro Asn Gly Leu Ile Thr Phe
565 570 575
Val Asn Pro Ser Ala Tyr Leu Ala Asp Asp Ile Lys Asn Gly Asn Ser
580 585 590
His Ile Val Pro Gly Val Gln Phe Tyr Ala Ser Asp Ala Cys Thr Leu
595 600 605
Ile Asp Ile Pro His Asp Leu Asp Thr Thr Ser Val Gly Leu Ser Val
610 615 620
Leu His Lys Phe Gly Lys Val Asp Lys Asp Thr Leu Asn Lys Val Leu
625 630 635 640
Asp Arg Met Leu Glu Gln Val Ser Glu Asp Asp Gly Ile Leu Gln Val
645 650 655
Tyr Phe Asp Val Glu Arg Pro Arg Ile Asp Pro Val Val Val Ala Asn
660 665 670
Thr Val Phe Leu Phe His Leu Gly Lys Arg Gly His Glu Val Ala Arg
675 680 685
Ser Glu Lys Phe Val Glu Ser Val Leu Leu Gln Arg Ala Tyr Glu Glu
690 695 700
Gly Thr Leu Tyr Tyr Asn Leu Gly Glu Ala Phe Leu Val Ser Val Ala
705 710 715 720
Arg Leu Val His Glu Phe Lys Glu His Phe Thr Arg Ser Gly Met Arg
725 730 735
Arg Ala Leu Glu Glu Arg Leu Arg Glu Arg Ala Arg Ala Gly Met Gln
740 745 750
Glu Arg Asp Asp Ala Leu Ala Leu Ala Met Arg Ile Arg Ala Cys Ala
755 760 765
Leu Cys Gly Leu Ala Gly Glu Gly Leu Thr Lys Ala Ala Glu Gln Glu
770 775 780
Leu Leu Arg Leu Gln Cys Lys Ser Lys Gly Cys Trp Gly Cys His Pro
785 790 795 800
Phe Tyr Arg Asn Gly Ser Asn Val Leu Ser Trp Ile Gly Ser Glu Ala
805 810 815
Leu Thr Thr Ala Tyr Ala Ile Ala Ala Leu Gln Pro Ile Asp Ile
820 825 830
<210> 38
<211> 526
<212> PRT
<213> 隐孔菌(Cryptoporus volvatus)_CvTps1蛋白
<400> 38
Met Thr Thr Ile His Arg Arg His Thr Thr Leu Ile Leu Asp Leu Gly
1 5 10 15
Asp Val Leu Phe Arg Trp Ser Pro Lys Thr Glu Thr Ala Ile Pro Pro
20 25 30
Arg Gln Leu Lys Glu Ile Leu Thr Ser Val Thr Trp Phe Glu Tyr Glu
35 40 45
Arg Gly Gln Ile Ser Gln Thr Glu Cys Tyr Glu Arg Cys Ala Ala Glu
50 55 60
Phe Lys Val Asp Pro Leu Val Ile Ala Glu Ala Phe Lys Gln Ala Arg
65 70 75 80
Glu Ser Leu Arg Pro Asn Lys Ala Phe Ile Ala Leu Ile Arg Glu Leu
85 90 95
Arg His Gln Met His Gly Asp Leu Thr Val Leu Ala Leu Ser Asn Ile
100 105 110
Ser Leu Pro Asp Tyr Glu Tyr Ile Met Ser Leu Ser Ser Asp Trp Ala
115 120 125
Thr Val Phe Asn Arg Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys
130 135 140
Pro His Leu Gly Cys Tyr Arg Lys Val Ile Ser Glu Met Ser Leu Glu
145 150 155 160
Pro Gln Thr Thr Val Phe Val Asp Asp Lys Leu Asp Asn Val Ala Ser
165 170 175
Ala Arg Ser Leu Gly Met His Gly Ile Val Phe Asp Asn Glu Ala Asn
180 185 190
Val Phe Arg Gln Leu Arg Asn Ile Phe Gly Asn Pro Val Ser Arg Gly
195 200 205
Gln Gly Tyr Leu Arg Lys His Ala Gly Lys Leu Glu Ser Ser Thr Asp
210 215 220
Asn Gly Leu Thr Phe Glu Glu Asn Phe Thr Gln Leu Ile Ile Tyr Glu
225 230 235 240
Val Thr Gln Asp Arg Ser Leu Ile Thr Leu Ser Glu Cys Pro Arg Thr
245 250 255
Trp Asn Phe Phe Arg Gly Gln Pro Leu Phe Ser Glu Ser Phe Pro Asp
260 265 270
Asp Val Asp Thr Thr Ser Val Ala Leu Thr Val Leu Gln Pro Asp Arg
275 280 285
Ala Leu Val Asp Ser Ile Leu Asp Gln Met Leu Glu Tyr Val Asp Ala
290 295 300
Asp Gly Ile Met Gln Thr Tyr Phe Asp Ser Ser Arg Pro Arg Ile Asp
305 310 315 320
Pro Phe Val Cys Val Asn Val Leu Ser Leu Phe Tyr Ala Asn Gly Arg
325 330 335
Gly Arg Glu Leu Pro His Thr Leu Glu Trp Val Tyr Glu Val Leu Leu
340 345 350
His Arg Ala Tyr His Gly Gly Ser Arg Tyr Tyr Leu Ser Pro Asp Cys
355 360 365
Phe Leu Phe Phe Met Ser Arg Leu Leu Lys Arg Ala Asn Asp Ser Ala
370 375 380
Leu Gln Ala Arg Phe Arg Pro Leu Phe Met Glu Arg Val Lys Glu Arg
385 390 395 400
Val Gly Ala Ala Gly Asp Ser Met Asp Leu Ala Phe Arg Ile Leu Ala
405 410 415
Ala Ala Thr Ile Gly Val His Cys Pro Gln Asp Leu Glu Arg Leu Ala
420 425 430
Ala Ala Gln Cys Glu Asp Gly Gly Trp Asp Met Cys Trp Phe Tyr Ala
435 440 445
Phe Gly Ser Thr Gly Ile Lys Ala Gly Asn Arg Gly Leu Thr Thr Ala
450 455 460
Leu Ala Val Ala Ala Ile Arg Thr Ala Leu Gly Arg Pro Pro Ser Pro
465 470 475 480
Ser Pro Ser Asn Ile Ser Ser Ser Ser Lys Leu Asp Ala Pro Asn Ser
485 490 495
Phe Leu Gly Ile Pro Arg Pro Thr Ser Pro Ile Arg Phe Gly Glu Leu
500 505 510
Phe Arg Ser Trp Arg Lys Asn Lys Pro Thr Ala Lys Ser Gln
515 520 525
<210> 39
<211> 1956
<212> DNA
<213> 隐孔菌(Cryptoporus volvatus)_CvTps1转录物(包括非编码序列)
<400> 39
catcccgcct tttgagcatg gcacacaaac agcctttaag gagctccttg gttgcctagt 60
catgcctcca cctgccccct cctcactcat cccctcgcat cctaaaacat gaccacgatt 120
caccgtcggc acaccactct catcttggac ctcggcgacg tcctcttccg ctggtcacca 180
aagaccgaga ccgccatccc ccctcggcag cttaaggaga tacttacctc cgtcacctgg 240
ttcgagtacg aacgaggcca gatatcccaa acagaatgtt acgaacgatg cgctgcagaa 300
ttcaaagtcg accccttagt gatcgctgaa gccttcaagc aagctcgcga gtcattacgg 360
cccaacaaag cgttcatcgc cttgattcgc gaacttcgcc atcaaatgca tggagacctc 420
acggtcctcg ccctttccaa catttccctc cccgattacg aatatatcat gtctctgagc 480
tcggattggg caaccgtctt caatcgcgta ttcccttctg cacttgttgg cgagcgaaaa 540
ccccatctgg ggtgctaccg caaggtcatt tcggagatga gcttggaacc ccagacaacc 600
gtatttgtcg atgataagct agacaacgtc gcctctgctc gctcacttgg catgcacggc 660
atcgtattcg acaacgaagc caatgtcttc cggcaactgc gcaatatctt cgggaatccg 720
gttagccgcg gtcaaggcta tcttcgcaag catgccggaa agcttgagtc ttctaccgac 780
aatggcttga cctttgagga gaacttcacc cagctcatca tctacgaggt gacacaagac 840
aggagtctca tcacgctctc agaatgtccc cgtacctgga atttctttcg aggtcaaccg 900
ctcttctcgg agtctttccc ggatgatgtg gacacaacat ccgtggcatt gacagtacta 960
caacccgata gagcgctcgt tgattctatt ctagaccaaa tgcttgaata tgttgacgcc 1020
gacggcatca tgcagacata cttcgacagc tcgcgaccac gcatagaccc ttttgtttgc 1080
gtcaatgtgc tttctctgtt ctacgcaaac ggccggggtc gggagctccc tcacacactg 1140
gagtgggtct atgaagtact cctgcatcgc gcctaccatg gaggctcacg ttactaccta 1200
tcaccggact gctttttatt cttcatgagc cgcttgctca agcgcgccaa cgactcggcc 1260
ctccaggctc ggttccgccc actgttcatg gagagagtga aagaacgagt aggggcagcc 1320
ggagactcaa tggacctggc cttccgcatc ctcgccgcgg ctaccattgg cgtccattgc 1380
ccccaagatc tagaaagatt ggccgccgcg caatgcgagg acggtggatg ggacatgtgc 1440
tggttctacg cgttcgggtc gacaggtatc aaggcgggca accgcggcct caccacggcc 1500
cttgccgtcg cagctatacg aaccgccctc gggcgccccc cctctcccag cccctccaac 1560
atctcgtcgt cgtcgaagct cgacgctccc aacagcttct tgggcatccc gcgcccaacc 1620
agccccattc gctttggcga acttttccgt tcctggcgaa agaacaaacc gaccgcaaaa 1680
tctcaatgaa tctcaggttc tcgtgctctc gtgctatctt cgtacttatg ctactcgaca 1740
ttacccgtcg ctgtctacaa tgatacgggt actttgatga aactgtagat gtatttgtat 1800
catattgacc tccatccata gtcacctagc tactgttcgt gttatcacct gttgctgtta 1860
tatgatacaa gatgcccaaa cgagaatgta gaaatgttcc gtacacttgt gtacctgtga 1920
tgaagctaca taggccttca atcgatcact tggtcc 1956
<210> 40
<211> 1581
<212> DNA
<213> 隐孔菌(Cryptoporus volvatus)_CvTps1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 40
atgaccacga ttcaccgtcg gcacaccact ctcatcttgg acctcggcga cgtcctcttc 60
cgctggtcac caaagaccga gaccgccatc ccccctcggc agcttaagga gatacttacc 120
tccgtcacct ggttcgagta cgaacgaggc cagatatccc aaacagaatg ttacgaacga 180
tgcgctgcag aattcaaagt cgacccctta gtgatcgctg aagccttcaa gcaagctcgc 240
gagtcattac ggcccaacaa agcgttcatc gccttgattc gcgaacttcg ccatcaaatg 300
catggagacc tcacggtcct cgccctttcc aacatttccc tccccgatta cgaatatatc 360
atgtctctga gctcggattg ggcaaccgtc ttcaatcgcg tattcccttc tgcacttgtt 420
ggcgagcgaa aaccccatct ggggtgctac cgcaaggtca tttcggagat gagcttggaa 480
ccccagacaa ccgtatttgt cgatgataag ctagacaacg tcgcctctgc tcgctcactt 540
ggcatgcacg gcatcgtatt cgacaacgaa gccaatgtct tccggcaact gcgcaatatc 600
ttcgggaatc cggttagccg cggtcaaggc tatcttcgca agcatgccgg aaagcttgag 660
tcttctaccg acaatggctt gacctttgag gagaacttca cccagctcat catctacgag 720
gtgacacaag acaggagtct catcacgctc tcagaatgtc cccgtacctg gaatttcttt 780
cgaggtcaac cgctcttctc ggagtctttc ccggatgatg tggacacaac atccgtggca 840
ttgacagtac tacaacccga tagagcgctc gttgattcta ttctagacca aatgcttgaa 900
tatgttgacg ccgacggcat catgcagaca tacttcgaca gctcgcgacc acgcatagac 960
ccttttgttt gcgtcaatgt gctttctctg ttctacgcaa acggccgggg tcgggagctc 1020
cctcacacac tggagtgggt ctatgaagta ctcctgcatc gcgcctacca tggaggctca 1080
cgttactacc tatcaccgga ctgcttttta ttcttcatga gccgcttgct caagcgcgcc 1140
aacgactcgg ccctccaggc tcggttccgc ccactgttca tggagagagt gaaagaacga 1200
gtaggggcag ccggagactc aatggacctg gccttccgca tcctcgccgc ggctaccatt 1260
ggcgtccatt gcccccaaga tctagaaaga ttggccgccg cgcaatgcga ggacggtgga 1320
tgggacatgt gctggttcta cgcgttcggg tcgacaggta tcaaggcggg caaccgcggc 1380
ctcaccacgg cccttgccgt cgcagctata cgaaccgccc tcgggcgccc cccctctccc 1440
agcccctcca acatctcgtc gtcgtcgaag ctcgacgctc ccaacagctt cttgggcatc 1500
ccgcgcccaa ccagccccat tcgctttggc gaacttttcc gttcctggcg aaagaacaaa 1560
ccgaccgcaa aatctcaatg a 1581
<210> 41
<211> 1581
<212> DNA
<213> 人工序列_CvTps1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 41
atgactacga tccaccgccg ccatactacg ctgatcctgg acctgggtga tgttctgttc 60
cgctggtccc cgaaaaccga aaccgcaatt ccgcctcgtc agctgaaaga aatcttgacc 120
agcgttacct ggttcgagta tgagcgtggc caaattagcc agaccgaatg ctacgagcgt 180
tgtgctgccg agtttaaagt tgatccgctg gttattgccg aagcgtttaa acaagcgcgt 240
gaaagcctgc gtccgaacaa agcgtttatc gcgttgatcc gtgagttgcg ccaccagatg 300
catggtgacc tgacggtcct ggcactgagc aacattagcc tgcctgatta tgagtacatt 360
atgtcgctga gctccgattg ggcgacggtc tttaatcgcg tgtttccgag cgcactggtg 420
ggtgagcgta agccacacct gggttgctac cgcaaggtca tcagcgagat gtctctggag 480
ccgcagacca cggttttcgt cgatgacaaa ctggacaatg tcgcaagcgc tcgtagcctg 540
ggcatgcatg gcatcgtgtt cgacaacgaa gcgaacgttt ttcgtcagct gcgtaatatc 600
ttcggtaacc cggttagccg cggtcaaggt tacttgcgta aacacgccgg taaactggaa 660
tctagcacgg ataatggtct gaccttcgaa gagaacttca ctcaattaat tatttacgaa 720
gtcacgcaag accgcagcct gatcaccctg agcgagtgcc cgcgtacctg gaacttcttc 780
cgcggtcaac cactgttttc tgagagcttt ccggacgacg tggacaccac ctctgtggcg 840
ttgaccgttc tgcagccgga tcgtgcgttg gtggatagca tcctggacca gatgttggaa 900
tatgttgacg cggatggtat tatgcaaacc tactttgatt catcccgtcc gcgcattgac 960
ccgttcgtgt gcgtgaatgt cctgagcctg ttctacgcca atggcagagg ccgcgagctg 1020
ccacacacgc tggaatgggt ctatgaagtt ctgctgcacc gtgcgtacca cggcggtagc 1080
cgttattacc tgagcccgga ctgtttcctg ttctttatga gccgtctgct gaagcgcgcg 1140
aatgactcgg cgctgcaggc ccgttttcgc ccgcttttca tggaacgtgt gaaagagcgt 1200
gtgggcgcag ccggcgatag catggacctg gcgttccgca ttctggccgc tgcaaccatc 1260
ggcgttcatt gcccacaaga tctggagcgt ctggcagcag cgcagtgcga agatggtggc 1320
tgggatatgt gttggtttta tgcgtttggc agcacgggta tcaaggctgg caaccgcggt 1380
ctgaccaccg cgttggctgt cgccgcaatt cgtaccgcgc tgggtcgtcc gccttccccg 1440
agcccgagca atatttctag ctccagcaaa ctggacgcgc cgaactcctt cctgggcatc 1500
ccgcgtccga ccagcccgat ccgtttcggt gaactgtttc gtagctggcg taagaacaag 1560
ccgaccgcga aaagccagta a 1581
<210> 42
<211> 521
<212> PRT
<213> 药用拟层孔菌(Laricifomes officinalis)_LoTps1蛋白
<400> 42
Met Tyr Thr Ala Leu Ile Leu Asp Leu Gly Asp Val Leu Phe Ser Trp
1 5 10 15
Ser Ser Thr Thr Asn Thr Thr Ile Pro Pro Arg Gln Leu Lys Glu Ile
20 25 30
Leu Ser Ser Pro Ala Trp Phe Glu Tyr Glu Arg Gly Arg Ile Thr Gln
35 40 45
Ala Glu Cys Tyr Glu Arg Val Ser Ala Glu Phe Ser Leu Asp Ala Thr
50 55 60
Ala Val Ala Glu Ala Phe Arg Gln Ala Arg Asp Ser Leu Arg Pro Asn
65 70 75 80
Asp Lys Phe Leu Thr Leu Ile Arg Glu Leu Arg Gln Gln Ser His Gly
85 90 95
Glu Leu Thr Val Leu Ala Leu Ser Asn Ile Ser Leu Pro Asp Tyr Glu
100 105 110
Phe Ile Met Ala Leu Asp Ser Lys Trp Thr Ser Val Phe Asp Arg Val
115 120 125
Phe Pro Ser Ala Leu Val Gly Glu Arg Lys Pro His Leu Gly Ala Phe
130 135 140
Arg Gln Val Leu Ser Glu Met Asn Leu Asp Pro His Thr Thr Val Phe
145 150 155 160
Val Asp Asp Lys Leu Asp Asn Val Val Ser Ala Arg Ser Leu Gly Met
165 170 175
His Gly Val Val Phe Asp Ser Gln Asp Asn Val Phe Arg Met Leu Arg
180 185 190
Asn Ile Phe Gly Asp Pro Ile His Arg Gly Arg Asp Tyr Leu Arg Gln
195 200 205
His Ala Gly Arg Leu Glu Thr Ser Thr Asp Ala Gly Val Val Phe Glu
210 215 220
Glu Asn Phe Thr Gln Leu Ile Ile Tyr Glu Leu Thr Asn Asp Lys Ser
225 230 235 240
Leu Ile Thr Thr Ser Asn Cys Ala Arg Thr Trp Asn Phe Phe Arg Gly
245 250 255
Lys Pro Leu Phe Ser Ala Ser Phe Pro Asp Asp Met Asp Thr Thr Ser
260 265 270
Val Ala Leu Thr Val Leu Arg Leu Asp His Ala Leu Val Asn Ser Val
275 280 285
Leu Asp Glu Met Leu Lys Tyr Val Asp Ala Asp Gly Ile Met Gln Thr
290 295 300
Tyr Phe Asp His Thr Arg Pro Arg Met Asp Pro Phe Val Cys Val Asn
305 310 315 320
Val Leu Ser Leu Phe His Glu Gln Gly Arg Gly His Glu Leu Pro Asn
325 330 335
Thr Leu Glu Trp Val His Glu Val Leu Leu His Arg Ala Tyr Ile Gly
340 345 350
Gly Ser Arg Tyr Tyr Leu Ser Ala Asp Cys Phe Leu Phe Phe Met Ser
355 360 365
Arg Leu Leu Gln Arg Ile Thr Asp Pro Ser Val Leu Gly Arg Phe Arg
370 375 380
Pro Leu Phe Ile Glu Arg Val Arg Glu Arg Val Gly Ala Thr Gly Asp
385 390 395 400
Ser Ile Asp Leu Ala Phe Arg Ile Ile Ala Ala Ser Thr Val Gly Ile
405 410 415
Gln Cys Pro Arg Asp Leu Glu Ser Leu Leu Ala Ala Gln Cys Glu Asp
420 425 430
Gly Gly Trp Asp Leu Cys Trp Phe Tyr Gln Tyr Gly Ser Thr Gly Val
435 440 445
Lys Ala Gly Asn Arg Gly Leu Thr Thr Ala Leu Ala Ile Lys Ala Ile
450 455 460
Asp Ser Ala Ile Ala Arg Pro Pro Ser Pro Ala Leu Ser Val Ala Ser
465 470 475 480
Ser Ser Lys Ser Glu Ile Pro Lys Pro Ile Gln Arg Ser Leu Arg Pro
485 490 495
Leu Ser Pro Arg Arg Phe Gly Gly Phe Leu Met Pro Trp Arg Arg Ser
500 505 510
Gln Arg Asn Gly Val Ala Val Ser Ser
515 520
<210> 43
<211> 2291
<212> DNA
<213> 药用拟层孔菌(Laricifomes officinalis)_LoTps1转录物(包括非编码序列)
<400> 43
gcgtctgctg cggtctctca ccgcgccgag cgacgggaag cggaggcttt ttgatgcagc 60
cagctcagcg ccatcctctc acgcaggggg tttgatccag atctgatcgc ctccgggttc 120
tcatctagaa cgcacggcgg ctcccaggaa gttctatcga ccctctgcgc gctggtcggc 180
ggcacgatgt ggctacacca gtcccaatca tatctcacac ccagcaccat catctcgggc 240
ctcttcgtca tgtaaccctc ccaagcctat ttttcagggc gttccccctc accggcgcgc 300
ttcttaaaga atcccgaaat gtatacggct cttatccttg acctcggcga cgttctgttc 360
tcttggtcgt cgacgaccaa cacgactatt ccccctcggc agctaaagga gatcctctca 420
tctcctgcct ggtttgagta cgagcgtggt cgcataacgc aagccgaatg ctacgagcgt 480
gtcagcgccg agttcagcct agacgccacc gccgtcgcgg aagcattccg gcaagctcgc 540
gactccttgc gcccgaacga caagttcctc acgttaattc gcgagcttcg acaacaatct 600
catggggagc tcacggtgct tgcgctgtcc aacatatccc ttcccgacta tgaattcatc 660
atggccctcg actcgaagtg gacttctgtc tttgaccgcg tcttcccttc tgccctcgtg 720
ggcgaacgga agccacacct tggagcgttt cgccaggttc tgtccgagat gaatcttgac 780
ccgcacacaa ctgtgttcgt cgatgacaag ctggacaatg tcgtctccgc acggtccctc 840
gggatgcacg gcgtcgtgtt cgactcccaa gacaatgtct ttcggatgct gagaaacatc 900
tttggcgatc ccattcatcg gggacgtgac tatctccgac agcacgccgg acgtctggag 960
acctccacgg atgccggtgt ggtcttcgaa gagaatttca cgcaactcat catctacgaa 1020
ctgacgaatg acaagtctct catcacgaca tcaaactgtg ctcgtacttg gaatttcttt 1080
cgtgggaagc ctttgttctc agcatcgttc cctgacgaca tggacacgac ctcggttgcc 1140
ttgactgtat tacgtttaga ccacgccctc gtgaactcgg ttttggacga gatgctaaag 1200
tatgtcgacg cagacggcat catgcagacc tacttcgacc atacacgccc acgcatggat 1260
ccatttgtct gcgtcaatgt gctctcgttg tttcacgaac aaggtcgtgg ccacgagctt 1320
ccgaacaccc tcgaatgggt ccatgaggtc ctcctccacc gcgcgtacat cgggggctcg 1380
cggtactacc tctccgcgga ctgcttcctc tttttcatga gccgcctcct gcagcgcatc 1440
accgacccgt ccgtccttgg ccgcttccgt ccactattca tagagcgcgt tcgggagcgt 1500
gtaggtgcga ccggggactc catcgatctc gcattccgca tcatcgccgc gtccacagta 1560
ggcatccagt gtccacgcga cttggaaagt ctcctcgccg cacagtgtga agacggtggc 1620
tgggacctgt gctggttcta ccagtacgga tcgaccggtg tcaaggcggg caaccgcggg 1680
ctcaccaccg ctctggcgat caaagctatt gactccgcca ttgcgaggcc accttcgcct 1740
gccctctcag tcgcttcgtc gtccaaatcg gagataccga aacccataca acggtccctt 1800
aggcccctta gcccccgccg gtttggcggt ttcctgatgc cgtggcgcag gtcacagcgc 1860
aatggcgtgg cggtctctag ttgaacactt gacccttgac acttcgcttt gcactgcctg 1920
ctcccctgcc aatcctcccc tacgatcgta tcatccctct cttgccctcg cctccccctc 1980
gtaccccctc tcatggggtg ccatttgtag atatgtacgt agcgtgatgt agcggtactc 2040
ggatcgttct cgtactcgtc ttgctctgcc gtcgcttcca gcccgtgctg ttctctcgtt 2100
caggctattc gttggttacg cgtatatcgt aatagaccgc cccggttcct cgcctacaga 2160
cactcgcccg tctcgccacg gactcggcta cggattcaga ctacatgagt ggcagttatc 2220
acacgcagat ccctccttgg tcgttctgta gtacccacat atgtaattgt accagtccac 2280
tgttgcagat c 2291
<210> 44
<211> 1566
<212> DNA
<213> 药用拟层孔菌(Laricifomes officinalis)_LoTps1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 44
atgtatacgg ctcttatcct tgacctcggc gacgttctgt tctcttggtc gtcgacgacc 60
aacacgacta ttccccctcg gcagctaaag gagatcctct catctcctgc ctggtttgag 120
tacgagcgtg gtcgcataac gcaagccgaa tgctacgagc gtgtcagcgc cgagttcagc 180
ctagacgcca ccgccgtcgc ggaagcattc cggcaagctc gcgactcctt gcgcccgaac 240
gacaagttcc tcacgttaat tcgcgagctt cgacaacaat ctcatgggga gctcacggtg 300
cttgcgctgt ccaacatatc ccttcccgac tatgaattca tcatggccct cgactcgaag 360
tggacttctg tctttgaccg cgtcttccct tctgccctcg tgggcgaacg gaagccacac 420
cttggagcgt ttcgccaggt tctgtccgag atgaatcttg acccgcacac aactgtgttc 480
gtcgatgaca agctggacaa tgtcgtctcc gcacggtccc tcgggatgca cggcgtcgtg 540
ttcgactccc aagacaatgt ctttcggatg ctgagaaaca tctttggcga tcccattcat 600
cggggacgtg actatctccg acagcacgcc ggacgtctgg agacctccac ggatgccggt 660
gtggtcttcg aagagaattt cacgcaactc atcatctacg aactgacgaa tgacaagtct 720
ctcatcacga catcaaactg tgctcgtact tggaatttct ttcgtgggaa gcctttgttc 780
tcagcatcgt tccctgacga catggacacg acctcggttg ccttgactgt attacgttta 840
gaccacgccc tcgtgaactc ggttttggac gagatgctaa agtatgtcga cgcagacggc 900
atcatgcaga cctacttcga ccatacacgc ccacgcatgg atccatttgt ctgcgtcaat 960
gtgctctcgt tgtttcacga acaaggtcgt ggccacgagc ttccgaacac cctcgaatgg 1020
gtccatgagg tcctcctcca ccgcgcgtac atcgggggct cgcggtacta cctctccgcg 1080
gactgcttcc tctttttcat gagccgcctc ctgcagcgca tcaccgaccc gtccgtcctt 1140
ggccgcttcc gtccactatt catagagcgc gttcgggagc gtgtaggtgc gaccggggac 1200
tccatcgatc tcgcattccg catcatcgcc gcgtccacag taggcatcca gtgtccacgc 1260
gacttggaaa gtctcctcgc cgcacagtgt gaagacggtg gctgggacct gtgctggttc 1320
taccagtacg gatcgaccgg tgtcaaggcg ggcaaccgcg ggctcaccac cgctctggcg 1380
atcaaagcta ttgactccgc cattgcgagg ccaccttcgc ctgccctctc agtcgcttcg 1440
tcgtccaaat cggagatacc gaaacccata caacggtccc ttaggcccct tagcccccgc 1500
cggtttggcg gtttcctgat gccgtggcgc aggtcacagc gcaatggcgt ggcggtctct 1560
agttga 1566
<210> 45
<211> 1566
<212> DNA
<213> 人工序列_LoTps1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 45
atgtacacgg cgctgatttt ggatttgggt gatgttctgt ttagctggag ctcaacgact 60
aacaccacca ttccgccgcg tcagctgaaa gaaatcttga gctccccggc gtggttcgag 120
tacgagcgtg gccgtatcac ccaggcagag tgttatgagc gtgtcagcgc agagtttagc 180
ctggatgcga cggccgtggc tgaggctttt cgtcaggcac gtgatagcct gcgtccgaac 240
gacaaatttc tgaccctgat ccgtgagctg cgtcaacaga gccacggtga attgaccgtt 300
ctggccttgt ctaacatcag cctgccggat tacgaattta ttatggcact ggactcgaag 360
tggaccagcg tgtttgatcg tgtgttcccg agcgccctgg tgggcgaacg caagccgcac 420
ctgggcgcgt tccgccaagt cctgtccgag atgaatttgg acccgcatac caccgttttt 480
gtggacgaca aactggacaa tgttgtcagc gcacgcagcc tgggtatgca cggtgtcgtg 540
ttcgacagcc aagacaatgt ttttcgtatg ctgcgtaaca ttttcggtga cccaattcac 600
cgcggtcgtg actatctgcg ccagcacgct ggtcgtcttg aaacgtccac cgatgcgggc 660
gttgtgttcg aagagaactt cacccaactg atcatttacg aactgaccaa cgataagagc 720
ctgatcacca cctctaattg cgcccgcacc tggaacttct tccgcggcaa acctctgttc 780
tccgcgagct ttccggacga tatggacact acgtcggtag cgctgaccgt gctgcgtctg 840
gaccatgcgc tggtgaatag cgttctggat gaaatgctga aatacgtcga tgctgacggt 900
attatgcaga cctactttga tcatacgcgt cctcgtatgg acccgttcgt ttgcgtcaat 960
gtgctgagcc tgtttcacga gcaaggtcgc ggtcatgaac tgccgaatac gctggaatgg 1020
gtgcatgaag tcctgctgca ccgtgcgtat atcggtggca gccgctatta tctgagcgcg 1080
gattgtttcc tgttctttat gagccgtctg ttgcaacgta ttaccgaccc gagcgtttta 1140
ggtagatttc gcccgctgtt catcgagcgt gttcgcgagc gcgttggcgc gactggcgac 1200
agcatcgacc tggcattccg tatcatcgcg gccagcacgg tcggcattca atgcccgcgt 1260
gacctggagt ctctgctggc agcacagtgc gaagatggtg gctgggatct gtgttggttt 1320
taccagtacg gcagcacggg tgttaaggcc ggtaaccgtg gtctgaccac ggcgttggcg 1380
atcaaagcga ttgacagcgc catcgcgcgt ccgccaagcc cggccctgtc cgttgcaagc 1440
tccagcaaga gcgagattcc gaagccgatt cagcgtagcc tccgcccgtt gagcccgcgt 1500
cgcttcggtg gcttcctgat gccgtggcgt cgtagccaac gcaatggtgt cgcggtgagc 1560
tcttaa 1566
<210> 46
<211> 527
<212> PRT
<213> 白腐菌(Obba rivulosa)_OCH93767.1蛋白
<400> 46
Met Ser Ala Ala Val Arg Tyr Thr Thr Leu Ile Leu Asp Leu Gly Asp
1 5 10 15
Val Leu Phe Thr Trp Ser Pro Lys Thr Lys Thr Ser Ile Ser Pro Arg
20 25 30
Ile Leu Lys Glu Ile Leu Asn Ser Ala Thr Trp Tyr Glu Tyr Glu Arg
35 40 45
Gly Ser Ile Thr Gln His Glu Cys Tyr Glu Arg Val Gly Val Glu Phe
50 55 60
Gly Ile Ala Pro Ser Glu Ile His Asn Ala Phe Lys Gln Ala Arg Asp
65 70 75 80
Ser Met Glu Ser Asn Asp Glu Leu Ile Ala Leu Val Arg Glu Leu Lys
85 90 95
Glu Gln Ser Asp Gly Glu Leu Leu Val Phe Ala Leu Ser Asn Ile Ser
100 105 110
Leu Pro Asp Tyr Glu Tyr Val Leu Thr Lys Pro Ala Asp Trp Ser Ile
115 120 125
Phe Asp Lys Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys Pro His
130 135 140
Leu Gly Ile Tyr Lys His Val Ile Ala Glu Thr Gly Val Asp Pro Arg
145 150 155 160
Thr Thr Val Phe Val Asp Asp Lys Ile Asp Asn Val Leu Ser Ala Arg
165 170 175
Ser Leu Gly Met His Gly Ile Val Phe Asp Lys His Glu Asp Val Met
180 185 190
Arg Ala Leu Arg Asn Ile Phe Gly Asp Pro Val Arg Arg Gly Arg Glu
195 200 205
Tyr Leu Arg Arg Asn Ala Arg Lys Leu Glu Ser Ile Thr Asp His Gly
210 215 220
Val Ala Phe Gly Glu Asn Phe Thr Gln Leu Leu Ile Leu Glu Leu Thr
225 230 235 240
Ser Asp Ala Ser Leu Val Thr Leu Pro Asp Arg Pro Arg Thr Trp Asn
245 250 255
Phe Phe Arg Gly Lys Pro Leu Phe Ser Glu Ala Phe Pro Asp Asp Leu
260 265 270
Asp Thr Thr Ser Leu Ala Leu Thr Val Leu Lys Arg Asp Ala Ala Thr
275 280 285
Val Ser Ser Val Met Asp Glu Met Leu Lys Tyr Arg Asp Ala Asp Gly
290 295 300
Ile Met Gln Thr Tyr Phe Asp Asn Gly Arg Gln Arg Leu Asp Pro Phe
305 310 315 320
Val Asn Ala Asn Val Leu Thr Leu Phe Tyr Ala Asn Gly Arg Gly His
325 330 335
Glu Leu Asp Gln Ser Leu Ser Trp Val Arg Glu Val Leu Leu Tyr Arg
340 345 350
Ala Tyr Leu Gly Gly Ser Arg Tyr Tyr Pro Ser Ala Asp Cys Phe Leu
355 360 365
Tyr Phe Ile Ser Arg Leu Phe Ala Cys Thr Ser Asp Pro Val Leu His
370 375 380
His Gln Leu Lys Pro Leu Phe Val Glu Arg Val His Glu Arg Ile Gly
385 390 395 400
Val Gln Gly Asp Ala Leu Glu Leu Ala Phe Arg Leu Leu Val Cys Ala
405 410 415
Ser Phe Asn Ile Ser Asn Gln Pro Asp Met Arg Lys Leu Leu Glu Met
420 425 430
Gln Cys Gln Asp Gly Gly Trp Asp Gly Gly Asn Leu Tyr Arg Phe Gly
435 440 445
Thr Thr Gly Leu Lys Val Thr Asn Arg Gly Leu Thr Thr Ala Ala Ala
450 455 460
Val Gln Ala Ile Glu Ala Thr Gln Leu Arg Pro Pro Ser Pro Ala Phe
465 470 475 480
Ser Val Glu Ser Pro Lys Ser Pro Val Thr Pro Val Thr Pro Met Leu
485 490 495
Glu Ile Pro Ala Leu Gly Leu Ser Ile Ser Arg Pro Ser Ser Pro Leu
500 505 510
Leu Gly Tyr Phe Lys Leu Pro Trp Lys Lys Ser Ala Glu Val His
515 520 525
<210> 47
<211> 1584
<212> DNA
<213> 白腐菌(Obba rivulosa)_OCH93767.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 47
atgtccgcag cagttcggta cacgaccctc atcctcgacc ttggcgacgt cttgttcact 60
tggtcaccga agacgaagac cagcatctcg cctcgtattc tgaaggagat cctgaattcc 120
gcgacctggt atgagtacga gcgcggtagt atcactcagc acgaatgtta cgaacgcgtt 180
ggcgtggagt tcggtattgc gccgagcgag atccacaacg cgttcaagca ggctcgggac 240
tctatggagt cgaatgacga gctgatcgcc cttgttcggg aactgaagga gcagtcagat 300
ggagagcttc tcgtcttcgc attatcgaac atctcactgc cggactacga atacgtcctg 360
acgaagcccg cggactggtc catcttcgac aaagtctttc cttccgctct cgtcggcgag 420
cgcaagcccc atctcggcat ctacaaacac gtcatcgcag agacgggcgt tgatccgcga 480
acaaccgtct tcgtggacga caagatcgac aatgtgcttt cggcgcggtc gctcggtatg 540
cacggcattg tcttcgacaa acacgaagac gtaatgcgcg ctctgcgaaa cattttcggt 600
gaccccgtgc gaagaggacg agaatatttg cgtcgaaatg caaggaaatt ggaatccatc 660
acagatcacg gcgtcgcctt cggggagaac ttcacccagc ttctgatcct cgaacttact 720
agtgatgcgt ccctcgttac tctccctgat cgtcctcgga catggaattt tttccgaggg 780
aagccgctct tttcggaggc cttccccgat gaccttgata ctacttcctt ggcactcact 840
gtcctgaaaa gagatgccgc cactgtatcg tccgtgatgg acgagatgct gaaatacagg 900
gacgcggacg gcatcatgca gacatacttc gacaacggtc ggcaacgact cgatccgttc 960
gtcaacgcca acgttttgac cctcttctac gccaacggtc gcggacacga gctggatcag 1020
agcctcagct gggttcgcga agtcttgctc taccgcgctt acctcggcgg ttcccgctac 1080
tacccctccg ccgactgctt cctatatttc atcagccgcc tcttcgcctg caccagcgac 1140
ccggtcctcc atcatcaact taagcccctc tttgttgagc gtgtgcacga gcggatagga 1200
gtgcagggcg acgcgctgga gctcgccttc cgcctgcttg tatgcgcgag cttcaacatc 1260
tcgaaccagc ctgacatgcg caagctgctc gagatgcagt gccaggacgg aggctgggat 1320
ggcggaaacc tgtatcgttt cggcaccacg ggcctcaagg tcacgaaccg gggtctgacc 1380
accgcagcag ccgtgcaagc catcgaggcg acgcagctgc gtccaccatc accggcgttc 1440
tctgtcgagt cgcctaagag cccggtgacg ccggtgacgc ccatgctgga gattccagcg 1500
ctgggtctca gcatctcgcg gccctccagt cctctgttgg ggtatttcaa gctcccgtgg 1560
aagaagtcag ccgaggttca ttga 1584
<210> 48
<211> 1584
<212> DNA
<213> 人工序列_OCH93767 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 48
atgtctgcag ctgttcgtta tactactctg atcctggatt tgggcgatgt tctgttcacc 60
tggtccccga aaaccaagac ctctatcagc ccacgtatcc tgaaagaaat cctgaacagc 120
gcgacctggt acgagtatga gcgtggcagc atcacccagc acgagtgcta cgagcgtgtt 180
ggcgtcgaat ttggtattgc gccgagcgag attcacaacg cgttcaaaca agcccgcgac 240
agcatggaat ccaacgacga actgattgct ctggtgcgtg agctgaaaga acagagcgat 300
ggtgagctgc tggtctttgc cctgagcaat atctctctgc cggattacga atacgttctg 360
accaaaccag cggactggtc aatcttcgat aaagtctttc cgagcgcttt ggtcggtgag 420
cgtaaaccgc atctgggtat ttacaaacac gttattgcgg aaaccggtgt tgacccgaga 480
acgaccgttt ttgttgacga taagattgac aacgtcctga gcgcacgcag cctgggtatg 540
catggtattg tctttgataa acacgaagat gtgatgcgtg ctctgcgcaa tatctttggc 600
gacccggtgc gtcgcggtcg tgagtatttg cgccgcaacg cgcgcaaatt ggagtccatt 660
accgatcatg gtgtcgcatt tggtgagaat ttcacccagc tcctgattct ggaactgacc 720
agcgacgcgt ccctggtgac gctgccggat cgtccgcgta cgtggaactt cttccgcggc 780
aagccgctgt ttagcgaagc gttcccggat gacctggaca ccacgagcct ggcactgacg 840
gtgctgaaac gcgatgcagc aactgtgagc tccgtcatgg acgaaatgct gaagtaccgc 900
gacgcggatg gcatcatgca gacgtatttc gacaacggtc gtcagcgtct ggacccgttt 960
gtcaacgcca atgttctgac gctgttttac gcgaatggcc gtggtcatga actggaccag 1020
agcttatcat gggtgcgtga agtgctgctg tatcgcgcct atctgggtgg cagccgctac 1080
tatccgagcg cggactgttt tctgtacttc attagccgct tgttcgcctg caccagcgat 1140
ccggttctgc atcaccaact gaagccattg ttcgtcgagc gtgtgcacga gcgtattggt 1200
gttcagggcg acgcactgga actggcgttc cgtctgttgg tgtgtgcgag cttcaacatt 1260
agcaatcagc cggatatgcg taagctgctg gaaatgcaat gccaagatgg cggctgggac 1320
ggtggtaatc tgtaccgttt tggcaccacc ggtttaaaag tgacgaatcg tggtttgacc 1380
accgctgcgg ccgttcaagc aattgaagca acgcaactgc gtccgccgag cccagcattt 1440
agcgtagagt cgcctaagag cccggttacg ccggtgacgc cgatgctgga aatcccggcg 1500
ctgggtctgt ctatcagccg tccgtcgagc ccgctgctgg gctatttcaa gttgccgtgg 1560
aagaaaagcg ccgaagtgca ctaa 1584
<210> 49
<211> 533
<212> PRT
<213> 弯孢胶化孔菌(Gelatoporia subvermispora)_EMD37666.1蛋白
<400> 49
Met Ser Ala Ala Ala Gln Tyr Thr Thr Leu Ile Leu Asp Leu Gly Asp
1 5 10 15
Val Leu Phe Thr Trp Ser Pro Lys Thr Lys Thr Ser Ile Pro Pro Arg
20 25 30
Thr Leu Lys Glu Ile Leu Asn Ser Ala Thr Trp Tyr Glu Tyr Glu Arg
35 40 45
Gly Arg Ile Ser Gln Asp Glu Cys Tyr Glu Arg Val Gly Thr Glu Phe
50 55 60
Gly Ile Ala Pro Ser Glu Ile Asp Asn Ala Phe Lys Gln Ala Arg Asp
65 70 75 80
Ser Met Glu Ser Asn Asp Glu Leu Ile Ala Leu Val Arg Glu Leu Lys
85 90 95
Thr Gln Leu Asp Gly Glu Leu Leu Val Phe Ala Leu Ser Asn Ile Ser
100 105 110
Leu Pro Asp Tyr Glu Tyr Val Leu Thr Lys Pro Ala Asp Trp Ser Ile
115 120 125
Phe Asp Lys Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys Pro His
130 135 140
Leu Gly Val Tyr Lys His Val Ile Ala Glu Thr Gly Ile Asp Pro Arg
145 150 155 160
Thr Thr Val Phe Val Asp Asp Lys Ile Asp Asn Val Leu Ser Ala Arg
165 170 175
Ser Val Gly Met His Gly Ile Val Phe Glu Lys Gln Glu Asp Val Met
180 185 190
Arg Ala Leu Arg Asn Ile Phe Gly Asp Pro Val Arg Arg Gly Arg Glu
195 200 205
Tyr Leu Arg Arg Asn Ala Met Arg Leu Glu Ser Val Thr Asp His Gly
210 215 220
Val Ala Phe Gly Glu Asn Phe Thr Gln Leu Leu Ile Leu Glu Leu Thr
225 230 235 240
Asn Asp Pro Ser Leu Val Thr Leu Pro Asp Arg Pro Arg Thr Trp Asn
245 250 255
Phe Phe Arg Gly Asn Gly Gly Arg Pro Ser Lys Pro Leu Phe Ser Glu
260 265 270
Ala Phe Pro Asp Asp Leu Asp Thr Thr Ser Leu Ala Leu Thr Val Leu
275 280 285
Gln Arg Asp Pro Gly Val Ile Ser Ser Val Met Asp Glu Met Leu Asn
290 295 300
Tyr Arg Asp Pro Asp Gly Ile Met Gln Thr Tyr Phe Asp Asp Gly Arg
305 310 315 320
Gln Arg Leu Asp Pro Phe Val Asn Val Asn Val Leu Thr Phe Phe Tyr
325 330 335
Thr Asn Gly Arg Gly His Glu Leu Asp Gln Cys Leu Thr Trp Val Arg
340 345 350
Glu Val Leu Leu Tyr Arg Ala Tyr Leu Gly Gly Ser Arg Tyr Tyr Pro
355 360 365
Ser Ala Asp Cys Phe Leu Tyr Phe Ile Ser Arg Leu Phe Ala Cys Thr
370 375 380
Asn Asp Pro Val Leu His His Gln Leu Lys Pro Leu Phe Val Glu Arg
385 390 395 400
Val Gln Glu Gln Ile Gly Val Glu Gly Asp Ala Leu Glu Leu Ala Phe
405 410 415
Arg Leu Leu Val Cys Ala Ser Leu Asp Val Gln Asn Ala Ile Asp Met
420 425 430
Arg Arg Leu Leu Glu Met Gln Cys Glu Asp Gly Gly Trp Glu Gly Gly
435 440 445
Asn Leu Tyr Arg Phe Gly Thr Thr Gly Leu Lys Val Thr Asn Arg Gly
450 455 460
Leu Thr Thr Ala Ala Ala Val Gln Ala Ile Glu Ala Ser Gln Arg Arg
465 470 475 480
Pro Pro Ser Pro Ser Pro Ser Val Glu Ser Thr Lys Ser Pro Ile Thr
485 490 495
Pro Val Thr Pro Met Leu Glu Val Pro Ser Leu Gly Leu Ser Ile Ser
500 505 510
Arg Pro Ser Ser Pro Leu Leu Gly Tyr Phe Arg Leu Pro Trp Lys Lys
515 520 525
Ser Ala Glu Val His
530
<210> 50
<211> 1602
<212> DNA
<213> 弯孢胶化孔菌(Gelatoporia subvermispora)_EMD37666.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 50
atgtccgcgg cagctcaata cacgaccctc attctcgacc ttggcgacgt cctgttcacc 60
tggtcaccga aaaccaagac gagcatcccc cctcggactc tgaaggagat tctcaattcc 120
gcgacatggt atgagtatga gcgcggccgc atctctcagg acgaatgtta cgaacgcgtt 180
ggcacggagt tcggaatcgc gcctagcgaa atcgacaacg cgttcaagca agctcgggat 240
tccatggaat ccaacgacga actgatcgcc cttgttcggg aactcaagac gcagttggac 300
ggcgaactcc ttgtcttcgc actctcaaat atctcgttgc ctgactacga gtacgtcctc 360
acgaaaccgg ccgactggtc catcttcgac aaggtcttcc cttccgccct cgtgggcgag 420
cgcaagccgc acctcggcgt ttacaagcac gtcattgcag aaacgggcat tgatccgcga 480
accaccgttt tcgtggacga caagatcgac aacgtgctct cagcgcggtc tgtaggtatg 540
catgggatcg ttttcgagaa gcaggaagac gtaatgcgcg ctctccgaaa catcttcgga 600
gacccggttc ggcgagggcg cgagtacttg cgccgtaatg ccatgaggct tgaatcggtt 660
acagaccatg gtgtggcgtt tggcgagaac ttcacacaac tccttatcct cgaactaacg 720
aacgatccct ccctcgttac gctccctgat cgtcctcgaa catggaattt cttccgaggt 780
aacgggggac gaccaagcaa accattattc tcggaggcct tccccgatga cttggacact 840
acttcactag cgttgactgt cctccaaaga gatcccggcg tcatctcttc tgtgatggac 900
gaaatgttga actacaggga tccggacggc attatgcaga catacttcga cgatggtcgg 960
caaagactcg atccatttgt caatgtcaat gtcttaacgt tcttctacac caacggacgt 1020
ggtcatgaac tggaccaatg ccttacatgg gtccgcgaag ttttgctcta tcgcgcctat 1080
ctcggcggct cacgttatta cccctccgcc gactgctttc tctacttcat cagccgcctt 1140
ttcgcatgca cgaatgaccc cgtgctacac caccaactca aaccgctctt cgtcgagcgc 1200
gtgcaggagc aaatcggcgt ggagggcgat gcgctcgagt tggcgttccg attgctcgtc 1260
tgtgcaagcc tggacgtcca aaacgcgatc gacatgcgca ggctgctcga gatgcaatgc 1320
gaagatggcg gctgggaggg cgggaacctt tataggtttg gcacgaccgg gctcaaggtg 1380
actaaccggg gcctgacgac tgcagcggcc gtacaggcca tcgaggcgtc ccaacggcgc 1440
ccaccatcac cgtccccctc cgtcgaatct acaaagagcc caataacccc tgtgacgccc 1500
atgctggagg tcccctcgct cggcctgagc atctcgaggc cgtccagccc tttactcggc 1560
tacttcaggc tcccgtggaa gaagtcggcc gaagtacact ga 1602
<210> 51
<211> 1602
<212> DNA
<213> 人工序列_EMD37666.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 51
atgtctgcgg cggctcaata cacgactttg attctggatc tgggtgatgt tctgttcact 60
tggtccccga aaaccaagac cagcatccct ccgcgtaccc tgaaagaaat cctgaatagc 120
gctacctggt atgagtacga gcgtggtcgc atttcccaag acgagtgtta cgaacgtgtg 180
ggcaccgagt tcggcattgc gccgagcgag attgacaacg cgttcaaaca agcgcgcgat 240
tcgatggaaa gcaatgatga actgatcgca ctggtccgtg agctgaaaac gcagctggac 300
ggtgagctgc tggttttcgc actgtccaat attagcctgc cggattacga atacgtcttg 360
accaaaccgg cggactggag catctttgac aaagtgttcc ctagcgcctt ggtgggcgag 420
cgtaagccgc atctgggcgt ttataaacac gttattgcgg aaacgggcat tgatccgcgc 480
acgacggttt tcgtggacga caagattgac aatgtgttaa gcgcacgcag cgtcggtatg 540
catggtatcg tgtttgagaa acaagaagat gtcatgcgtg cactgcgtaa catctttggt 600
gatccggtcc gtcgtggtcg tgagtatctg cgtagaaacg caatgcgtct ggagtccgtg 660
accgaccacg gcgtggcgtt tggtgagaac tttacccagt tgctgattct ggaattgacg 720
aacgacccga gcctggtcac cctgcctgat cgtccgcgta cctggaactt ttttcgcggc 780
aatggtggcc gcccgagcaa gccgctgttc agcgaagcgt tcccggatga tctggatacc 840
acgagcctgg cgctgaccgt gctgcagcgc gacccgggtg ttatcagcag cgttatggac 900
gaaatgctga attaccgtga cccggacggt atcatgcaga cttatttcga tgacggtcgc 960
caacgcttgg acccatttgt gaacgtcaat gttctgacct ttttctatac gaacggccgt 1020
ggtcacgaac tggaccagtg tctgacgtgg gtgcgtgaag tcctcttgta tcgtgcgtac 1080
cttggtggct cacgctacta cccatcggcg gattgcttcc tgtacttcat ctctcgtctg 1140
tttgcgtgta ccaatgaccc ggtgctgcac catcagctga agccactgtt tgttgagcgt 1200
gtccaagagc aaattggtgt cgagggtgat gcactggaac tggcttttcg tctgctggtc 1260
tgcgccagcc tggatgtcca gaatgccatc gacatgcgcc gtctgctgga aatgcagtgc 1320
gaagatggcg gttgggaggg tggtaacctc taccgcttcg gcaccacggg cctgaaagtt 1380
accaaccgcg gtctgacgac cgcagccgcc gttcaagcga tcgaagcgag ccaacgccgt 1440
ccgccgagcc cgagcccgtc tgtagagagc acgaaaagcc cgattacccc ggtgaccccg 1500
atgctggaag ttccaagcct gggcttatct atcagccgtc cgtccagccc gctgctgggt 1560
tatttccgtt tgccgtggaa gaaaagcgca gaagtgcact aa 1602
<210> 52
<211> 527
<212> PRT
<213> 弯孢胶化孔菌(Gelatoporia subvermispora)_EMD37666-B蛋白
<400> 52
Met Ser Ala Ala Ala Gln Tyr Thr Thr Leu Ile Leu Asp Leu Gly Asp
1 5 10 15
Val Leu Phe Thr Trp Ser Pro Lys Thr Lys Thr Ser Ile Pro Pro Arg
20 25 30
Thr Leu Lys Glu Ile Leu Asn Ser Ala Thr Trp Tyr Glu Tyr Glu Arg
35 40 45
Gly Arg Ile Ser Gln Asp Glu Cys Tyr Glu Arg Val Gly Thr Glu Phe
50 55 60
Gly Ile Ala Pro Ser Glu Ile Asp Asn Ala Phe Lys Gln Ala Arg Asp
65 70 75 80
Ser Met Glu Ser Asn Asp Glu Leu Ile Ala Leu Val Arg Glu Leu Lys
85 90 95
Thr Gln Leu Asp Gly Glu Leu Leu Val Phe Ala Leu Ser Asn Ile Ser
100 105 110
Leu Pro Asp Tyr Glu Tyr Val Leu Thr Lys Pro Ala Asp Trp Ser Ile
115 120 125
Phe Asp Lys Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys Pro His
130 135 140
Leu Gly Val Tyr Lys His Val Ile Ala Glu Thr Gly Ile Asp Pro Arg
145 150 155 160
Thr Thr Val Phe Val Asp Asp Lys Ile Asp Asn Val Leu Ser Ala Arg
165 170 175
Ser Val Gly Met His Gly Ile Val Phe Glu Lys Gln Glu Asp Val Met
180 185 190
Arg Ala Leu Arg Asn Ile Phe Gly Asp Pro Val Arg Arg Gly Arg Glu
195 200 205
Tyr Leu Arg Arg Asn Ala Met Arg Leu Glu Ser Val Thr Asp His Gly
210 215 220
Val Ala Phe Gly Glu Asn Phe Thr Gln Leu Leu Ile Leu Glu Leu Thr
225 230 235 240
Asn Asp Pro Ser Leu Val Thr Leu Pro Asp Arg Pro Arg Thr Trp Asn
245 250 255
Phe Phe Arg Gly Lys Pro Leu Phe Ser Glu Ala Phe Pro Asp Asp Leu
260 265 270
Asp Thr Thr Ser Leu Ala Leu Thr Val Leu Gln Arg Asp Pro Gly Val
275 280 285
Ile Ser Ser Val Met Asp Glu Met Leu Asn Tyr Arg Asp Pro Asp Gly
290 295 300
Ile Met Gln Thr Tyr Phe Asp Asp Gly Arg Gln Arg Leu Asp Pro Phe
305 310 315 320
Val Asn Val Asn Val Leu Thr Phe Phe Tyr Thr Asn Gly Arg Gly His
325 330 335
Glu Leu Asp Gln Cys Leu Thr Trp Val Arg Glu Val Leu Leu Tyr Arg
340 345 350
Ala Tyr Leu Gly Gly Ser Arg Tyr Tyr Pro Ser Ala Asp Cys Phe Leu
355 360 365
Tyr Phe Ile Ser Arg Leu Phe Ala Cys Thr Asn Asp Pro Val Leu His
370 375 380
His Gln Leu Lys Pro Leu Phe Val Glu Arg Val Gln Glu Gln Ile Gly
385 390 395 400
Val Glu Gly Asp Ala Leu Glu Leu Ala Phe Arg Leu Leu Val Cys Ala
405 410 415
Ser Leu Asp Val Gln Asn Ala Ile Asp Met Arg Arg Leu Leu Glu Met
420 425 430
Gln Cys Glu Asp Gly Gly Trp Glu Gly Gly Asn Leu Tyr Arg Phe Gly
435 440 445
Thr Thr Gly Leu Lys Val Thr Asn Arg Gly Leu Thr Thr Ala Ala Ala
450 455 460
Val Gln Ala Ile Glu Ala Ser Gln Arg Arg Pro Pro Ser Pro Ser Pro
465 470 475 480
Ser Val Glu Ser Thr Lys Ser Pro Ile Thr Pro Val Thr Pro Met Leu
485 490 495
Glu Val Pro Ser Leu Gly Leu Ser Ile Ser Arg Pro Ser Ser Pro Leu
500 505 510
Leu Gly Tyr Phe Arg Leu Pro Trp Lys Lys Ser Ala Glu Val His
515 520 525
<210> 53
<211> 1584
<212> DNA
<213> 人工序列_EMD37666-B 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 53
atgtctgcgg ctgctcaata tactactttg attctggatc tgggcgacgt tctgttcacg 60
tggagcccga aaaccaagac cagcattcca ccgcgtaccc tgaaggagat cctcaatagc 120
gcgacttggt acgagtatga gcgtggccgc atcagccaag acgagtgcta cgaacgcgtc 180
ggtacggaat ttggcattgc accaagcgag attgacaatg cgtttaaaca agcgcgtgac 240
agcatggaaa gcaatgacga actgatcgcg ctggtccgtg agctgaaaac ccagctggat 300
ggtgagctgt tggtgtttgc gctgtcgaac atctctctgc cggactacga gtatgttctg 360
accaaaccgg cggattggag catttttgat aaagtgtttc cgagcgcgct ggttggtgag 420
cgcaagccgc acctgggtgt gtacaaacac gttattgcag agactggcat cgacccgcgt 480
acgacggttt tcgttgacga caagatcgat aacgttctga gcgcacgtag cgtcggtatg 540
cacggtattg ttttcgaaaa acaagaagat gttatgcgcg cactgcgtaa tatcttcggc 600
gatccggtca gacgtggccg tgagtatctg cgccgcaatg cgatgcgtct ggaatcggtg 660
accgatcatg gtgtcgcctt tggcgagaat ttcacccagc tgctgatttt agagctgacc 720
aatgatccta gcctggtgac gctgccggat cgtccgcgta cctggaactt tttccgcggc 780
aagccgttgt tctccgaagc cttcccggac gacctggaca cgaccagcct ggcgctgacc 840
gtgctgcaac gtgatccggg tgtgatctct tccgtaatgg acgaaatgct gaactaccgt 900
gacccggacg gtatcatgca gacctatttt gacgacggtc gtcagcgtct ggacccgttt 960
gtgaacgtga atgtcctgac gttcttttac accaatggtc gcggtcacga actggatcag 1020
tgtctgacct gggtccgcga agtgctgctg tatcgtgcat acctgggtgg cagccgttat 1080
tacccgagcg ccgattgctt tctgtacttt atcagccgtc tgttcgcgtg cacgaacgat 1140
ccggttctgc atcaccagct gaagccgtta tttgttgagc gcgttcagga acaaattggt 1200
gtcgagggtg atgcgctgga attggcattc cgcctgttgg tctgcgccag ccttgatgtc 1260
cagaacgcca ttgacatgcg tcgcttgctc gaaatgcagt gtgaggacgg cggttgggag 1320
ggtggcaacc tgtaccgttt cggtacgacc ggcctgaaag tcacgaaccg tggtctgacg 1380
acggcagctg cggtgcaagc aattgaagcc agccaacgtc gtccgccatc cccgtcaccg 1440
agcgttgagt ccaccaagag cccgattacc cctgtgaccc cgatgcttga agttccgagc 1500
ctgggtctga gcatctcccg tcctagcagc ccgctgttgg gttacttccg cctgccgtgg 1560
aagaaaagcg ctgaggtgca ttaa 1584
<210> 54
<211> 486
<212> PRT
<213> 土曲霉(Aspergillus terreus)_XP_001217376.1蛋白
<400> 54
Met Ala Ile Thr Lys Gly Pro Val Lys Ala Leu Ile Leu Asp Phe Ser
1 5 10 15
Asn Val Leu Cys Ser Trp Lys Pro Pro Ser Asn Val Ala Val Pro Pro
20 25 30
Gln Ile Leu Lys Met Ile Met Ser Ser Asp Ile Trp His Asp Tyr Glu
35 40 45
Cys Gly Arg Tyr Ser Arg Glu Asp Cys Tyr Ala Arg Val Ala Asp Arg
50 55 60
Phe His Ile Ser Ala Ala Asp Met Glu Asp Thr Leu Lys Gln Ala Arg
65 70 75 80
Lys Ser Leu Gln Val His His Glu Thr Leu Leu Phe Ile Gln Gln Val
85 90 95
Lys Lys Asp Ala Gly Gly Glu Leu Met Val Cys Gly Met Thr Asn Thr
100 105 110
Pro Arg Pro Glu Gln Asp Val Met His Ser Ile Asn Ala Glu Tyr Pro
115 120 125
Val Phe Asp Arg Ile Tyr Ile Ser Gly Leu Met Gly Met Arg Lys Pro
130 135 140
Ser Ile Cys Phe Tyr Gln Arg Val Met Glu Glu Ile Gly Leu Ser Gly
145 150 155 160
Asp Ala Ile Met Phe Ile Asp Asp Lys Leu Glu Asn Val Ile Ala Ala
165 170 175
Gln Ser Val Gly Ile Arg Gly Val Leu Phe Gln Ser Gln Gln Asp Leu
180 185 190
Arg Arg Val Val Leu Asn Phe Leu Gly Asp Pro Val His Arg Gly Leu
195 200 205
Gln Phe Leu Ala Ala Asn Ala Lys Lys Met Asp Ser Val Thr Asn Thr
210 215 220
Gly Asp Thr Ile Gln Asp Asn Phe Ala Gln Leu Leu Ile Leu Glu Leu
225 230 235 240
Ala Gln Asp Arg Glu Leu Val Lys Leu Gln Ala Gly Lys Arg Thr Trp
245 250 255
Asn Tyr Phe Ile Gly Pro Pro Lys Leu Thr Thr Ala Thr Phe Pro Asp
260 265 270
Asp Met Asp Thr Thr Ser Met Ala Leu Ser Val Leu Pro Val Ala Glu
275 280 285
Asp Val Val Ser Ser Val Leu Asp Glu Met Leu Lys Phe Val Thr Asp
290 295 300
Asp Gly Ile Phe Met Thr Tyr Phe Asp Ser Ser Arg Pro Arg Val Asp
305 310 315 320
Pro Val Val Cys Ile Asn Val Leu Gly Val Phe Cys Arg His Asn Arg
325 330 335
Glu Arg Asp Val Leu Pro Thr Phe His Trp Ile Arg Asp Ile Leu Ile
340 345 350
Asn Arg Ala Tyr Leu Ser Gly Thr Arg Tyr Tyr Pro Ser Pro Asp Leu
355 360 365
Phe Leu Phe Phe Leu Ala Arg Leu Cys Leu Ala Val Arg Asn Gln Ser
370 375 380
Leu Arg Glu Gln Leu Val Leu Pro Leu Val Asp Arg Leu Arg Glu Arg
385 390 395 400
Val Gly Ala Pro Gly Glu Ala Val Ser Leu Ala Ala Arg Ile Leu Ala
405 410 415
Cys Arg Ser Phe Gly Ile Asp Ser Ala Arg Asp Met Asp Ser Leu Arg
420 425 430
Gly Lys Gln Cys Glu Asp Gly Gly Trp Pro Val Glu Trp Val Tyr Arg
435 440 445
Phe Ala Ser Phe Gly Leu Asn Val Gly Asn Arg Gly Leu Ala Thr Ala
450 455 460
Phe Ala Val Arg Ala Leu Glu Ser Pro Tyr Gly Glu Ser Ala Val Lys
465 470 475 480
Val Met Arg Arg Ile Val
485
<210> 55
<211> 1461
<212> DNA
<213> 土曲霉(Aspergillus terreus)_XP_001217376.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 55
atggctatca ccaagggtcc agttaaggcg cttattcttg acttttccaa tgttctctgc 60
tcgtggaagc ctcccagcaa tgttgcggtg ccgccccaga tactcaaaat gatcatgtcc 120
tctgacatat ggcatgacta cgagtgcgga cggtactcga gagaggactg ctatgccaga 180
gtggcagacc gttttcatat cagcgccgcg gacatggaag acacgctgaa acaggcgcgc 240
aagagcctgc aggttcacca tgagacactg ttgtttatcc agcaagtcaa gaaggatgcc 300
gggggcgagt tgatggtgtg tgggatgacc aacacgcccc ggccagagca agacgtaatg 360
cattcaatca acgcggagta tcctgtgttt gataggatat atatatccgg tctcatgggc 420
atgaggaagc cgagcatctg cttctaccag cgggtgatgg aggagattgg cctatcaggc 480
gatgcgatca tgtttataga tgacaagttg gagaatgtca tcgccgccca gtcggtaggg 540
atccgaggcg ttctatttca gagtcagcaa gatctccgtc gggttgtatt aaatttcttg 600
ggcgatccgg tccatcgcgg cctgcagttc ctagcggcca atgcgaaaaa gatggatagt 660
gtgaccaaca ccggcgatac tatccaagat aattttgctc agctcctcat cttggagctg 720
gcccaggaca gggaattggt gaagcttcag gctggaaaaa ggacttggaa ttacttcata 780
gggcctccca agctcacaac agccacgttc cccgatgaca tggacaccac atctatggct 840
ctctcggtcc ttcctgtggc cgaggatgtg gtctcttctg tcctggatga gatgcttaaa 900
ttcgtcaccg atgacggtat ctttatgact tacttcgatt cctcgcgccc tcgagtcgac 960
ccagtcgtat gtatcaacgt cttgggtgtt ttctgcaggc ataaccgaga gcgagacgtc 1020
cttccaacgt tccattggat tcgagacatc ctgatcaacc gggcatatct ctcgggcacc 1080
cgatactacc catcgcccga tttgtttttg tttttccttg cacgcctctg cctggcagtc 1140
cggaatcaga gcctacggga acaacttgtc ttgcctctgg tagaccgact gcgtgagcgg 1200
gtgggcgcac ctggagaagc ggtctcattg gcagcgcgga tccttgcctg ccgtagcttt 1260
ggtatcgaca gtgcgagaga catggacagc ttgaggggaa aacaatgcga ggatggcggc 1320
tggccagtgg agtgggttta ccggtttgcc tctttcggcc tgaacgtagg caatcggggt 1380
cttgctactg ccttcgcggt cagggcgctc gaaagcccct atggtgagtc ggcggtgaag 1440
gttatgagac gcatcgtctg a 1461
<210> 56
<211> 1461
<212> DNA
<213> 人工序列_XP_001217376.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 56
atggcaatca ctaagggccc agttaaagcg ctgattcttg atttttctaa cgttctgtgt 60
agctggaagc cgccgagcaa tgttgcggtc ccgcctcaaa ttctgaagat gattatgtcg 120
agcgacatct ggcatgatta tgagtgtggc cgttacagcc gtgaggactg ctacgcccgt 180
gttgctgacc gttttcatat cagcgcagcg gacatggaag ataccctgaa acaggcacgt 240
aagtccctgc aagtgcacca cgaaacgctg ctgttcatcc aacaggtgaa gaaagacgcg 300
ggtggtgagc tgatggtttg cggcatgacc aacacgccgc gtccggaaca agacgtgatg 360
cattccatca atgctgagta tccggtgttc gaccgtattt acattagcgg cctgatgggc 420
atgcgtaaac cgagcatttg tttctaccaa cgcgtaatgg aagagattgg tctgagcggt 480
gacgccatca tgttcattga cgataaactg gaaaatgtga ttgccgcaca gagcgtgggt 540
atccgcggtg tgctgttcca aagccagcaa gatctgcgtc gtgtcgtgct gaactttctg 600
ggcgatccgg tccaccgtgg tctgcagttc ttggcggcga acgcaaagaa aatggacagc 660
gtcacgaata ccggcgacac tatccaagac aatttcgcac agctgttgat cttagagctg 720
gcgcaggatc gcgaattggt gaaattgcag gccggtaaac gtacctggaa ctactttatt 780
ggtccgccga agctgaccac ggcgacgttt ccggatgata tggacacgac cagcatggcg 840
ctgtcggtgc tgcctgtcgc ggaagatgtc gtgagctctg ttctggacga gatgctgaag 900
ttcgtgaccg atgatggtat ctttatgacc tatttcgact ctagccgtcc gcgtgtcgat 960
ccggttgtct gcattaatgt gttgggtgtt ttctgccgcc acaatcgtga gcgcgacgtg 1020
ttgccgacct ttcactggat tcgtgatatt ctgatcaacc gcgcatatct gagcggcacg 1080
cgctattacc cgtccccgga tctgtttctg tttttcctgg ctcgtctgtg cctggccgtt 1140
cgcaaccaga gcctgcgcga acaactggtt ctcccgctgg ttgatcgtct gcgcgagcgt 1200
gttggtgctc cgggtgaggc tgtgagcctg gcggcacgta tcctggcgtg ccgtagcttc 1260
ggtatcgact cagcccgcga catggactcc ttgcgtggca aacagtgtga agatggtggt 1320
tggccggtcg aatgggtcta tcgcttcgcg agctttggtc tgaacgttgg caaccgtggt 1380
ttggccaccg cgtttgcggt tagagcgctg gagtccccat acggcgagag cgcagttaag 1440
gttatgcgcc gtatcgtgta a 1461
<210> 57
<211> 483
<212> PRT
<213> 棘孢曲霉(Aspergillus aculeatus)_OJJ98394.1蛋白
<400> 57
Met Pro Ser Val Lys Ala Leu Val Leu Asp Phe Ala Gly Val Leu Cys
1 5 10 15
Ser Trp Thr Pro Pro Ala Glu Ser Pro Leu Ser Pro Ala Gln Leu Lys
20 25 30
Gln Leu Met Ser Ser Glu Ile Trp Phe Glu Tyr Glu Arg Gly Arg Tyr
35 40 45
Ser Glu Glu Glu Cys Tyr Ala Lys Leu Val Glu Arg Phe Ser Ile Ser
50 55 60
Ala Ala Asp Met Ala Ser Thr Met Glu Gln Ala Arg Gln Ser Leu Glu
65 70 75 80
Leu Asn His Ala Val Leu Gln Leu Val Ser Glu Ile Arg Lys Arg Asn
85 90 95
Pro Gly Leu Lys Val Tyr Gly Met Thr Asn Thr Pro His Ala Glu Gln
100 105 110
Asp Cys Val Asn Arg Ile Val Asn Ser Tyr Pro Val Phe Asp His Val
115 120 125
Tyr Leu Ser Gly Leu Val Gly Met Arg Lys Pro Asp Leu Gly Phe Tyr
130 135 140
Arg Phe Val Leu Ala Glu Thr Gly Leu Arg Pro Asp Glu Val Val Phe
145 150 155 160
Val Asp Asp Lys Thr Glu Asn Val Leu Val Ala Gln Ser Val Gly Met
165 170 175
His Gly Val Val Phe Gln Asn Val Thr Asp Phe Lys Gln Gln Ile Ile
180 185 190
Asn Val Thr Gly Asp Pro Val Ser Arg Gly Leu Arg Tyr Leu Arg Ser
195 200 205
Asn Ala Lys Ser Leu Leu Thr Val Thr Ser Asn Asn Ser Val Ile His
210 215 220
Glu Asn Phe Ala Gln Leu Leu Ile Leu Glu Leu Thr Gly Asp Arg Asp
225 230 235 240
Leu Ile Glu Leu Glu Pro Trp Asp Arg Thr Trp Asn Tyr Phe Ile Gly
245 250 255
Val Pro Gln Ser Pro Thr Ser Thr Phe Pro Asn Asp Leu Asp Thr Thr
260 265 270
Ser Ile Ala Leu Ser Val Leu Pro Ile His Lys Asp Val Val Ala Asp
275 280 285
Val Met Asp Glu Ile Met Leu Leu Leu Asp Asn Asp Gly Ile Val Pro
290 295 300
Thr Tyr Phe Asp Pro Thr Arg Pro Arg Val Asp Pro Val Val Cys Val
305 310 315 320
Asn Val Leu Ser Leu Phe Ala Gln Asn Gly Arg Glu Ser Glu Leu Leu
325 330 335
Ala Thr Phe Asn Trp Val Leu Asp Val Leu Arg His Arg Ala Tyr Leu
340 345 350
Gln Gly Thr Arg Tyr Tyr Ile Ser Pro Asp Ala Phe Leu Tyr Phe Leu
355 360 365
Ala Arg Leu Ser Val Phe Leu Arg Met Ser Pro Leu Arg Ala Arg Leu
370 375 380
Met Pro Leu Leu Glu Glu Arg Val Tyr Glu Arg Ile Gly Ala His Gly
385 390 395 400
Asp Ala Ile Ser Leu Ala Met Arg Ile Tyr Thr Cys Lys Leu Leu Gly
405 410 415
Met Ser Asn Met Leu Asp Glu Arg Ala Leu Arg Asp Met Gln Cys Glu
420 425 430
Asp Gly Gly Phe Pro Thr Ser Trp Val Tyr Arg Phe Gly Ser Thr Gly
435 440 445
Val Lys Ile Gly Asn Arg Gly Leu Thr Thr Ala Leu Ala Ile Lys Ala
450 455 460
Ile Glu Met Pro Leu Ala Ser Leu Trp Lys Ser Trp Gly Leu Thr Thr
465 470 475 480
Asp Ile Arg
<210> 58
<211> 1452
<212> DNA
<213> 棘孢曲霉(Aspergillus aculeatus)_OJJ98394.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 58
atgccctccg tcaaagcact ggtcctggac ttcgccggag ttctatgctc atggaccccg 60
ccagccgaga gcccgctctc cccagcccag ctcaaacaac tcatgtcctc cgagatatgg 120
ttcgaatacg agcgcgggag atattccgaa gaagaatgtt atgcgaagct cgtcgaacgg 180
ttctccatca gcgctgcgga catggcttcc accatggaac aggcccgtca gagcctggaa 240
ctgaaccacg ccgtacttca gcttgtcagc gagataagga agcggaaccc cgggctcaaa 300
gtttatggca tgacgaacac gccccatgcg gaacaggatt gtgtgaatcg catcgtgaac 360
agctatcctg ttttcgacca tgtgtatctc tccgggctcg ttgggatgcg caaaccagat 420
cttggattct atcggtttgt tctcgcagag accgggttga ggcctgacga ggtcgtgttc 480
gtcgacgaca aaacggagaa tgtgttggtc gcgcagtccg tggggatgca cggcgtggtg 540
ttccagaacg ttacggattt caagcagcag atcataaacg tgacgggaga ccctgtctct 600
cggggcttga ggtatctccg ctcgaatgca aagagcctcc tcactgtgac tagcaataac 660
tccgtgatcc acgaaaactt tgcgcagttg ctgattctgg agctgacggg cgaccgagac 720
ttgatcgaac tcgagccttg ggatcgaaca tggaactact tcatcggggt tcctcagtcg 780
ccgacgagca ccttccccaa cgacctggac accacctcta tcgcgctctc ggtccttccc 840
attcataagg acgtcgttgc cgatgtgatg gacgagatta tgcttctcct agacaacgac 900
gggatagtcc caacatattt tgatcccact cgccctcgag tcgacccagt cgtgtgtgtg 960
aatgtactca gcctgtttgc ccaaaacggc cgagaatccg agttactcgc caccttcaac 1020
tgggtgctgg acgtgctgcg acatagagcc tacctgcagg gcacgagata ttacatcagt 1080
ccggacgcct tcttgtactt tctagccaga ctctcggtct ttctgaggat gagtccactc 1140
cgcgctcggc taatgcctct cctggaagaa agagtgtatg agcgaattgg tgcccatggc 1200
gacgccattt cgctggctat gcggatctat acgtgtaagc tgctcgggat gtcgaatatg 1260
ctcgatgaaa gagcattgcg ggacatgcag tgtgaggatg gcggcttccc tacaagttgg 1320
gtctatagat ttggatcgac cggagtgaag attgggaaca gggggttgac tactgcactt 1380
gcaataaagg ccattgagat gcctctcgct tcgctttgga agtcgtgggg attgacgact 1440
gacattcgat aa 1452
<210> 59
<211> 1452
<212> DNA
<213> 人工序列_OJJ98394.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 59
atgccgtcgg ttaaagcgtt ggttctggat tttgcgggtg tgttgtgttc ttggactcca 60
ccggcggaaa gcccgttgtc cccagcgcag ctgaagcagc tgatgagcag cgagatctgg 120
tttgagtatg agcgtggccg ctatagcgaa gaagagtgtt atgcaaaatt ggtggagcgt 180
ttctctatct cggccgcaga tatggcgagc acgatggaac aggcccgtca atcgctggag 240
ttgaaccacg ccgtgctgca attagtttcc gagattcgta aacgtaatcc gggcttaaag 300
gtttacggta tgactaatac cccgcatgca gagcaagatt gtgtgaaccg tattgtcaat 360
agctatccgg tttttgatca tgtctacctg agcggtctgg tgggtatgcg caaaccggat 420
ctgggctttt accgtttcgt tctggcagag actggtctgc gcccggatga agtcgtgttc 480
gttgacgaca agaccgaaaa tgtcctggtg gctcaatccg ttggcatgca tggtgtggtg 540
ttccaaaatg taaccgactt caaacaacag attatcaatg tcacgggtga tcctgtcagc 600
cgtggtttgc gctacttgcg ttccaacgcg aagtctctgc tcactgttac cagcaataac 660
agcgttatcc atgagaattt cgcgcagctg ctgatcctgg aactgacggg cgaccgtgac 720
ctgattgaac tggaaccgtg ggaccgtacg tggaactact ttatcggcgt gccgcaaagc 780
ccgaccagca cctttccgaa cgacctggat acgaccagca ttgccctgag cgttctgccg 840
attcacaaag atgtggttgc ggacgtgatg gatgagatta tgctgctgct ggacaatgac 900
ggtattgtcc cgacctactt cgatccaacc cgtccgcgtg ttgatcctgt tgtgtgcgtc 960
aacgttctga gcctgttcgc acagaacggt cgcgagtccg aattgctggc gacgttcaac 1020
tgggttttgg acgttctgag acaccgtgcg tatttgcagg gtacgcgcta ttatatcagc 1080
ccggatgcct ttctgtattt tctggcgcgc ctgtctgtgt ttctgcgtat gtctccgttg 1140
cgcgctcgtc tgatgccgct gctggaagaa cgcgtttatg agcgtatcgg cgcacacggc 1200
gatgctatta gcctggcgat gcgcatttac acctgtaagc tgctgggcat gagcaatatg 1260
ctggacgagc gtgcactgcg tgacatgcag tgtgaagatg gtggtttccc aaccagctgg 1320
gtgtaccgtt ttggtagcac gggcgtgaaa attggtaacc gtggcttgac gaccgcactg 1380
gccattaagg ccatcgaaat gccgctggcc agcctttgga aaagctgggg cctgaccacc 1440
gatattcgct aa 1452
<210> 60
<211> 485
<212> PRT
<213> 乌达加瓦曲霉(Aspergillus udagawae)_GAO87501.1蛋白
<400> 60
Met Thr Arg Gln Lys Ser Pro Gln Tyr Lys Ala Ile Ile Phe Asp Leu
1 5 10 15
Gly Asp Val Phe Phe Thr Trp Asp Ala Pro Lys Asp Thr Ala Val Leu
20 25 30
Pro Asn Leu Phe Lys Lys Met Leu Thr Ser Pro Thr Trp Ser Asp Tyr
35 40 45
Glu Arg Gly Lys Leu Ser Glu Glu Ser Cys Tyr Glu Arg Leu Ala Glu
50 55 60
Gln Phe Asp Val Asp Ser Ser Glu Ile Ala Arg Ser Leu Arg Lys Ala
65 70 75 80
Gln Gln Ser Leu Thr Thr Asp Ala Ala Ile Val Ser Leu Ile Ser Glu
85 90 95
Ile Arg Ala Leu Ala Gly His Ile Ala Ile Tyr Ala Met Ser Asn Ile
100 105 110
Ser Ala Pro Ala Tyr Ala Ala Val Leu Gln Thr Gln Pro Glu Met Gly
115 120 125
Ile Phe Asp Gly Val Phe Pro Ser Gly Cys Tyr Gly Thr Arg Lys Pro
130 135 140
Glu Leu Leu Phe Tyr Lys Lys Val Leu Gln Glu Ile Ala Val Pro Pro
145 150 155 160
Asn Gln Ile Ile Phe Ile Asp Asp Gln Leu Glu Asn Val Val Ser Ala
165 170 175
Gln Ser Thr Gly Met His Gly Ile Val Tyr Thr Gly Ala Gly Glu Leu
180 185 190
Ser Arg Gln Leu Arg Asn Leu Val Leu Asp Pro Val Gln Arg Gly Arg
195 200 205
Glu Phe Leu Arg Arg Asn Ala Gly Ala Leu Tyr Ser Ile Cys Glu Thr
210 215 220
Gly Gln Val Ile Arg Glu Asn Phe Ser Gln Leu Leu Ile Leu Glu Ala
225 230 235 240
Thr Gly Asp Arg Ser Leu Val Asn Leu Glu Tyr Gln Gln Arg Ser Trp
245 250 255
Asn Phe Phe Gln Gly Gly Pro Pro Ser Thr Ser Glu Thr Phe Pro Asp
260 265 270
Asp Val Asp Thr Thr Ser Ile Ala Leu Met Ile Leu Pro Ala Asp Asp
275 280 285
Asn Thr Val Asn Ser Val Leu Gly Glu Ile Ser Glu Val Ala Asn Asp
290 295 300
Glu Gly Ile Val Asn Thr Tyr Phe Asp Gln Thr Arg Gln Arg Ile Asp
305 310 315 320
Pro Ala Val Cys Val Asn Val Leu Arg Leu Phe Tyr Thr Tyr Gly Arg
325 330 335
Gly Ala Thr Leu Pro Leu Thr Leu Gln Trp Val Ser Asp Val Leu Glu
340 345 350
His Arg Ala His Leu His Gly Thr Arg Tyr Tyr Pro Ser Pro Glu Val
355 360 365
Phe Leu Tyr Phe Val Ser Gln Leu Cys Arg Phe Ser Lys Arg Glu Pro
370 375 380
Thr Leu Gln Leu Leu Glu Thr Leu Leu Thr Asp Arg Leu Lys Glu Arg
385 390 395 400
Ile Gln Val Lys Ala Asp Thr Leu Ser Leu Ala Met Arg Ile Leu Ala
405 410 415
Cys Leu Ser Val Gly Ile Ser Gln Val Glu Val Asp Val Arg Glu Leu
420 425 430
Leu Ala Leu Gln Cys Lys Asp Gly Ser Trp Glu Pro Gly Ser Phe Tyr
435 440 445
Arg Phe Gly Ser Ser Lys Met Asn Val Gly Asn Arg Gly Leu Thr Thr
450 455 460
Ala Leu Ala Thr Arg Ala Val Glu Leu Tyr Gln Gly Thr Arg Ile Arg
465 470 475 480
Ser Lys Gly Thr Glu
485
<210> 61
<211> 1458
<212> DNA
<213> 乌达加瓦曲霉(Aspergillus udagawae)_GAO87501.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 61
atgacccgac agaaatcgcc tcaatacaaa gcaatcatct ttgacctagg ggatgtcttt 60
ttcacctggg acgcccccaa agacactgct gtcttgccca acctcttcaa gaaaatgctt 120
acctcgccaa cctggtcaga ttacgagcgc ggcaagttga gcgaagaaag ctgctacgag 180
agactggccg aacagtttga cgttgactcg tcggaaatcg cgcgcagctt aaggaaagca 240
cagcagtctc ttaccacaga cgcagcaatc gtgagcctga tatcagagat cagagcgttg 300
gccggacata ttgccatcta cgccatgtcc aacatttccg ccccagctta tgcagctgtg 360
ctccagactc agcccgaaat gggcatcttt gacggagtgt tcccgtctgg atgctatggg 420
acgaggaagc cggagctgtt gttctataag aaagtcttgc aggagattgc agtgccgcca 480
aatcagatca tctttattga tgatcagcta gagaatgtag tttctgcgca gtcaacaggt 540
atgcacggca ttgtctacac cggtgcgggt gagctcagtc gacagctcag aaatctggtg 600
ttggaccctg tacaaagggg tcgagagttt ctacggcgca atgctggggc attgtatagt 660
atctgcgaga ctggtcaagt catccgggaa aacttctcgc agctgctcat cctagaggcg 720
acgggtgata gaagcctggt caaccttgaa tatcagcagc ggagctggaa tttctttcaa 780
ggaggtcccc cttctacgtc ggaaacattc ccagatgatg tcgacacaac atccattgcc 840
ttgatgattc tccctgccga tgataacaca gtcaactcgg ttctcggcga gatttccgag 900
gtagctaatg acgagggcat tgtaaatacg tactttgacc agacccgaca gcgaatcgac 960
ccagcagtct gcgtcaatgt cctccgtctc ttttatacct acggccgggg cgccactctc 1020
ccattgaccc tccagtgggt gtccgacgtt cttgagcatc gtgcgcactt acatggtacg 1080
cgatactacc ccagcccgga ggttttcctc tactttgtca gtcaactctg ccggttctcc 1140
aagagggaac cgacgctgca gctgctggag acgttgctca cggatcgcct caaggagcgc 1200
attcaggtca aggcagacac tctgtcactg gctatgcgga tcctggcatg cttgtctgtg 1260
ggtatatcac aagttgaagt ggatgtccga gagctgctcg ccttgcaatg caaggatgga 1320
tcgtgggaac ccggctcgtt ttaccggttt gggtcgtcca agatgaacgt tggtaatcga 1380
ggtcttacga ctgcgttggc gactagggcg gttgagttgt accaggggac tagaatacgc 1440
tctaagggca ccgagtag 1458
<210> 62
<211> 1458
<212> DNA
<213> 人工序列_GAO87501.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 62
atgactcgcc aaaaaagccc tcaatacaaa gcaattatct tcgatctggg tgacgttttc 60
ttcacctggg atgcgccgaa agatacggcc gtactgccga acctgttcaa gaaaatgctg 120
acctcgccga cctggagcga ctatgagcgt ggtaagctgt ctgaggaaag ctgttacgaa 180
cgcttggccg agcaatttga cgtggacagc agcgagatcg cgcgtagcct ccgtaaagcg 240
cagcaaagcc tgacgaccga cgcagccatc gtgagcctga tcagcgagat ccgcgcattg 300
gcgggtcaca ttgctatcta tgctatgtct aacatttctg cgccagcata cgcagcggtg 360
ttacagaccc agccggaaat gggtatcttt gatggtgttt ttccgagcgg ctgctatggt 420
acgcgtaaac cggaactgct gttttacaaa aaagtgcttc aagaaattgc ggttccgccg 480
aatcagatta tcttcattga cgatcagctg gaaaacgtcg tcagcgcaca gtccacgggc 540
atgcatggca ttgtttacac cggtgccggt gagctgagcc gtcaactgcg taatctggtc 600
ctggacccgg tgcagcgtgg tcgtgagttc ctgcgccgta atgctggcgc cctgtacagc 660
atttgtgaga ctggccaagt tatccgtgag aacttcagcc agctgctgat tctggaagca 720
accggcgatc gttcgctggt gaacctggag tatcaacaac gttcctggaa cttctttcag 780
ggtggccctc catccacgag cgaaactttt ccggatgatg ttgacacgac ctcaatcgcg 840
ctgatgattt taccggcgga cgataatacc gtcaatagcg tcctgggtga aatcagcgaa 900
gtcgcgaatg acgagggcat tgtgaatacc tatttcgatc agacccgcca acgtatcgat 960
ccggccgtgt gtgtcaacgt gttgcgcctg ttttacacct atggtcgtgg cgctacgctg 1020
ccgttgaccc tgcaatgggt tagcgacgtg ctggagcacc gtgcgcatct gcacggcacc 1080
cgctactatc cgtccccaga ggttttcctg tactttgtct ctcagctgtg ccgtttttcc 1140
aagcgcgaac cgaccctgca gctgctggaa acgctgttga ccgacagact gaaggaacgc 1200
atccaagtta aggcagatac gctgagcttg gcaatgcgta ttttggcgtg cctgagcgtg 1260
ggcatcagcc aggttgaggt tgacgtccgc gaactgctgg cgctgcagtg caaggacggt 1320
agctgggagc cgggtagctt ctaccgtttc ggtagcagca agatgaatgt cggtaaccgc 1380
ggtctgacga ccgctttggc gacccgtgcg gttgagctgt accagggtac gcgtattcgt 1440
agcaagggca ccgagtaa 1458
<210> 63
<211> 524
<212> PRT
<213> 云芝栓孔菌(Trametes versicolor)_XP_008034151.1蛋白
<400> 63
Met Ala Ser Pro His Arg Arg Tyr Thr Thr Leu Ile Leu Asp Leu Gly
1 5 10 15
Asp Val Leu Phe Ser Trp Ser Ser Lys Thr Asn Thr Pro Ile Pro Pro
20 25 30
Lys Lys Leu Lys Glu Ile Leu Ser Ser Leu Thr Trp Phe Glu Tyr Glu
35 40 45
Arg Gly Arg Ile Ser Gln Ala Glu Cys Tyr Asp Arg Val Ser Ser Glu
50 55 60
Phe Ser Leu Asp Ala Ala Thr Ile Ala Glu Ala Phe Gln Gln Ala Arg
65 70 75 80
Asp Ser Leu Arg Pro Asn Glu Glu Phe Leu Ala Leu Ile Arg Glu Leu
85 90 95
Arg Gln Gln Thr His Gly Gln Leu Thr Val Leu Ala Leu Ser Asn Ile
100 105 110
Ser Leu Pro Asp Tyr Glu Tyr Ile Met Ala Leu Asp Ser Asp Trp Thr
115 120 125
Ser Val Phe Asp Arg Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys
130 135 140
Pro His Leu Gly Ala Tyr Arg Arg Val Ile Ser Glu Met His Leu Asp
145 150 155 160
Pro Glu Thr Thr Val Phe Val Asp Asp Lys Leu Asp Asn Val Val Ser
165 170 175
Ala Arg Ser Leu Gly Met His Gly Val Val Phe Asp Ser Gln Glu Asn
180 185 190
Val Phe Gln Thr Leu Arg Asn Ile Phe Gly Asp Pro Ile His Arg Gly
195 200 205
Arg Asp Tyr Leu Arg Arg His Ala Gly Arg Leu Glu Thr Ser Thr Asp
210 215 220
Ala Gly Val Val Phe Glu Glu Asn Phe Thr Gln Leu Ile Ile Tyr Glu
225 230 235 240
Leu Thr Asn Asp Lys Ser Leu Ile Thr Thr Ser Asp Cys Pro Arg Thr
245 250 255
Trp Asn Phe Phe Arg Gly Lys Pro Leu Phe Ser Ala Ser Phe Pro Asp
260 265 270
Asp Val Asp Thr Thr Ser Val Ala Leu Thr Val Leu Arg Pro Pro Arg
275 280 285
Thr Leu Val Asn Ser Ile Leu Asp Glu Met Leu Glu Tyr Val Asp Ala
290 295 300
Asp Gly Ile Met Gln Thr Tyr Phe Asp His Ser Arg Pro Arg Met Asp
305 310 315 320
Pro Phe Val Cys Val Asn Val Leu Ser Leu Phe Tyr Glu Tyr Gly Arg
325 330 335
Gly Gln Asp Leu Pro Lys Thr Leu Glu Trp Val Tyr Glu Val Leu Leu
340 345 350
His Arg Ala Tyr Ile Gly Gly Ser Arg Tyr Tyr Met Ser Ala Asp Cys
355 360 365
Phe Leu Phe Phe Met Ser Arg Leu Leu Gln Arg Ile Thr Asp Pro Ala
370 375 380
Val Leu Asn Arg Leu Arg Pro Leu Phe Val Glu Arg Met His Glu Arg
385 390 395 400
Val Ser Ala Pro Gly Asp Ser Met Glu Leu Ala Phe Arg Ile Leu Ala
405 410 415
Gly Ser Ser Val Gly Ile Gln Phe Pro Arg Asp Leu Glu Lys Leu Leu
420 425 430
Ala Ala Gln Cys Ala Asp Gly Gly Trp Asp Leu Cys Trp Phe Tyr Gln
435 440 445
Tyr Gly Ser Thr Gly Val Lys Ala Gly Asn Arg Gly Leu Thr Thr Ala
450 455 460
Leu Ala Ile Lys Ala Ile Glu Ser Ala Ile Ala Arg Pro Pro Ser Pro
465 470 475 480
Ala Leu Ser Ala Val Ser Ser Ser Lys Leu Glu Val Pro Lys Pro Ile
485 490 495
Leu Gln Arg Pro Leu Ser Pro Arg Arg Leu Gly Asp Phe Leu Met Pro
500 505 510
Trp Arg Arg Ala Gln Arg Glu Val Ala Val Ser Ser
515 520
<210> 64
<211> 1575
<212> DNA
<213> 云芝栓孔菌(Trametes versicolor)_XP_008034151.1 - cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 64
atggcttcac ctcaccgcag gtatacgaca ctcatcctag acctgggcga cgtcctcttc 60
tcttggtcat ccaagaccaa cacacctatc cctcccaaga agctgaagga gatcctctcg 120
tccctgacct ggttcgagta cgagcgcggt cggatatcac aggccgagtg ctatgaccgg 180
gtcagctccg agttcagtct tgacgctgcc accatcgcag aagcgttcca gcaggctcgc 240
gactctctgc gaccgaacga agagttcctg gcgttgattc gcgaactccg ccaacaaacg 300
catggtcagc ttaccgtcct cgcgctctcg aacatctcac tccccgacta tgaatacatc 360
atggctctcg actcggactg gacgtcggtc ttcgaccgcg tcttcccttc tgccctcgtc 420
ggcgagcgca agccacatct gggggcgtac cgccgtgtca tctctgagat gcacctagac 480
ccagaaacga ccgtctttgt ggacgacaag ctggacaacg tggtgtccgc gcgatcgctc 540
gggatgcacg gcgtggtctt cgactcccag gagaacgtct tccagacgct gaggaatatc 600
ttcggcgacc cgatacatcg cggacgtgac tatctccgca ggcatgccgg tcgtctggag 660
acatctacgg acgccggcgt tgtcttcgag gaaaacttta cgcagctcat catctacgaa 720
ctaacaaatg acaaatccct catcacgaca tcagactgtc cccgcacttg gaacttcttc 780
cgcgggaagc ccttgttctc ggcctcgttt cccgacgatg tggacacgac gtcggttgcc 840
ctgacagtgt tgcgcccacc ccgcacgctt gtcaactcga tcttggacga gatgctagag 900
tatgtcgacg ccgacggcat catgcagacc tacttcgacc actcgcgccc gcggatggat 960
ccgttcgtct gtgtcaacgt cctgtcgctg ttctacgagt acggccgggg acaggacctc 1020
ccgaagaccc tcgaatgggt atacgaggtt ctgctgcacc gcgcctacat cggcggctcg 1080
cggtactaca tgtccgcgga ctgcttcctc ttcttcatga gccgccttct ccaacgtatc 1140
accgacccag ccgtcctgaa ccgcctccgc ccgttgttcg tcgagcgcat gcacgaacgt 1200
gtcagcgcac cgggcgactc catggagctc gcgttccgca tcctcgctgg ctcgtccgtc 1260
ggcatccagt tcccacgtga cctggagaag ctcctcgccg cgcagtgcgc cgacggcggc 1320
tgggacctgt gctggttcta ccagtatggg tccaccggcg tgaaggcagg caaccgcggg 1380
ctcaccaccg cgctcgccat caaggctatc gagagcgcta tcgcgcgccc tccgtccccc 1440
gctctatcag ctgtatcgtc gtcgaaactg gaagtgccga aaccaattct ccagcgtccc 1500
ctcagcccgc gccggcttgg cgacttcctg atgccctgga ggagagcaca gcgcgaggtc 1560
gcggtttcca gctag 1575
<210> 65
<211> 1575
<212> DNA
<213> 人工序列_XP_008034151 - 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 65
atggctagcc cgcaccgtcg ctatactact ctgattctgg atttgggtga tgttttgttt 60
agctggagca gcaaaaccaa tacgcctatt ccgccgaaaa agctgaaaga aatcctgtct 120
agcctgacct ggttcgagta cgagcgcggt cgcatttctc aagccgagtg ctatgaccgt 180
gtgagctctg agtttagcct ggacgcagcg accattgcag aggcattcca acaggctcgt 240
gactcgctgc gcccgaacga agaatttctg gcgttgattc gtgagctgcg ccagcagacc 300
cacggccaac tcaccgttct ggcactgagc aacatctccc tgccggatta cgagtacatc 360
atggctctgg atagcgattg gaccagcgtc tttgatagag ttttcccgag cgcgctggtt 420
ggtgagcgta agccgcatct gggtgcttac cgtcgtgtca ttagcgagat gcatctggac 480
ccggagacta cggtgtttgt ggacgacaaa ctggacaacg ttgtctccgc gcgcagcctg 540
ggtatgcacg gcgtcgtttt tgactcacaa gaaaatgttt tccagacgct gcgtaacatt 600
ttcggtgacc ctatccaccg tggccgcgac tatttgcgtc gtcatgccgg tcgtttggaa 660
accagcaccg acgcgggcgt tgtttttgaa gaaaacttca cccagctgat catctacgaa 720
ctgacgaatg acaagagcct gatcaccacg agcgattgtc cgcgcacctg gaacttcttc 780
cgtggtaagc cgctgtttag cgcgtccttc ccagacgatg tcgatacgac ttcggtggcc 840
ctgaccgttc tgcgcccacc gcgcaccctg gtaaacagca tcctggacga aatgttagaa 900
tacgtcgatg cggatggtat tatgcagacc tatttcgacc acagccgtcc gcgcatggac 960
ccgtttgtgt gtgtgaatgt gttgagcctg ttctatgagt acggccgtgg tcaagatctg 1020
ccaaaaaccc tggaatgggt ctacgaagtc cttctgcatc gtgcctacat cggtggctcc 1080
cgttattaca tgagcgcaga ttgctttttg ttctttatgt ctcgtctgct gcagcgcatc 1140
acggaccctg ccgtgctgaa tcgtctgcgt ccgctgttcg tggagcgtat gcacgagcgc 1200
gtgtctgccc cgggtgacag catggaactg gcgttccgta tcctggcggg cagcagcgtg 1260
ggtattcaat ttccgcgtga tttggagaaa ctgctggctg cgcagtgtgc ggacggtggc 1320
tgggatctgt gctggtttta tcaatacggt agcaccggcg ttaaggccgg caatcgtggc 1380
ctgacgacgg cactggcaat taaggccatt gagtccgcga ttgcgcgtcc gccgagcccg 1440
gcattgagcg cggtcagcag cagcaaactg gaagtgccga agccgatctt gcagcgtcca 1500
ctgagcccgc gtcgtctggg tgacttcctg atgccgtggc gccgtgcgca acgcgaagtc 1560
gcggttagct cctaa 1575
<210> 66
<211> 527
<212> PRT
<213> 污叉丝孔菌(Dichomitus squalens)_XP_007369631.1蛋白
<400> 66
Met Ala Ser Ile His Arg Arg Tyr Thr Thr Leu Ile Leu Asp Leu Gly
1 5 10 15
Asp Val Leu Phe Arg Trp Ser Pro Lys Thr Glu Thr Ala Ile Pro Pro
20 25 30
Gln Gln Leu Lys Asp Ile Leu Ser Ser Val Thr Trp Phe Glu Tyr Glu
35 40 45
Arg Gly Arg Leu Ser Gln Glu Ala Cys Tyr Glu Arg Cys Ala Glu Glu
50 55 60
Phe Lys Ile Glu Ala Ser Val Ile Ala Glu Ala Phe Lys Gln Ala Arg
65 70 75 80
Gly Ser Leu Arg Pro Asn Glu Glu Phe Ile Ala Leu Ile Arg Asp Leu
85 90 95
Arg Arg Glu Met His Gly Asp Leu Thr Val Leu Ala Leu Ser Asn Ile
100 105 110
Ser Leu Pro Asp Tyr Glu Tyr Ile Met Ser Leu Ser Ser Asp Trp Thr
115 120 125
Thr Val Phe Asp Arg Val Phe Pro Ser Ala Leu Val Gly Glu Arg Lys
130 135 140
Pro His Leu Gly Cys Tyr Arg Lys Val Ile Ser Glu Met Asn Leu Glu
145 150 155 160
Pro Gln Thr Thr Val Phe Val Asp Asp Lys Leu Asp Asn Val Ala Ser
165 170 175
Ala Arg Ser Leu Gly Met His Gly Ile Val Phe Asp Asn Gln Ala Asn
180 185 190
Val Phe Arg Gln Leu Arg Asn Ile Phe Gly Asp Pro Ile Arg Arg Gly
195 200 205
Gln Glu Tyr Leu Arg Gly His Ala Gly Lys Leu Glu Ser Ser Thr Asp
210 215 220
Asn Gly Leu Ile Phe Glu Glu Asn Phe Thr Gln Leu Ile Ile Tyr Glu
225 230 235 240
Leu Thr Gln Asp Arg Thr Leu Ile Ser Leu Ser Glu Cys Pro Arg Thr
245 250 255
Trp Asn Phe Phe Arg Gly Glu Pro Leu Phe Ser Glu Thr Phe Pro Asp
260 265 270
Asp Val Asp Thr Thr Ser Val Ala Leu Thr Val Leu Gln Pro Asp Arg
275 280 285
Ala Leu Val Asn Ser Val Leu Asp Glu Met Leu Glu Tyr Val Asp Ala
290 295 300
Asp Gly Ile Met Gln Thr Tyr Phe Asp Arg Ser Arg Pro Arg Met Asp
305 310 315 320
Pro Phe Val Cys Val Asn Val Leu Ser Leu Phe Tyr Glu Asn Gly Arg
325 330 335
Gly His Glu Leu Pro Arg Thr Leu Asp Trp Val Tyr Glu Val Leu Leu
340 345 350
His Arg Ala Tyr His Gly Gly Ser Arg Tyr Tyr Leu Ser Pro Asp Cys
355 360 365
Phe Leu Phe Phe Met Ser Arg Leu Leu Lys Arg Ala Asp Asp Pro Ala
370 375 380
Val Gln Ala Arg Leu Arg Pro Leu Phe Val Glu Arg Val Asn Glu Arg
385 390 395 400
Val Gly Ala Ala Gly Asp Ser Met Asp Leu Ala Phe Arg Ile Leu Ala
405 410 415
Ala Ala Ser Val Gly Val Gln Cys Pro Arg Asp Leu Glu Arg Leu Thr
420 425 430
Ala Gly Gln Cys Asp Asp Gly Gly Trp Asp Leu Cys Trp Phe Tyr Val
435 440 445
Phe Gly Ser Thr Gly Val Lys Ala Gly Asn Arg Gly Leu Thr Thr Ala
450 455 460
Leu Ala Val Thr Ala Ile Gln Thr Ala Ile Gly Arg Pro Pro Ser Pro
465 470 475 480
Ser Pro Ser Ala Ala Ser Ser Ser Phe Arg Pro Ser Ser Pro Tyr Lys
485 490 495
Phe Leu Gly Ile Ser Arg Pro Ala Ser Pro Ile Arg Phe Gly Asp Leu
500 505 510
Leu Arg Pro Trp Arg Lys Met Ser Arg Ser Asn Leu Lys Ser Gln
515 520 525
<210> 67
<211> 1584
<212> DNA
<213> 污叉丝孔菌(Dichomitus squalens)_XP_007369631.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 67
atggcctcaa tccaccgtcg atacactact ctcatcctcg acctcggcga cgtactcttt 60
cgttggtctc caaagactga gaccgccatt ccacctcaac aactcaagga tatcctctcc 120
tctgtcacct ggtttgagta cgaacgcggc agactatccc aggaagcatg ctacgagcgc 180
tgcgccgagg agttcaagat agaggcctcg gtcattgcag aagcctttaa gcaggctcgc 240
gggtcactgc ggcccaacga ggagttcatc gccttgatcc gtgacctccg ccgtgagatg 300
cacggtgacc ttaccgttct tgccctctcc aacatctccc tccccgacta cgaatacatc 360
atgtcgctaa gctcagattg gacgaccgtc ttcgatcgcg tattcccctc tgcactcgtt 420
ggcgagcgca agcctcatct gggatgctat cgcaaggtca tctcggagat gaacctagaa 480
cctcagacga ctgtgttcgt ggatgacaag cttgacaacg tcgcgtctgc tcgctcactt 540
ggtatgcacg gcatcgtgtt tgacaaccaa gccaacgtct tccgccaact ccgcaatatc 600
ttcggagacc ccatccgccg tggccaagag tatctccgtg ggcatgctgg caaactcgag 660
tcttcgaccg acaacgggtt gatcttcgag gagaacttca cacagctgat catctacgag 720
ttgacgcaag acaggactct catctcgctt tcagaatgtc ctcgtacttg gaatttcttc 780
cgaggcgaac cgctattctc ggagaccttc ccggatgatg tcgacacaac atctgtggcg 840
ttgacggtat tgcaaccgga cagagcactg gtcaactccg ttctagacga gatgctggag 900
tatgtcgacg ccgatggcat catgcagaca tacttcgatc gttcacgacc acgcatggac 960
cccttcgtct gcgtgaacgt actctccctg ttctacgaga acggtcgtgg tcacgagctc 1020
cctcgcacat tggactgggt ctacgaggtg ctcctccatc gcgcgtacca cggcggttcg 1080
cgttattacc tgtcgcccga ctgctttcta ttcttcatga gccgcctact caagcgcgca 1140
gacgatccag cagtccaggc tcggctccgc ccgctcttcg tcgagcgggt gaacgagcga 1200
gtaggcgccg ctggcgactc gatggacctc gccttccgca tcctcgccgc agcgtctgtt 1260
ggcgtccagt gcccccgcga tctggaaagg ttgactgccg ggcaatgcga cgacggtgga 1320
tgggacctct gctggttcta cgtgttcggc tcgacgggcg tgaaggcggg caaccgcggc 1380
ctcacaacgg ccctcgctgt cacggccata cagacggcca tcggacgccc cccttcgccc 1440
agtccctccg cggcctcctc gtctttcaga cctagttccc cttacaaatt cctaggcatt 1500
tcgcgcccag ctagccccat tcgctttggc gacttacttc gcccatggcg gaagatgagc 1560
aggtcgaact tgaagtctca atga 1584
<210> 68
<211> 1584
<212> DNA
<213> 人工序列_XP_007369631.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 68
atggcaagca ttcatcgtcg ctatactacg ctgattctgg acctgggtga tgttttgttc 60
cgctggagcc cgaaaaccga gactgcgatt cctccgcaac aactgaaaga catcctgagc 120
agcgtcacct ggttcgagta cgagcgtggc cgtctgagcc aagaggcttg ctacgagcgt 180
tgcgccgaag agttcaagat tgaagccagc gtgattgcgg aagcgttcaa acaagcgcgt 240
ggtagcctgc gtccgaacga agaatttatc gcactgatcc gtgatctgcg tcgcgagatg 300
catggtgacc tgaccgttct ggctctgagc aatatctcgt tgccggatta cgagtatatt 360
atgtctctga gcagcgactg gacgacggtc tttgatcgtg tgttcccgtc agctctggtg 420
ggcgagcgta aaccgcactt gggttgctat cgcaaggtca tcagcgagat gaacctggaa 480
cctcagacca cggtctttgt ggacgataaa ctggataatg tcgcaagcgc gcgtagcctg 540
ggtatgcacg gtatcgtgtt tgataatcaa gcgaatgtgt ttcgccagct gcgtaatatt 600
ttcggtgatc caatccgtcg cggtcaagag tatctgcgtg gccatgccgg taaattggag 660
agcagcacgg acaatggttt gatctttgaa gagaacttca cccagctgat catttatgaa 720
ctgacccagg accgcacgtt gatcagcctg tcggagtgtc cgcgtacctg gaacttcttc 780
cgtggcgagc cgttgttttc tgaaaccttc ccggacgacg tggacaccac gtccgttgca 840
ctgacggttc tgcaaccgga tcgcgcactg gttaacagcg tgctggacga aatgctggaa 900
tatgtcgatg cggatggcat catgcagacg tatttcgacc gctcgcgtcc gcgtatggac 960
ccgtttgttt gcgtcaacgt actgagcctg ttttacgaga acggtcgtgg tcacgaactg 1020
ccgcgcactc tggattgggt gtacgaagtc ctgctccacc gcgcctacca cggtggttcc 1080
cgttactacc tgagcccgga ctgtttcttg ttttttatga gccgtctgct gaaacgtgca 1140
gacgacccag cggttcaggc gagattgcgt ccgctgtttg tggaacgcgt taacgaacgt 1200
gttggcgcgg ccggtgatag catggacctg gcgtttcgca ttctggccgc agcgagcgtg 1260
ggtgtgcagt gtccgcgcga cctggagcgt ctgaccgctg gtcaatgcga tgatggcggc 1320
tgggatctgt gttggttcta cgttttcggc agcaccggcg ttaaggccgg taatcgtggt 1380
ctgaccacgg cgctggcagt caccgcgatc cagaccgcca tcggccgtcc gcctagcccg 1440
agcccgtccg cggcaagctc cagcttccgc ccgagcagcc cgtacaagtt tctgggtatt 1500
agccgtccgg cgtccccaat tcgcttcggt gaccttctgc gtccgtggcg taaaatgtct 1560
cgctctaacc tgaagtccca gtaa 1584
<210> 69
<211> 495
<212> PRT
<213> 牛樟芝(Antrodia cinnamomea)_ACg006372蛋白
<400> 69
Met Arg Arg Asn Val Leu Asn Lys Ala Thr His Ser Gln Ser Pro Leu
1 5 10 15
Lys Pro Asn Ile Thr Thr Leu Ile Phe Asp Leu Gly Asp Val Leu Leu
20 25 30
Thr Trp Ser Asp Ser Thr Pro Lys Ser Pro Leu Pro Pro Lys Ile Val
35 40 45
Lys Gly Ile Leu Arg Ser Leu Thr Trp Phe Glu Tyr Glu Lys Gly Asn
50 55 60
Leu Thr Glu Ser Gln Thr Tyr Gly Gln Val Ala Gln Glu Phe Gly Val
65 70 75 80
Asp Ala Ser Glu Val Lys Ala Ser Phe Glu Ala Ala Arg Asp Ser Leu
85 90 95
Lys Ser Asn Pro Met Leu Leu Gln Leu Ile Arg Ser Leu Lys Asp Ser
100 105 110
Gly His Val Ile Tyr Ala Met Ser Asn Ile Ser Ala Pro Asp Trp Glu
115 120 125
Phe Leu Lys Thr Arg Ala Asp Leu Ser Asp Trp Ala Leu Phe Asp Arg
130 135 140
Val Phe Pro Ser Ala Glu Ala His Asp Arg Lys Pro Asn Ile Gly Phe
145 150 155 160
Tyr Gln His Val Ile Asn Glu Thr Gly Leu Asn Pro Ser Asn Thr Val
165 170 175
Phe Val Asp Asp Arg Ile Glu Asn Val Val Ser Ala Arg Ser Ala Gly
180 185 190
Met His Gly Ile Val Phe Asp Asp Ile Asn Asn Val Ile Arg Gln Leu
195 200 205
Lys Asn Leu Cys Glu Asp Pro Ile His Arg Ala Arg Ser Phe Leu Tyr
210 215 220
Ala Asn Lys Lys Cys Leu Asn Thr Val Ser Thr Asp Gly Thr Ile Val
225 230 235 240
Ser Glu Asn Phe Ser Gln Leu Leu Ile Leu Glu Ala Ile Gly Asp Glu
245 250 255
Ser Leu Val Asp Phe Val Arg His Glu Gly Arg Phe Asn Phe Phe Gln
260 265 270
Gly Glu Ala Lys Leu Ile Met Thr Asn His Tyr Pro Asp Asp Phe Asp
275 280 285
Thr Thr Ser Ile Gly Leu Thr Val Val Pro Tyr Ile Asp Asp Lys Thr
290 295 300
Arg Asn Arg Val Met Asp Glu Ile Leu Ala Tyr Gln Ser Glu Asp Gly
305 310 315 320
Ile Val Leu Val Tyr Phe Asp His Lys Arg Pro Arg Ile Asp Pro Val
325 330 335
Val Cys Val Asn Val Leu Thr Leu Phe Tyr Arg Tyr Gly Arg Gly His
340 345 350
Gln Leu Gln Lys Thr Leu Asp Trp Val Glu Gln Val Leu Ile Asn Arg
355 360 365
Ala Cys Ala Ser Gly Thr Phe Tyr Tyr Ala Thr Glu Glu Gln Phe Leu
370 375 380
Phe Phe Leu Ser Arg Leu Ile Gln Ser Ser Pro Asp Val Arg Gln Arg
385 390 395 400
Leu Glu Gly Val Phe Lys Arg Arg Val Val Glu Arg Phe Gly Ala Asp
405 410 415
Gly Asp Ala Leu Ala Met Ala Met Arg Ile His Thr Ala Ala Ser Val
420 425 430
Gly Leu Val Asp His Val Asp Leu Asp Lys Leu Phe Ala Leu Gln Gln
435 440 445
Asn Asp Gly Ser Trp Arg Asp Ser Ala Phe Tyr Arg Phe Pro Ser Ala
450 455 460
Arg Gln Leu Ala Ser Asn Asp Gly Leu Thr Thr Ala Ile Ala Ile Gln
465 470 475 480
Ala Ile Gln Ala Ala Glu Arg Leu Arg Glu Asp Gly Asn Val Leu
485 490 495
<210> 70
<211> 1488
<212> DNA
<213> 牛樟芝(Antrodia cinnamomea)_ACg006372 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 70
atgaggcgaa acgtactcaa caaagcaaca cattctcagt caccattgaa gcccaacatc 60
acgacgctca tatttgactt gggcgacgta cttctcacgt ggtccgactc aacacctaaa 120
tctccactgc ccccaaaaat tgtcaaggga atactacgtt cactgacctg gtttgagtac 180
gagaaaggga acttgacaga gtcccagacc tacgggcaag ttgctcagga atttggagtg 240
gatgcttccg aagtcaaagc ttccttcgaa gcagctcgcg actcgctcaa gagcaaccca 300
atgcttctcc agttgatccg tagcctcaaa gactctggcc acgtcattta cgcaatgtct 360
aacatatctg ctcccgactg ggaatttttg aagacgcggg cagacctctc agattgggct 420
ctttttgaca gagtcttccc ttctgccgaa gcgcatgacc gcaagccgaa cattggtttc 480
tatcagcacg tcataaacga gactggtctg aacccgtcca acactgtctt tgtcgatgac 540
aggatcgaga atgttgtatc cgcacgctca gcaggaatgc acgggatcgt gtttgacgac 600
ataaataatg tgatccgaca gttgaaaaac ctctgcgagg atccgattca ccgcgcacga 660
tcttttcttt atgcaaataa gaagtgtttg aatacggtta gcacagatgg cacaattgtg 720
agcgagaact tctcgcaatt gttgatcctt gaggccattg gcgacgaaag cctagtcgac 780
tttgtgaggc atgagggccg attcaacttc ttccaggggg aggccaaact catcatgacg 840
aatcactacc ccgatgattt cgatactaca tccataggtt taaccgttgt tccatatatt 900
gacgacaaga ctagaaatag agttatggat gagatcctgg cctaccaaag cgaagacggc 960
attgtgctgg tatactttga ccacaagcgc cccaggattg atcctgttgt ctgtgtcaat 1020
gtcctcaccc tcttctatag gtatggccgt gggcaccagc ttcaaaagac actggattgg 1080
gtcgaacagg tcctgatcaa ccgtgcgtgt gcgtccggca cgttctatta cgcaacagag 1140
gaacaattcc tctttttcct ctcccgcctg atccaaagct ctccggacgt acgacagcgg 1200
ttggaagggg tctttaaaag aagagtagtc gagcggtttg gtgcagacgg cgacgctctc 1260
gctatggcga tgcgcattca caccgcggcg agcgtgggcc tcgttgacca tgtcgatctt 1320
gacaagctgt tcgcattgca gcaaaatgac ggttcttgga gagacagcgc tttctacaga 1380
tttccgtcgg ccaggcaact ggctagtaac gacggcttga cgactgcaat cgctattcag 1440
gccattcaag ctgcggagag gctcagggag gatgggaacg tgctttga 1488
<210> 71
<211> 1488
<212> DNA
<213> 人工序列_ACg006372 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 71
atgcgccgta atgtcctgaa caaagcaacc catagccagt caccgttgaa accgaatatc 60
accacgctga tttttgactt gggcgatgtc ctgctgacct ggagcgacag cactccgaaa 120
tctccgttgc cgccgaagat cgtcaagggc atcctgcgta gcctgacttg gttcgagtac 180
gaaaagggca atttgaccga aagccaaacg tatggtcagg tcgcgcaaga atttggtgtg 240
gatgcctctg aagtgaaggc cagctttgag gctgcgcgtg atagcttgaa atcgaatccg 300
atgctgctgc agctgattcg cagcctgaaa gattccggtc acgtgatcta cgccatgagc 360
aacatcagcg cgcctgattg ggaatttctg aaaacccgcg ctgacctgtc tgactgggcc 420
ctgtttgacc gcgtgttccc gtctgccgag gcacatgacc gcaaaccgaa cattggcttt 480
taccaacacg tgatcaatga aacgggtctg aatccatcca ataccgtgtt cgttgacgac 540
cgtattgaaa acgttgttag cgcacgtagc gctggtatgc acggtatcgt tttcgatgac 600
attaacaacg tcattcgcca gctgaagaat ctgtgcgagg acccaattca ccgtgcacgt 660
tcctttttgt atgcgaacaa aaagtgcctg aataccgtga gcaccgatgg tacgatcgtc 720
agcgagaact ttagccagct tctgattctg gaagccattg gtgacgagtc cctggtagac 780
ttcgtccgcc atgagggccg ttttaacttc ttccagggtg aggcaaagct gatcatgacc 840
aatcactacc cggacgattt cgataccacg agcattggtc tgaccgttgt cccgtatatc 900
gatgacaaaa cgcgtaatcg tgtgatggat gaaatcctgg cgtatcagtc cgaggatggt 960
atcgttctgg tgtacttcga tcacaagcgt ccgcgcattg acccggtcgt ttgtgtgaac 1020
gttctgacgc tgttctaccg ctatggtcgt ggccatcaac tgcagaaaac cctggactgg 1080
gttgagcaag tcctgattaa tcgtgcgtgt gcgagcggca cgttctacta cgcgaccgaa 1140
gaacagttcc tgtttttcct gagccgtctg attcagtcga gccctgacgt gcgccaacgt 1200
ctggaaggcg tgttcaagcg tcgtgtcgtt gagcgctttg gtgcggacgg tgatgccctg 1260
gcaatggcga tgcgtatcca taccgcagcg agcgttggcc tggtggacca cgtggatctg 1320
gataagctgt tcgcgctgca acagaacgac ggtagctggc gcgatagcgc gttttatcgt 1380
tttccgagcg cgcgtcaact cgcgagcaac gacggcttga ccacggcaat tgctattcag 1440
gccatccaag cggctgagag attacgtgag gatggtaacg ttctgtaa 1488
<210> 72
<211> 543
<212> PRT
<213> 焦曲霉(Aspergillus ustus)_KIA75676.1蛋白
<400> 72
Met Val Arg Ala Leu Ile Leu Asp Leu Gly Asp Val Leu Phe Asn Trp
1 5 10 15
Asp Ala Pro Lys Ser Thr Pro Val Ser Arg Lys Thr Leu Ser Gln Met
20 25 30
Leu His Ser Asp Ile Trp Gly Glu Tyr Glu Cys Gly Gln Leu Thr Glu
35 40 45
Pro Glu Ser Tyr Lys Ala Leu Ala Ser Arg Tyr Ser Cys Gln Ala Gln
50 55 60
Asp Val Ala Asp Thr Phe Tyr Leu Ala Arg Glu Ser Leu Arg Leu Asp
65 70 75 80
Ala Thr Phe Lys Thr Phe Leu Gln Asp Leu Lys Gln Arg Ala Asn Gly
85 90 95
Ser Leu Arg Val Tyr Gly Met Ser Asn Ile Ser Gln Pro Asp Tyr Glu
100 105 110
Val Leu Leu Ser Lys Ala Asp Asp Leu Ser Leu Phe Asp Lys Ile Phe
115 120 125
Pro Ser Gly His Val Gly Met Arg Lys Pro Asp Leu Ala Phe Phe Arg
130 135 140
His Val Leu Arg Glu Ile Ser Thr Ala Ser Glu Asp Ile Val Phe Val
145 150 155 160
Asp Asp Asn Leu Glu Asn Val Thr Ser Ala Arg Ser Leu Gly Met Gln
165 170 175
Gly Ile Val Phe Arg Asp Lys Glu Asp Val Gln Arg Gln Leu Arg Asn
180 185 190
Leu Phe Gly Ser Pro Ala Glu Arg Gly Arg Glu Tyr Leu Ser Ile Asn
195 200 205
Lys Thr Lys Leu Gln Ser Val Thr Thr Thr Asn Ile Pro Ile Leu Asp
210 215 220
Asn Phe Gly Gln Leu Leu Ile Leu Glu Ala Thr Arg Asp Pro Asp Leu
225 230 235 240
Val Ser Met His Pro Gly Gln Arg Thr Trp Asn Phe Phe Ile Gly Ser
245 250 255
Pro Thr Leu Thr Thr Asp Ala Phe Pro Asp Asp Met Asp Thr Thr Ser
260 265 270
Leu Gly Leu Ser Ile Ile Pro Pro Ser Pro Glu Ile Ala Ala Ser Val
275 280 285
Met Asp Glu Ile Val Thr Arg Leu Asn Lys Asp Gly Ile Val Pro Thr
290 295 300
Tyr Phe Asp Ser Thr Arg Pro Arg Val Asp Pro Ile Val Cys Val Asn
305 310 315 320
Val Leu Thr Leu Phe Ala Lys Tyr Gly Arg Glu Asp Glu Leu Ser Gly
325 330 335
Thr Ile Ala Trp Val Arg Asp Val Leu Tyr His Arg Ala Tyr Leu Ala
340 345 350
Gly Thr Arg Tyr Tyr Ala Ser Pro Glu Ala Phe Leu Phe Phe Phe Thr
355 360 365
Arg Phe Thr Arg Asn Leu Arg Pro Gly Pro Arg Lys Gln Glu Leu Thr
370 375 380
Ala Leu Leu Ser Gln Arg Leu Gln Glu Arg Asn Lys Thr Pro Val Asp
385 390 395 400
Ala Leu Ala Leu Ser Met Arg Ile Ile Ala Cys Leu Thr Leu Gly Ile
405 410 415
Glu Ser Pro Ala Asp Asp Val Ala Thr Leu Thr Gly Met Gln Cys Gly
420 425 430
Asp Gly Gly Trp Pro Ala Cys Val Ile Tyr Lys Tyr Gly Ala Gly Gly
435 440 445
Leu Gly Ile Thr Asn Arg Gly Val Ser Thr Ala Phe Ala Val Lys Ala
450 455 460
Ile Thr Thr Thr Pro Leu Ala Val Gln Pro Glu Val Ser Val Ser Ala
465 470 475 480
Gly Ala Gly Gly Ser Ser Arg Pro Val Gly Ala Asp Ala Ala Ala Val
485 490 495
Ser Leu Arg Pro Arg Trp Arg Ala Val Val Gln Ser Leu His Pro Leu
500 505 510
Ser Arg Val Gly Gly Leu Val Ala Val Ile Phe Ala Ala Leu His Phe
515 520 525
Asn Leu Ala Trp Leu Tyr Asn Val Ser Leu Ala Ser Arg Ile Val
530 535 540
<210> 73
<211> 1632
<212> DNA
<213> 焦曲霉(Aspergillus ustus)_KIA75676.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 73
atggtccgcg cactgattct cgatctcggc gacgtcctct tcaactggga cgccccaaag 60
tcaacccccg tttcccgcaa gacactcagc cagatgctgc atagcgacat ctggggcgaa 120
tacgaatgtg gccaactgac agagccggaa agctacaagg cgcttgccag ccgctattct 180
tgccaggctc aagatgttgc agataccttc tatctagccc gcgaatcgct gaggctcgat 240
gcgaccttca agaccttcct gcaggacttg aagcagaggg ccaacggctc acttcgcgta 300
tatgggatgt ccaacatctc ccagcccgat tatgaggtcc tgctgtccaa ggcggatgac 360
ttgagcctgt ttgacaagat cttcccatcc ggccacgtcg ggatgcgtaa gcctgacctt 420
gcgttttttc gacatgtcct gcgtgagatc tcgacggcca gcgaggatat tgtgtttgtt 480
gacgacaacc tggagaacgt gacatctgcc cggtctctgg gcatgcaggg gattgtcttt 540
cgcgacaagg aggatgtaca gagacagctg cggaacctct ttggcagtcc tgctgaacgt 600
ggaagggagt atttgtccat caacaagaca aagctccaga gcgtcacgac gaccaatatc 660
cccattctcg acaactttgg ccagctcctt atcctcgaag ccaccagaga cccagacctg 720
gtgtccatgc atcctggaca gaggacctgg aactttttca tcggatctcc aactctgaca 780
acggacgcct tcccagacga tatggacacc acctcacttg gcctttctat tataccccca 840
agtcccgaga ttgcagcgtc cgtgatggat gagattgtga cccgcctgaa caaggacggc 900
attgtcccaa catattttga cagcaccaga ccccgcgtcg acccgatcgt ctgcgtcaac 960
gttctcaccc tcttcgctaa atacggccgc gaagacgagc tgtccgggac catagcctgg 1020
gtgcgcgatg tgctgtatca cagggcctac cttgcaggga ccagatacta cgcatcccca 1080
gaagcattcc ttttcttctt cacgcgcttc acccgaaacc tgcgcccggg cccgcgcaag 1140
caggagctca cggcgctgct gtcccagcgc ctgcaggagc gcaacaagac gcccgttgac 1200
gcacttgcgc tctcgatgcg gattattgcg tgcctcacgc tgggtattga atcccccgct 1260
gacgacgtgg ctaccctcac gggcatgcag tgtggggatg gcgggtggcc ggcctgtgtc 1320
atctacaagt acggcgccgg tgggctgggg atcacgaaca ggggggtctc gaccgcgttt 1380
gctgtcaagg caatcactac tactcctttg gcggtgcagc ctgaagttag tgtcagcgca 1440
ggtgcaggag gcagcagtcg ccctgtgggt gccgatgctg ctgcagtctc gctccgcccg 1500
agatggcgag ctgttgtgca gagtctccat ccgctctctc gggttggtgg gttggtggcc 1560
gtcatttttg ctgcactgca tttcaacttg gcctggcttt ataatgtgtc ccttgctagt 1620
aggatcgttt ag 1632
<210> 74
<211> 1632
<212> DNA
<213> 人工序列_KIA75676.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 74
atggttcgtg cattgatttt ggatttgggt gatgtgttgt ttaactggga tgcgcctaag 60
agcaccccgg tttcccgcaa gactctgagc caaatgctgc actcggatat ttggggcgag 120
tacgagtgtg gtcaactgac tgagccggag tcctataaag ccctggcgag ccgctatagc 180
tgccaggcgc aagatgtcgc tgacaccttt tacctggcgc gtgagagcct gcgtctggac 240
gcaacgttta agaccttcct gcaagatctg aagcaacgcg ccaacggttc tctgcgtgtc 300
tatggtatga gcaatatcag ccagccggat tacgaagtcc tgctgagcaa agctgacgat 360
ctcagcctgt ttgacaaaat ctttccgtcg ggtcacgttg gtatgagaaa gcctgacctg 420
gcgtttttcc gtcacgttct gcgtgagatc agcacggcta gcgaagatat tgtgtttgtt 480
gacgacaatt tggaaaacgt cacgtctgca cgctccctgg gtatgcaagg catcgtcttt 540
cgtgataagg aagatgtcca gcgccagctg cgcaatctgt tcggttcccc ggcagagcgc 600
ggtcgtgagt atctgagcat taataagacc aaactgcaga gcgtgaccac caccaatatc 660
ccgattctgg acaacttcgg tcagttgctg atcctggaag ctacccgtga cccggattta 720
gtcagcatgc atccaggcca acgtacgtgg aacttcttca ttggcagccc gaccttgacg 780
accgacgcgt ttccggacga tatggacacg acttctctgg gcctgagcat catcccgccg 840
agcccggaaa ttgcagcaag cgttatggac gaaatcgtca cccgtctgaa taaagatggt 900
attgtgccga cctacttcga cagcacgcgt ccacgtgtgg acccgatcgt ctgcgttaac 960
gtcctgacct tgtttgcgaa atatggtcgt gaagatgaac tgagcggcac gattgcgtgg 1020
gtccgcgacg ttctgtatca tcgcgcatac ctggcgggca cgcgctacta cgcgtcccca 1080
gaggccttcc tgttcttctt tacgcgtttc acccgcaatc tgcgtccggg tccgcgtaaa 1140
caagaactta cggcgctgct gagccagcgt ctgcaggaac gcaacaagac gccggttgac 1200
gctctggccc tgagcatgcg tatcatcgcc tgtctgaccc tgggcattga gagcccggca 1260
gacgacgtgg ccaccctgac cggtatgcag tgtggtgatg gtggctggcc ggcgtgcgtg 1320
atctacaaat atggtgcggg tggcttgggt atcacgaatc gtggcgttag cactgccttc 1380
gcggtgaaag cgattacgac caccccgctg gcagtgcagc cagaagtcag cgtcagcgct 1440
ggtgccggcg gctccagccg cccggttggt gcggatgcgg cagcggttag cttgcgtccg 1500
cgttggcgtg cggttgtgca gagcctgcat ccgctgagcc gcgtgggtgg cctggttgcc 1560
gtgatcttcg cggcactgca ctttaacctg gcgtggctgt acaacgtaag cctggctagc 1620
cgtattgtgt aa 1632
<210> 75
<211> 477
<212> PRT
<213> 米曲霉(Aspergillus oryzae)_XP_001820867.2蛋白
<400> 75
Met Thr Arg Trp Lys Ser Ser Gln Tyr Gln Ala Ile Ile Phe Asp Leu
1 5 10 15
Gly Gly Val Ile Leu Thr Trp Asp Leu Pro Glu Asp Thr Val Ile Ser
20 25 30
Ala Gln Ile Phe Lys Arg Met Leu Thr Ser Gln Thr Trp Ser Asp Tyr
35 40 45
Glu Arg Gly Asn Leu Ser Glu Asn Gly Cys Tyr Gln Arg Leu Ala Glu
50 55 60
Asp Phe Gly Ile Asp Ser Ala Asp Ile Ala His Thr Val Arg Gln Ala
65 70 75 80
Arg Glu Ser Leu Val Thr Asp Thr Ala Ile Met Asn Ile Ile Ser Glu
85 90 95
Ile Arg Ala Gly Ala Asn His Ile Ala Ile Phe Ala Met Ser Asn Ile
100 105 110
Ser Gln Pro Asp Tyr Ala Ala Leu Leu Leu Asp His Arg Gly Met Cys
115 120 125
Ser Phe Asp Arg Val Phe Pro Ser Gly Cys Tyr Gly Thr Arg Lys Pro
130 135 140
Glu Leu Ser Phe Tyr Asn Lys Val Leu Arg Glu Ile Asp Thr Pro Pro
145 150 155 160
Glu Asn Val Ile Phe Val Asp Asp Gln Leu Glu Asn Val Ile Ser Ala
165 170 175
Gln Ser Ile Gly Ile His Gly Ile Ala Tyr Thr Asn Ala Ala Glu Leu
180 185 190
Gly Arg Gln Leu Arg Asn Leu Ile Phe Asp Pro Val Glu Arg Gly Arg
195 200 205
Glu Phe Leu Arg Arg Asn Ala Gly Glu Phe His Ser Ile Thr Glu Thr
210 215 220
Asp Gln Ile Val Arg Glu Asn Phe Ser Gln Leu Leu Ile Leu Glu Ala
225 230 235 240
Thr Gly Asp Lys Ser Leu Val Ser Leu Glu Tyr His Gln Lys Ser Trp
245 250 255
Asn Phe Phe Gln Gly Asn Pro Ile Leu Thr Thr Glu Thr Phe Pro Asp
260 265 270
Asp Val Asp Thr Thr Ser Leu Ala Leu Met Thr Leu Pro Thr Asp Thr
275 280 285
Lys Thr Ala Asn Leu Leu Leu Asp Gln Ile Leu Gly Leu Val Asn Ala
290 295 300
Asp Glu Ile Val Thr Thr Tyr Phe Asp Gln Thr Arg Glu Arg Ile Asp
305 310 315 320
Pro Val Val Cys Val Asn Val Leu Arg Leu Phe Cys Thr Tyr Gly Arg
325 330 335
Gly Ile Ala Leu Pro Leu Thr Leu Gln Trp Val Tyr Asp Val Leu Ala
340 345 350
His Arg Ala Tyr Ile Asn Gly Thr Arg Tyr Tyr Thr Ser Pro Glu Ser
355 360 365
Phe Leu Tyr Phe Val Gly Gln Leu Cys Arg Phe Ser Thr Gly Val Leu
370 375 380
Ala Leu Arg Pro Leu Glu Thr Leu Leu Ile Asp Arg Leu Lys Glu Arg
385 390 395 400
Leu Gln Val Lys Ala Asp Pro Leu Ser Leu Ala Met Arg Ile Leu Thr
405 410 415
Cys Leu Ser Val Gly Val Ser Gln Val Glu Val Asp Leu Arg Glu Leu
420 425 430
Leu Ser Met Gln Cys Glu Asp Gly Ser Trp Glu His Cys Pro Phe Thr
435 440 445
Arg Tyr Gly Leu Ser Lys Val Ser Ile Gly Asn Arg Gly Leu Thr Thr
450 455 460
Ala Phe Val Val Lys Ala Val Glu Met Cys Arg Gly Ser
465 470 475
<210> 76
<211> 1434
<212> DNA
<213> 米曲霉(Aspergillus oryzae)_XP_001820867.2 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 76
atgactcgat ggaaatcgtc ccaataccaa gcaattatct ttgacctagg cggtgtcatt 60
ttaacatggg acctcccgga agacactgtg atatcggccc agatctttaa gagaatgctc 120
acatcgcaga catggtcaga ttatgagcgc ggaaatctca gcgaaaatgg ttgctaccag 180
aggttggccg aggattttgg cattgactct gccgacattg cacataccgt tagacaagca 240
cgggaatccc ttgtcactga taccgctatc atgaacatta tatctgagat cagagctggg 300
gctaaccata ttgctatctt cgctatgtcg aacatctccc aaccagatta tgcggctctg 360
ctccttgatc atcgcgggat gtgcagtttt gaccgggtgt tcccatctgg atgctacggg 420
acaaggaaac cagagctctc attctataac aaagtcttgc gggagattga cacgccaccg 480
gaaaacgtca tctttgtcga tgatcagctg gaaaatgtga tctctgcgca gtccattggc 540
atacacggga ttgcctatac gaatgctgct gaactcggtc gacagcttag gaacctaata 600
tttgaccctg tagagagggg tagggaattc ttacggcgca atgctggaga gttccatagc 660
atcactgaaa ccgatcaaat tgttcgggaa aatttctcac agttgctcat tctagaagcg 720
actggtgata agagtctggt atctcttgaa tatcaccaga agagctggaa tttcttccaa 780
ggaaacccta ttctcacgac agagacattc ccagatgatg ttgacacaac atctcttgcc 840
ttgatgactc tacctacaga cacaaaaact gcaaatttgt tactcgacca gattttgggg 900
ctagtcaacg ctgatgaaat cgtaacaaca tactttgacc agacccgaga acggatcgat 960
ccagtagtct gcgtcaatgt ccttcgtctc ttttgcacct acggccgggg cattgcgctc 1020
cctttgactc ttcagtgggt gtacgacgtc ctcgctcatc gggcatatat aaacggtaca 1080
cgttactaca caagtcccga aagcttccta tacttcgtcg gtcaactttg tcgattctca 1140
acaggggtac tggcacttcg gccgctggaa acgttgctta tagatcgtct caaggaacgt 1200
cttcaggtca aagcagatcc tctatcactc gctatgcgga tcttgacctg tttgtccgtt 1260
ggtgtgtctc aagttgaagt cgatctccga gagttgctct cgatgcagtg tgaagatggc 1320
tcgtgggaac attgtccatt cacccggtat ggtttgtcca aagtgagcat tggcaatcgg 1380
ggccttacaa ctgcttttgt ggtcaaggcg gttgaaatgt gtcgaggcag ttag 1434
<210> 77
<211> 1434
<212> DNA
<213> 人工序列_XP_001820867.2 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 77
atgactcgtt ggaaaagctc tcaatatcag gcaatcattt tcgatctggg cggtgttatt 60
ctgacctggg acttgccgga agatacggtt atctccgcgc aaatctttaa gcgtatgctg 120
accagccaga cctggtccga ttatgagcgc ggtaatctga gcgagaacgg ctgctatcaa 180
cgtttggcgg aagatttcgg catcgatagc gccgatattg cccacaccgt ccgtcaggca 240
cgtgagtccc tggtgaccga caccgccatc atgaatatca tctccgagat ccgtgcaggc 300
gcgaaccaca tcgcaatttt cgcgatgagc aacatctcac agccggatta cgctgcgctg 360
ctgctggacc atcgcggtat gtgcagcttt gaccgcgtct ttccgagcgg ttgttacggc 420
acccgtaagc ctgagctgag cttctacaat aaagtgctgc gtgaaattga caccccgccg 480
gaaaatgtta ttttcgttga cgatcaattg gaaaatgtga ttagcgcgca aagcattggt 540
attcatggca ttgcgtatac gaatgccgcg gaactgggcc gccagctgag aaacctgatc 600
ttcgatccgg tggagcgcgg tcgtgagttc ctgcgtcgta acgctggtga gtttcactct 660
attacggaaa cggaccagat tgtgcgcgag aacttcagcc agctgctgat tctggaagcg 720
accggtgaca aaagcctggt tagcctggaa taccaccaaa agtcgtggaa cttcttccaa 780
ggtaacccaa tcctgacgac ggaaaccttc ccggacgatg ttgacactac tagcctggct 840
ctgatgacgc tgccgacgga caccaagacc gcgaatctgt tgctggacca gattctgggt 900
ttggttaatg ccgatgaaat tgtgactacg tacttcgacc agacccgtga gcgtatcgat 960
ccagtggtct gtgtgaatgt cctgcgcctg ttctgtacgt acggccgcgg catcgcgctg 1020
ccgctgaccc tgcaatgggt ctacgatgtg ctggcgcacc gcgcatacat taacggtacg 1080
cgttattaca ccagcccgga gagctttctg tattttgtcg gtcagctctg tcgttttagc 1140
accggtgtgc tggcactgcg tccgctggag actctgctga ttgatcgtct gaaagagcgc 1200
ctgcaagtta aagctgaccc gctgagcctg gcaatgcgca tccttacgtg cttatctgtc 1260
ggtgtcagcc aggttgaagt ggacttgcgt gagttgttga gcatgcagtg cgaggacggt 1320
agctgggagc attgcccgtt cacccgctac ggcctgagca aggtttccat cggtaaccgt 1380
ggcctgacca cggcgtttgt ggttaaagcc gtcgagatgt gccgtggcag ctaa 1434
<210> 78
<211> 528
<212> PRT
<213> 热焦曲霉(Aspergillus calidoustus)_CEN60542.1蛋白
<400> 78
Met Val Arg Ala Leu Ile Leu Asp Leu Gly Asp Val Leu Phe Asn Trp
1 5 10 15
Asp Ala Pro Ala Ser Thr Pro Ile Ser Arg Lys Thr Leu Gly Gln Met
20 25 30
Leu His Ser Glu Ile Trp Gly Glu Tyr Glu Arg Gly His Leu Thr Glu
35 40 45
Asp Glu Ala Tyr Asn Ala Leu Ala Lys Arg Tyr Ser Cys Glu Ala Lys
50 55 60
Asp Val Ala His Thr Phe Val Leu Ala Arg Glu Ser Leu Arg Leu Asp
65 70 75 80
Thr Lys Phe Lys Thr Phe Leu Gln Thr Leu Lys Gln Asn Ala Asn Gly
85 90 95
Ser Leu Arg Val Tyr Gly Met Ser Asn Ile Ser Lys Pro Asp Phe Glu
100 105 110
Val Leu Leu Gly Lys Ala Asp Asp Trp Thr Leu Phe Asp Lys Ile Phe
115 120 125
Pro Ser Gly His Val Gly Met Arg Lys Pro Asp Leu Ala Phe Phe Arg
130 135 140
Tyr Val Leu Lys Asp Ile Ser Thr Pro Val Glu Asp Val Val Phe Val
145 150 155 160
Asp Asp Asn Leu Asp Asn Val Thr Ser Ala Arg Ser Leu Gly Met Arg
165 170 175
Ser Val Leu Phe His Lys Lys Asp Glu Val Gln Arg Gln Leu Thr Asn
180 185 190
Ile Phe Gly Ser Pro Ala Glu Arg Gly Leu Glu Tyr Leu Ser Ala Asn
195 200 205
Lys Thr Asn Leu Gln Ser Ala Thr Thr Thr Asp Ile Pro Ile Gln Asp
210 215 220
Asn Phe Gly Gln Leu Leu Ile Leu Glu Ala Thr Glu Asp Pro Ser Leu
225 230 235 240
Val Arg Met Glu Pro Gly Lys Arg Thr Trp Asn Phe Phe Ile Gly Ser
245 250 255
Pro Ser Leu Thr Thr Asp Thr Phe Pro Asp Asp Leu Asp Thr Thr Ser
260 265 270
Leu Ala Leu Ser Ile Val Pro Thr Ser Pro Asp Val Val Asn Ser Val
275 280 285
Ile Asp Glu Ile Ile Ser Arg Arg Asp Lys Asp Gly Ile Val Pro Thr
290 295 300
Tyr Phe Asp Asn Thr Arg Pro Arg Val Asp Pro Ile Val Cys Val Asn
305 310 315 320
Val Leu Ser Met Phe Ala Lys Tyr Gly Arg Glu His Asp Leu Pro Ala
325 330 335
Thr Val Ala Trp Val Arg Asp Val Leu Tyr His Arg Ala Tyr Leu Gly
340 345 350
Gly Thr Arg Tyr Tyr Gly Ser Ala Glu Ala Phe Leu Phe Phe Phe Thr
355 360 365
Arg Phe Val Arg Asn Leu Arg Pro Gly Thr Leu Lys Gln Asp Leu His
370 375 380
Ala Leu Leu Ser Glu Arg Val Arg Glu Arg Leu Asn Thr Pro Val Asp
385 390 395 400
Ala Leu Ala Leu Ser Met Arg Ile Gln Ala Cys His Ala Leu Gly Phe
405 410 415
Asp Ala Pro Ala Asp Ile Ala Thr Leu Ile Thr Met Gln Asp Glu Asp
420 425 430
Gly Gly Trp Pro Ala Ala Val Ile Tyr Lys Tyr Gly Ala Gly Gly Leu
435 440 445
Gly Ile Thr Asn Arg Gly Val Ser Thr Ala Phe Ala Val Lys Ala Ile
450 455 460
Thr Gly Ser Pro Val Lys Thr Glu Thr Asn Ile Gly Gly Asp Gly Ala
465 470 475 480
Arg Ala Val Ser Ala Met Ser Ser Leu Glu Ala Arg Arg Leu Gln Pro
485 490 495
Ile Ser Ser Val Gly Asp Trp Val Arg Phe Ile Ile Ala Ser Leu His
500 505 510
Val His Leu Ala Trp Leu Trp Asn Val Leu Leu Leu Ser Lys Val Val
515 520 525
<210> 79
<211> 1587
<212> DNA
<213> 热焦曲霉(Aspergillus calidoustus)_CEN60542.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 79
atggtccgcg cactcatcct cgatctcggc gatgtcctct tcaactggga cgcgcctgcg 60
tccaccccca tttcacgcaa gaccctcggc cagatgctgc atagtgagat ctggggtgag 120
tatgaacgtg gccatttgac agaagacgag gcatacaacg cactcgcgaa gcggtattcc 180
tgcgaggcca aggatgtcgc acataccttt gtcctggcac gagaatcgct gcggctcgac 240
acgaaattca aaacgtttct gcagactcta aagcagaatg ccaacggctc ccttcgtgtc 300
tatggcatgt cgaatatatc gaaaccggat ttcgaagtcc tgctgggcaa ggccgatgac 360
tggactctgt ttgacaagat cttcccctct ggccatgtcg gtatgcgcaa gccagatctt 420
gccttcttcc gctatgtgct caaggacatt tcaacgcctg tcgaggatgt ggtgtttgtt 480
gacgataacc tggacaacgt gacgagtgct cggtctctgg gcatgcgcag cgtcctcttt 540
cataagaaag acgaggtcca gcgacagctc accaacatct ttggcagccc tgctgagcgg 600
ggcttggagt atctctccgc caacaagacg aatctgcaga gtgctaccac gacagatatc 660
ccaatccagg ataactttgg ccaacttctg attctcgagg ccactgaaga cccatcgctg 720
gtccgcatgg agcccggtaa gcgaacctgg aatttcttca tcggttctcc atccctcaca 780
accgacacct tccccgacga tctcgacacc acatcccttg ccctctccat cgtacccaca 840
agccccgacg tcgtcaactc ggtcatcgac gagattatca gccgtcgcga caaggacggt 900
atcgtcccga cttacttcga caacacccgc ccccgcgtgg acccaatcgt ctgcgtaaac 960
gtcctctcca tgttcgcaaa gtacggccgc gagcacgacc tccccgcaac agttgcgtgg 1020
gtccgcgacg tcttgtatca tcgagcatac ctcggcggaa cacggtacta cgggtcagct 1080
gaggccttcc tcttcttctt cactcgcttc gttcgcaacc tccgaccggg aactctcaag 1140
caggatctac acgcattgct atcagagcgc gtgcgcgagc gactcaatac ccccgtcgac 1200
gcactcgccc tgtcaatgcg catccaggcc tgtcatgcgc tgggctttga cgcccccgca 1260
gacattgcga cgctcatcac aatgcaggac gaggacggcg ggtggccggc agccgtcatc 1320
tacaagtacg gggccggggg gttggggatc acgaaccggg gtgtttcgac tgcgtttgcc 1380
gtaaaggcga ttacagggtc gcccgtgaag actgaaacca acataggcgg cgatggagct 1440
cgcgctgtct cggccatgtc ctccttggag gcgaggaggc tacagccgat ctcgtcggtt 1500
ggggactggg tgcggtttat cattgcgtcg ttgcatgtcc atctggcttg gctttggaat 1560
gttttgcttt tgagcaaggt tgtttga 1587
<210> 80
<211> 1587
<212> DNA
<213> 人工序列_CEN60542.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 80
atggttcgtg cgttgatttt ggatttgggt gatgtgttgt ttaattggga cgcccctgca 60
agcactccga tcagccgtaa gaccctgggc cagatgctgc attccgagat ttggggtgag 120
tatgagcgtg gtcacctgac cgaagatgaa gcgtacaacg cgctggcaaa gcgctacagc 180
tgcgaggcaa aagacgtggc gcatactttt gttttggcgc gtgaaagcct gcgcctggat 240
accaagttta agacttttct gcagaccctg aaacagaacg cgaacggctc gctgcgtgtt 300
tatggtatgt ccaatatcag caaaccggat tttgaagtgc tgctgggtaa agctgacgac 360
tggaccttgt tcgacaagat cttcccgagc ggtcatgtcg gtatgcgcaa accggacctg 420
gctttctttc gttacgtgct gaaagacatc agcaccccgg ttgaggatgt tgtgtttgtt 480
gacgataacc tggataatgt gacgtctgcc cgttccctgg gtatgcgtag cgtcctgttc 540
cacaaaaaag acgaagtcca acgtcagctg accaacattt tcggtagccc tgctgagcgc 600
ggtctggagt atctgtccgc gaacaagacc aatctgcaaa gcgcaaccac caccgacatc 660
cctatccaag acaactttgg tcaattactg attctggaag ccaccgaaga tccgagcctg 720
gtacgcatgg aaccgggcaa gcgtacctgg aatttcttca ttggctctcc gagcctgacg 780
acggatacct tcccggatga cctggacacg acgagcctcg cactgtccat cgtgccgacc 840
agcccagatg ttgttaatag cgtgatcgat gagatcatca gccgtcgcga caaggacggt 900
attgtgccga cgtactttga taacacgcgc ccgcgtgtgg acccgattgt ttgtgttaac 960
gttctgtcta tgttcgcgaa atatggccgt gagcacgatc tgccggcgac ggtcgcgtgg 1020
gtccgcgacg tcctctatca tcgcgcatac ctgggtggca ccagatacta cggtagcgcg 1080
gaagccttcc ttttcttctt tacgcgcttt gtgcgtaatc tgcgtccggg cacgctgaaa 1140
caagatctgc acgcgttgct gagcgagcgt gtccgtgagc gcctgaatac cccggtggat 1200
gcgctggcgc tgagcatgcg cattcaggct tgccacgcac tgggctttga cgccccagct 1260
gacatcgcga cgctgattac catgcaagat gaagatggtg gctggccggc ggcagttatc 1320
tacaaatatg gtgcgggtgg cctgggcatt acgaaccgtg gtgtgtccac ggcattcgcg 1380
gtgaaggcaa tcacgggtag cccggttaaa accgaaacca acatcggcgg cgacggtgcc 1440
cgtgcagtgt cggccatgag cagcctggaa gcccgtcgtt tgcagccgat ttctagcgtc 1500
ggcgactggg tccgtttcat catcgcatca ctgcacgtcc acctggcgtg gctgtggaat 1560
gtcctgctgc tgagcaaagt cgtttaa 1587
<210> 81
<211> 531
<212> PRT
<213> 不整异担子菌(Heterobasidion irregulare)_XP_009547469.1蛋白
<400> 81
Met Ser Met Ile Pro Arg Cys Ser Asn Leu Ile Leu Asp Ile Gly Asp
1 5 10 15
Val Leu Phe Thr Trp Ser Pro Lys Thr Ser Thr Ser Ile Ser Pro Arg
20 25 30
Thr Met Lys Ser Ile Leu Ser Ser Thr Thr Trp His Gln Tyr Glu Thr
35 40 45
Gly His Ile Ser Gln Gly Asp Cys Tyr Arg Leu Ile Gly Asn Gln Phe
50 55 60
Ser Ile Asp Pro Gln Glu Val Gly Leu Ala Phe Gln Gln Ala Arg Asp
65 70 75 80
Ser Leu Gln Pro Asn Val Asp Phe Ile His Phe Ile Arg Ala Leu Lys
85 90 95
Ala Glu Ser His Gly Thr Leu Arg Val Phe Ala Met Ser Asn Ile Ser
100 105 110
Gln Pro Asp Tyr Ala Val Leu Arg Thr Lys Asp Ala Asp Trp Ala Val
115 120 125
Phe Asp Asp Ile Phe Thr Ser Ala Asp Ala Gly Val Arg Lys Pro His
130 135 140
Leu Gly Phe Tyr Lys Leu Val Leu Gly Lys Ile Gly Ala Asp Pro Asn
145 150 155 160
Asp Thr Val Phe Val Asp Asp Lys Gly Asp Asn Val Leu Ser Ala Arg
165 170 175
Ser Leu Gly Leu His Gly Ile Val Phe Asp Ser Met Asp Asn Val Lys
180 185 190
Arg Ala Leu Arg Tyr Leu Ile Ser Asp Pro Ile Arg Arg Gly Arg Glu
195 200 205
Phe Leu Gln Ala Arg Ala Gly His Leu Glu Ser Glu Thr Asn Thr Gly
210 215 220
Ile Glu Ile Gly Asp Asn Phe Ala Gln Leu Leu Ile Leu Glu Ala Thr
225 230 235 240
Lys Asp Arg Thr Leu Val Asn Tyr Met Asp His Pro Asn Lys Trp Asn
245 250 255
Phe Phe Arg Asp Gln Pro Leu Leu Thr Thr Glu Glu Phe Pro Phe Asp
260 265 270
Leu Asp Thr Thr Ser Ile Gly Thr Leu Ala Thr Gln Arg Asp Asp Gly
275 280 285
Thr Ala Asn Leu Val Met Asp Glu Met Leu Gln Tyr Arg Asp Glu Asp
290 295 300
Gly Ile Ile Gln Thr Tyr Phe Asp His Glu Arg Pro Arg Ile Asp Pro
305 310 315 320
Ile Val Cys Val Asn Val Leu Ser Leu Phe Tyr Ser Arg Gly Arg Gly
325 330 335
Ser Glu Leu Ala Pro Thr Leu Glu Trp Val Arg Gly Val Leu Lys His
340 345 350
Arg Ala Tyr Leu Asp Gly Thr Arg Tyr Tyr Glu Thr Gly Glu Cys Phe
355 360 365
Leu Phe Phe Leu Ser Arg Leu Leu Gln Ser Thr Lys Asp Ala Ala Leu
370 375 380
His Ala Ser Leu Lys Ser Leu Phe Ala Glu Arg Val Lys Glu Arg Ile
385 390 395 400
Gly Ala Pro Gly Asp Ala Leu Ala Leu Ala Met Arg Ile Leu Ala Cys
405 410 415
Ala Ala Val Gly Val Arg Asp Glu Ile Asp Leu Arg Ser Leu Leu Pro
420 425 430
Leu Gln Cys Glu Asp Gly Gly Trp Glu Ala Gly Trp Val Tyr Lys Tyr
435 440 445
Gly Ser Ser Gly Val Lys Ile Gly Asn Arg Gly Leu Thr Thr Ala Leu
450 455 460
Ala Leu Asn Ala Ile Glu Ala Val Glu Gly Arg Arg Thr Arg Pro Lys
465 470 475 480
Ser Gly Lys Ile Ser Arg Val Ser Arg His Ser Glu Val Ala Ala Ala
485 490 495
Pro Arg Ser Ser Thr Ser Ser His Arg Ser Asn Arg Ser Ile Ser Arg
500 505 510
Thr Phe Gln Ala Tyr Phe Lys Ala Ser Trp Thr Ser Met Lys Gln Val
515 520 525
Ala Val Ala
530
<210> 82
<211> 1596
<212> DNA
<213> 不整异担子菌(Heterobasidion irregulare)_XP_009547469.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 82
atgtccatga tacccagatg ctcgaatctc atcctcgaca tcggggatgt tctcttcaca 60
tggtctccga agacgtccac ttcgatctcc ccccgcacca tgaagagcat actgtcatcg 120
acgacctggc accaatacga gaccgggcac atttcacagg gcgactgcta ccgcctcata 180
ggcaaccagt tctccatcga tcctcaggaa gtcggacttg cattccaaca agctcgggac 240
tcattgcagc ctaatgttga cttcattcac ttcatccgcg ccctcaaggc ggaatcacac 300
gggacgctgc gcgtcttcgc tatgtccaac atctctcagc ccgattacgc agttcttcgg 360
actaaggacg ccgactgggc cgtttttgac gatatattca cgtctgcaga tgctggggtt 420
cgaaagccac accttgggtt ctacaagttg gtactcggaa agatcggcgc cgatccaaac 480
gataccgtct tcgtcgatga caagggggac aatgtcctct ctgcacggtc tctcggcctt 540
catggaatcg tctttgacag tatggacaac gtcaagcgag ccctgcgcta cttgatcagc 600
gaccccatac ggcgaggacg agagtttctc caagcgcgag ccggccattt ggagtcggag 660
accaatacgg gcatcgaaat cggtgataat tttgcccagc tccttattct cgaggccacg 720
aaggatagga cactcgtcaa ttatatggac catccgaaca aatggaattt cttccgagat 780
caaccgctcc tcacaacgga ggagttccct ttcgatctcg atacgacatc tattggaacg 840
cttgcgacgc agcgcgatga tgggactgcc aatctagtaa tggatgagat gcttcagtac 900
cgtgatgagg atggcataat acaaacatat ttcgatcatg aacgaccgag gatagatccc 960
atcgtctgtg tcaacgtctt gagccttttc tactcccggg gtcgtggttc ggagctagca 1020
ccgacactag agtgggtgcg tggtgtcctc aagcaccgcg cgtatctcga tggaacgcga 1080
tactacgaga caggcgaatg cttccttttc ttcctcagcc ggctcttgca atcaaccaag 1140
gacgccgcct tgcacgcatc gttgaaatct ttgttcgccg aacgggtcaa ggagcgcata 1200
ggggcaccag gggacgcgct ggcgctggcg atgcgtatac tggcatgcgc agcagtgggc 1260
gtgcgggacg agatcgatct tcgatcacta ttacctctgc agtgcgagga tggggggtgg 1320
gaggcaggct gggtgtacaa gtatgggtct tcgggagtca agatcggcaa tcgtggcctc 1380
acgactgcgc ttgcgctcaa tgccatcgag gctgtggagg gacgtcgcac gaggccgaag 1440
tcgggtaaga tcagccgagt cagccgtcat tctgaggtcg cagcagcgcc acggtcttcc 1500
accagcagtc atcgttctaa tcgctcgatc tcaaggacat tccaggcgta cttcaaggcg 1560
tcgtggacat cgatgaaaca ggtggccgtg gcgtga 1596
<210> 83
<211> 1596
<212> DNA
<213> 人工序列_XP_009547469.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 83
atgagcatga ttccacgttg tagcaatctg attctcgaca tcggtgatgt gttgtttacg 60
tggagcccga aaaccagcac cagcattagc ccgcgtacca tgaaatctat cctgagctct 120
accacctggc atcaatatga gactggccac atcagccagg gtgattgcta ccgcctgatc 180
ggtaatcagt tctccatcga cccgcaagag gtcggtttgg ccttccagca agccagagac 240
agcctgcaac cgaatgttga tttcatccat ttcattcgtg ccctgaaagc tgagtcgcac 300
ggcaccctgc gcgtttttgc gatgagcaat atcagccaac ctgactatgc agtcctgcgt 360
acgaaagacg cggactgggc tgtttttgat gatatcttca cgagcgcgga tgctggtgtt 420
cgtaaaccgc acctgggttt ttataaactg gtcttaggca agattggcgc ggaccctaac 480
gacaccgttt ttgtggatga taagggtgac aacgtcctct ctgcacgttc cctgggtctg 540
cacggtatcg tttttgattc aatggacaac gtgaagcgcg cactgcgcta cctgattagc 600
gacccgatcc gccgcggccg tgaatttctg caggcccgtg cgggtcacct ggagtccgaa 660
acgaacacgg gtattgagat tggtgataat ttcgcgcaat tgctgatcct ggaagcgacc 720
aaagatcgta ctctggtgaa ctacatggac cacccgaaca agtggaactt cttccgtgac 780
cagccgctgc tgaccaccga agaatttccg ttcgacctgg acacgaccag cattggcacg 840
ctggccaccc aacgtgacga tggtacggcg aatctggtaa tggacgaaat gttgcagtat 900
cgtgacgaag atggcatcat tcagacctat ttcgatcatg agcgcccgcg tattgatccg 960
attgtttgtg tgaatgtgct gtctctgttc tacagccgtg gccgtggctc tgagttggcg 1020
ccgacgctgg aatgggtgcg cggtgtgttg aaacatcgtg cgtacctgga tggtacgcgt 1080
tattacgaga ctggtgagtg tttcctgttt ttcctgagcc gtctgctgca gagcaccaaa 1140
gacgcagccc tgcacgcgag cctgaagtcc ctgtttgcag agcgtgttaa agagcgcatc 1200
ggtgcgccgg gcgatgctct ggcgctggct atgcgcatcc tggcgtgcgc cgctgttggt 1260
gtgcgcgatg aaattgattt gcgtagcctg ctgccgctgc aatgcgaaga tggcggctgg 1320
gaagcgggct gggtctacaa atacggcagc agcggtgtga agattggcaa tcgcggtctt 1380
accacggcgc tggcattgaa tgctatcgaa gccgttgagg gccgtcgcac ccgcccaaag 1440
tccggtaaga tcagccgtgt tagccgtcat agcgaagtcg cagcggcacc gcgttcctcg 1500
acgagcagcc accgtagcaa ccgtagcatt agccgcacct tccaggcata ttttaaagcg 1560
agctggacca gcatgaaaca agtcgcagtg gcgtaa 1596
<210> 84
<211> 518
<212> PRT
<213> 近光彩裂孔菌(Schizopora paradoxa)_KLO09124.1蛋白
<400> 84
Met Ser Ile His Gly Ser Ser Met Ser Ser Tyr Ser Ser Thr Val Pro
1 5 10 15
Ser Met Thr Ser Ser Pro Ala Ser Thr Ser Thr Pro Ser Ser Pro Ala
20 25 30
Ser Ser Ile His Glu Ile Gly Pro Val Pro Glu Ala Arg Arg Lys Gly
35 40 45
Gln Cys Asn Ala Leu Ile Phe Asp Leu Gly Asp Val Leu Phe Thr Trp
50 55 60
Ser Ala Glu Thr Lys Thr Thr Ile Ser Pro Lys Leu Leu Lys Lys Ile
65 70 75 80
Leu Asn Ser Leu Thr Trp Phe Glu Tyr Glu Lys Gly Asn Ile Gly Glu
85 90 95
Gln Glu Ala Tyr Asp Ala Val Ala Lys Glu Phe Gly Val Pro Ser Ser
100 105 110
Glu Val Gly Ala Ala Phe Gln Cys Ala Arg Asp Ser Leu Gln Ser Asn
115 120 125
Pro Arg Leu Val Ser Leu Ile Arg Glu Leu Lys Ser Gln Tyr Asp Leu
130 135 140
Lys Val Tyr Ala Met Ser Asn Ile Ser Ala Pro Asp Trp Glu Val Leu
145 150 155 160
Arg Thr Lys Ala Thr Pro Glu Glu Trp Ala Met Phe Asp Arg Val Phe
165 170 175
Thr Ser Ala Ala Ala Arg Glu Arg Lys Pro Asn Leu Gly Phe Tyr Arg
180 185 190
Gln Val Val Glu Ala Thr Gly Val Asp Pro Ala Arg Ser Val Phe Val
195 200 205
Asp Asp Lys Leu Asp Asn Val Ile Ser Ala Arg Ser Val Gly Leu Asn
210 215 220
Ala Ile Ile Phe Asp Ser Phe Glu Asn Val Ala Arg Gln Leu Lys Asn
225 230 235 240
Tyr Val Ala Asp Pro Ile Gly Arg Ala Glu Ala Trp Leu Arg Asp Asn
245 250 255
Ala Lys Lys Met Leu Ser Ile Thr Asp Ala Gly Val Val Val Tyr Glu
260 265 270
Asn Phe Gly Gln Met Leu Ile Leu Glu Ala Thr Gly Asp Arg Ser Leu
275 280 285
Val Asp Tyr Val Glu Tyr Pro Arg Leu Phe Asn Phe Phe Gln Gly Asn
290 295 300
Gly Val Phe Thr Thr Glu Ser Phe Pro Cys Asp Leu Asp Ser Thr Ser
305 310 315 320
Ile Gly Leu Thr Val Thr Asn His Val Asp Glu Lys Thr Arg His Ser
325 330 335
Val Met Asp Glu Met Leu Thr Tyr Lys Asn Glu Asp Gly Ile Ile Ala
340 345 350
Thr Tyr Phe Asp Ala Thr Arg Pro Arg Ile Asp Pro Val Val Cys Ala
355 360 365
Asn Val Leu Thr Phe Phe Tyr Lys Asn Gly Arg Gly Glu Glu Leu Asn
370 375 380
Glu Thr Leu Asp Trp Val Tyr Asp Ile Leu Leu His Arg Ala Tyr Leu
385 390 395 400
Asp Gly Thr Arg Tyr Tyr Phe Gly Ser Asp Thr Phe Leu Phe Phe Leu
405 410 415
Ser Arg Leu Leu Ser Glu Ser Pro Ser Val Tyr Ala Arg Phe Ala Pro
420 425 430
Val Phe Gln Glu Arg Val Lys Glu Arg Met Gly Ala Thr Gly Asp Ala
435 440 445
Met Ser Leu Ala Met Arg Ile Ile Ala Ala Ala Thr Val Lys Ile Gln
450 455 460
Asp Arg Val Asp Cys Asp Ala Leu Leu Gln Thr Gln Glu Asp Asp Gly
465 470 475 480
Gly Phe Pro Ile Gly Trp Met Tyr Lys Tyr Gly Ala Thr Gly Met Leu
485 490 495
Leu Gly Asn Lys Gly Leu Ser Thr Ala Leu Ala Ile Gln Ala Ile Lys
500 505 510
Ala Val Glu Ser Phe Pro
515
<210> 85
<211> 1557
<212> DNA
<213> 近光彩裂孔菌(Schizopora paradoxa)_KLO09124.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 85
atgtcgattc acggttcttc tatgtcctcc tattcctcga ctgtgccgtc aatgacttcc 60
tctcccgcgt ccacttctac tccgtcgtct cctgcatcgt cgatccatga gattggtcct 120
gtcccagaag ctcgacgaaa gggacagtgc aacgcgctga tcttcgacct cggagacgtc 180
ctcttcacct ggtcggcaga gactaagacc accatttccc cgaaactcct gaaaaagatc 240
cttaactcct taacatggtt cgaatacgag aagggaaaca tcggggagca ggaggcgtat 300
gacgcagtcg caaaggagtt tggcgtcccg tcgtccgagg tcggggccgc tttccagtgc 360
gcgcgcgatt cgctacagag caatccccgc ctcgtctcgc tcatccgtga gctgaagtcg 420
caatatgatc tcaaggtgta cgccatgtcc aacatctctg cgccggactg ggaagtccta 480
aggacgaagg cgacccctga ggagtgggca atgtttgacc gcgtcttcac gagcgcggcc 540
gcgcgcgagc gtaagccaaa cctcggattc tacagacagg ttgttgaggc gaccggcgtc 600
gaccccgctc gctccgtgtt cgtcgacgat aaactcgaca atgtcatctc tgcgcgttca 660
gtcggattaa atgcgatcat cttcgactca tttgagaacg tcgcccggca gctcaaaaac 720
tatgtcgctg atcctatcgg acgggcggag gcgtggttgc gcgataacgc aaagaagatg 780
ttgtcaatta cggatgccgg ggtggtcgta tacgagaatt tcggccagat gctgatcttg 840
gaggcaacag gcgataggtc gcttgtggac tacgtcgagt accctcgtct cttcaacttc 900
ttccaaggca atggcgtctt tacgaccgag tcattccctt gcgaccttga ttcgacttcc 960
atcggcttaa ccgtcacgaa ccacgtcgat gagaaaacaa ggcacagcgt catggatgag 1020
atgctgacct acaaaaatga ggatggtatc attgcgactt actttgatgc cacgcgtccc 1080
cgaattgacc ccgtcgtctg cgccaatgtc ttgacgttct tctacaagaa cggccgaggg 1140
gaggagctca atgaaacact tgactgggtc tacgacatcc tccttcatcg cgcgtacctc 1200
gatggcacac gctattattt cggctcagac accttcctct tcttcctttc tcgacttctc 1260
tccgaatcgc catccgttta cgcccgtttc gctccggtgt tccaggagag agtcaaggag 1320
cgcatggggg cgacgggaga tgcgatgtcc cttgcgatgc gcatcatcgc ggccgcaact 1380
gtcaagatcc aagaccgagt cgactgcgac gctctgctgc agacgcagga agacgacggt 1440
ggattcccga taggttggat gtacaagtac ggggcgaccg ggatgcttct gggtaacaag 1500
ggcttgtcga cagctctggc aatccaagct atcaaagcgg tcgaatcttt cccttga 1557
<210> 86
<211> 1557
<212> DNA
<213> 人工序列_KLO09124.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 86
atgtcgattc acggtagcag catgtcgtct tatagcagca cggttccatc tatgactagc 60
agcccggctt ccacgagcac gccgtccagc ccggccagca gcatccacga aatcggcccg 120
gtccctgagg cgcgtcgcaa gggccaatgc aatgcactga tcttcgacct gggtgatgtt 180
ctgtttacct ggagcgcaga aaccaagacc acgatcagcc cgaagctgct gaaaaagatt 240
ctgaacagct tgacctggtt tgagtatgag aaaggcaaca tcggtgaaca agaagcctat 300
gacgccgttg cgaaagagtt cggtgtgccg agctctgagg ttggcgctgc gtttcaatgt 360
gcgcgtgact ccctgcaaag caatccgcgt ttggttagcc tgattcgtga gctgaagtcc 420
cagtacgacc tgaaagtgta cgctatgagc aatattagcg cgccagactg ggaagtgctg 480
cgtactaaag cgaccccgga agagtgggca atgttcgatc gtgtctttac ttctgcggcg 540
gcgcgtgagc gtaagccgaa cttgggcttt taccgccaag tcgtggaagc aaccggtgtc 600
gatccggcgc gtagcgtttt cgtcgatgat aaactggaca atgtgatcag cgcgcgctct 660
gtcggtctga acgctattat cttcgactcc ttcgaaaacg tcgcccgtca gctgaagaat 720
tacgtcgcag acccgattgg tcgcgctgag gcgtggctgc gcgacaacgc aaagaaaatg 780
ctgagcatca ccgatgcggg tgttgtggtt tacgagaatt ttggccagat gctgatcctg 840
gaagctaccg gtgaccgtag cctggtggac tatgtggagt atccgcgcct ctttaacttc 900
ttccagggta acggcgtttt tacgaccgag agctttccat gcgatctgga cagcaccagc 960
atcggtctga ctgtgaccaa tcatgtggac gaaaagactc gccacagcgt catggacgaa 1020
atgctgacct acaaaaatga agatggtatt attgcgacgt actttgacgc gacgcgcccg 1080
cgcattgacc ctgttgtctg tgccaatgtt ctgaccttct tctacaaaaa cggtcgtggt 1140
gaagaattga acgaaaccct ggattgggtg tacgacattc tgctgcatcg cgcgtatctg 1200
gacggtacgc gttattattt cggctccgat acgttcctgt ttttcctgag ccgtctgctg 1260
agcgagtctc cgagcgttta cgcgcgtttt gccccggtgt ttcaagagcg cgtgaaagag 1320
cgtatgggcg cgaccggtga tgcgatgagc ctggccatgc gtatcattgc agcagcaacc 1380
gtaaagatcc aggatcgtgt ggattgcgac gcactgttgc agacccaaga agatgatggc 1440
ggtttcccga ttggttggat gtacaaatat ggtgcgaccg gtatgttgct gggcaacaaa 1500
ggcctgagca cggccctggc gatccaggca attaaagccg tcgagtcgtt cccgtaa 1557
<210> 87
<211> 507
<212> PRT
<213> 杂色曲霉(Aspergillus versicolor)_OJI95797.1蛋白
<400> 87
Met Gly Ser Thr Lys Ala Leu Val Val Asp Phe Gly Asn Val Leu Cys
1 5 10 15
Thr Trp Thr Pro Pro Arg Glu Leu Ser Ile Pro Pro Lys Lys Leu Lys
20 25 30
Gln Ile Met Ser Ser Asp Ile Trp Leu Asp Tyr Glu Arg Gly Ile Tyr
35 40 45
Lys Ser Glu Asp Glu Cys Tyr Leu Ala Val Ala Thr Arg Phe Gly Val
50 55 60
Ser Pro Ser Asp Leu Ser Ser Val Met Lys Lys Ala Arg Glu Ser Leu
65 70 75 80
Gln Pro Asn Thr Ala Thr Leu Asn His Leu Ser His Leu Lys Lys Thr
85 90 95
Gln Pro Gly Leu Arg Ile Tyr Gly Leu Thr Asn Thr Pro Leu Pro Glu
100 105 110
Gln Ser Ser Val Arg Ser Ile Ala Gln Glu Trp Pro Ile Phe Asp His
115 120 125
Ile Tyr Ile Ser Gly Ile Leu Gly Met Arg Lys Pro Asp Ile Gly Cys
130 135 140
Tyr Arg Leu Val Leu Arg Lys Ile Gly Leu Pro Ala Glu Ser Val Val
145 150 155 160
Phe Ile Asp Asp Ser Pro Glu Asn Ile Leu Ala Ala Gln Ser Leu Gly
165 170 175
Val His Ser Ile Leu Phe Gln Ser His Asp Gln Leu Ser Arg Gln Leu
180 185 190
Gly Asn Val Leu Gly Asp Pro Ile Gln Arg Gly His Asn Phe Leu Leu
195 200 205
Ser Asn Ala Lys Gln Met Asn Ser Thr Thr Asp Lys Gly Val Ile Ile
210 215 220
Arg Asp Asn Phe Ala Gln Leu Leu Ile Ile Glu Leu Thr Gln Asn Pro
225 230 235 240
Asp Leu Val Ala Leu Glu Thr Trp Asp Arg Thr Trp Asn Phe Phe Ile
245 250 255
Gly Pro Pro Gln Leu Thr Thr Glu Ser Phe Pro Asn Asp Leu Asp Thr
260 265 270
Thr Ser Ile Ala Leu Ser Val Leu Pro Val Asp Lys Glu Val Val Trp
275 280 285
Ser Val Met Asp Glu Met Leu Thr Phe Thr Asn Ala Asp Gly Ile Phe
290 295 300
Met Thr Tyr Phe Asp Arg Ser Arg Pro Arg Val Asp Pro Val Val Cys
305 310 315 320
Thr Asn Val Leu Asn Leu Phe Cys Met His Gly Arg Glu Ser Glu Val
325 330 335
Ala Ala Thr Phe Asp Trp Val Leu Asp Val Leu Arg Asn Ser Ala Tyr
340 345 350
Leu Ser Gly Ser Arg Tyr Tyr Ser Ser Pro Asp Cys Phe Leu Tyr Phe
355 360 365
Leu Ser Arg Leu Ser Cys Val Val Arg Asp Gly Thr Arg Arg Arg Glu
370 375 380
Leu Lys Ser Leu Leu Lys Gln Gln Val Ser Gln Arg Ile Gly Ala Asp
385 390 395 400
Gly Asp Ser Val Ser Leu Ala Thr Arg Leu Leu Ala Ser Asn Ile Leu
405 410 415
Gly Ile Thr Asn Gly Arg Asp Arg Ser Arg Leu Leu Ala Leu Gln Glu
420 425 430
Thr Asp Gly Gly Trp Pro Ala Gly Trp Val Tyr Lys Phe Gly Ser Ser
435 440 445
Gly Val Gln Ile Gly Asn Arg Gly Leu Ser Thr Ala Leu Ala Leu Lys
450 455 460
Ser Ile Glu Arg Gln Lys Gly Pro Val Glu Ala Ile Ser Ser Glu Pro
465 470 475 480
Glu Ala Trp Trp Pro Ser Leu Arg Leu Asp Arg Leu Leu Asn Val Trp
485 490 495
Pro Phe Ile Asp Trp Lys Gly Tyr Ser Pro Ser
500 505
<210> 88
<211> 1524
<212> DNA
<213> 杂色曲霉(Aspergillus versicolor)_OJI95797.1 cDNA
<220>
<221> misc_feature
<223> cDNA
<400> 88
atgggttcca ccaaggctct tgttgttgac tttgggaatg ttttgtgtac ctggacacca 60
cccagggagt tatccatccc gcccaagaag ctgaaacaaa tcatgtcttc tgacatttgg 120
ctcgactatg aacggggtat ctataagtcg gaggacgagt gctacttggc ggttgcaact 180
cgcttcggcg tctctcccag cgacctctcc tcggtgatga aaaaggcccg cgagagcctg 240
caaccaaaca ccgcaaccct gaatcatctg tctcatctca aaaagaccca gcctggcctc 300
aggatatacg gtttgaccaa cacccctctc ccagaacaaa gcagtgtacg atccatcgcc 360
caggaatggc ctatcttcga ccatatctac atatcaggca tcctcggaat gcgcaagccg 420
gacattggct gctacaggct ggtgctgcga aagattgggc ttccagcgga gtccgtggtc 480
ttcattgatg attcacccga gaacatcctg gccgcgcagt cactgggagt acacagcata 540
ctgttccaaa gccacgacca gctctctcgt cagcttggca atgtgctggg tgatccaatc 600
cagcggggcc ataacttcct actctcgaac gcaaagcaaa tgaatagtac gaccgacaag 660
ggagttatta tccgggacaa ctttgcgcaa ctgctgatca tcgagctgac gcagaaccca 720
gaccttgtgg cgttagaaac atgggaccgt acctggaatt tttttattgg acctccacaa 780
ttgacaactg aaagctttcc caatgatctt gacactacct ccatcgctct ctcggttctt 840
ccggttgaca aagaagtggt atggtctgtg atggacgaga tgctaacgtt taccaatgcg 900
gatgggattt ttatgaccta tttcgaccga tcacgccctc gagttgatcc ggtagtttgc 960
accaatgtcc tgaatctttt ctgcatgcat ggacgggaaa gcgaagttgc agccacattt 1020
gactgggtgc tggacgttct tcgaaattcg gcctatttat caggatccag atactattct 1080
tcgcctgatt gctttctata ctttctttca cggctgagct gtgtggtccg agacggcacg 1140
cgacgcaggg agctcaagtc actgttgaaa caacaagtga gccagcgtat tggcgctgat 1200
ggtgattccg tctctctcgc cactaggcta cttgcatcga acattttagg aatcacaaat 1260
ggccgtgatc gctccaggct tcttgctctg caggaaactg acggtggatg gcctgctggg 1320
tgggtttata aattcggaag ctcgggggta cagattggca atcgggggct cagtacagcc 1380
ttggcgttaa aatcaattga gcgtcagaag gggcctgttg aggcgatatc cagtgagcca 1440
gaagcgtggt ggccatccct caggcttgac cgacttctca acgtttggcc tttcatcgac 1500
tggaagggat attcgccgag ttga 1524
<210> 89
<211> 1524
<212> DNA
<213> 人工序列_OJI95797.1 优化的cDNA
<220>
<223> 密码子优化的cDNA
<400> 89
atgggttcta cgaaagcgtt ggttgttgat tttggtaatg ttctgtgcac ttggacgcca 60
ccacgtgaat tgtccatccc gccgaagaaa ctgaagcaaa tcatgagcag cgacatttgg 120
ctggactatg agcgtggtat ctacaaatcg gaagatgagt gctacctggc agttgcgacg 180
cgctttggtg tcagcccgtc cgacctgagc tccgttatga aaaaagcccg tgagagcctg 240
cagccgaata ccgcaacgct gaaccacttg agccatctga agaaaaccca gcctggcctt 300
cgtatctacg gcctgacgaa caccccgttg ccggaacaga gctcagtccg tagcattgcg 360
caggaatggc cgatttttga ccacatctac attagcggca tcttgggtat gcgcaaaccg 420
gatattggtt gttaccgtct ggttctgcgt aagatcggtc tgccagcgga gtccgtcgta 480
ttcatcgacg acagcccgga gaacattctg gcagctcaat cgttgggtgt ccatagcatc 540
ctgttccagt cccacgatca gctgagccgt cagctgggca atgtgctggg tgatccgatt 600
cagcgcggtc acaacttcct cctgtccaac gcgaagcaaa tgaacagcac caccgataag 660
ggtgtgatta tccgcgacaa cttcgcccag ctgctgatta ttgagctgac ccaaaatccg 720
gatctggttg cgctggagac ttgggaccgt acgtggaatt tctttattgg tccgccgcaa 780
ctgaccaccg agagctttcc gaacgacctg gacaccacga gcattgccct gagcgtgttg 840
ccggtggata aagaagtcgt ttggtctgtg atggatgaga tgctgacctt caccaacgca 900
gacggcatct tcatgaccta tttcgatcgt agccgtccgc gtgttgaccc ggtcgtttgt 960
accaatgtcc tgaatctgtt ttgcatgcat ggtcgcgaga gcgaagtggc cgcgacgttc 1020
gactgggtgc tggacgtgct gcgcaacagc gcgtacctga gcggttcccg ttattacagc 1080
agcccggatt gttttctgta tttcctgtct cgtctgagct gcgtcgtccg tgatggcacg 1140
cgtcgtcgtg aactgaaaag cctgctgaag caacaagttt ctcaacgtat cggcgctgac 1200
ggtgattccg tcagcctggc cacccgtttg ctggcgagca acatcctggg cattactaac 1260
ggtcgtgacc gcagccgtct gctggcattg caagaaaccg atggtggctg gcctgcaggc 1320
tgggtctata agtttggtag cagcggcgtg caaattggca atcgcggtct gagcaccgcg 1380
ctggctctga agtctatcga gcgccagaaa ggtccggtgg aagcaatcag cagcgagccg 1440
gaagcgtggt ggcctagctt acgcttggac cgcttgctga atgtttggcc atttatcgac 1500
tggaagggct actccccgag ctaa 1524
<210> 90
<211> 540
<212> PRT
<213> 双孢蘑菇(Agaricus bisporus)_XP_006461126.1蛋白
<400> 90
Met Ala Pro Pro Gln Arg Pro Phe Thr Ala Ile Val Phe Asp Ile Gly
1 5 10 15
Asp Val Leu Phe Gln Trp Ser Ala Thr Thr Lys Thr Ser Ile Ser Pro
20 25 30
Lys Thr Leu Arg Ser Ile Leu Asn Cys Pro Thr Trp Phe Asp Tyr Glu
35 40 45
Arg Gly Arg Leu Ala Glu Asn Ala Cys Tyr Ala Ala Ile Ser Gln Glu
50 55 60
Phe Asn Val Asn Pro Asp Glu Val Arg Asp Ala Phe Ser Gln Ala Arg
65 70 75 80
Asp Ser Leu Gln Ala Asn His Asp Phe Ile Ser Leu Ile Arg Glu Leu
85 90 95
Lys Ala Gln Ala Asn Gly Arg Leu Arg Val Tyr Ala Met Ser Asn Ile
100 105 110
Ser Leu Pro Asp Trp Glu Val Leu Arg Met Lys Pro Ala Asp Trp Asp
115 120 125
Ile Phe Asp His Val Phe Thr Ser Gly Ala Val Gly Glu Arg Lys Pro
130 135 140
Asn Leu Ala Phe Tyr Arg His Val Ile Ala Ala Thr Asp Leu Gln Pro
145 150 155 160
His Gln Thr Ile Phe Val Asp Asp Lys Leu Glu Asn Val Leu Ser Ala
165 170 175
Arg Ser Leu Gly Phe Thr Gly Ile Val Phe Asp Glu Pro Ser Glu Val
180 185 190
Lys Arg Ala Leu Arg Asn Leu Ile Gly Asp Pro Val Gln Arg Gly Gly
195 200 205
Glu Phe Leu Val Arg Asn Ala Gly Lys Leu Gly Ser Ile Thr Arg Thr
210 215 220
Thr Ala Lys His Glu Ser Ile Pro Leu Asp Glu Asn Phe Ala Gln Leu
225 230 235 240
Leu Ile Leu Glu Ile Thr Gly Asn Arg Ala Leu Val Asn Leu Val Glu
245 250 255
His Pro Gln Thr Trp Asn Phe Phe Gln Gly Lys Gly Gln Leu Thr Thr
260 265 270
Glu Glu Phe Pro Phe Asp Leu Asp Thr Thr Ser Leu Gly Leu Thr Ile
275 280 285
Leu Lys Arg Ser Arg Glu Ile Ala Asp Ser Val Met Asp Glu Met Leu
290 295 300
Glu Tyr Val Asp Pro Asp Gly Ile Ile Gln Thr Tyr Phe Asp His Arg
305 310 315 320
Arg Pro Arg Phe Asp Pro Val Val Cys Val Asn Ala Leu Ser Leu Phe
325 330 335
Tyr Ala Tyr Gly Arg Gly Glu Gln Leu Arg Ser Thr Leu Thr Trp Val
340 345 350
His Glu Val Leu Leu Asn Arg Ala Tyr Leu Asp Gly Thr Arg Tyr Tyr
355 360 365
Glu Thr Ala Glu Cys Phe Leu Tyr Phe Met Ser Arg Leu Leu Ala Thr
370 375 380
Ser Gly Asp Pro Asp Leu His Ser Leu Leu Lys Pro Leu Leu Lys Glu
385 390 395 400
Arg Val Gln Glu Arg Ile Gly Ala Asp Gly Asp Ser Leu Ala Leu Ala
405 410 415
Met Arg Ile Leu Ala Cys Asp Phe Val Gly Ile Arg Asp Glu Val Asp
420 425 430
Leu Arg Thr Leu Leu Thr Leu Gln Cys Glu Asp Gly Gly Trp Glu Val
435 440 445
Gly Trp Met Tyr Lys Tyr Gly Ser Ser Gly Ile Ser Ile Gly Asn Arg
450 455 460
Gly Leu Ala Thr Ala Leu Ala Ile Lys Ala Val Asp Thr Met Phe Gln
465 470 475 480
Pro Gln Ile Arg Phe Ser Glu Ser Pro Thr Asp Thr Leu Val Glu Asn
485 490 495
Ala Ile His Lys Arg Arg Pro Ser Phe Ser Glu Lys Phe Leu Gly Lys
500 505 510
Arg Pro Arg Ser Gly Ser Phe Arg Lys Pro Leu Gln Trp Ile Leu Gln
515 520 525
Gly Ser Lys Leu Arg Lys Ser Val Glu Ile Gly Ser
530 535 540
<210> 91
<211> 1808
<212> DNA
<213> 双孢蘑菇(Agaricus bisporus)_XP_006461126.1 cDNA
<400> 91
atggctccgc ctcagcgacc ctttactgcg attgtctttg acatcgggga tgttctattc 60
caatggtctg caaccaccaa aacctctatc tcaccaaaga cactccgctc tattctcaac 120
tgtccgacat ggtttgacta tgaacgtgga cgcctggcag aaaacgcttg ttatgccgct 180
atctcacaag aattcaacgt caacccagac gaagttcgcg acgctttcag ccaagcgcgc 240
gactctctcc aagcaaacca cgacttcatc agtctcatcc gtgagctgaa ggcacaagca 300
aatggtcgtt tacgtgtgta cgccatgtcg aacatatctc ttcctgattg ggaagtgctg 360
cggatgaaac ctgctgattg ggatattttc gaccacgtct tcacatccgg tgcggttggg 420
gaacgcaagc ccaatctcgc cttttatcgc catgttatcg cggccaccga tctgcagcct 480
catcagacaa tatttgttga cgataagctg gagaatgttc tctcagcacg ttccctcggg 540
ttcacaggca tcgtgtttga cgagccctcc gaggtcaaac gtgcgcttcg taacctcatt 600
ggggatcctg ttcaacgagg aggtgaattc ttggttcgga atgccggaaa gcttggctct 660
atcacaagga ctactgcaaa gcacgagtca atccccctcg acgagaattt tgctcagctt 720
cttattctcg agataacggg gaacaggtgc gttagcttct tgtagggtct tctgtcgtaa 780
tactaaattt tttctggtgt ttagggcttt ggtcaacctc gttgagcatc ctcaaacgtg 840
gaatttcttc caaggtgcgc tgctaaaata aacatccagt tgcgtttcga agctcattgt 900
gggcgtcccg tcacaggcaa gggccagctg acaacagaag aatttccatt cgatctcgat 960
acaacttctc ttggtctcac gatcctcaag cgaagcaggg aaatcgccga ttcagtcatg 1020
gatgaaatgc tggagtatgt cgatcctgat ggtatcattc aggcaagttt catttatcgg 1080
cttgagaaaa taaagacaaa aacgttctga tggggggatg tttctagacg tatttcgatc 1140
atcggagacc acgttttgat ccagtcgtgt gtgtcaatgc attaagcctc ttctatgctt 1200
acggccgcgg ggagcaactg cggtcgactt tgacatgggt acatgaagtc cttctcaatc 1260
gagcctactt ggatggcaca cggtactacg aaacagccga atgcttcctc tatttcatga 1320
gccgacttct cgccacttca ggcgaccctg accttcactc ccttcttaaa cctcttctca 1380
aagaacgggt gcaagaacgc attggagctg atggagactc tcttgcactc gcaatgcgta 1440
ttctcgcctg tgatttcgtc ggaatcagag atgaagtgga tttacgcaca cttctgactt 1500
tgcaatgtga agatggaggt tgggaagtgg gttggatgta caagtatgga tcttccggta 1560
tcagtatcgg aaatcgtgga ctggccaccg cgctcgctat caaggccgtc gacacgatgt 1620
ttcaacccca aattcggttc tctgaatcac ccacagatac tttggttgaa aacgctatcc 1680
acaaacgccg tccctcattt tccgaaaaat tcctcggcaa acgtcctcgc agcggatcgt 1740
tcaggaaacc tttacagtgg atactgcaag gttccaagct tcgcaaatct gtcgaaatag 1800
gaagctaa 1808
<210> 92
<211> 1623
<212> DNA
<213> 双孢蘑菇(Agaricus bisporus)_XP_006461126.1 优化的cDNA
<400> 92
atggcaccac cgcaacgtcc gttcactgca attgttttcg atattggcga tgttttgttc 60
caatggtctg cgaccacgaa aaccagcatt agcccgaaaa ccctgcgcag cattctgaat 120
tgtccgacct ggtttgatta tgagcgcggc cgtctggcgg aaaatgcgtg ttacgctgcg 180
atcagccaag aatttaacgt caacccggac gaagttcgcg acgccttcag ccaagcgcgc 240
gacagcctgc aggcgaatca cgacttcatc agcctgattc gtgagctgaa agctcaggcg 300
aacggtcgtc tgcgtgtcta cgccatgtct aatatcagcc tgccggattg ggaagtcctg 360
cgtatgaagc cagccgattg ggacatcttt gaccatgtat ttaccagcgg tgcggtgggt 420
gagcgcaagc cgaacctggc cttttatcgt cacgtcatcg cggccacgga tctgcagccg 480
caccagacga tcttcgtgga tgacaaactg gaaaacgtgc tgtctgcgcg ctcgctgggc 540
ttcacgggta tcgtgttcga cgagccaagc gaagtcaaac gtgcgctgcg taatctgatc 600
ggcgacccgg tgcagcgtgg tggcgagttc ctggttcgta atgctggcaa actgggttct 660
atcacccgta cgaccgcaaa acatgagagc atcccgctgg atgagaattt tgcacaactg 720
ttgattctgg aaattactgg taaccgcgca ctggtcaatc tggttgagca cccgcagacg 780
tggaacttct tccagggtaa gggccagctg acgaccgaag aatttccttt tgacctggat 840
acgacgagcc tgggtctgac gatcctgaag cgtagccgcg agattgccga ctccgtcatg 900
gacgaaatgt tggaatacgt ggaccctgac ggcatcattc agacctactt cgatcatcgt 960
cgcccgcgct ttgacccggt tgtttgcgtt aatgccctga gcctgttcta tgcatacggc 1020
cgtggtgagc aactgcgttc caccttgacc tgggtgcacg aagttctgtt gaaccgtgcg 1080
tatttggatg gtacgcgtta ctatgaaacg gccgagtgct ttctgtattt catgtcccgt 1140
ctgctggcaa ccagcggtga cccggatctg cattccctgc tgaagccgtt gctgaaggaa 1200
cgcgtgcaag agcgcatcgg cgctgacggt gacagcctgg cgctggcgat gcgcattttg 1260
gcatgtgatt ttgttggcat ccgtgatgaa gtggatctgc gtaccctgct gaccttacag 1320
tgcgaggatg gcggttggga agtgggctgg atgtacaaat acggtagcag cggtattagc 1380
attggtaacc gtggtctggc aaccgcattg gcgatcaaag ctgttgacac catgtttcaa 1440
ccgcaaatcc gtttcagcga gagcccgacc gacactctgg tggagaacgc gattcacaag 1500
cgccgcccga gcttttcaga gaaattttta ggtaagcgtc cgcgttccgg ttcgttccgt 1560
aaaccgctgc aatggattct gcagggcagc aagctgcgca agagcgtcga gatcggtagc 1620
taa 1623
<210> 93
<211> 1584
<212> DNA
<213> XP_007369631.1 针对酿酒酵母表达而优化的cDNA
<400> 93
atggcttcta tccacagaag atacactact ttgatcttgg acttgggtga cgttttgttc 60
agatggtctc caaagactga aactgctatc ccaccacaac aattgaagga catcttgtct 120
tctgttactt ggttcgaata cgaaagaggt agattgtctc aagaagcttg ttacgaaaga 180
tgtgctgaag aattcaagat cgaagcttct gttatcgctg aagctttcaa gcaagctaga 240
ggttctttga gaccaaacga agaattcatc gctttgatca gagacttgag aagagaaatg 300
cacggtgact tgactgtttt ggctttgtct aacatctctt tgccagacta cgaatacatc 360
atgtctttgt cttctgactg gactactgtt ttcgacagag ttttcccatc tgctttggtt 420
ggtgaaagaa agccacactt gggttgttac agaaaggtta tctctgaaat gaacttggaa 480
ccacaaacta ctgttttcgt tgacgacaag ttggacaacg ttgcttctgc tagatctttg 540
ggtatgcacg gtatcgtttt cgacaaccaa gctaacgttt tcagacaatt gagaaacatc 600
ttcggtgacc caatcagaag aggtcaagaa tacttgagag gtcacgctgg taagttggaa 660
tcttctactg acaacggttt gatcttcgaa gaaaacttca ctcaattgat catctacgaa 720
ttgactcaag acagaacttt gatctctttg tctgaatgtc caagaacttg gaacttcttc 780
agaggtgaac cattgttctc tgaaactttc ccagacgacg ttgacactac ttctgttgct 840
ttgactgttt tgcaaccaga cagagctttg gttaactctg ttttggacga aatgttggaa 900
tacgttgacg ctgacggtat catgcaaact tacttcgaca gatctagacc aagaatggac 960
ccattcgttt gtgttaacgt tttgtctttg ttctacgaaa acggtagagg tcacgaattg 1020
ccaagaactt tggactgggt ttacgaagtt ttgttgcaca gagcttacca cggtggttct 1080
agatactact tgtctccaga ctgtttcttg ttcttcatgt ctagattgtt gaagagagct 1140
gacgacccag ctgttcaagc tagattgaga ccattgttcg ttgaaagagt taacgaaaga 1200
gttggtgctg ctggtgactc tatggacttg gctttcagaa tcttggctgc tgcttctgtt 1260
ggtgttcaat gtccaagaga cttggaaaga ttgactgctg gtcaatgtga cgacggtggt 1320
tgggacttgt gttggttcta cgttttcggt tctactggtg ttaaggctgg taacagaggt 1380
ttgactactg ctttggctgt tactgctatc caaactgcta tcggtagacc accatctcca 1440
tctccatctg ctgcttcttc ttctttcaga ccatcttctc catacaagtt cttgggtatc 1500
tctagaccag cttctccaat cagattcggt gacttgttga gaccatggag aaagatgtct 1560
agatctaact tgaagtctca ataa 1584
<210> 94
<211> 1623
<212> DNA
<213> XP_006461126 针对酿酒酵母表达而优化的cDNA
<400> 94
atggctccac cacaaagacc attcactgct atcgttttcg acatcggtga cgttttgttc 60
caatggtctg ctactactaa gacttctatc tctccaaaga ctttgagatc tatcttgaac 120
tgtccaactt ggttcgacta cgaaagaggt agattggctg aaaacgcttg ttacgctgct 180
atctctcaag aattcaacgt taacccagac gaagttagag acgctttctc tcaagctaga 240
gactctttgc aagctaacca cgacttcatc tctttgatca gagaattgaa ggctcaagct 300
aacggtagat tgagagttta cgctatgtct aacatctctt tgccagactg ggaagttttg 360
agaatgaagc cagctgactg ggacatcttc gaccacgttt tcacttctgg tgctgttggt 420
gaaagaaagc caaacttggc tttctacaga cacgttatcg ctgctactga cttgcaacca 480
caccaaacta tcttcgttga cgacaagttg gaaaacgttt tgtctgctag atctttgggt 540
ttcactggta tcgttttcga cgaaccatct gaagttaaga gagctttgag aaacttgatc 600
ggtgacccag ttcaaagagg tggtgaattc ttggttagaa acgctggtaa gttgggttct 660
atcactagaa ctactgctaa gcacgaatct atcccattgg acgaaaactt cgctcaattg 720
ttgatcttgg aaatcactgg taacagagct ttggttaact tggttgaaca cccacaaact 780
tggaacttct tccaaggtaa gggtcaattg actactgaag aattcccatt cgacttggac 840
actacttctt tgggtttgac tatcttgaag agatctagag aaatcgctga ctctgttatg 900
gacgaaatgt tggaatacgt tgacccagac ggtatcatcc aaacttactt cgaccacaga 960
agaccaagat tcgacccagt tgtttgtgtt aacgctttgt ctttgttcta cgcttacggt 1020
agaggtgaac aattgagatc tactttgact tgggttcacg aagttttgtt gaacagagct 1080
tacttggacg gtactagata ctacgaaact gctgaatgtt tcttgtactt catgtctaga 1140
ttgttggcta cttctggtga cccagacttg cactctttgt tgaagccatt gttgaaggaa 1200
agagttcaag aaagaatcgg tgctgacggt gactctttgg ctttggctat gagaatcttg 1260
gcttgtgact tcgttggtat cagagacgaa gttgacttga gaactttgtt gactttgcaa 1320
tgtgaagacg gtggttggga agttggttgg atgtacaagt acggttcttc tggtatctct 1380
atcggtaaca gaggtttggc tactgctttg gctatcaagg ctgttgacac tatgttccaa 1440
ccacaaatca gattctctga atctccaact gacactttgg ttgaaaacgc tatccacaag 1500
agaagaccat ctttctctga aaagttcttg ggtaagagac caagatctgg ttctttcaga 1560
aagccattgc aatggatctt gcaaggttct aagttgagaa agtctgttga aatcggttct 1620
taa 1623
<210> 95
<211> 1566
<212> DNA
<213> SEQ ID NO: 68 - LoTps1 针对酿酒酵母表达而优化的cDNA
<400> 95
atgtacactg ctttgatctt ggacttgggt gacgttttgt tctcttggtc ttctactact 60
aacactacta tcccaccaag acaattgaag gaaatcttgt cttctccagc ttggttcgaa 120
tacgaaagag gtagaatcac tcaagctgaa tgttacgaaa gagtttctgc tgaattctct 180
ttggacgcta ctgctgttgc tgaagctttc agacaagcta gagactcttt gagaccaaac 240
gacaagttct tgactttgat cagagaattg agacaacaat ctcacggtga attgactgtt 300
ttggctttgt ctaacatctc tttgccagac tacgaattca tcatggcttt ggactctaag 360
tggacttctg ttttcgacag agttttccca tctgctttgg ttggtgaaag aaagccacac 420
ttgggtgctt tcagacaagt tttgtctgaa atgaacttgg acccacacac tactgttttc 480
gttgacgaca agttggacaa cgttgtttct gctagatctt tgggtatgca cggtgttgtt 540
ttcgactctc aagacaacgt tttcagaatg ttgagaaaca tcttcggtga cccaatccac 600
agaggtagag actacttgag acaacacgct ggtagattgg aaacttctac tgacgctggt 660
gttgttttcg aagaaaactt cactcaattg atcatctacg aattgactaa cgacaagtct 720
ttgatcacta cttctaactg tgctagaact tggaacttct tcagaggtaa gccattgttc 780
tctgcttctt tcccagacga catggacact acttctgttg ctttgactgt tttgagattg 840
gaccacgctt tggttaactc tgttttggac gaaatgttga agtacgttga cgctgacggt 900
atcatgcaaa cttacttcga ccacactaga ccaagaatgg acccattcgt ttgtgttaac 960
gttttgtctt tgttccacga acaaggtaga ggtcacgaat tgccaaacac tttggaatgg 1020
gttcacgaag ttttgttgca cagagcttac atcggtggtt ctagatacta cttgtctgct 1080
gactgtttct tgttcttcat gtctagattg ttgcaaagaa tcactgaccc atctgttttg 1140
ggtagattca gaccattgtt catcgaaaga gttagagaaa gagttggtgc tactggtgac 1200
tctatcgact tggctttcag aatcatcgct gcttctactg ttggtatcca atgtccaaga 1260
gacttggaat ctttgttggc tgctcaatgt gaagacggtg gttgggactt gtgttggttc 1320
taccaatacg gttctactgg tgttaaggct ggtaacagag gtttgactac tgctttggct 1380
atcaaggcta tcgactctgc tatcgctaga ccaccatctc cagctttgtc tgttgcttct 1440
tcttctaagt ctgaaatccc aaagccaatc caaagatctt tgagaccatt gtctccaaga 1500
agattcggtg gtttcttgat gccatggaga agatctcaaa gaaacggtgt tgctgtttct 1560
tcttaa 1566
<210> 96
<211> 1602
<212> DNA
<213> EMD37666.1 针对酿酒酵母表达而优化的cDNA
<400> 96
atgtctgctg ctgctcaata cactactttg atcttggact tgggtgacgt tttgttcact 60
tggtctccaa agactaagac ttctatccca ccaagaactt tgaaggaaat cttgaactct 120
gctacttggt acgaatacga aagaggtaga atctctcaag acgaatgtta cgaaagagtt 180
ggtactgaat tcggtatcgc tccatctgaa atcgacaacg ctttcaagca agctagagac 240
tctatggaat ctaacgacga attgatcgct ttggttagag aattgaagac tcaattggac 300
ggtgaattgt tggttttcgc tttgtctaac atctctttgc cagactacga atacgttttg 360
actaagccag ctgactggtc tatcttcgac aaggttttcc catctgcttt ggttggtgaa 420
agaaagccac acttgggtgt ttacaagcac gttatcgctg aaactggtat cgacccaaga 480
actactgttt tcgttgacga caagatcgac aacgttttgt ctgctagatc tgttggtatg 540
cacggtatcg ttttcgaaaa gcaagaagac gttatgagag ctttgagaaa catcttcggt 600
gacccagtta gaagaggtag agaatacttg agaagaaacg ctatgagatt ggaatctgtt 660
actgaccacg gtgttgcttt cggtgaaaac ttcactcaat tgttgatctt ggaattgact 720
aacgacccat ctttggttac tttgccagac agaccaagaa cttggaactt cttcagaggt 780
aacggtggta gaccatctaa gccattgttc tctgaagctt tcccagacga cttggacact 840
acttctttgg ctttgactgt tttgcaaaga gacccaggtg ttatctcttc tgttatggac 900
gaaatgttga actacagaga cccagacggt atcatgcaaa cttacttcga cgacggtaga 960
caaagattgg acccattcgt taacgttaac gttttgactt tcttctacac taacggtaga 1020
ggtcacgaat tggaccaatg tttgacttgg gttagagaag ttttgttgta cagagcttac 1080
ttgggtggtt ctagatacta cccatctgct gactgtttct tgtacttcat ctctagattg 1140
ttcgcttgta ctaacgaccc agttttgcac caccaattga agccattgtt cgttgaaaga 1200
gttcaagaac aaatcggtgt tgaaggtgac gctttggaat tggctttcag attgttggtt 1260
tgtgcttctt tggacgttca aaacgctatc gacatgagaa gattgttgga aatgcaatgt 1320
gaagacggtg gttgggaagg tggtaacttg tacagattcg gtactactgg tttgaaggtt 1380
actaacagag gtttgactac tgctgctgct gttcaagcta tcgaagcttc tcaaagaaga 1440
ccaccatctc catctccatc tgttgaatct actaagtctc caatcactcc agttactcca 1500
atgttggaag ttccatcttt gggtttgtct atctctagac catcttctcc attgttgggt 1560
tacttcagat tgccatggaa gaagtctgct gaagttcact aa 1602
<210> 97
<211> 1461
<212> DNA
<213> XP_001217376.1 针对酿酒酵母表达而优化的cDNA
<400> 97
atggctatca ctaagggtcc agttaaggct ttgatcttgg acttctctaa cgttttgtgt 60
tcttggaagc caccatctaa cgttgctgtt ccaccacaaa tcttgaagat gatcatgtct 120
tctgacatct ggcacgacta cgaatgtggt agatactcta gagaagactg ttacgctaga 180
gttgctgaca gattccacat ctctgctgct gacatggaag acactttgaa gcaagctaga 240
aagtctttgc aagttcacca cgaaactttg ttgttcatcc aacaagttaa gaaggacgct 300
ggtggtgaat tgatggtttg tggtatgact aacactccaa gaccagaaca agacgttatg 360
cactctatca acgctgaata cccagttttc gacagaatct acatctctgg tttgatgggt 420
atgagaaagc catctatctg tttctaccaa agagttatgg aagaaatcgg tttgtctggt 480
gacgctatca tgttcatcga cgacaagttg gaaaacgtta tcgctgctca atctgttggt 540
atcagaggtg ttttgttcca atctcaacaa gacttgagaa gagttgtttt gaacttcttg 600
ggtgacccag ttcacagagg tttgcaattc ttggctgcta acgctaagaa gatggactct 660
gttactaaca ctggtgacac tatccaagac aacttcgctc aattgttgat cttggaattg 720
gctcaagaca gagaattggt taagttgcaa gctggtaaga gaacttggaa ctacttcatc 780
ggtccaccaa agttgactac tgctactttc ccagacgaca tggacactac ttctatggct 840
ttgtctgttt tgccagttgc tgaagacgtt gtttcttctg ttttggacga aatgttgaag 900
ttcgttactg acgacggtat cttcatgact tacttcgact cttctagacc aagagttgac 960
ccagttgttt gtatcaacgt tttgggtgtt ttctgtagac acaacagaga aagagacgtt 1020
ttgccaactt tccactggat cagagacatc ttgatcaaca gagcttactt gtctggtact 1080
agatactacc catctccaga cttgttcttg ttcttcttgg ctagattgtg tttggctgtt 1140
agaaaccaat ctttgagaga acaattggtt ttgccattgg ttgacagatt gagagaaaga 1200
gttggtgctc caggtgaagc tgtttctttg gctgctagaa tcttggcttg tagatctttc 1260
ggtatcgact ctgctagaga catggactct ttgagaggta agcaatgtga agacggtggt 1320
tggccagttg aatgggttta cagattcgct tctttcggtt tgaacgttgg taacagaggt 1380
ttggctactg ctttcgctgt tagagctttg gaatctccat acggtgaatc tgctgttaag 1440
gttatgagaa gaatcgttta a 1461
<210> 98
<211> 1680
<212> DNA
<213> 胡椒莓(Drimys lanceolata)_DlTps589 野生型DNA序列
<400> 98
atggatctta ttaatccctc cccagcggct tccaccctcc ctctcccagt tgatggagat 60
tcagaagttg ttaggcgatc tgccgggttt catccgacta tctggggcga tcacttcctc 120
tcctacaagc ccgatccaaa gaaaatagat gcatggaata aaagggttga agagctgaag 180
gaagaagtga agaagatatt aagcaatgca aaagggacgg tggaagagct gaatttgatt 240
gatgatctcg tacaccttgg gattagttat cattttgaga aggagattga tgatgctcta 300
caacacatct ttgataccca tcttgatgat tttcctaagg atgatctata tgtcgccgct 360
ctccgatttg gcgtcttaag gaaacagggg caccgtgttt ctccagatgt attcaaaaaa 420
ttcaaagatg agcaggggaa tttcaaggca gagttgagca ccgatgcgaa aggtttgcta 480
tgtttaaatg atgtggctta tctcagcaca agaggggaag atatcttgga tgaagccatt 540
cctttcactg aggagcacct taggtcttgt attagccatg tagattctca tatggcagca 600
aaaattgaac attctctcga gcttcccctt catcatcgca taccaaggct agagaacagg 660
cactacatct cagtctatga aggagacaag gaaaggaacg aagttgtcct tgagcttgcc 720
aatttagatt tcaatctgat tcaaatcttg caccaaagag agctgagaga catcacaatg 780
tggtggaagg agattgacct tgcagcaaag ctgcctttta ttagggatag gttggtggag 840
tgctactact ggatcatggg ggtctatttt gaaccaatat actcgagggc tagggttttt 900
tccaccaaaa tgacaatgtt ggtctcagtt gtggacgaca tatatgatgt gtatgctacc 960
gaggatgagc ttcaactatt cactgatgcc atctataggt gggatgctga tgacattgat 1020
cagctgcctc agtacttgaa agatgctttt atggtactct acaacactgt gaagactcta 1080
gaagaagaac ttgaaccaga aggaaactct tatcgtggat tctatgtaaa agatgcaatg 1140
aaggttttgg caagggatta ctttgtggag cacaaatggt ataacagaaa aattgtgcca 1200
tccgtagagg aatacttgaa aatttcttgc atcagtgtgg ccgttcatat ggctacagtt 1260
cactgtattg ctgggatgta tgaaattgca accaaagagg cattcgaatg gttgatgact 1320
gagcccaaac ttgttattga tgcatctctg attggtcgtc tccttgatga catgcagtcc 1380
acctcgtttg agcaacagag aggccacgtg tcatcagcag tacagtgtta catggctgaa 1440
tatggtgtaa cagcggaaga agcatgtgaa aagctccgag atatggctgc aattgcttgg 1500
aaagatgtga acgaggcatg ccttaggccc acggttttcc ctatgcctat ccttttgcct 1560
tctatcaact tggcacgtgt ggcagaagtc atctacctac gtggagatgg atacacgcac 1620
gctgggggtg agaccaagaa acacatcacg gccatgcttg ttaagccaat tgaagtctga 1680
<210> 99
<211> 559
<212> PRT
<213> 胡椒莓(Drimys lanceolata)_DlTps589蛋白
<400> 99
Met Asp Leu Ile Asn Pro Ser Pro Ala Ala Ser Thr Leu Pro Leu Pro
1 5 10 15
Val Asp Gly Asp Ser Glu Val Val Arg Arg Ser Ala Gly Phe His Pro
20 25 30
Thr Ile Trp Gly Asp His Phe Leu Ser Tyr Lys Pro Asp Pro Lys Lys
35 40 45
Ile Asp Ala Trp Asn Lys Arg Val Glu Glu Leu Lys Glu Glu Val Lys
50 55 60
Lys Ile Leu Ser Asn Ala Lys Gly Thr Val Glu Glu Leu Asn Leu Ile
65 70 75 80
Asp Asp Leu Val His Leu Gly Ile Ser Tyr His Phe Glu Lys Glu Ile
85 90 95
Asp Asp Ala Leu Gln His Ile Phe Asp Thr His Leu Asp Asp Phe Pro
100 105 110
Lys Asp Asp Leu Tyr Val Ala Ala Leu Arg Phe Gly Val Leu Arg Lys
115 120 125
Gln Gly His Arg Val Ser Pro Asp Val Phe Lys Lys Phe Lys Asp Glu
130 135 140
Gln Gly Asn Phe Lys Ala Glu Leu Ser Thr Asp Ala Lys Gly Leu Leu
145 150 155 160
Cys Leu Asn Asp Val Ala Tyr Leu Ser Thr Arg Gly Glu Asp Ile Leu
165 170 175
Asp Glu Ala Ile Pro Phe Thr Glu Glu His Leu Arg Ser Cys Ile Ser
180 185 190
His Val Asp Ser His Met Ala Ala Lys Ile Glu His Ser Leu Glu Leu
195 200 205
Pro Leu His His Arg Ile Pro Arg Leu Glu Asn Arg His Tyr Ile Ser
210 215 220
Val Tyr Glu Gly Asp Lys Glu Arg Asn Glu Val Val Leu Glu Leu Ala
225 230 235 240
Asn Leu Asp Phe Asn Leu Ile Gln Ile Leu His Gln Arg Glu Leu Arg
245 250 255
Asp Ile Thr Met Trp Trp Lys Glu Ile Asp Leu Ala Ala Lys Leu Pro
260 265 270
Phe Ile Arg Asp Arg Leu Val Glu Cys Tyr Tyr Trp Ile Met Gly Val
275 280 285
Tyr Phe Glu Pro Ile Tyr Ser Arg Ala Arg Val Phe Ser Thr Lys Met
290 295 300
Thr Met Leu Val Ser Val Val Asp Asp Ile Tyr Asp Val Tyr Ala Thr
305 310 315 320
Glu Asp Glu Leu Gln Leu Phe Thr Asp Ala Ile Tyr Arg Trp Asp Ala
325 330 335
Asp Asp Ile Asp Gln Leu Pro Gln Tyr Leu Lys Asp Ala Phe Met Val
340 345 350
Leu Tyr Asn Thr Val Lys Thr Leu Glu Glu Glu Leu Glu Pro Glu Gly
355 360 365
Asn Ser Tyr Arg Gly Phe Tyr Val Lys Asp Ala Met Lys Val Leu Ala
370 375 380
Arg Asp Tyr Phe Val Glu His Lys Trp Tyr Asn Arg Lys Ile Val Pro
385 390 395 400
Ser Val Glu Glu Tyr Leu Lys Ile Ser Cys Ile Ser Val Ala Val His
405 410 415
Met Ala Thr Val His Cys Ile Ala Gly Met Tyr Glu Ile Ala Thr Lys
420 425 430
Glu Ala Phe Glu Trp Leu Met Thr Glu Pro Lys Leu Val Ile Asp Ala
435 440 445
Ser Leu Ile Gly Arg Leu Leu Asp Asp Met Gln Ser Thr Ser Phe Glu
450 455 460
Gln Gln Arg Gly His Val Ser Ser Ala Val Gln Cys Tyr Met Ala Glu
465 470 475 480
Tyr Gly Val Thr Ala Glu Glu Ala Cys Glu Lys Leu Arg Asp Met Ala
485 490 495
Ala Ile Ala Trp Lys Asp Val Asn Glu Ala Cys Leu Arg Pro Thr Val
500 505 510
Phe Pro Met Pro Ile Leu Leu Pro Ser Ile Asn Leu Ala Arg Val Ala
515 520 525
Glu Val Ile Tyr Leu Arg Gly Asp Gly Tyr Thr His Ala Gly Gly Glu
530 535 540
Thr Lys Lys His Ile Thr Ala Met Leu Val Lys Pro Ile Glu Val
545 550 555
<210> 100
<211> 1680
<212> DNA
<213> 人工序列_来自胡椒莓(D. lanceolata)的DlTps589的密码子优化的DNA序列
<220>
<223> 来自胡椒莓(D. lanceolata)的DlTps589的密码子优化的DNA序列
<400> 100
atggacctga ttaacccgag ccctgctgca tccaccctgc cactgccagt cgatggtgat 60
agcgaagttg tgcgccgtag cgcgggtttc catccgacca tctggggtga ccactttctg 120
tcttataagc cggacccgaa aaagattgat gcgtggaaca agcgtgttga ggaactgaaa 180
gaagaggtca aaaagatttt gagcaatgcg aaaggcacgg ttgaggaact gaatttgatt 240
gacgacctgg tacacctggg tattagctat cactttgaga aagaaatcga cgacgcgctg 300
cagcatatct tcgatacgca cctggatgat ttcccgaaag atgacctcta cgtggctgcg 360
ctgcgttttg gcgtcctgcg taagcaaggc catcgtgtca gcccggacgt ctttaagaaa 420
ttcaaagacg agcaaggcaa cttcaaagcg gagctgtcaa ccgatgcaaa gggcctgttg 480
tgcctgaacg atgtggcgta cctgagcacc cgtggtgagg atatcctgga cgaagcgatc 540
ccgttcacgg aagaacattt gcgctcgtgc attagccacg ttgatagcca catggcagcg 600
aagattgagc actctctgga gctgccgctg caccatcgca ttccgcgttt agagaatcgc 660
cattacatct ccgtgtacga gggtgacaaa gagcgtaatg aagtcgttct ggagttggct 720
aacttggact ttaatcttat ccagatcctg caccagcgcg agctgcgcga catcacgatg 780
tggtggaaag aaattgatct ggccgcaaag ctgccgttta ttcgtgaccg tctggtggag 840
tgttactatt ggattatggg cgtgtacttc gagccgatct acagccgtgc gcgcgtgttt 900
agcaccaaga tgaccatgct ggttagcgtg gtggatgaca tctatgatgt ctacgctacg 960
gaagatgagt tgcagctgtt taccgacgcc atttacagat gggacgccga tgacattgat 1020
caactgccgc aatatctgaa agacgccttt atggttctgt acaacaccgt caaaaccctg 1080
gaagaagaac tggagccgga aggtaactct tatcgtggtt tctacgttaa agatgcgatg 1140
aaagttctgg cgcgtgacta tttcgttgag cataagtggt acaatcgtaa gatcgtcccg 1200
tccgttgaag agtacttgaa gattagctgt atcagcgtcg cagtccacat ggcgaccgtg 1260
cactgtatcg ccggcatgta tgagatcgcc acgaaagaag cattcgagtg gctgatgacc 1320
gagccgaaac tggtgattga cgcaagcctg attggtcgcc tgctggacga tatgcagagc 1380
acgagctttg agcagcagcg cggtcatgtt agctccgcag ttcaatgcta catggctgag 1440
tacggtgtga ctgccgaaga agcatgcgag aagctgcgtg atatggcggc cattgcgtgg 1500
aaagatgtga atgaagcatg cctgcgcccg accgttttcc cgatgccgat tttactgcct 1560
agcatcaacc tggcacgtgt ggcggaagtt atctatctgc gtggcgacgg ttatacgcac 1620
gcgggtggtg agactaagaa gcacatcacc gcgatgctgg tcaagccgat cgaagtgtaa 1680
<210> 101
<211> 1656
<212> DNA
<213> 辛酸八角木(Drimys winteri)_SCH51_3228_9 野生型DNA序列
<400> 101
atggcttcca ccctccctct cccagcttat ggagattcag aagttgttag gcgatctgcc 60
gggtttcatc cgacgatctg gggcgatcac ttcctctcct acaagcctga tccaacgaaa 120
atagatgaat ggaataaaag ggttgaagag ctgaaggaag aagtgaagaa gatattaagc 180
aatgcaaaag ggacagtgga agagctgaat ttgcttgatg atctcgtaca ccttgggatt 240
agttatcatt ttgagaagga gattgatgat gctttacaac aaatctttga tacccatctt 300
gatgtttttc ctaaggatga tctatatgcc accgctctcc gatttggcgt cttaaggaaa 360
caggggcacc gtgtttctcc agatgtattc aaaaaattca aagatgagca ggggaatttc 420
aaggcagagt tgagcaccga tgcgaagggt ttgctatgtt tatatgatgt ggcttatctc 480
agcacaagag gggaagatat cttggatgaa gccattcctt tcactaagga gcaccttagg 540
tcttgtatta gccatgtcga ttctcatatg gcagcaaaaa ttgagcattc tctagagctt 600
ccccttcatc atcgcatacc aaggctagag aacaggcact acatctcagt ctatgaagga 660
gacaaggaaa ggaatgaagt tgtccttgag cttgccaaat tagatttcaa tctgattcaa 720
atcttgcacc aaagagagct gagggacatc acaacgtggt ggaaggagat tgaccttgca 780
gcaaagctac cttttattag ggataggttg gtggagtgct actattggat catgggagtc 840
tattttgaac caatatactc aagggctaga gttttttcga ccaaaatgac aatcttggtc 900
tcagttgtgg acgacatata tgatgtatat gctacagagg atgagctcca acttttcact 960
gatgcaatct ataggtggga tgctgaggac attgagcagc ttccacagta cttgaaagat 1020
gcttttcttg tactctataa cactgtgaag gacctagaag aggaattgga accagaagga 1080
aactcttatc gtggatacta tgtaaaagat gcgatgaagg ttttggcaag ggattacttt 1140
gtggagcaca aatggtataa cagaaaaatt gtgccatcag tagaggacta cctgcgaatt 1200
tcttgcatta gtgttgccgt tcatatggcc acagttcatt gtattgctgg gatgtatgaa 1260
attgcaacca aagaggcatt cgaatggttg aagacggaac ctaaacttgt tatagatgca 1320
tcactgattg ggcgtctcct cgatgacatg cagtccacct cgtttgagca acagagaggt 1380
catgtgtcat cagcggtaca gtgttacatg atccaatatg gggtatcaca cgaagaagcg 1440
tgtgagaagt tgcgagaaat ggctgcaatt gcgtggaaag atgtaaacca agcatgcctt 1500
aggcccactg ttttccctat gcctattctt ctgccctcca tcaaccttgc acgtgtggca 1560
gaagtgattt acctacgcgg agatggatat acacatgcgg gtggtgagac caaaaaacat 1620
atcacggcca tgcttgttga tccaatcaaa gtctga 1656
<210> 102
<211> 551
<212> PRT
<213> 辛酸八角木(Drimys winteri)_SCH51_3228_9蛋白
<400> 102
Met Ala Ser Thr Leu Pro Leu Pro Ala Tyr Gly Asp Ser Glu Val Val
1 5 10 15
Arg Arg Ser Ala Gly Phe His Pro Thr Ile Trp Gly Asp His Phe Leu
20 25 30
Ser Tyr Lys Pro Asp Pro Thr Lys Ile Asp Glu Trp Asn Lys Arg Val
35 40 45
Glu Glu Leu Lys Glu Glu Val Lys Lys Ile Leu Ser Asn Ala Lys Gly
50 55 60
Thr Val Glu Glu Leu Asn Leu Leu Asp Asp Leu Val His Leu Gly Ile
65 70 75 80
Ser Tyr His Phe Glu Lys Glu Ile Asp Asp Ala Leu Gln Gln Ile Phe
85 90 95
Asp Thr His Leu Asp Val Phe Pro Lys Asp Asp Leu Tyr Ala Thr Ala
100 105 110
Leu Arg Phe Gly Val Leu Arg Lys Gln Gly His Arg Val Ser Pro Asp
115 120 125
Val Phe Lys Lys Phe Lys Asp Glu Gln Gly Asn Phe Lys Ala Glu Leu
130 135 140
Ser Thr Asp Ala Lys Gly Leu Leu Cys Leu Tyr Asp Val Ala Tyr Leu
145 150 155 160
Ser Thr Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Pro Phe Thr Lys
165 170 175
Glu His Leu Arg Ser Cys Ile Ser His Val Asp Ser His Met Ala Ala
180 185 190
Lys Ile Glu His Ser Leu Glu Leu Pro Leu His His Arg Ile Pro Arg
195 200 205
Leu Glu Asn Arg His Tyr Ile Ser Val Tyr Glu Gly Asp Lys Glu Arg
210 215 220
Asn Glu Val Val Leu Glu Leu Ala Lys Leu Asp Phe Asn Leu Ile Gln
225 230 235 240
Ile Leu His Gln Arg Glu Leu Arg Asp Ile Thr Thr Trp Trp Lys Glu
245 250 255
Ile Asp Leu Ala Ala Lys Leu Pro Phe Ile Arg Asp Arg Leu Val Glu
260 265 270
Cys Tyr Tyr Trp Ile Met Gly Val Tyr Phe Glu Pro Ile Tyr Ser Arg
275 280 285
Ala Arg Val Phe Ser Thr Lys Met Thr Ile Leu Val Ser Val Val Asp
290 295 300
Asp Ile Tyr Asp Val Tyr Ala Thr Glu Asp Glu Leu Gln Leu Phe Thr
305 310 315 320
Asp Ala Ile Tyr Arg Trp Asp Ala Glu Asp Ile Glu Gln Leu Pro Gln
325 330 335
Tyr Leu Lys Asp Ala Phe Leu Val Leu Tyr Asn Thr Val Lys Asp Leu
340 345 350
Glu Glu Glu Leu Glu Pro Glu Gly Asn Ser Tyr Arg Gly Tyr Tyr Val
355 360 365
Lys Asp Ala Met Lys Val Leu Ala Arg Asp Tyr Phe Val Glu His Lys
370 375 380
Trp Tyr Asn Arg Lys Ile Val Pro Ser Val Glu Asp Tyr Leu Arg Ile
385 390 395 400
Ser Cys Ile Ser Val Ala Val His Met Ala Thr Val His Cys Cys Ala
405 410 415
Gly Met Asp Glu Ile Ala Thr Lys Glu Ala Phe Glu Trp Leu Lys Thr
420 425 430
Glu Pro Lys Leu Val Ile Asp Ala Ser Leu Ile Gly Arg Leu Leu Asp
435 440 445
Asp Met Gln Ser Thr Ser Phe Glu Gln Gln Arg Gly His Val Ser Ser
450 455 460
Ala Val Gln Cys Tyr Met Ile Gln Tyr Gly Val Ser His Glu Glu Ala
465 470 475 480
Cys Glu Lys Leu Arg Glu Met Ala Ala Ile Ala Trp Lys Asp Val Asn
485 490 495
Gln Ala Cys Leu Arg Pro Thr Val Phe Pro Met Pro Ile Leu Leu Pro
500 505 510
Ser Ile Asn Leu Ala Arg Val Ala Glu Val Ile Tyr Leu Arg Gly Asp
515 520 525
Gly Tyr Thr His Ala Gly Gly Glu Thr Lys Lys His Ile Thr Ala Met
530 535 540
Leu Val Asp Pro Ile Lys Val
545 550
<210> 103
<211> 1656
<212> DNA
<213> 人工序列_SCH51-3228-9的密码子优化的DNA序列
<220>
<223> SCH51-3228-9的密码子优化的DNA序列
<400> 103
atggcaagca ccctgccgct gcctgcctat ggtgatagcg aagttgttcg tcgtagcgca 60
ggttttcatc cgaccatttg gggtgatcat tttctgagct ataaaccgga tccgaccaaa 120
attgatgaat ggaataaacg tgtcgaagaa ctgaaagaag aagtgaaaaa aatcctgagc 180
aatgccaaag gcaccgttga ggaactgaat ctgctggatg atctggttca tctgggtatc 240
agctatcact ttgagaaaga aatcgatgat gcactgcagc agatttttga tacccatctg 300
gatgttttcc cgaaagatga tctgtatgca accgcactgc gttttggtgt tctgcgtaaa 360
cagggtcatc gtgttagtcc ggatgtgttc aaaaaattca aagatgaaca gggcaacttc 420
aaagcagaac tgagcaccga tgcaaaaggt ctgctgtgtc tgtatgatgt tgcatatctg 480
agcacccgtg gtgaagatat tctggatgaa gcaattccgt ttaccaaaga acatctgcgt 540
agctgtatta gccatgttga tagccacatg gcagcgaaaa ttgaacatag cctggaactg 600
cctctgcatc accgtattcc gcgtctggaa aatcgtcact atattagcgt ttatgagggc 660
gataaagaac gcaatgaagt tgtgctggaa ctggcaaaac tggattttaa cctgattcag 720
attctgcatc agcgtgaact gcgtgatatt accacctggt ggaaagaaat tgatctggca 780
gcaaaactgc cgtttattcg tgatcgtctg gttgaatgct attattggat tatgggcgtg 840
tatttcgaac cgatttatag ccgtgcacgt gtttttagca ccaaaatgac cattctggtt 900
agcgtggtgg atgatatcta tgatgtttat gccaccgaag atgaactgca gctgtttacc 960
gatgccattt atcgttggga tgcagaagat attgaacagc tgccgcagta tctgaaagat 1020
gcatttctgg ttctgtacaa caccgtgaaa gatctggaag aagaactgga accggaaggt 1080
aatagctatc gtggttatta tgttaaagat gccatgaaag ttctggcacg cgattatttt 1140
gttgagcaca aatggtataa ccgcaaaatt gttccgagcg tggaagatta tctgcgtatt 1200
agctgcatta gcgttgcagt tcacatggca accgttcatt gttgtgcagg tatggatgaa 1260
attgcaacca aagaagcatt tgagtggctg aaaaccgaac cgaaactggt tattgatgca 1320
agcctgattg gtcgtctgct ggacgatatg cagagcacca gctttgaaca gcagcgtggt 1380
catgttagca gcgcagttca gtgttatatg attcagtatg gtgttagcca tgaagaagca 1440
tgcgaaaaac tgcgcgaaat ggcagcaatt gcatggaaag atgttaatca ggcatgtctg 1500
cgtccgaccg tttttccgat gccgattctg ctgccgagca ttaatctggc acgtgttgcc 1560
gaagttatct atctgcgtgg tgatggttat acccatgccg gtggtgaaac caaaaaacat 1620
attaccgcaa tgctggtcga tccgattaaa gtttaa 1656
<210> 104
<211> 1656
<212> DNA
<213> 辛酸八角木(Drimys winteri)_SCH51_3228_11 野生型DNA序列
<400> 104
atggcttcca ccctccctct cccagcttat ggagattcag aagttgttag gcgatctgcc 60
gggtttcatc cgacgatctg gggcgatcac ttcctctcct acaagcctga tccaacgaaa 120
atagatgaat ggaataaaag ggttgaagag ctgaaggaag aagtgaagaa gatattaagc 180
aatgcaaaag ggacagtgga agagctgaat ttgcttgatg atctcgtaca ccttgggatt 240
agttatcatt ttgagaagga gattgatgat gctttacaac aaatctttga tacccatctt 300
gatgtttttc ctaaggatga tctatatgcc accgctctcc gatttggcgt cttaaggaaa 360
caggggcacc gtgtttctcc agatgtattc aaaaaattca aagatgagca ggggaatttc 420
aaggcagagt tgagcaccga tgcgaagggt ttgctatgtt tatatgatgt ggcttatctc 480
agcacaagag gggaagatat cttggatgaa gccattcctt tcactaagga gcaccttagg 540
tcttgtatta gccatgtcga ttctcatatg gcagcaaaaa ttgagcattc tctagagctt 600
ccccttcatc atcgcatacc aaggctagag aacaggcact acatctcagt ctatgaagga 660
gacaaggaaa ggaatgaagt tgtccttgag cttgccaaat tagatttcaa tctgattcaa 720
atcttgcacc aaagagagct gagggacatc acaatgtggt ggaaggagat tgaccttgca 780
gcaaagctac cttttattag agataggttg gtggagtgct actactggat catgggggtc 840
tattttgaac caatatactc cagggctagg gttttttcca ctaaaatgac aatcttggtc 900
tcagttgtgg acgacatata tgatgtctat gctacggagg atgagcttca actattcact 960
gatgcaatct ataggtggga tgctgatgac attgatcagc tgcctcagta cttgaaagat 1020
gcttttatgg tactctataa cactgtgaag actctagaag aagaacttga accagaagga 1080
aactcttatc gtggatacta cgtaaaagat gcaatgaagg ttttggcaag agattacttt 1140
gtggaacaca aatggtataa cagacaaatt gtgccatccg tagaggaata cttgaaaatt 1200
tcttgcatta gtgtggctgt tcatatggct acagttcatt gtattgctgg gatgtatgaa 1260
attgctacca aagaggcatt cgaatggttg aagactgaac ccaaacttgt tatcgatgca 1320
tctctgatcg gtcgtcttct tgatgacatg cagtctacct cgtttgagca acaaagaggg 1380
cacgtgtcat cagcagtaca gtgttacatg gcccaatatg gagtaacagc agaagaagca 1440
tgtgaaaagc tacgagaaat ggctgcaatt gcttggaaag atgtgaatga agcatgcctt 1500
aggcccacgg tattccctat gcctatcctc ttgccttcta tcaacttggc acgtgtggca 1560
gaagtgatct acctacgtgg agatggatac acgcacgctg ggggtgagac caaaaaacac 1620
atcacggcca tgcttgttaa gccaattgaa gtctga 1656
<210> 105
<211> 551
<212> PRT
<213> 辛酸八角木(Drimys winteri)_SCH51_3228_11蛋白
<400> 105
Met Ala Ser Thr Leu Pro Leu Pro Ala Tyr Gly Asp Ser Glu Val Val
1 5 10 15
Arg Arg Ser Ala Gly Phe His Pro Thr Ile Trp Gly Asp His Phe Leu
20 25 30
Ser Tyr Lys Pro Asp Pro Thr Lys Ile Asp Glu Trp Asn Lys Arg Val
35 40 45
Glu Glu Leu Lys Glu Glu Val Lys Lys Ile Leu Ser Asn Ala Lys Gly
50 55 60
Thr Val Glu Glu Leu Asn Leu Leu Asp Asp Leu Val His Leu Gly Ile
65 70 75 80
Ser Tyr His Phe Glu Lys Glu Ile Asp Asp Ala Leu Gln Gln Ile Phe
85 90 95
Asp Thr His Leu Asp Val Phe Pro Lys Asp Asp Leu Tyr Ala Thr Ala
100 105 110
Leu Arg Phe Gly Val Leu Arg Lys Gln Gly His Arg Val Ser Pro Asp
115 120 125
Val Phe Lys Lys Phe Lys Asp Glu Gln Gly Asn Phe Lys Ala Glu Leu
130 135 140
Ser Thr Asp Ala Lys Gly Leu Leu Cys Leu Tyr Asp Val Ala Tyr Leu
145 150 155 160
Ser Thr Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Pro Phe Thr Lys
165 170 175
Glu His Leu Arg Ser Cys Ile Ser His Val Asp Ser His Met Ala Ala
180 185 190
Lys Ile Glu His Ser Leu Glu Leu Pro Leu His His Arg Ile Pro Arg
195 200 205
Leu Glu Asn Arg His Tyr Ile Ser Val Tyr Glu Gly Asp Lys Glu Arg
210 215 220
Asn Glu Val Val Leu Glu Leu Ala Lys Leu Asp Phe Asn Leu Ile Gln
225 230 235 240
Ile Leu His Gln Arg Glu Leu Arg Asp Ile Thr Met Trp Trp Lys Glu
245 250 255
Ile Asp Leu Ala Ala Lys Leu Pro Phe Ile Arg Asp Arg Leu Val Glu
260 265 270
Cys Tyr Tyr Trp Ile Met Gly Val Tyr Phe Glu Pro Ile Tyr Ser Arg
275 280 285
Ala Arg Val Phe Ser Thr Lys Met Thr Ile Leu Val Ser Val Val Asp
290 295 300
Asp Ile Tyr Asp Val Tyr Ala Thr Glu Asp Glu Leu Gln Leu Phe Thr
305 310 315 320
Asp Ala Ile Tyr Arg Trp Asp Ala Asp Asp Ile Asp Gln Leu Pro Gln
325 330 335
Tyr Leu Lys Asp Ala Phe Met Val Leu Tyr Asn Thr Val Lys Thr Leu
340 345 350
Glu Glu Glu Leu Glu Pro Glu Gly Asn Ser Tyr Arg Gly Tyr Tyr Val
355 360 365
Lys Asp Ala Met Lys Val Leu Ala Arg Asp Tyr Phe Val Glu His Lys
370 375 380
Trp Tyr Asn Arg Gln Ile Val Pro Ser Val Glu Glu Tyr Leu Lys Ile
385 390 395 400
Ser Cys Ile Ser Val Ala Val His Met Ala Thr Val His Cys Ile Ala
405 410 415
Gly Met Tyr Glu Ile Ala Thr Lys Glu Ala Phe Glu Trp Leu Lys Thr
420 425 430
Glu Pro Lys Leu Val Ile Asp Ala Ser Leu Ile Gly Arg Leu Leu Asp
435 440 445
Asp Met Gln Ser Thr Ser Phe Glu Gln Gln Arg Gly His Val Ser Ser
450 455 460
Ala Val Gln Cys Tyr Met Ala Gln Tyr Gly Val Thr Ala Glu Glu Ala
465 470 475 480
Cys Glu Lys Leu Arg Glu Met Ala Ala Ile Ala Trp Lys Asp Val Asn
485 490 495
Glu Ala Cys Leu Arg Pro Thr Val Phe Pro Met Pro Ile Leu Leu Pro
500 505 510
Ser Ile Asn Leu Ala Arg Val Ala Glu Val Ile Tyr Leu Arg Gly Asp
515 520 525
Gly Tyr Thr His Ala Gly Gly Glu Thr Lys Lys His Ile Thr Ala Met
530 535 540
Leu Val Lys Pro Ile Glu Val
545 550
<210> 106
<211> 1656
<212> DNA
<213> 人工序列_SCH51-3228-11的密码子优化的DNA序列
<220>
<223> SCH51-3228-11的密码子优化的DNA序列
<400> 106
atggcatcta ctcttccact gccggcttat ggtgattctg aggttgttcg tcgttccgcg 60
ggttttcacc ctaccatctg gggcgatcac tttctgtcct ataagccaga cccgaccaag 120
attgacgagt ggaataagcg tgtcgaggaa ctgaaagaag aagtgaaaaa gatcctgtcc 180
aacgcaaaag gtactgtcga ggagctgaat ctgctggatg acctggtgca tctgggcatc 240
agctatcact tcgaaaagga aattgacgac gctttgcagc aaatttttga tacgcacctg 300
gacgtctttc cgaaagatga cctgtatgcg accgcgctgc gctttggtgt gctgcgtaaa 360
cagggtcatc gcgtgtctcc tgatgtgttc aagaaattta aagatgaaca gggcaatttc 420
aaggccgagt tgagcacgga cgccaaaggt ttgctctgcc tgtacgacgt tgcatatctg 480
agcacccgtg gtgaagatat cctggacgaa gcgattccgt tcaccaagga acatctgcgc 540
tcgtgcattt cccatgtaga tagccacatg gcggccaaga tcgagcacag cctggagctg 600
cctttgcacc atcgtattcc gcgcctggag aatcgccatt acattagcgt ctatgagggt 660
gacaaagagc gcaacgaagt cgtgttagag ctggcgaagc tggacttcaa cctgattcaa 720
attctgcatc aacgcgagct gcgcgacatt accatgtggt ggaaagagat tgatctggca 780
gcgaagctgc cgttcatccg cgatcgtctg gttgagtgct actactggat catgggcgtc 840
tacttcgagc cgatctacag ccgcgctcgt gtgttttcga cgaagatgac catcctggtt 900
agcgttgttg atgacattta tgacgtttac gcgaccgaag atgaactgca gctgtttacg 960
gacgcaatct accgttggga cgcggatgat atcgaccagc tgccgcaata cttgaaagat 1020
gcgttcatgg ttttgtacaa caccgtcaaa acgctggaag aagaactgga gccggaaggc 1080
aacagctacc gtggttacta tgttaaagat gcgatgaaag ttctggcgcg cgactacttc 1140
gtcgagcaca agtggtataa ccgtcagatt gtgccgagcg tcgaggaata cctgaagatt 1200
agctgtatca gcgttgccgt tcacatggca acggtgcact gcatcgccgg tatgtacgag 1260
attgcgacga aagaagcctt cgaatggttg aaaaccgagc cgaagctggt tatcgacgcc 1320
agcctgatcg gtcgtttgct ggacgacatg caaagcacga gcttcgagca gcagcgcggc 1380
catgtgagca gcgctgttca gtgttatatg gcgcaatatg gcgtgaccgc agaagaagcg 1440
tgcgagaagc tgcgtgagat ggcagcaatt gcgtggaaag atgtgaatga agcctgtctg 1500
cgtccgactg tgtttccgat gccgatcctg ctgccgagca ttaacctggc gcgtgtggca 1560
gaggtcatct atctgcgtgg tgacggttac acccacgcgg gtggcgaaac caagaaacat 1620
atcaccgcaa tgctggttaa gccgattgaa gtgtaa 1656
<210> 107
<211> 1677
<212> DNA
<213> 辛酸八角木(Drimys winteri)_SCH51_998_28 野生型DNA序列
<400> 107
atggatctta gtacttcacc tgttctttct tcctcccccc ttccggtgga agacggaaaa 60
aatccggccg ttcgccgttc agctggattt caccccagta tttggggtga tcatttcctc 120
tcctacactg aagatcacaa gaagctggat gcatggagcg aaaggactca agtgttgaag 180
gaagaggtga ggagaatttt aatcaatgcc aaggggtcac tagaagagtt ggatttgttg 240
gatgcaatcc aacgccttgg ggtgaaatat cactttgaga aagagattga agaggcatta 300
caccatattt atgttgcaga aactcatgtt tctactgatg acttatattc cgtttctctc 360
cggtttcgac ttcttagaca acaagggtac aatgtatctg ctgatgtatt taaaaagttc 420
aaagatgaga ggggcaactt caaggcaagc ttaagtactg atgccagggg gttgctaagc 480
ttgtatgaag ctgcatttct cagcatacga ggagatgata tcttagatga agccataact 540
ttcacaagag agcagcttaa gtcttctatg acccatgttg atgcccctct tgccaaacaa 600
atagcccatg ccttagaggt accagcgcac aagcgcatac aaagactaga gaacattcgc 660
tacctcacaa tctaccaaga agagaaagga aggaatgatg tgttgcttga gcttgccaag 720
ttggatttca atatcttaca acaattgcat aagaaagaac tgagagacct tacaaagtgg 780
tggaaggaca cagacgttgc aggaaagcta cctttcatca gagataggtt ggtggaatgc 840
tattattgga tcttgggtgt gtattatgag ccagaatact ccagagctag aattttttct 900
accaaaatga caatcatggt ctcagttgtt gatgacatat atgacgtata tgctactgaa 960
gatgagctcc aactattcac tgatgcaatc tataggtggg atctggaggg cctagatcaa 1020
ctcccacagt tcttgaaaga ctgttttctt gtactctatg acaccgtcaa ggaattagaa 1080
gacgaactag aaccggaagg aaaatcctat cgtggatact atgtaaagga tgcgatgaag 1140
gttttggcta gagattactt cgttgagcac aaatggtata acagaaacat agtgccaagt 1200
gtagaagaat atctccgtgt ttcttgcatc agtgttgcag tccatatggc taacgtccat 1260
tgctgtgctg ggatgggaga tgtaatgagc aaagaggcat tcgaatggtt gaagagtgaa 1320
ccaaaggttg taatggatgc atcactaatt ggccgactgc tcgatgacat gcagtccacc 1380
gagtttgagc aaaagagagg ccatgttgca tcggctgtcc aatgttacat gaatgagtat 1440
ggagtgactt acaaagaagc gtgtgaaaag ctgcatgaaa tggctgccct tgcatggaaa 1500
gacgtaaacc aggcttgcct taaaccaact gttttccctc tccctgtatt tatgcctgca 1560
atcaaccttg cgcgagtggc tgaagtcatc taccttcgtg gagatgggta tactcattca 1620
ggaggagaga ctaaagaaaa tatcacgttg atgcttgtca atccaatctc tgtgtga 1677
<210> 108
<211> 558
<212> PRT
<213> 辛酸八角木(Drimys winteri)_SCH51_998_28蛋白
<400> 108
Met Asp Leu Ser Thr Ser Pro Val Leu Ser Ser Ser Pro Leu Pro Val
1 5 10 15
Glu Asp Gly Lys Asn Pro Ala Val Arg Arg Ser Ala Gly Phe His Pro
20 25 30
Ser Ile Trp Gly Asp His Phe Leu Ser Tyr Thr Glu Asp His Lys Lys
35 40 45
Leu Asp Ala Trp Ser Glu Arg Thr Gln Val Leu Lys Glu Glu Val Arg
50 55 60
Arg Ile Leu Ile Asn Ala Lys Gly Ser Leu Glu Glu Leu Asp Leu Leu
65 70 75 80
Asp Ala Ile Gln Arg Leu Gly Val Lys Tyr His Phe Glu Lys Glu Ile
85 90 95
Glu Glu Ala Leu His His Ile Tyr Val Ala Glu Thr His Val Ser Thr
100 105 110
Asp Asp Leu Tyr Ser Val Ser Leu Arg Phe Arg Leu Leu Arg Gln Gln
115 120 125
Gly Tyr Asn Val Ser Ala Asp Val Phe Lys Lys Phe Lys Asp Glu Arg
130 135 140
Gly Asn Phe Lys Ala Ser Leu Ser Thr Asp Ala Arg Gly Leu Leu Ser
145 150 155 160
Leu Tyr Glu Ala Ala Phe Leu Ser Ile Arg Gly Asp Asp Ile Leu Asp
165 170 175
Glu Ala Ile Thr Phe Thr Arg Glu Gln Leu Lys Ser Ser Met Thr His
180 185 190
Val Asp Ala Pro Leu Ala Lys Gln Ile Ala His Ala Leu Glu Val Pro
195 200 205
Ala His Lys Arg Ile Gln Arg Leu Glu Asn Ile Arg Tyr Leu Thr Ile
210 215 220
Tyr Gln Glu Glu Lys Gly Arg Asn Asp Val Leu Leu Glu Leu Ala Lys
225 230 235 240
Leu Asp Phe Asn Ile Leu Gln Gln Leu His Lys Lys Glu Leu Arg Asp
245 250 255
Leu Thr Lys Trp Trp Lys Asp Thr Asp Val Ala Gly Lys Leu Pro Phe
260 265 270
Ile Arg Asp Arg Leu Val Glu Cys Tyr Tyr Trp Ile Leu Gly Val Tyr
275 280 285
Tyr Glu Pro Glu Tyr Ser Arg Ala Arg Ile Phe Ser Thr Lys Met Thr
290 295 300
Ile Met Val Ser Val Val Asp Asp Ile Tyr Asp Val Tyr Ala Thr Glu
305 310 315 320
Asp Glu Leu Gln Leu Phe Thr Asp Ala Ile Tyr Arg Trp Asp Leu Glu
325 330 335
Gly Leu Asp Gln Leu Pro Gln Phe Leu Lys Asp Cys Phe Leu Val Leu
340 345 350
Tyr Asp Thr Val Lys Glu Leu Glu Asp Glu Leu Glu Pro Glu Gly Lys
355 360 365
Ser Tyr Arg Gly Tyr Tyr Val Lys Asp Ala Met Lys Val Leu Ala Arg
370 375 380
Asp Tyr Phe Val Glu His Lys Trp Tyr Asn Arg Asn Ile Val Pro Ser
385 390 395 400
Val Glu Glu Tyr Leu Arg Val Ser Cys Ile Ser Val Ala Val His Met
405 410 415
Ala Asn Val His Cys Cys Ala Gly Met Gly Asp Val Met Ser Lys Glu
420 425 430
Ala Phe Glu Trp Leu Lys Ser Glu Pro Lys Val Val Met Asp Ala Ser
435 440 445
Leu Ile Gly Arg Leu Leu Asp Asp Met Gln Ser Thr Glu Phe Glu Gln
450 455 460
Lys Arg Gly His Val Ala Ser Ala Val Gln Cys Tyr Met Asn Glu Tyr
465 470 475 480
Gly Val Thr Tyr Lys Glu Ala Cys Glu Lys Leu His Glu Met Ala Ala
485 490 495
Leu Ala Trp Lys Asp Val Asn Gln Ala Cys Leu Lys Pro Thr Val Phe
500 505 510
Pro Leu Pro Val Phe Met Pro Ala Ile Asn Leu Ala Arg Val Ala Glu
515 520 525
Val Ile Tyr Leu Arg Gly Asp Gly Tyr Thr His Ser Gly Gly Glu Thr
530 535 540
Lys Glu Asn Ile Thr Leu Met Leu Val Asn Pro Ile Ser Val
545 550 555
<210> 109
<211> 1677
<212> DNA
<213> 人工序列_SCH51-998-28的密码子优化的DNA序列
<220>
<223> SCH51-998-28的密码子优化的DNA序列
<400> 109
atggatctga gcaccagtcc ggttctgagc agctcaccgc tgccggttga agatggtaaa 60
aatccggcag ttcgtcgtag cgcaggtttt catccgagca tttggggtga tcattttctg 120
agctataccg aggatcacaa aaaactggat gcatggtcag aacgtaccca ggttctgaaa 180
gaagaagtgc gtcgtattct gattaatgca aaaggtagcc tggaagaact ggatctgctg 240
gatgcaattc agcgtctggg tgttaaatat cactttgaga aagaaatcga agaagccctg 300
catcatattt atgttgcaga aacccatgtg tcaaccgatg atctgtatag cgttagcctg 360
cgttttcgtc tgctgcgtca gcagggttat aatgttagcg cagatgtgtt caaaaaattc 420
aaagatgaac gcggtaactt caaagcaagc ctgagcaccg atgcacgtgg tctgctgagc 480
ctgtatgaag cagcatttct gagcattcgt ggtgatgata ttctggatga agcaattacc 540
tttacccgtg aacagctgaa aagcagcatg acccatgttg atgcaccgct ggcaaaacaa 600
attgcacatg cactggaagt tccggcacat aaacgtattc agcgcctgga aaatattcgc 660
tatctgacca tttaccaaga agagaaaggt cgtaacgatg ttctgctgga actggccaaa 720
ctggatttta acattctgca gcagctgcat aaaaaagaac tgcgtgatct gaccaaatgg 780
tggaaagata ccgatgttgc aggtaaactg ccgtttattc gtgatcgtct ggttgaatgc 840
tattattgga ttctgggcgt ttattatgag ccggaatata gccgtgcacg tatttttagc 900
accaaaatga ccattatggt tagcgtggtg gatgacatct atgatgttta tgcaaccgaa 960
gatgaactgc agctgtttac cgatgcaatt tatcgttggg atctggaagg tctggatcag 1020
ctgccgcagt tcctgaaaga ttgttttctg gttctgtatg ataccgtgaa agaactggaa 1080
gatgagctgg aaccggaagg taaaagctat cgtggttatt atgttaaaga tgccatgaaa 1140
gttctggcac gcgattattt tgttgagcac aaatggtata accgcaatat tgttccgagc 1200
gtggaagaat atctgcgtgt tagctgtatt agcgttgcag ttcacatggc aaatgttcat 1260
tgttgtgcag gtatgggtga tgtgatgagc aaagaagcat ttgaatggct gaaaagtgaa 1320
ccgaaagttg ttatggatgc cagcctgatt ggtcgcctgc tggacgatat gcagagcacc 1380
gaatttgaac agaaacgtgg tcatgttgca agcgcagttc agtgttatat gaatgaatat 1440
ggcgtgacct ataaagaggc atgcgaaaaa ctgcatgaaa tggcagcact ggcatggaaa 1500
gatgttaatc aggcatgtct gaaaccgacc gtttttccgc tgcctgtttt tatgcctgca 1560
attaatctgg cacgtgttgc cgaagttatt tacctgcgtg gggatggtta tacccatagc 1620
ggtggtgaaa ccaaagaaaa cattaccctg atgctggtta atccgattag cgtttaa 1677
<210> 110
<211> 1680
<212> DNA
<213> 胡椒莓(Drimys lanceolata)_SCH52_13163_6
<400> 110
atggatgttc taattccctc ccctgtggct tccactctcc ctctgcccga agatggaaac 60
ttggacgtcg ttcgcagatc cgccgggttt catccgacgg tctggggcga tcacttcctc 120
gcttactcgc ccgatccaac caaaatagat gcttggacta aaagagttga agagctgaag 180
caagaagtga agaggattct aagcaatgtg aaagggtcac tggaagagct gaacttgctt 240
gatgctatcc aacaccttgg gattggttat cattttgaga aagagattga tgatgcttta 300
caactaatct ttgattccca tattgatgct tttcctactg atgatctata tgtggctgcc 360
ctccgattta gcctactaag gcgacaaggg cactgtgttt cttcagatgt attcaaaaaa 420
ttcaaagatg agcaggggaa tttcaaggca gagctgagca ccgatgcgaa aggtttgctg 480
agtctctatg acgcggcgta tctcagtgta agaggggaag atatattgga tgaggccatt 540
cctttcacta gggagcacct taggacttgt attagccatg tagattctca tttggcagca 600
aaaattgagc attctctaga gcttcccctg catcatcgca taccaaggct agagaacagg 660
cactacatct cagtgtacga aggagagaag gaaaggaatg aagttgtact agagcttgcc 720
aaattagatt tcaatctgat tcaaatcttg caccaaagag agctgaggga catcacaacg 780
tggtggaatg agattgacct cgcagcaaag ctaccattta ttagggatag gttggtggag 840
tgctactatt ggatcatggg tgtctatttt gaaccaatat tctcaagggc tagagttttt 900
tcgaccaaaa tgacaatttt ggtctcagtt gtcgacgaca tatatgatgt ctacgctaca 960
gaggatgagc tccaactttt cactgacgca atctataggt gggatgccga ggacattgag 1020
cagcttccac agtacttgaa agattctttt cttgtactct ataacaccgt gaaggactta 1080
gaagaggagc tgaaaccaga aggaaactca tatcgtggag actatgtaaa agatgcgatg 1140
aaggttttgg caagagatta ctttgtggag cacaaatggt ataacagaaa aattgtaccg 1200
tcagtagagg actacctacg aatttcttgc attagtgttg ccgttcatat ggctacagtt 1260
cattgttgtg ctgggatgga tgaaattgca accaaagagg cattcgaatg gttgaagacc 1320
gaacctaaac ttgttataga tgcatcactg attgggcgtc tcctcgatga catgcagtcc 1380
acctcgtttg agcaacagag aggtcatgtg tcatcggcgg tacagtgtta catgatccaa 1440
tatggcgtat cacacgaaga agcgtgtgag aagttgacag aaatggctgc aattgcatgg 1500
aaagatgtaa accaagcatg ccttaggccc actgttttcc caatgcctat tcttctgcct 1560
tcaatcaacc ttgcacgtgt ggcagaagtc atctacctgc gcggagatgg atatacacat 1620
gctggtggtg agaccaaaaa acatatcacg gccatgcttg ttgaaccaat ccaagtctga 1680
<210> 111
<211> 559
<212> PRT
<213> 胡椒莓(Drimys lanceolata)_SCH52_13163_6蛋白
<400> 111
Met Asp Val Leu Ile Pro Ser Pro Val Ala Ser Thr Leu Pro Leu Pro
1 5 10 15
Glu Asp Gly Asn Leu Asp Val Val Arg Arg Ser Ala Gly Phe His Pro
20 25 30
Thr Val Trp Gly Asp His Phe Leu Ala Tyr Ser Pro Asp Pro Thr Lys
35 40 45
Ile Asp Ala Trp Thr Lys Arg Val Glu Glu Leu Lys Gln Glu Val Lys
50 55 60
Arg Ile Leu Ser Asn Val Lys Gly Ser Leu Glu Glu Leu Asn Leu Leu
65 70 75 80
Asp Ala Ile Gln His Leu Gly Ile Gly Tyr His Phe Glu Lys Glu Ile
85 90 95
Asp Asp Ala Leu Gln Leu Ile Phe Asp Ser His Ile Asp Ala Phe Pro
100 105 110
Thr Asp Asp Leu Tyr Val Ala Ala Leu Arg Phe Ser Leu Leu Arg Arg
115 120 125
Gln Gly His Cys Val Ser Ser Asp Val Phe Lys Lys Phe Lys Asp Glu
130 135 140
Gln Gly Asn Phe Lys Ala Glu Leu Ser Thr Asp Ala Lys Gly Leu Leu
145 150 155 160
Ser Leu Tyr Asp Ala Ala Tyr Leu Ser Val Arg Gly Glu Asp Ile Leu
165 170 175
Asp Glu Ala Ile Pro Phe Thr Arg Glu His Leu Arg Thr Cys Ile Ser
180 185 190
His Val Asp Ser His Leu Ala Ala Lys Ile Glu His Ser Leu Glu Leu
195 200 205
Pro Leu His His Arg Ile Pro Arg Leu Glu Asn Arg His Tyr Ile Ser
210 215 220
Val Tyr Glu Gly Glu Lys Glu Arg Asn Glu Val Val Leu Glu Leu Ala
225 230 235 240
Lys Leu Asp Phe Asn Leu Ile Gln Ile Leu His Gln Arg Glu Leu Arg
245 250 255
Asp Ile Thr Thr Trp Trp Asn Glu Ile Asp Leu Ala Ala Lys Leu Pro
260 265 270
Phe Ile Arg Asp Arg Leu Val Glu Cys Tyr Tyr Trp Ile Met Gly Val
275 280 285
Tyr Phe Glu Pro Ile Phe Ser Arg Ala Arg Val Phe Ser Thr Lys Met
290 295 300
Thr Ile Leu Val Ser Val Val Asp Asp Ile Tyr Asp Val Tyr Ala Thr
305 310 315 320
Glu Asp Glu Leu Gln Leu Phe Thr Asp Ala Ile Tyr Arg Trp Asp Ala
325 330 335
Glu Asp Ile Glu Gln Leu Pro Gln Tyr Leu Lys Asp Ser Phe Leu Val
340 345 350
Leu Tyr Asn Thr Val Lys Asp Leu Glu Glu Glu Leu Lys Pro Glu Gly
355 360 365
Asn Ser Tyr Arg Gly Asp Tyr Val Lys Asp Ala Met Lys Val Leu Ala
370 375 380
Arg Asp Tyr Phe Val Glu His Lys Trp Tyr Asn Arg Lys Ile Val Pro
385 390 395 400
Ser Val Glu Asp Tyr Leu Arg Ile Ser Cys Ile Ser Val Ala Val His
405 410 415
Met Ala Thr Val His Cys Cys Ala Gly Met Asp Glu Ile Ala Thr Lys
420 425 430
Glu Ala Phe Glu Trp Leu Lys Thr Glu Pro Lys Leu Val Ile Asp Ala
435 440 445
Ser Leu Ile Gly Arg Leu Leu Asp Asp Met Gln Ser Thr Ser Phe Glu
450 455 460
Gln Gln Arg Gly His Val Ser Ser Ala Val Gln Cys Tyr Met Ile Gln
465 470 475 480
Tyr Gly Val Ser His Glu Glu Ala Cys Glu Lys Leu Thr Glu Met Ala
485 490 495
Ala Ile Ala Trp Lys Asp Val Asn Gln Ala Cys Leu Arg Pro Thr Val
500 505 510
Phe Pro Met Pro Ile Leu Leu Pro Ser Ile Asn Leu Ala Arg Val Ala
515 520 525
Glu Val Ile Tyr Leu Arg Gly Asp Gly Tyr Thr His Ala Gly Gly Glu
530 535 540
Thr Lys Lys His Ile Thr Ala Met Leu Val Glu Pro Ile Gln Val
545 550 555
<210> 112
<211> 1680
<212> DNA
<213> 人工序列_SCH51-13163-6的密码子优化的DNA序列
<220>
<223> SCH51-13163-6的密码子优化的DNA序列
<400> 112
atggatgttc tgattccgag tccggttgca agcaccctgc cgctgccgga agatggtaat 60
ctggatgttg ttcgtcgtag cgcaggtttt catccgaccg tttggggtga tcattttctg 120
gcatatagtc cggatccgac caaaattgat gcatggacca aacgtgttga ggaactgaaa 180
caagaagtga aacgtattct gagcaatgtg aaaggtagcc tggaagaact gaatctgctg 240
gatgcaattc agcatctggg tattggttat cacttcgaga aagaaattga tgatgcactg 300
cagctgatct ttgatagcca tattgatgcc tttccgaccg atgatctgta tgttgcagca 360
ctgcgtttta gcctgctgcg tcgtcagggt cattgtgtta gcagtgatgt tttcaaaaaa 420
ttcaaagacg agcagggcaa ctttaaagca gaactgagca ccgatgcaaa aggtctgctg 480
agcctgtatg atgccgcata tctgagcgtt cgtggtgaag atattctgga tgaagcaatt 540
ccgtttaccc gtgaacatct gcgtacctgt attagccatg tggatagcca tctggcagca 600
aaaattgaac atagtctgga actgcctctg catcatcgta ttccgcgtct ggaaaatcgt 660
cactatatta gcgtttatga aggcgaaaaa gaacgcaatg aagttgtgct ggaactggca 720
aaactggatt ttaacctgat tcagattctg catcagcgtg aactgcgtga tattaccacc 780
tggtggaatg aaattgacct ggcagccaaa ctgccgttta ttcgtgatcg tctggttgaa 840
tgctattatt ggattatggg cgtgtatttt gaaccgattt ttagccgtgc acgtgtgttt 900
agcaccaaaa tgaccattct ggttagcgtg gtggatgata tctatgatgt ttatgcaacc 960
gaagatgagc tgcaactgtt taccgatgcc atttatcgtt gggatgcaga agatattgaa 1020
cagctgcctc agtatctgaa agatagcttt ctggttctgt acaacaccgt gaaagatctg 1080
gaagaagaac tgaaaccgga aggtaatagc tatcgtggtg attatgttaa agacgccatg 1140
aaagttctgg cacgcgatta ttttgttgag cacaaatggt ataaccgcaa aattgttccg 1200
agcgtggaag attatctgcg tattagctgc attagcgttg cagttcacat ggcaaccgtt 1260
cattgttgtg caggtatgga tgaaattgca accaaagaag catttgagtg gctgaaaacc 1320
gaaccgaaac tggttattga tgcaagcctg attggtcgtc tgctggacga tatgcagtca 1380
accagctttg aacagcagcg tggtcatgtt agcagcgcag ttcagtgtta tatgattcag 1440
tatggtgtta gccatgaaga agcatgcgaa aaactgaccg aaatggcagc aattgcatgg 1500
aaagatgtta atcaggcatg tctgcgtccg accgtgtttc ctatgccgat tctgctgccg 1560
agcattaatc tggcacgtgt tgccgaagtt atctatctgc gtggtgatgg ttatacccat 1620
gccggtggtg aaaccaaaaa acatattacc gcaatgctgg tagaaccgat tcaggtttaa 1680
<210> 113
<211> 1671
<212> DNA
<213> 黑水缬草(Valeriana amurensis)_VaTPS3
<400> 113
atgtctactg cattaaacag tgagcatgaa actgttcgtc cattagcaag ttttaaaccg 60
agtacatggg gcgatctttt catctcttat tctgaagata gccagcttaa ggaagtatat 120
ggtaaagagc acgaatgtct gaaacaacaa gtgaaaacaa tgttgttgga tctgacaaat 180
tatagaattt cggagaaaat cgctttcata aatacgttgg agagattagg ggtatctcat 240
gagtttgaga atgagattga agggctgctt catcaaatgt ttgatgctca ttctaaattc 300
caagatggca ttcaacactt tgatttgttc acattgggga tttactttag gattctcagg 360
caacatggct atagaatctc ttgtgatgtt ttcaacaagt tgaaagatag caacaatgaa 420
ttcaagaagg aacttaaaga ggacgctatt ggtttgctaa gtttgtacga agcgacacaa 480
gtaagagcac acgctgaaga aattttagac gaagccctca ttttcacaaa ggctcaactt 540
gaatccatag ccgcaacctc gagcttaagc ccatttgtcg agaagcaaat tactcatgct 600
ttggtccaag ctctccacaa aggaatccca agagtcgaat cgcgccattt catctctgtt 660
tatgaagaag atcctgacaa aaatgatttg ttgttgaggt tctcaaagat tgattacaat 720
cttgtacaaa tgcttcacaa gcaagaattg tgccatatct caaagtggtg gagagattcg 780
gagctcgaaa caaaactaac ttatgtgagg aatagagtgg cggaatgctt tttatggact 840
ctttgtgtgt accacgaacc aaagtactct ccggctcggc ttctgttagg caaactcata 900
aatatcatat cttgcactga tgacacatat gatgcgtatg gtacattaga ggaagttcag 960
atctttacag atgtcataca aaggttggat aggagttcta tggagcagct gccggattac 1020
atgaaaatcc tctacaaagc tgtccttgat ctttttgacg aagtagaagt tcagctatcg 1080
aaccatgaaa ctaataatac ttatcgtatg gcttatgcga aggaagagtt aaaagctatc 1140
gccaagtgct acgaaaagga gcacatatgg ttcagaaaat gtcacgtgcc cccattcgaa 1200
gaatatctag agaatgcggt agtgtcaatc ggtaatcgtt tggccgtacc tttttctttt 1260
ctgggaatgg atcaagtagc aggtgttgaa gcgttcgagt gggccaaaac tgatcccaaa 1320
atggtaaaat cgtgcggtaa agtcttacga cttgttgacg atgtaatgag ccacgaggag 1380
gaagatgtaa gaggacacgt ggcaacggga gtcgaatgct acatgaaaga acacggagtg 1440
agtagggaag aggccatcgt ggagttctac aagagggtcg agtacgcgtg gaaggatgtg 1500
aacgaggaat ttataacgcc gaaccatctg catatcgacc tcctcaaccg cgttcttaac 1560
cttacaagaa ttgcagacgt tgtttacaag tttgaagacg gctacacgca tcccgagaag 1620
actctgaaac atcatatcat ggcgttgttc gtcgaccccg tccccatata g 1671
<210> 114
<211> 556
<212> PRT
<213> 黑水缬草(Valeriana amurensis)_VaTPS3蛋白
<400> 114
Met Ser Thr Ala Leu Asn Ser Glu His Glu Thr Val Arg Pro Leu Ala
1 5 10 15
Ser Phe Lys Pro Ser Thr Trp Gly Asp Leu Phe Ile Ser Tyr Ser Glu
20 25 30
Asp Ser Gln Leu Lys Glu Val Tyr Gly Lys Glu His Glu Cys Leu Lys
35 40 45
Gln Gln Val Lys Thr Met Leu Leu Asp Leu Thr Asn Tyr Arg Ile Ser
50 55 60
Glu Lys Ile Ala Phe Ile Asn Thr Leu Glu Arg Leu Gly Val Ser His
65 70 75 80
Glu Phe Glu Asn Glu Ile Glu Gly Leu Leu His Gln Met Phe Asp Ala
85 90 95
His Ser Lys Phe Gln Asp Gly Ile Gln His Phe Asp Leu Phe Thr Leu
100 105 110
Gly Ile Tyr Phe Arg Ile Leu Arg Gln His Gly Tyr Arg Ile Ser Cys
115 120 125
Asp Val Phe Asn Lys Leu Lys Asp Ser Asn Asn Glu Phe Lys Lys Glu
130 135 140
Leu Lys Glu Asp Ala Ile Gly Leu Leu Ser Leu Tyr Glu Ala Thr Gln
145 150 155 160
Val Arg Ala His Ala Glu Glu Ile Leu Asp Glu Ala Leu Ile Phe Thr
165 170 175
Lys Ala Gln Leu Glu Ser Ile Ala Ala Thr Ser Ser Leu Ser Pro Phe
180 185 190
Val Glu Lys Gln Ile Thr His Ala Leu Val Gln Ala Leu His Lys Gly
195 200 205
Ile Pro Arg Val Glu Ser Arg His Phe Ile Ser Val Tyr Glu Glu Asp
210 215 220
Pro Asp Lys Asn Asp Leu Leu Leu Arg Phe Ser Lys Ile Asp Tyr Asn
225 230 235 240
Leu Val Gln Met Leu His Lys Gln Glu Leu Cys His Ile Ser Lys Trp
245 250 255
Trp Arg Asp Ser Glu Leu Glu Thr Lys Leu Thr Tyr Val Arg Asn Arg
260 265 270
Val Ala Glu Cys Phe Leu Trp Thr Leu Cys Val Tyr His Glu Pro Lys
275 280 285
Tyr Ser Pro Ala Arg Leu Leu Leu Gly Lys Leu Ile Asn Ile Ile Ser
290 295 300
Cys Thr Asp Asp Thr Tyr Asp Ala Tyr Gly Thr Leu Glu Glu Val Gln
305 310 315 320
Ile Phe Thr Asp Val Ile Gln Arg Leu Asp Arg Ser Ser Met Glu Gln
325 330 335
Leu Pro Asp Tyr Met Lys Ile Leu Tyr Lys Ala Val Leu Asp Leu Phe
340 345 350
Asp Glu Val Glu Val Gln Leu Ser Asn His Glu Thr Asn Asn Thr Tyr
355 360 365
Arg Met Ala Tyr Ala Lys Glu Glu Leu Lys Ala Ile Ala Lys Cys Tyr
370 375 380
Glu Lys Glu His Ile Trp Phe Arg Lys Cys His Val Pro Pro Phe Glu
385 390 395 400
Glu Tyr Leu Glu Asn Ala Val Val Ser Ile Gly Asn Arg Leu Ala Val
405 410 415
Pro Phe Ser Phe Leu Gly Met Asp Gln Val Ala Gly Val Glu Ala Phe
420 425 430
Glu Trp Ala Lys Thr Asp Pro Lys Met Val Lys Ser Cys Gly Lys Val
435 440 445
Leu Arg Leu Val Asp Asp Val Met Ser His Glu Glu Glu Asp Val Arg
450 455 460
Gly His Val Ala Thr Gly Val Glu Cys Tyr Met Lys Glu His Gly Val
465 470 475 480
Ser Arg Glu Glu Ala Ile Val Glu Phe Tyr Lys Arg Val Glu Tyr Ala
485 490 495
Trp Lys Asp Val Asn Glu Glu Phe Ile Thr Pro Asn His Leu His Ile
500 505 510
Asp Leu Leu Asn Arg Val Leu Asn Leu Thr Arg Ile Ala Asp Val Val
515 520 525
Tyr Lys Phe Glu Asp Gly Tyr Thr His Pro Glu Lys Thr Leu Lys His
530 535 540
His Ile Met Ala Leu Phe Val Asp Pro Val Pro Ile
545 550 555
<210> 115
<211> 1671
<212> DNA
<213> 人工序列_VaTPS3 nt序列,经密码子优化
<220>
<223> 密码子优化的序列
<400> 115
atgagcaccg cgttgaactc cgagcatgaa accgtccgtc cgctggctag ctttaaaccg 60
agcacgtggg gtgacctgtt catcagctac agcgaggaca gccagctgaa agaagtgtat 120
ggtaaagagc atgaatgtct taagcaacaa gttaagacca tgctgctgga cctgacgaat 180
taccgtatca gcgagaagat tgccttcatc aatacgctgg agcgcctggg tgtttctcac 240
gagttcgaga atgaaatcga aggcctcctg catcagatgt tcgacgcgca ctccaagttt 300
caagatggca ttcagcactt tgacctgttt accctgggca tttacttccg tattttgcgc 360
cagcacggtt atcgtatctc gtgcgatgtg tttaacaagc tgaaggactc taataacgaa 420
ttcaagaaag aactgaaaga agatgcaatt ggtctgctgt ctctgtatga agcgacccaa 480
gtgcgtgccc atgcagaaga gattttggac gaagcgctga tcttcaccaa ggctcagctg 540
gagagcatcg cggcgacgag cagcctgagc ccgtttgtcg agaaacagat tacccacgcc 600
ttggtgcaag cgttgcataa aggcatccca cgcgtggaga gccgccactt cattagcgtg 660
tacgaagagg acccggacaa gaacgatttg ctgctgcgtt tttccaagat tgactacaat 720
ttagttcaaa tgctgcacaa acaagagttg tgtcatatta gcaaatggtg gcgtgactcc 780
gagctggaga ctaaactgac ctacgtccgt aatcgcgtgg cagagtgttt tctgtggacc 840
ctgtgtgttt accacgagcc gaagtatagc ccggcacgtc tgctgctggg taaactgatc 900
aacatcattt cttgcacgga cgacacctat gatgcatacg gtacgctgga agaagtccaa 960
atctttaccg acgtgatcca gcgtttggac cgtagctcga tggagcagct gccggattac 1020
atgaagattc tgtataaagc tgttctggat ctgttcgatg aagttgaggt tcagctgagc 1080
aaccatgaga ctaacaatac ctaccgcatg gcgtacgcaa aagaagaact gaaggctatt 1140
gcgaaatgct acgagaaaga gcacatctgg tttcgcaagt gtcatgttcc accgttcgaa 1200
gagtatctgg agaacgccgt ggtgagcatc ggtaatcgtc tggcggtccc gttcagcttc 1260
ttgggtatgg accaggttgc gggcgtcgag gcctttgagt gggcaaagac cgatcctaaa 1320
atggttaaaa gctgcggtaa ggttctgcgc ctggtcgatg atgtcatgag ccatgaagaa 1380
gaagatgtgc gtggtcacgt ggcgacgggc gttgagtgct acatgaaaga gcacggtgtc 1440
agccgtgaag aggcgatcgt tgaattctat aagcgtgtcg agtatgcatg gaaagacgtc 1500
aacgaagagt tcattactcc gaatcacttg cacattgatc tgctgaaccg tgttctgaac 1560
ttaacccgca ttgccgatgt cgtatacaag tttgaagatg gctataccca cccggaaaag 1620
acgctgaaac accatatcat ggcgctgttc gtggacccgg tgccgatcta a 1671
<210> 116
<211> 1332
<212> DNA
<213> 香鳞毛蕨(Dryopteris fragrans)_DfACT13天然核苷酸序列
<400> 116
atggcctgtg gaggcggcgt gaggaatatt gaggtgaagg cgcaggagcc cgttttggtg 60
cagcctttat ctacagcaca aagctctgcc tataatcttc ttacaaaact ggaccagacg 120
ctcgcacaat tggtggtgca gattgtcttt gtcttcgatg tgaaaaaccc cgtaacccgc 180
cgaccccacg atggcgccga tcctgccaag cttctgaagg aggctctgag gaaggtcctt 240
gtgcccttct acccgctcgc tgggcgtctt tgcctctcgc ccgatgacgg cagtctgttc 300
attgactgca atgctcaggg ggtttccttt gttgaggcca atgccgacgc ggatatctcc 360
gaactcggcg acttctcgca gcccgatttc gcgactctgg gctcccttgt cttcccctta 420
cctcctatag cttcggatga tggtcctctt ctatctgcgc aggtgaccag attcaagtgt 480
ggaggatttg tgctgggctt cgtattccat cattgtttat ttgatggatt cgcactttcg 540
gaattcctaa atgcgtgggc ggagactgca tgcggcgtgc ctctttctac acctcctgtc 600
ctcgacagaa cttttccgag ggcgcgttct cccttgcaaa tcaagtatcc ccacaccgag 660
ttcctggaag tcgaagatgt ctctttgact caaaatatct ctaacgatgc catcaaccgg 720
tctttctgct ttacttcggc aagtctagag atactcaaga agaaagcctt ggaggatggg 780
gtgctatcaa aatgcactac ttttgaagct ttatctgggc tgatatggag ggcccgaact 840
agagccctgt ggagcgatta cccggagcac aaactaaagg tgctcattgt cgtcgacccg 900
agagcacgtt ttgagcctcg tgtggtgcca aaagggtatg tgggcaatgc ggtgcttttt 960
acatgtgctt tcgcaagcgc aagggagctg gaagaaaatc ccttgtcaca tgcagtgaag 1020
catgtgcaac atgccatcgg gcgtatgacg gaggagtaca tgttgtcgca aatcgactac 1080
atggagcatc agaaggtatg gtgcccaccg ctaggagcta gtacatcttt catgaccaaa 1140
tggtctaggt tggccttcaa tattctagac tttgggtggg gcaggccgaa gtatgtgggg 1200
ccggccacgt cgctgtcgat ggagacgact acttttgtat cttatggaaa gggcatgagt 1260
gtggtgttgg ctcttcctcc agaagcaatg cgcaaatttg aaaaaatcgt acatccctac 1320
ctcaatccat ga 1332
<210> 117
<211> 1332
<212> DNA
<213> 人工_DfACT13_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1332)
<400> 117
atg gca tgt ggc ggt ggg gtc agg aat atc gaa gtg aag gct caa gag 48
Met Ala Cys Gly Gly Gly Val Arg Asn Ile Glu Val Lys Ala Gln Glu
1 5 10 15
cca gtc ttg gtg caa ccg tta agc act gct caa tcg tca gcc tac aat 96
Pro Val Leu Val Gln Pro Leu Ser Thr Ala Gln Ser Ser Ala Tyr Asn
20 25 30
ttg tta aca aag tta gac cag act ttg gcg caa ttg gtt gtc caa ata 144
Leu Leu Thr Lys Leu Asp Gln Thr Leu Ala Gln Leu Val Val Gln Ile
35 40 45
gtg ttt gtt ttc gat gtg aag aac cct gtg aca aga agg cct cat gat 192
Val Phe Val Phe Asp Val Lys Asn Pro Val Thr Arg Arg Pro His Asp
50 55 60
gga gcc gat cca gct aag tta ttg aaa gag gcc tta agg aaa gta cta 240
Gly Ala Asp Pro Ala Lys Leu Leu Lys Glu Ala Leu Arg Lys Val Leu
65 70 75 80
gta cca ttc tat ccc ctt gcc gga aga cta tgt ctt tcc cca gac gat 288
Val Pro Phe Tyr Pro Leu Ala Gly Arg Leu Cys Leu Ser Pro Asp Asp
85 90 95
ggt agt cta ttc ata gac tgc aac gct caa ggt gtt tca ttt gtc gaa 336
Gly Ser Leu Phe Ile Asp Cys Asn Ala Gln Gly Val Ser Phe Val Glu
100 105 110
gca aac gct gat gcc gac atc agt gag tta ggt gat ttc tcc caa ccg 384
Ala Asn Ala Asp Ala Asp Ile Ser Glu Leu Gly Asp Phe Ser Gln Pro
115 120 125
gat ttt gcg acc tta ggt tct tta gtc ttc cct ctt cca cca att gcg 432
Asp Phe Ala Thr Leu Gly Ser Leu Val Phe Pro Leu Pro Pro Ile Ala
130 135 140
tct gac gat gga cca ttg tta agt gct caa gtc aca aga ttc aaa tgt 480
Ser Asp Asp Gly Pro Leu Leu Ser Ala Gln Val Thr Arg Phe Lys Cys
145 150 155 160
ggt ggc ttc gtt cta ggt ttc gtc ttc cat cac tgt cta ttt gat ggt 528
Gly Gly Phe Val Leu Gly Phe Val Phe His His Cys Leu Phe Asp Gly
165 170 175
ttt gcc tta tct gaa ttt ctt aat gca tgg gct gag aca gcg tgt gga 576
Phe Ala Leu Ser Glu Phe Leu Asn Ala Trp Ala Glu Thr Ala Cys Gly
180 185 190
gtc ccg tta tca acc cca cct gtt tta gat aga aca ttt cct cgt gct 624
Val Pro Leu Ser Thr Pro Pro Val Leu Asp Arg Thr Phe Pro Arg Ala
195 200 205
aga tct ccc ctg caa ata aag tac cca cat aca gag ttc cta gag gta 672
Arg Ser Pro Leu Gln Ile Lys Tyr Pro His Thr Glu Phe Leu Glu Val
210 215 220
gaa gat gtt tca tta act cag aac atc tct aac gat gct atc aac aga 720
Glu Asp Val Ser Leu Thr Gln Asn Ile Ser Asn Asp Ala Ile Asn Arg
225 230 235 240
tcc ttc tgt ttt aca tca gcc tct ttg gaa att ttg aag aag aag gcg 768
Ser Phe Cys Phe Thr Ser Ala Ser Leu Glu Ile Leu Lys Lys Lys Ala
245 250 255
cta gaa gac ggg gtg tta agc aaa tgt acg acc ttc gaa gct cta tct 816
Leu Glu Asp Gly Val Leu Ser Lys Cys Thr Thr Phe Glu Ala Leu Ser
260 265 270
ggc tta ata tgg aga gca cgt acc aga gcc tta tgg agc gac tac ccc 864
Gly Leu Ile Trp Arg Ala Arg Thr Arg Ala Leu Trp Ser Asp Tyr Pro
275 280 285
gag cac aag tta aag gtc ctg att gtc gtg gac cct cgt gcc aga ttt 912
Glu His Lys Leu Lys Val Leu Ile Val Val Asp Pro Arg Ala Arg Phe
290 295 300
gaa ccg aga gtg gtg cca aaa ggc tac gta ggg aat gca gtc ttg ttt 960
Glu Pro Arg Val Val Pro Lys Gly Tyr Val Gly Asn Ala Val Leu Phe
305 310 315 320
act tgt gca ttt gct tca gcc aga gaa cta gaa gaa aat cca tta tct 1008
Thr Cys Ala Phe Ala Ser Ala Arg Glu Leu Glu Glu Asn Pro Leu Ser
325 330 335
cat gct gtt aaa cac gta cag cac gcg atc ggc cgt atg act gag gaa 1056
His Ala Val Lys His Val Gln His Ala Ile Gly Arg Met Thr Glu Glu
340 345 350
tat atg cta tca caa att gat tac atg gag cac cag aag gtt tgg tgt 1104
Tyr Met Leu Ser Gln Ile Asp Tyr Met Glu His Gln Lys Val Trp Cys
355 360 365
ccc cca cta ggt gcg tcg act tct ttt atg act aag tgg tca agg ctt 1152
Pro Pro Leu Gly Ala Ser Thr Ser Phe Met Thr Lys Trp Ser Arg Leu
370 375 380
gcc ttt aac atc tta gat ttt ggt tgg ggt aga cct aag tat gtt ggt 1200
Ala Phe Asn Ile Leu Asp Phe Gly Trp Gly Arg Pro Lys Tyr Val Gly
385 390 395 400
ccg gct act tca ttg tct atg gaa aca act aca ttc gtc tcc tat gga 1248
Pro Ala Thr Ser Leu Ser Met Glu Thr Thr Thr Phe Val Ser Tyr Gly
405 410 415
aag ggt atg agc gtg gtg tta gcc cta ccc cca gag gca atg aga aag 1296
Lys Gly Met Ser Val Val Leu Ala Leu Pro Pro Glu Ala Met Arg Lys
420 425 430
ttc gaa aag att gta cac cct tat ttg aac cct taa 1332
Phe Glu Lys Ile Val His Pro Tyr Leu Asn Pro
435 440
<210> 118
<211> 443
<212> PRT
<213> 人工_DfACT13_针对其在酿酒酵母中的表达经密码子优化
<400> 118
Met Ala Cys Gly Gly Gly Val Arg Asn Ile Glu Val Lys Ala Gln Glu
1 5 10 15
Pro Val Leu Val Gln Pro Leu Ser Thr Ala Gln Ser Ser Ala Tyr Asn
20 25 30
Leu Leu Thr Lys Leu Asp Gln Thr Leu Ala Gln Leu Val Val Gln Ile
35 40 45
Val Phe Val Phe Asp Val Lys Asn Pro Val Thr Arg Arg Pro His Asp
50 55 60
Gly Ala Asp Pro Ala Lys Leu Leu Lys Glu Ala Leu Arg Lys Val Leu
65 70 75 80
Val Pro Phe Tyr Pro Leu Ala Gly Arg Leu Cys Leu Ser Pro Asp Asp
85 90 95
Gly Ser Leu Phe Ile Asp Cys Asn Ala Gln Gly Val Ser Phe Val Glu
100 105 110
Ala Asn Ala Asp Ala Asp Ile Ser Glu Leu Gly Asp Phe Ser Gln Pro
115 120 125
Asp Phe Ala Thr Leu Gly Ser Leu Val Phe Pro Leu Pro Pro Ile Ala
130 135 140
Ser Asp Asp Gly Pro Leu Leu Ser Ala Gln Val Thr Arg Phe Lys Cys
145 150 155 160
Gly Gly Phe Val Leu Gly Phe Val Phe His His Cys Leu Phe Asp Gly
165 170 175
Phe Ala Leu Ser Glu Phe Leu Asn Ala Trp Ala Glu Thr Ala Cys Gly
180 185 190
Val Pro Leu Ser Thr Pro Pro Val Leu Asp Arg Thr Phe Pro Arg Ala
195 200 205
Arg Ser Pro Leu Gln Ile Lys Tyr Pro His Thr Glu Phe Leu Glu Val
210 215 220
Glu Asp Val Ser Leu Thr Gln Asn Ile Ser Asn Asp Ala Ile Asn Arg
225 230 235 240
Ser Phe Cys Phe Thr Ser Ala Ser Leu Glu Ile Leu Lys Lys Lys Ala
245 250 255
Leu Glu Asp Gly Val Leu Ser Lys Cys Thr Thr Phe Glu Ala Leu Ser
260 265 270
Gly Leu Ile Trp Arg Ala Arg Thr Arg Ala Leu Trp Ser Asp Tyr Pro
275 280 285
Glu His Lys Leu Lys Val Leu Ile Val Val Asp Pro Arg Ala Arg Phe
290 295 300
Glu Pro Arg Val Val Pro Lys Gly Tyr Val Gly Asn Ala Val Leu Phe
305 310 315 320
Thr Cys Ala Phe Ala Ser Ala Arg Glu Leu Glu Glu Asn Pro Leu Ser
325 330 335
His Ala Val Lys His Val Gln His Ala Ile Gly Arg Met Thr Glu Glu
340 345 350
Tyr Met Leu Ser Gln Ile Asp Tyr Met Glu His Gln Lys Val Trp Cys
355 360 365
Pro Pro Leu Gly Ala Ser Thr Ser Phe Met Thr Lys Trp Ser Arg Leu
370 375 380
Ala Phe Asn Ile Leu Asp Phe Gly Trp Gly Arg Pro Lys Tyr Val Gly
385 390 395 400
Pro Ala Thr Ser Leu Ser Met Glu Thr Thr Thr Phe Val Ser Tyr Gly
405 410 415
Lys Gly Met Ser Val Val Leu Ala Leu Pro Pro Glu Ala Met Arg Lys
420 425 430
Phe Glu Lys Ile Val His Pro Tyr Leu Asn Pro
435 440
<210> 119
<211> 1440
<212> DNA
<213> 碳核曲霉(Aspergillus sclerotiicarbonarius)_PYI04555.1天然核苷酸序列
<400> 119
atgggtgcca gcgtctcttt ccagcccttt gtgcccactc ccctggacca tgctatgccc 60
ccaatttacg tgtctcaatt tctatgcttt cctaccacaa ctccgcaatc cgctattcaa 120
agtctccaag tgggaatcga gagattattc gaacgcctgc cattcctggc gggagagatt 180
ctcatcaatg aacacaccgg agccatcaaa gtccaggctc ccagtgcttt gatccgggaa 240
attccctaca tggccctccg agcccatcct gatctttacc ttccagctaa gcaatgtgca 300
accacaccaa tcgagagaca gttgaagacc aacagccttg atgaatccta ccatccactc 360
ccggcggcac ttccactctc ccaaccccag cccgtcatcc gattccaagc aaacaccctc 420
gcagacggca ttctctttgc agtcagctat catcattgca tattcgatgg cactggatgc 480
ggacagattc tggaaatgtt ggctcagtgc tgttcggcct ccgacgataa gatctccctg 540
ccgactgatt gccacaccga tgtgctcctc cgcgaataca tctccaatct aagccctact 600
accaacatcc cccacgatta cacgcaagcg tatagcacta cggtgcaacc ggaccccgat 660
gcctcagacc cagacacgtc ccccgccata ccctcctcac tctacacaga agcattcacc 720
tttccctccc aacaaatcac cactctccgt gatgcatgca accacctctt gcccaaatta 780
cccagcacca gcaacgcaca tccccacaaa ccaacaccga atcccctatc atcaaatgac 840
gtcctcaccg cactaatagc cctatgcatc acacgcgcca ccaacaccac caccccaccc 900
ctccaaccca acaatcacag tctctcaatg gccgtcaacc tccgaacccg catccagccc 960
caagtaccag atcactttct gggaaacttc gccacgctac tcccaataca ctttaccagt 1020
ccagtccaca cccaacagtc tgatctcctc ctcaccacag aaccccccga ccccgccctc 1080
atccacctaa ccaccctcgc ctcccaaatc cggtccagcc tatccacagt caacaccgat 1140
tacatctgcg gtctcatgac ggatctccga acccggcgca atgcaggaga aaacagcagt 1200
cttctaattg aaggcattaa gatttccagt tggagacatc tatccgtcta caagccggac 1260
tttggccctg ggttggggaa aatagcgggg tttgagttcc aggcggggct tatggataat 1320
ttggtggtga ttttgccttg gagaaatggg gattgggatg tgcgtgttac gttgttggag 1380
agggatatgc gcgggtttag ggaggatcgg ttggttaggt gggcgttggg gtctgggtag 1440
<210> 120
<211> 1470
<212> DNA
<213> 人工_PYI04555.1_针对其在酿酒酵母中的表达经密码子优化_C-末端扩展
<220>
<221> CDS
<222> (1)..(1470)
<400> 120
atg ggt gcg tca gtt agt ttt caa cct ttc gtt cct acc ccc ttg gat 48
Met Gly Ala Ser Val Ser Phe Gln Pro Phe Val Pro Thr Pro Leu Asp
1 5 10 15
cac gcc atg cca ccg ata tac gtg tca cag ttc ttg tgt ttc cct act 96
His Ala Met Pro Pro Ile Tyr Val Ser Gln Phe Leu Cys Phe Pro Thr
20 25 30
acg acg ccg cag tca gca ata caa tcc cta cag gtc ggt att gag cgt 144
Thr Thr Pro Gln Ser Ala Ile Gln Ser Leu Gln Val Gly Ile Glu Arg
35 40 45
ttg ttt gaa aga cta cct ttt cta gcc ggg gaa att ctg ata aat gag 192
Leu Phe Glu Arg Leu Pro Phe Leu Ala Gly Glu Ile Leu Ile Asn Glu
50 55 60
cac aca ggt gca ata aaa gtt caa gct ccc tct gct ctg att aga gaa 240
His Thr Gly Ala Ile Lys Val Gln Ala Pro Ser Ala Leu Ile Arg Glu
65 70 75 80
att ccg tac atg gcc tta agg gct cat ccg gat ttg tat ctg cct gct 288
Ile Pro Tyr Met Ala Leu Arg Ala His Pro Asp Leu Tyr Leu Pro Ala
85 90 95
aag caa tgt gca act act cca ata gag agg cag ttg aaa act aac agc 336
Lys Gln Cys Ala Thr Thr Pro Ile Glu Arg Gln Leu Lys Thr Asn Ser
100 105 110
ctt gat gag tct tat cat cca tta cct gcg gca cta cca ttg tct caa 384
Leu Asp Glu Ser Tyr His Pro Leu Pro Ala Ala Leu Pro Leu Ser Gln
115 120 125
cca caa cca gtc atc aga ttc caa gct aat aca ctt gct gac gga ata 432
Pro Gln Pro Val Ile Arg Phe Gln Ala Asn Thr Leu Ala Asp Gly Ile
130 135 140
ctt ttc gct gtc agt tac cac cat tgt ata ttt gat gga acg ggc tgc 480
Leu Phe Ala Val Ser Tyr His His Cys Ile Phe Asp Gly Thr Gly Cys
145 150 155 160
gga cag atc ctt gaa atg tta gcc cag tgt tgt tcg gcc tca gac gat 528
Gly Gln Ile Leu Glu Met Leu Ala Gln Cys Cys Ser Ala Ser Asp Asp
165 170 175
aag atc agc ttg cct acg gac tgt cat aca gat gta ttg ttg aga gaa 576
Lys Ile Ser Leu Pro Thr Asp Cys His Thr Asp Val Leu Leu Arg Glu
180 185 190
tat att tcg aat cta tct cca acc acg aat atc ccg cat gac tac act 624
Tyr Ile Ser Asn Leu Ser Pro Thr Thr Asn Ile Pro His Asp Tyr Thr
195 200 205
caa gct tat tct act aca gtt caa cca gat cct gac gca tcc gat cct 672
Gln Ala Tyr Ser Thr Thr Val Gln Pro Asp Pro Asp Ala Ser Asp Pro
210 215 220
gat act agc cca gcc att ccg agt tcg ctg tac act gaa gca ttt acg 720
Asp Thr Ser Pro Ala Ile Pro Ser Ser Leu Tyr Thr Glu Ala Phe Thr
225 230 235 240
ttt cct tca caa cag att acc aca cta aga gat gcg tgc aac cat tta 768
Phe Pro Ser Gln Gln Ile Thr Thr Leu Arg Asp Ala Cys Asn His Leu
245 250 255
ttg cct aaa tta cca tct act tca aac gca cat ccc cac aaa cca aca 816
Leu Pro Lys Leu Pro Ser Thr Ser Asn Ala His Pro His Lys Pro Thr
260 265 270
cca aac ccg tta tcc tca aat gac gtt ttg acg gca ctg atc gca ttg 864
Pro Asn Pro Leu Ser Ser Asn Asp Val Leu Thr Ala Leu Ile Ala Leu
275 280 285
tgc atc acc aga gct act aat acg acg aca ccc cca tta caa cca aac 912
Cys Ile Thr Arg Ala Thr Asn Thr Thr Thr Pro Pro Leu Gln Pro Asn
290 295 300
aac cat tct ctt agc atg gcc gtc aat cta agg act cgt att caa ccc 960
Asn His Ser Leu Ser Met Ala Val Asn Leu Arg Thr Arg Ile Gln Pro
305 310 315 320
caa gtc ccg gac cac ttt ctt ggt aat ttt gcc acc ttg cta cct att 1008
Gln Val Pro Asp His Phe Leu Gly Asn Phe Ala Thr Leu Leu Pro Ile
325 330 335
cac ttc aca agt cca gtc cat acg cag caa tca gat tta tta ttg act 1056
His Phe Thr Ser Pro Val His Thr Gln Gln Ser Asp Leu Leu Leu Thr
340 345 350
act gag ccg cca gac cca gca ttg atc cac ctt aca acc ctt gca tcg 1104
Thr Glu Pro Pro Asp Pro Ala Leu Ile His Leu Thr Thr Leu Ala Ser
355 360 365
caa att agg tct agt tta tcg acc gtt aac acc gat tac ata tgt ggc 1152
Gln Ile Arg Ser Ser Leu Ser Thr Val Asn Thr Asp Tyr Ile Cys Gly
370 375 380
ttg atg aca gat ctg agg act agg aga aac gcc ggt gag aat tcc tct 1200
Leu Met Thr Asp Leu Arg Thr Arg Arg Asn Ala Gly Glu Asn Ser Ser
385 390 395 400
ttg tta ata gaa ggt att aag att tct tca tgg aga cat ctt agc gtt 1248
Leu Leu Ile Glu Gly Ile Lys Ile Ser Ser Trp Arg His Leu Ser Val
405 410 415
tac aag cca gat ttc gga cct gga tta ggg aag att gca gga ttt gag 1296
Tyr Lys Pro Asp Phe Gly Pro Gly Leu Gly Lys Ile Ala Gly Phe Glu
420 425 430
ttt caa gct ggt tta atg gac aac ctt gtt gtg ata ttg cct tgg aga 1344
Phe Gln Ala Gly Leu Met Asp Asn Leu Val Val Ile Leu Pro Trp Arg
435 440 445
aac ggt gat tgg gac gtg cgt gta act ctt ctg gag aga gat atg agg 1392
Asn Gly Asp Trp Asp Val Arg Val Thr Leu Leu Glu Arg Asp Met Arg
450 455 460
ggt ttt agg gaa gac aga tta gtc aga tgg gca tta ggt agc gga aca 1440
Gly Phe Arg Glu Asp Arg Leu Val Arg Trp Ala Leu Gly Ser Gly Thr
465 470 475 480
ggc ccc ttt tcc ttt gtc gat atc atg taa 1470
Gly Pro Phe Ser Phe Val Asp Ile Met
485
<210> 121
<211> 489
<212> PRT
<213> 人工_PYI04555.1_针对其在酿酒酵母中的表达经密码子优化_C-末端扩展
<400> 121
Met Gly Ala Ser Val Ser Phe Gln Pro Phe Val Pro Thr Pro Leu Asp
1 5 10 15
His Ala Met Pro Pro Ile Tyr Val Ser Gln Phe Leu Cys Phe Pro Thr
20 25 30
Thr Thr Pro Gln Ser Ala Ile Gln Ser Leu Gln Val Gly Ile Glu Arg
35 40 45
Leu Phe Glu Arg Leu Pro Phe Leu Ala Gly Glu Ile Leu Ile Asn Glu
50 55 60
His Thr Gly Ala Ile Lys Val Gln Ala Pro Ser Ala Leu Ile Arg Glu
65 70 75 80
Ile Pro Tyr Met Ala Leu Arg Ala His Pro Asp Leu Tyr Leu Pro Ala
85 90 95
Lys Gln Cys Ala Thr Thr Pro Ile Glu Arg Gln Leu Lys Thr Asn Ser
100 105 110
Leu Asp Glu Ser Tyr His Pro Leu Pro Ala Ala Leu Pro Leu Ser Gln
115 120 125
Pro Gln Pro Val Ile Arg Phe Gln Ala Asn Thr Leu Ala Asp Gly Ile
130 135 140
Leu Phe Ala Val Ser Tyr His His Cys Ile Phe Asp Gly Thr Gly Cys
145 150 155 160
Gly Gln Ile Leu Glu Met Leu Ala Gln Cys Cys Ser Ala Ser Asp Asp
165 170 175
Lys Ile Ser Leu Pro Thr Asp Cys His Thr Asp Val Leu Leu Arg Glu
180 185 190
Tyr Ile Ser Asn Leu Ser Pro Thr Thr Asn Ile Pro His Asp Tyr Thr
195 200 205
Gln Ala Tyr Ser Thr Thr Val Gln Pro Asp Pro Asp Ala Ser Asp Pro
210 215 220
Asp Thr Ser Pro Ala Ile Pro Ser Ser Leu Tyr Thr Glu Ala Phe Thr
225 230 235 240
Phe Pro Ser Gln Gln Ile Thr Thr Leu Arg Asp Ala Cys Asn His Leu
245 250 255
Leu Pro Lys Leu Pro Ser Thr Ser Asn Ala His Pro His Lys Pro Thr
260 265 270
Pro Asn Pro Leu Ser Ser Asn Asp Val Leu Thr Ala Leu Ile Ala Leu
275 280 285
Cys Ile Thr Arg Ala Thr Asn Thr Thr Thr Pro Pro Leu Gln Pro Asn
290 295 300
Asn His Ser Leu Ser Met Ala Val Asn Leu Arg Thr Arg Ile Gln Pro
305 310 315 320
Gln Val Pro Asp His Phe Leu Gly Asn Phe Ala Thr Leu Leu Pro Ile
325 330 335
His Phe Thr Ser Pro Val His Thr Gln Gln Ser Asp Leu Leu Leu Thr
340 345 350
Thr Glu Pro Pro Asp Pro Ala Leu Ile His Leu Thr Thr Leu Ala Ser
355 360 365
Gln Ile Arg Ser Ser Leu Ser Thr Val Asn Thr Asp Tyr Ile Cys Gly
370 375 380
Leu Met Thr Asp Leu Arg Thr Arg Arg Asn Ala Gly Glu Asn Ser Ser
385 390 395 400
Leu Leu Ile Glu Gly Ile Lys Ile Ser Ser Trp Arg His Leu Ser Val
405 410 415
Tyr Lys Pro Asp Phe Gly Pro Gly Leu Gly Lys Ile Ala Gly Phe Glu
420 425 430
Phe Gln Ala Gly Leu Met Asp Asn Leu Val Val Ile Leu Pro Trp Arg
435 440 445
Asn Gly Asp Trp Asp Val Arg Val Thr Leu Leu Glu Arg Asp Met Arg
450 455 460
Gly Phe Arg Glu Asp Arg Leu Val Arg Trp Ala Leu Gly Ser Gly Thr
465 470 475 480
Gly Pro Phe Ser Phe Val Asp Ile Met
485
<210> 122
<211> 1425
<212> DNA
<213> 鞭苔(Bazzania trilobata)_ERR364415-1_contig_8546天然核苷酸序列
<400> 122
atggcccggg caccaccacc tccgcctcct ggtctcagaa tgagagacac agtgctcagc 60
atcgtgaagc cgatacggaa gacacagcat ttggagacga tcgacgcaac gttcgtcgat 120
ttgatgagaa tggacagctt tatacccgtg attttcgcct acaggcctgc ggacaagtcc 180
gaggccgcgt actcgcgtct cgtgaatcgc atcaaggagt cgctccagaa ggttttggtc 240
ccctttttcg ggttcgcggg ccgctgggtt ccaagcagtg gcgggagcag gcggctcttg 300
tgcaacgatg agggcgttcc cttcattgaa gcgtttgtgg acgaagagtt ggactcggtg 360
gtgaaggctt ccgccgcatt ccagccggtt acggagctga atggcttggg cgtcctcgga 420
atggacatga cttcatacga tcaaaggatg ccaccggaag gtgggcaacc ttgcgtcgtt 480
gctcaagtca cacggttcaa atgtggggga gtggttctgg gggtggcttt caatcacact 540
cacactgacg gccagggatt ctacaccttc atgcgagcat ggtccgactt ctctcgaacc 600
aacggaacgg caatcaaggt ggaccacaac cgggccctgc cagaactggc ttccctctca 660
cagttcttca tcaaacagca cgaccgaata ggaggcaaaa cttctaccga tcgagtcaac 720
gatcattgtt ctaaagttcc ggaacggctg gctttgaaag ctttcgaggt tcgtgcgtct 780
aagatcaaag ccgcaaagct agcagccgaa gatggagggg ttgggtatgt cagcacggta 840
gattgcattg tggctcactt atggaaaact cttgccagat tgccgcccgt cgtgttggat 900
gggagggaga ttacggtctt ctcgcctgtg gaggggagga acagattctt ggacccgcca 960
agacccaata tgtgtggaaa ttgttttgca gcaatggtga cccccaaaat cccaacccag 1020
gagttgctgg agatgcctct cgctgcaatt gcaggcaagc aacgggagaa attatccaca 1080
acccgaaggg aggaatggtt tggacagcaa agctttaggg agctggcctc cgcgatgaac 1140
accagcaaat ctgctctact tattgtgacc tcgtggttca actttcccat gtatgagatc 1200
gactttggag ctggcaaacc attttttgca tccactacga acatgatttc tcctatcaac 1260
ggcgtgtgtt gtggagtcat tgcaccccca actcctggga gctgctcctc cattgccact 1320
ctgtacattt tgtgccttcc cgcggtacta gaggctcttg aaaatgttcc agatttccta 1380
tccttcttcg ttcctcaccc aaatcacaaa gataactcgc aatag 1425
<210> 123
<211> 1455
<212> DNA
<213> 人工_ERR364415-1_contig_8546_针对其在酿酒酵母中的表达经密码子优化_C-末端扩展
<220>
<221> CDS
<222> (1)..(1455)
<400> 123
atg gcc agg gct cca ccc cca cca cca ccg ggc ttg cgt atg aga gat 48
Met Ala Arg Ala Pro Pro Pro Pro Pro Pro Gly Leu Arg Met Arg Asp
1 5 10 15
acg gtc cta tca atc gtc aaa cct ata aga aag act caa cac cta gag 96
Thr Val Leu Ser Ile Val Lys Pro Ile Arg Lys Thr Gln His Leu Glu
20 25 30
aca att gat gca aca ttc gta gat tta atg aga atg gac tca ttt atc 144
Thr Ile Asp Ala Thr Phe Val Asp Leu Met Arg Met Asp Ser Phe Ile
35 40 45
cct gtg atc ttc gct tat aga cct gca gat aag agc gag gct gcg tac 192
Pro Val Ile Phe Ala Tyr Arg Pro Ala Asp Lys Ser Glu Ala Ala Tyr
50 55 60
tct aga tta gtt aat agg ata aaa gaa agc tta cag aaa gta cta gta 240
Ser Arg Leu Val Asn Arg Ile Lys Glu Ser Leu Gln Lys Val Leu Val
65 70 75 80
cca ttc ttt gga ttt gct gga agg tgg gtg cct tct tcc ggt ggc tca 288
Pro Phe Phe Gly Phe Ala Gly Arg Trp Val Pro Ser Ser Gly Gly Ser
85 90 95
agg cgt ctg cta tgc aat gac gaa ggt gtc cct ttt att gag gcg ttt 336
Arg Arg Leu Leu Cys Asn Asp Glu Gly Val Pro Phe Ile Glu Ala Phe
100 105 110
gta gac gaa gaa ctg gac tct gtc gtt aag gct tca gct gcc ttc caa 384
Val Asp Glu Glu Leu Asp Ser Val Val Lys Ala Ser Ala Ala Phe Gln
115 120 125
cct gta act gaa ctt aat ggt cta ggt gtg ttg ggt atg gat atg act 432
Pro Val Thr Glu Leu Asn Gly Leu Gly Val Leu Gly Met Asp Met Thr
130 135 140
agt tat gat caa aga atg ccc cct gaa ggc ggt caa ccg tgc gtc gta 480
Ser Tyr Asp Gln Arg Met Pro Pro Glu Gly Gly Gln Pro Cys Val Val
145 150 155 160
gct cag gta acg aga ttt aaa tgc ggc ggt gtg gta ttg ggt gta gca 528
Ala Gln Val Thr Arg Phe Lys Cys Gly Gly Val Val Leu Gly Val Ala
165 170 175
ttc aac cat acc cat act gac ggg cag ggc ttt tat act ttt atg cgt 576
Phe Asn His Thr His Thr Asp Gly Gln Gly Phe Tyr Thr Phe Met Arg
180 185 190
gca tgg tcg gat ttc agt aga acg aat ggt acc gca att aaa gtc gat 624
Ala Trp Ser Asp Phe Ser Arg Thr Asn Gly Thr Ala Ile Lys Val Asp
195 200 205
cac aac agg gca cta ccc gaa tta gca tct ttg agt cag ttc ttt att 672
His Asn Arg Ala Leu Pro Glu Leu Ala Ser Leu Ser Gln Phe Phe Ile
210 215 220
aag cag cat gat aga att ggt ggg aaa act tcc acc gat aga gtc aat 720
Lys Gln His Asp Arg Ile Gly Gly Lys Thr Ser Thr Asp Arg Val Asn
225 230 235 240
gac cac tgt agc aaa gtc cct gaa aga cta gca cta aag gct ttt gaa 768
Asp His Cys Ser Lys Val Pro Glu Arg Leu Ala Leu Lys Ala Phe Glu
245 250 255
gtt agg gcg tcc aaa atc aag gca gca aaa cta gcc gca gaa gat ggt 816
Val Arg Ala Ser Lys Ile Lys Ala Ala Lys Leu Ala Ala Glu Asp Gly
260 265 270
ggt gta ggc tac gtc tcg acg gtg gat tgt att gtt gct cat cta tgg 864
Gly Val Gly Tyr Val Ser Thr Val Asp Cys Ile Val Ala His Leu Trp
275 280 285
aag aca cta gct cgt ttg cca cca gtt gtg tta gac ggt cgt gag atc 912
Lys Thr Leu Ala Arg Leu Pro Pro Val Val Leu Asp Gly Arg Glu Ile
290 295 300
act gtg ttt agc cca gta gaa ggc aga aat agg ttt ctt gat ccg ccc 960
Thr Val Phe Ser Pro Val Glu Gly Arg Asn Arg Phe Leu Asp Pro Pro
305 310 315 320
cgt ccg aac atg tgt ggt aat tgt ttc gct gca atg gta acc cct aaa 1008
Arg Pro Asn Met Cys Gly Asn Cys Phe Ala Ala Met Val Thr Pro Lys
325 330 335
atc cca aca caa gaa ttg tta gag atg cca tta gcc gcc att gcc ggc 1056
Ile Pro Thr Gln Glu Leu Leu Glu Met Pro Leu Ala Ala Ile Ala Gly
340 345 350
aag cag aga gag aaa cta tct acc acg aga cgt gaa gag tgg ttc gga 1104
Lys Gln Arg Glu Lys Leu Ser Thr Thr Arg Arg Glu Glu Trp Phe Gly
355 360 365
cag caa tca ttc agg gag ttg gct tca gct atg aat act tct aaa tca 1152
Gln Gln Ser Phe Arg Glu Leu Ala Ser Ala Met Asn Thr Ser Lys Ser
370 375 380
gct ttg tta atc gtg aca tct tgg ttt aac ttc ccg atg tat gaa atc 1200
Ala Leu Leu Ile Val Thr Ser Trp Phe Asn Phe Pro Met Tyr Glu Ile
385 390 395 400
gat ttt ggt gcc ggt aag ccg ttc ttc gct agt acg act aat atg att 1248
Asp Phe Gly Ala Gly Lys Pro Phe Phe Ala Ser Thr Thr Asn Met Ile
405 410 415
tct cct ata aat gga gtt tgt tgt ggt gtc ata gcc ccg ccc acc ccc 1296
Ser Pro Ile Asn Gly Val Cys Cys Gly Val Ile Ala Pro Pro Thr Pro
420 425 430
ggt tcc tgt tca tcc ata gcg aca tta tac att tta tgt tta cca gcc 1344
Gly Ser Cys Ser Ser Ile Ala Thr Leu Tyr Ile Leu Cys Leu Pro Ala
435 440 445
gtg tta gaa gct ctt gaa aat gtc cca gat ttc ctt tcg ttc ttc gta 1392
Val Leu Glu Ala Leu Glu Asn Val Pro Asp Phe Leu Ser Phe Phe Val
450 455 460
ccg cat cca aac cat aaa gac aac agc caa aca ggc ccc ttt tcc ttt 1440
Pro His Pro Asn His Lys Asp Asn Ser Gln Thr Gly Pro Phe Ser Phe
465 470 475 480
gtc gat atc atg taa 1455
Val Asp Ile Met
<210> 124
<211> 484
<212> PRT
<213> 人工_ERR364415-1_contig_8546_针对其在酿酒酵母中的表达经密码子优化_C-末端扩展
<400> 124
Met Ala Arg Ala Pro Pro Pro Pro Pro Pro Gly Leu Arg Met Arg Asp
1 5 10 15
Thr Val Leu Ser Ile Val Lys Pro Ile Arg Lys Thr Gln His Leu Glu
20 25 30
Thr Ile Asp Ala Thr Phe Val Asp Leu Met Arg Met Asp Ser Phe Ile
35 40 45
Pro Val Ile Phe Ala Tyr Arg Pro Ala Asp Lys Ser Glu Ala Ala Tyr
50 55 60
Ser Arg Leu Val Asn Arg Ile Lys Glu Ser Leu Gln Lys Val Leu Val
65 70 75 80
Pro Phe Phe Gly Phe Ala Gly Arg Trp Val Pro Ser Ser Gly Gly Ser
85 90 95
Arg Arg Leu Leu Cys Asn Asp Glu Gly Val Pro Phe Ile Glu Ala Phe
100 105 110
Val Asp Glu Glu Leu Asp Ser Val Val Lys Ala Ser Ala Ala Phe Gln
115 120 125
Pro Val Thr Glu Leu Asn Gly Leu Gly Val Leu Gly Met Asp Met Thr
130 135 140
Ser Tyr Asp Gln Arg Met Pro Pro Glu Gly Gly Gln Pro Cys Val Val
145 150 155 160
Ala Gln Val Thr Arg Phe Lys Cys Gly Gly Val Val Leu Gly Val Ala
165 170 175
Phe Asn His Thr His Thr Asp Gly Gln Gly Phe Tyr Thr Phe Met Arg
180 185 190
Ala Trp Ser Asp Phe Ser Arg Thr Asn Gly Thr Ala Ile Lys Val Asp
195 200 205
His Asn Arg Ala Leu Pro Glu Leu Ala Ser Leu Ser Gln Phe Phe Ile
210 215 220
Lys Gln His Asp Arg Ile Gly Gly Lys Thr Ser Thr Asp Arg Val Asn
225 230 235 240
Asp His Cys Ser Lys Val Pro Glu Arg Leu Ala Leu Lys Ala Phe Glu
245 250 255
Val Arg Ala Ser Lys Ile Lys Ala Ala Lys Leu Ala Ala Glu Asp Gly
260 265 270
Gly Val Gly Tyr Val Ser Thr Val Asp Cys Ile Val Ala His Leu Trp
275 280 285
Lys Thr Leu Ala Arg Leu Pro Pro Val Val Leu Asp Gly Arg Glu Ile
290 295 300
Thr Val Phe Ser Pro Val Glu Gly Arg Asn Arg Phe Leu Asp Pro Pro
305 310 315 320
Arg Pro Asn Met Cys Gly Asn Cys Phe Ala Ala Met Val Thr Pro Lys
325 330 335
Ile Pro Thr Gln Glu Leu Leu Glu Met Pro Leu Ala Ala Ile Ala Gly
340 345 350
Lys Gln Arg Glu Lys Leu Ser Thr Thr Arg Arg Glu Glu Trp Phe Gly
355 360 365
Gln Gln Ser Phe Arg Glu Leu Ala Ser Ala Met Asn Thr Ser Lys Ser
370 375 380
Ala Leu Leu Ile Val Thr Ser Trp Phe Asn Phe Pro Met Tyr Glu Ile
385 390 395 400
Asp Phe Gly Ala Gly Lys Pro Phe Phe Ala Ser Thr Thr Asn Met Ile
405 410 415
Ser Pro Ile Asn Gly Val Cys Cys Gly Val Ile Ala Pro Pro Thr Pro
420 425 430
Gly Ser Cys Ser Ser Ile Ala Thr Leu Tyr Ile Leu Cys Leu Pro Ala
435 440 445
Val Leu Glu Ala Leu Glu Asn Val Pro Asp Phe Leu Ser Phe Phe Val
450 455 460
Pro His Pro Asn His Lys Asp Asn Ser Gln Thr Gly Pro Phe Ser Phe
465 470 475 480
Val Asp Ile Met
<210> 125
<211> 1530
<212> DNA
<213> 费希尔曲霉(Aspergillus fischeri)_XP_001258079天然核苷酸序列
<400> 125
atgaccgtga ccatcagttt cgagccatat gtgggctcct ctgtcgatgc tctaagcatc 60
cctctctatc ttcgatgtca actcgtcttc aaactttcta agccacttgc tgcggtgcct 120
ctgcttgagt ctggagttaa tcgtcttgta caagcgttac ccttcctctc gggcgagttc 180
acggccgtgc cagcatccga cggtgggaaa gaaattcttc tcgttcgccc tgtgctcaac 240
ttcgagctca gccgtatact caagatcaag taccatgaaa catccctacg acatgtatgc 300
aaacagatga acaggccaag cagccagggt ggtgaccttc cgcatgagcc atacatgccc 360
tacccacgac ttccagatcc ttcacgccct caacccatcg tcgggttcca agtcaacgtt 420
cacacggatg gcatcattct ctccgttgct acgcatcact gttcctttga cgcaacaggg 480
atgggatcaa tcgtccaaaa cctcgcggct tgttgccgtt ctcctccgag cgacgagcct 540
gacttgacca cgtcgccagc ccaggaagca gaagcaagaa aagtcctctc gcaagtccgc 600
gagacgcctt ttgatccaaa gatgttcccg gagtacagac ccttggacag tatgctgtcc 660
tattacaaag gcgtccagtc agcgctccag ggtcgtcaaa ccactatcgt caatcgatgt 720
ttcacaatcg ccgccgacaa gatcaacgcg ctcaaaaggc gctgcaatca actgataccg 780
gaaatggtga agaagtatgg gctttcaact gaggatgcca ttgggagcgc ctgggtctcg 840
agtaatgatg ttgttgctgc cctcttgtgg acatgtatca atcgagcgcg atatcccgaa 900
atccgcgagc gcagcgttca ccagctccca ccagacctcc tacatgcgac atctagcctg 960
ggtgtgccag tgaacgttcg ctctcgactg tcgccgccct tacccaaatc gactttaggt 1020
aacgccgtgt gtcttctccg ggagaaggtc ccgctgcaat ttttcgcttt gcctagtcat 1080
gccaacatgg aggccacttc tagcgtttgc gcagaccatt ccggagacga cgaatgggcc 1140
ttgtccttct gtcgagtcgc ctacggactc agagcgaagc tgaacgcaat tgatgacgac 1200
tatatccgcg actatatctc ctacgtgcaa aagtctccgt gccatctgtc agtgacactg 1260
gatacagaga acctgtacct cagtaactgg cgcgagatcg gtgtgtatga tgctgatttt 1320
ggaggcatgc tgggcaagcc gctacggatg agagctccgg atggatacac cgatggcctg 1380
atttttgtga tggcgcagcg gagcgaagat aagtctgcac cgtgggagtt taatatctcg 1440
ctggaggcat cgacaatgaa gcgtattgtg catgatcccc tctggtgcaa gtatgttgag 1500
ctggatgcgt tctggcatgg agaagaatga 1530
<210> 126
<211> 1530
<212> DNA
<213> 人工_XP_001258079_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1530)
<400> 126
atg acc gtc acg att tca ttc gaa cca tat gtg gga tct agc gtg gac 48
Met Thr Val Thr Ile Ser Phe Glu Pro Tyr Val Gly Ser Ser Val Asp
1 5 10 15
gcg ctg tcc ata ccc ttg tat cta agg tgc caa ctg gtg ttc aaa tta 96
Ala Leu Ser Ile Pro Leu Tyr Leu Arg Cys Gln Leu Val Phe Lys Leu
20 25 30
tcc aag cct ctt gca gcc gtg ccc ctg cta gaa tct ggg gta aac cgt 144
Ser Lys Pro Leu Ala Ala Val Pro Leu Leu Glu Ser Gly Val Asn Arg
35 40 45
cta gta caa gca tta cca ttc ttg tct ggt gaa ttt act gct gtc cct 192
Leu Val Gln Ala Leu Pro Phe Leu Ser Gly Glu Phe Thr Ala Val Pro
50 55 60
gca agc gat ggt ggg aag gaa atc tta ctt gtt aga cca gtc tta aat 240
Ala Ser Asp Gly Gly Lys Glu Ile Leu Leu Val Arg Pro Val Leu Asn
65 70 75 80
ttc gaa cta agt cgt ata ctg aag atc aaa tac cac gaa aca tcc tta 288
Phe Glu Leu Ser Arg Ile Leu Lys Ile Lys Tyr His Glu Thr Ser Leu
85 90 95
aga cac gta tgc aag cag atg aac aga cca tcc tcg caa ggt ggt gac 336
Arg His Val Cys Lys Gln Met Asn Arg Pro Ser Ser Gln Gly Gly Asp
100 105 110
ttg cca cac gaa ccg tat atg cca tac ccc agg tta cca gat cca tct 384
Leu Pro His Glu Pro Tyr Met Pro Tyr Pro Arg Leu Pro Asp Pro Ser
115 120 125
agg cct caa cca atc gtg ggt ttt caa gtc aat gtc cac act gac ggt 432
Arg Pro Gln Pro Ile Val Gly Phe Gln Val Asn Val His Thr Asp Gly
130 135 140
ata atc ctg agt gta gca act cac cat tgc agt ttt gac gcc acg ggg 480
Ile Ile Leu Ser Val Ala Thr His His Cys Ser Phe Asp Ala Thr Gly
145 150 155 160
atg gga agt att gta cag aac ttg gcc gca tgc tgt aga tct ccg cca 528
Met Gly Ser Ile Val Gln Asn Leu Ala Ala Cys Cys Arg Ser Pro Pro
165 170 175
tcg gac gag cca gat tta act aca agc cct gct caa gaa gca gaa gct 576
Ser Asp Glu Pro Asp Leu Thr Thr Ser Pro Ala Gln Glu Ala Glu Ala
180 185 190
agg aag gtc ctg agc caa gtt aga gaa aca cca ttc gac cca aag atg 624
Arg Lys Val Leu Ser Gln Val Arg Glu Thr Pro Phe Asp Pro Lys Met
195 200 205
ttt ccc gaa tat agg ccc tta gac tct atg tta tct tat tac aaa ggt 672
Phe Pro Glu Tyr Arg Pro Leu Asp Ser Met Leu Ser Tyr Tyr Lys Gly
210 215 220
gtc cag tct gct ttg cag ggt cgt caa act act atc gtt aac aga tgt 720
Val Gln Ser Ala Leu Gln Gly Arg Gln Thr Thr Ile Val Asn Arg Cys
225 230 235 240
ttc act atc gct gct gat aag ata aac gcc tta aag agg aga tgt aac 768
Phe Thr Ile Ala Ala Asp Lys Ile Asn Ala Leu Lys Arg Arg Cys Asn
245 250 255
caa ctt att ccg gaa atg gta aag aaa tac gga ttg agt aca gaa gat 816
Gln Leu Ile Pro Glu Met Val Lys Lys Tyr Gly Leu Ser Thr Glu Asp
260 265 270
gct atc ggt tct gca tgg gtc tct tct aat gac gtg gtt gcc gca ctt 864
Ala Ile Gly Ser Ala Trp Val Ser Ser Asn Asp Val Val Ala Ala Leu
275 280 285
ttg tgg acc tgt att aat cgt gct aga tac cca gag att aga gaa aga 912
Leu Trp Thr Cys Ile Asn Arg Ala Arg Tyr Pro Glu Ile Arg Glu Arg
290 295 300
agt gta cat cag ctt cca cca gac ttg ttg cat gct aca tct tca ttg 960
Ser Val His Gln Leu Pro Pro Asp Leu Leu His Ala Thr Ser Ser Leu
305 310 315 320
ggt gta cct gtt aat gtt aga tct aga ctt tca cca ccc tta ccc aaa 1008
Gly Val Pro Val Asn Val Arg Ser Arg Leu Ser Pro Pro Leu Pro Lys
325 330 335
tcc act ctt ggg aat gcg gtc tgc ctg cta agg gag aag gta cct ttg 1056
Ser Thr Leu Gly Asn Ala Val Cys Leu Leu Arg Glu Lys Val Pro Leu
340 345 350
caa ttc ttc gct ctt ccg agt cac gca aac atg gaa gca acc agt tca 1104
Gln Phe Phe Ala Leu Pro Ser His Ala Asn Met Glu Ala Thr Ser Ser
355 360 365
gtt tgc gct gac cat tca ggg gat gat gaa tgg gcg ttg agc ttt tgt 1152
Val Cys Ala Asp His Ser Gly Asp Asp Glu Trp Ala Leu Ser Phe Cys
370 375 380
agg gta gct tat ggt ctg cgt gca aaa ttg aac gcg atc gat gat gat 1200
Arg Val Ala Tyr Gly Leu Arg Ala Lys Leu Asn Ala Ile Asp Asp Asp
385 390 395 400
tac ata agg gat tac att agt tat gta cag aag tcg ccc tgt cat ctt 1248
Tyr Ile Arg Asp Tyr Ile Ser Tyr Val Gln Lys Ser Pro Cys His Leu
405 410 415
tca gtt aca cta gat act gag aac tta tac ctg tct aat tgg aga gaa 1296
Ser Val Thr Leu Asp Thr Glu Asn Leu Tyr Leu Ser Asn Trp Arg Glu
420 425 430
ata ggc gtc tac gat gca gac ttc ggt ggt atg cta ggg aaa cca ttg 1344
Ile Gly Val Tyr Asp Ala Asp Phe Gly Gly Met Leu Gly Lys Pro Leu
435 440 445
cgt atg aga gct ccg gat ggc tac act gac ggt ttg att ttc gtt atg 1392
Arg Met Arg Ala Pro Asp Gly Tyr Thr Asp Gly Leu Ile Phe Val Met
450 455 460
gcc caa aga tct gaa gac aag tca gct ccg tgg gaa ttc aac ata tcc 1440
Ala Gln Arg Ser Glu Asp Lys Ser Ala Pro Trp Glu Phe Asn Ile Ser
465 470 475 480
ctt gag gct tct aca atg aaa agg att gta cat gat ccg ctg tgg tgt 1488
Leu Glu Ala Ser Thr Met Lys Arg Ile Val His Asp Pro Leu Trp Cys
485 490 495
aaa tat gtt gaa ttg gat gcc ttt tgg cat ggg gaa gag taa 1530
Lys Tyr Val Glu Leu Asp Ala Phe Trp His Gly Glu Glu
500 505
<210> 127
<211> 509
<212> PRT
<213> 人工_XP_001258079_针对其在酿酒酵母中的表达经密码子优化
<400> 127
Met Thr Val Thr Ile Ser Phe Glu Pro Tyr Val Gly Ser Ser Val Asp
1 5 10 15
Ala Leu Ser Ile Pro Leu Tyr Leu Arg Cys Gln Leu Val Phe Lys Leu
20 25 30
Ser Lys Pro Leu Ala Ala Val Pro Leu Leu Glu Ser Gly Val Asn Arg
35 40 45
Leu Val Gln Ala Leu Pro Phe Leu Ser Gly Glu Phe Thr Ala Val Pro
50 55 60
Ala Ser Asp Gly Gly Lys Glu Ile Leu Leu Val Arg Pro Val Leu Asn
65 70 75 80
Phe Glu Leu Ser Arg Ile Leu Lys Ile Lys Tyr His Glu Thr Ser Leu
85 90 95
Arg His Val Cys Lys Gln Met Asn Arg Pro Ser Ser Gln Gly Gly Asp
100 105 110
Leu Pro His Glu Pro Tyr Met Pro Tyr Pro Arg Leu Pro Asp Pro Ser
115 120 125
Arg Pro Gln Pro Ile Val Gly Phe Gln Val Asn Val His Thr Asp Gly
130 135 140
Ile Ile Leu Ser Val Ala Thr His His Cys Ser Phe Asp Ala Thr Gly
145 150 155 160
Met Gly Ser Ile Val Gln Asn Leu Ala Ala Cys Cys Arg Ser Pro Pro
165 170 175
Ser Asp Glu Pro Asp Leu Thr Thr Ser Pro Ala Gln Glu Ala Glu Ala
180 185 190
Arg Lys Val Leu Ser Gln Val Arg Glu Thr Pro Phe Asp Pro Lys Met
195 200 205
Phe Pro Glu Tyr Arg Pro Leu Asp Ser Met Leu Ser Tyr Tyr Lys Gly
210 215 220
Val Gln Ser Ala Leu Gln Gly Arg Gln Thr Thr Ile Val Asn Arg Cys
225 230 235 240
Phe Thr Ile Ala Ala Asp Lys Ile Asn Ala Leu Lys Arg Arg Cys Asn
245 250 255
Gln Leu Ile Pro Glu Met Val Lys Lys Tyr Gly Leu Ser Thr Glu Asp
260 265 270
Ala Ile Gly Ser Ala Trp Val Ser Ser Asn Asp Val Val Ala Ala Leu
275 280 285
Leu Trp Thr Cys Ile Asn Arg Ala Arg Tyr Pro Glu Ile Arg Glu Arg
290 295 300
Ser Val His Gln Leu Pro Pro Asp Leu Leu His Ala Thr Ser Ser Leu
305 310 315 320
Gly Val Pro Val Asn Val Arg Ser Arg Leu Ser Pro Pro Leu Pro Lys
325 330 335
Ser Thr Leu Gly Asn Ala Val Cys Leu Leu Arg Glu Lys Val Pro Leu
340 345 350
Gln Phe Phe Ala Leu Pro Ser His Ala Asn Met Glu Ala Thr Ser Ser
355 360 365
Val Cys Ala Asp His Ser Gly Asp Asp Glu Trp Ala Leu Ser Phe Cys
370 375 380
Arg Val Ala Tyr Gly Leu Arg Ala Lys Leu Asn Ala Ile Asp Asp Asp
385 390 395 400
Tyr Ile Arg Asp Tyr Ile Ser Tyr Val Gln Lys Ser Pro Cys His Leu
405 410 415
Ser Val Thr Leu Asp Thr Glu Asn Leu Tyr Leu Ser Asn Trp Arg Glu
420 425 430
Ile Gly Val Tyr Asp Ala Asp Phe Gly Gly Met Leu Gly Lys Pro Leu
435 440 445
Arg Met Arg Ala Pro Asp Gly Tyr Thr Asp Gly Leu Ile Phe Val Met
450 455 460
Ala Gln Arg Ser Glu Asp Lys Ser Ala Pro Trp Glu Phe Asn Ile Ser
465 470 475 480
Leu Glu Ala Ser Thr Met Lys Arg Ile Val His Asp Pro Leu Trp Cys
485 490 495
Lys Tyr Val Glu Leu Asp Ala Phe Trp His Gly Glu Glu
500 505
<210> 128
<211> 1323
<212> DNA
<213> 土曲霉(Aspergillus terreus) _XP_001217250天然核苷酸序列
<400> 128
atggcaacct tcgaccatat cgaggatgtc atcggccagc tacctatgct gaagagctac 60
acccatatct tgctgtgctt tcccctcgct gagagccaac tcaatgaagc catcgagagc 120
cttgaatctg ctgtacgtca ggttataaaa accttttcgt tcttggctgg caaagtagtc 180
aacgaaggca agggacccaa cagctcgggt actttcaggg tcgctccctg tgagacgtgg 240
gagtccccag atcatcaatt tgtgcgagtc gttgatcgct ctttcatgct ggcctcctac 300
gatgagatcc gcggagcaca ggcacctgct tccatgctcc caggaagtca actcgggtat 360
cgggtggctt ttccagcgca ttatcacgag acagaggacg atcccgcgcc ggtcctggac 420
attcagtgca atttgatacg gggcggactg ctacttgaca tcgctgccca acacaacatt 480
atcgatgcga gcggtatctt tcaaattgcc agcttgatcg ctctttccat gcgaggcgag 540
tcaattcctg aggatgtcat caaggaaggg aaccgtgatc gacgcaatat cattccacta 600
ctagaggcag atgagcctct tcttgatcac agcgagctca aagccagcag cgcggtgcaa 660
aatccgccgc ccgtcaactt cctccagggg tataaatggc aaatcttcaa gctctctgcc 720
gaggtgttaa ctcgaattac cgctgaagga cgacgacagc cacaggagtt tgtcccctcc 780
gtcacatttg tctcggcaaa cgattgcttg acagcctttc tgtggcaacg ggtgatagcc 840
atgcgcctga agcggctcca tacgcccgag gccgtatcca aactgagccg tgctgttgat 900
cttcggcggg cgatgggcat taccccagca tacatgggcc atatgattcg tgtcgcaaat 960
actagtctca cttttcaaga aattgtggca tgctctttat ccaggcttgc atctctgctc 1020
cgcaagagca tcatcgatgt cagccagccg tatgcgattc ggagctacgt gacctttatt 1080
gcaaatgaga cggataaatc aaagattgcg tatgcgggtg ctttcaatcc ttgcaccgac 1140
atgtcgtgct cgtccattgc acacatcact gctcctgaat tcggtcgttt gggagcgcct 1200
gactttataa ggaggcctac ctacgggcct ctgccgtgct gcacctatgt cgctcccgat 1260
aagaacgatg gagctttgga tcaaaaccag gcatggtcag acgttgtgaa gcgcatcggt 1320
tga 1323
<210> 129
<211> 1323
<212> DNA
<213> 人工_XP_001217250_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1323)
<400> 129
atg gcc acc ttc gac cac att gag gac gtg ata gga caa ttg ccc atg 48
Met Ala Thr Phe Asp His Ile Glu Asp Val Ile Gly Gln Leu Pro Met
1 5 10 15
cta aag agc tac aca cat att tta ctt tgc ttt ccg ttg gca gaa tct 96
Leu Lys Ser Tyr Thr His Ile Leu Leu Cys Phe Pro Leu Ala Glu Ser
20 25 30
caa ctg aac gag gcg att gaa tct ttg gag tca gca gtc aga caa gta 144
Gln Leu Asn Glu Ala Ile Glu Ser Leu Glu Ser Ala Val Arg Gln Val
35 40 45
att aag acg ttt tca ttc ctt gca ggt aag gtt gtt aat gaa ggc aaa 192
Ile Lys Thr Phe Ser Phe Leu Ala Gly Lys Val Val Asn Glu Gly Lys
50 55 60
ggt ccg aat tcc tct ggt acg ttc cgt gtt gct ccg tgt gaa act tgg 240
Gly Pro Asn Ser Ser Gly Thr Phe Arg Val Ala Pro Cys Glu Thr Trp
65 70 75 80
gaa tct cct gat cat caa ttt gtt cgt gtt gta gat agg agc ttt atg 288
Glu Ser Pro Asp His Gln Phe Val Arg Val Val Asp Arg Ser Phe Met
85 90 95
ttg gcc tcg tat gat gag atc cgt ggt gct caa gcc ccc gct tcc atg 336
Leu Ala Ser Tyr Asp Glu Ile Arg Gly Ala Gln Ala Pro Ala Ser Met
100 105 110
tta cct ggg tca caa ctg ggg tat agg gta gca ttc cca gcc cat tat 384
Leu Pro Gly Ser Gln Leu Gly Tyr Arg Val Ala Phe Pro Ala His Tyr
115 120 125
cat gaa act gaa gac gat cca gct cca gtt cta gat atc cag tgt aat 432
His Glu Thr Glu Asp Asp Pro Ala Pro Val Leu Asp Ile Gln Cys Asn
130 135 140
ttg ata aga ggt ggt ctg ctt cta gat ata gcg gcc caa cac aac ata 480
Leu Ile Arg Gly Gly Leu Leu Leu Asp Ile Ala Ala Gln His Asn Ile
145 150 155 160
att gat gct tcc ggg ata ttt caa atc gct agt ctg atc gcc tta agc 528
Ile Asp Ala Ser Gly Ile Phe Gln Ile Ala Ser Leu Ile Ala Leu Ser
165 170 175
atg aga gga gaa agt atc cct gaa gat gtt atc aaa gaa gga aac aga 576
Met Arg Gly Glu Ser Ile Pro Glu Asp Val Ile Lys Glu Gly Asn Arg
180 185 190
gat aga agg aat atc att ccg tta ttg gaa gcc gat gag cct tta tta 624
Asp Arg Arg Asn Ile Ile Pro Leu Leu Glu Ala Asp Glu Pro Leu Leu
195 200 205
gac cat agt gaa ttg aag gca tcc agc gcc gtt cag aac ccg cca cca 672
Asp His Ser Glu Leu Lys Ala Ser Ser Ala Val Gln Asn Pro Pro Pro
210 215 220
gtt aat ttc ttg caa ggt tat aaa tgg cag att ttc aaa ctg tcc gct 720
Val Asn Phe Leu Gln Gly Tyr Lys Trp Gln Ile Phe Lys Leu Ser Ala
225 230 235 240
gag gta ttg acc cgt att act gcc gaa gga cgt aga caa cca caa gaa 768
Glu Val Leu Thr Arg Ile Thr Ala Glu Gly Arg Arg Gln Pro Gln Glu
245 250 255
ttt gtg ccc tca gtg acc ttt gtg tcc gca aac gac tgc ttg act gct 816
Phe Val Pro Ser Val Thr Phe Val Ser Ala Asn Asp Cys Leu Thr Ala
260 265 270
ttc ttg tgg cag cgt gtg ata gct atg agg cta aag aga ttg cat acc 864
Phe Leu Trp Gln Arg Val Ile Ala Met Arg Leu Lys Arg Leu His Thr
275 280 285
ccc gag gcc gtt tcc aag cta tct aga gcc gtg gat tta aga agg gcc 912
Pro Glu Ala Val Ser Lys Leu Ser Arg Ala Val Asp Leu Arg Arg Ala
290 295 300
atg ggt ata acc cct gca tac atg ggc cat atg ata aga gtc gca aac 960
Met Gly Ile Thr Pro Ala Tyr Met Gly His Met Ile Arg Val Ala Asn
305 310 315 320
aca tcc tta acc ttt caa gaa att gtc gca tgt agt tta tca cgt ctt 1008
Thr Ser Leu Thr Phe Gln Glu Ile Val Ala Cys Ser Leu Ser Arg Leu
325 330 335
gcg agc tta ctt aga aaa tct atc att gat gtg tcc caa cca tat gcg 1056
Ala Ser Leu Leu Arg Lys Ser Ile Ile Asp Val Ser Gln Pro Tyr Ala
340 345 350
atc aga tca tac gtc acc ttc ata gca aac gaa aca gac aaa tct aag 1104
Ile Arg Ser Tyr Val Thr Phe Ile Ala Asn Glu Thr Asp Lys Ser Lys
355 360 365
ata gcc tat gcc ggg gca ttt aac ccg tgt act gat atg tca tgt tca 1152
Ile Ala Tyr Ala Gly Ala Phe Asn Pro Cys Thr Asp Met Ser Cys Ser
370 375 380
tcc ata gct cat att acg gca ccg gag ttt ggt agg tta ggg gcg cct 1200
Ser Ile Ala His Ile Thr Ala Pro Glu Phe Gly Arg Leu Gly Ala Pro
385 390 395 400
gac ttc att agg aga ccc act tat ggc cca cta cct tgc tgt act tac 1248
Asp Phe Ile Arg Arg Pro Thr Tyr Gly Pro Leu Pro Cys Cys Thr Tyr
405 410 415
gta gct cct gac aag aat gac ggt gca ttg gat cag aat caa gca tgg 1296
Val Ala Pro Asp Lys Asn Asp Gly Ala Leu Asp Gln Asn Gln Ala Trp
420 425 430
tca gat gtc gta aag agg ata ggt taa 1323
Ser Asp Val Val Lys Arg Ile Gly
435 440
<210> 130
<211> 440
<212> PRT
<213> 人工_XP_001217250_针对其在酿酒酵母中的表达经密码子优化
<400> 130
Met Ala Thr Phe Asp His Ile Glu Asp Val Ile Gly Gln Leu Pro Met
1 5 10 15
Leu Lys Ser Tyr Thr His Ile Leu Leu Cys Phe Pro Leu Ala Glu Ser
20 25 30
Gln Leu Asn Glu Ala Ile Glu Ser Leu Glu Ser Ala Val Arg Gln Val
35 40 45
Ile Lys Thr Phe Ser Phe Leu Ala Gly Lys Val Val Asn Glu Gly Lys
50 55 60
Gly Pro Asn Ser Ser Gly Thr Phe Arg Val Ala Pro Cys Glu Thr Trp
65 70 75 80
Glu Ser Pro Asp His Gln Phe Val Arg Val Val Asp Arg Ser Phe Met
85 90 95
Leu Ala Ser Tyr Asp Glu Ile Arg Gly Ala Gln Ala Pro Ala Ser Met
100 105 110
Leu Pro Gly Ser Gln Leu Gly Tyr Arg Val Ala Phe Pro Ala His Tyr
115 120 125
His Glu Thr Glu Asp Asp Pro Ala Pro Val Leu Asp Ile Gln Cys Asn
130 135 140
Leu Ile Arg Gly Gly Leu Leu Leu Asp Ile Ala Ala Gln His Asn Ile
145 150 155 160
Ile Asp Ala Ser Gly Ile Phe Gln Ile Ala Ser Leu Ile Ala Leu Ser
165 170 175
Met Arg Gly Glu Ser Ile Pro Glu Asp Val Ile Lys Glu Gly Asn Arg
180 185 190
Asp Arg Arg Asn Ile Ile Pro Leu Leu Glu Ala Asp Glu Pro Leu Leu
195 200 205
Asp His Ser Glu Leu Lys Ala Ser Ser Ala Val Gln Asn Pro Pro Pro
210 215 220
Val Asn Phe Leu Gln Gly Tyr Lys Trp Gln Ile Phe Lys Leu Ser Ala
225 230 235 240
Glu Val Leu Thr Arg Ile Thr Ala Glu Gly Arg Arg Gln Pro Gln Glu
245 250 255
Phe Val Pro Ser Val Thr Phe Val Ser Ala Asn Asp Cys Leu Thr Ala
260 265 270
Phe Leu Trp Gln Arg Val Ile Ala Met Arg Leu Lys Arg Leu His Thr
275 280 285
Pro Glu Ala Val Ser Lys Leu Ser Arg Ala Val Asp Leu Arg Arg Ala
290 295 300
Met Gly Ile Thr Pro Ala Tyr Met Gly His Met Ile Arg Val Ala Asn
305 310 315 320
Thr Ser Leu Thr Phe Gln Glu Ile Val Ala Cys Ser Leu Ser Arg Leu
325 330 335
Ala Ser Leu Leu Arg Lys Ser Ile Ile Asp Val Ser Gln Pro Tyr Ala
340 345 350
Ile Arg Ser Tyr Val Thr Phe Ile Ala Asn Glu Thr Asp Lys Ser Lys
355 360 365
Ile Ala Tyr Ala Gly Ala Phe Asn Pro Cys Thr Asp Met Ser Cys Ser
370 375 380
Ser Ile Ala His Ile Thr Ala Pro Glu Phe Gly Arg Leu Gly Ala Pro
385 390 395 400
Asp Phe Ile Arg Arg Pro Thr Tyr Gly Pro Leu Pro Cys Cys Thr Tyr
405 410 415
Val Ala Pro Asp Lys Asn Asp Gly Ala Leu Asp Gln Asn Gln Ala Trp
420 425 430
Ser Asp Val Val Lys Arg Ile Gly
435 440
<210> 131
<211> 1485
<212> DNA
<213> 简青霉(Penicillium simplicissimum)_BAU61551天然核苷酸序列
<400> 131
atgtccaagc ccttattcga agcgtatcct ctcacagggc ttgatcatac gattcctcca 60
tgttatgttc gcttccttct aactttcccc gtgccggatg tggcattggc tgtcaatcag 120
ctgcaaaagg gagctgaaaa tttaatcgag aaacttcctt tcctggccgg atatttggct 180
tcatgcgaga cccccggcgt acgcccaggg cagctcgaga ttcgacctcc cgctggggaa 240
aggagacctg tctgcctcgt cgcacatcac tcgaactcct atctcgcaga ttccagtgcg 300
acgtcgacga cggaacagct gggcaccgcc aacgagaact atctccctgt cccgttcttc 360
ccggagctag acaagccggt gcccatcttc cgggttaagg tgaatgccat gacggacggc 420
atcattctgg gatttgcgtt ccaccatagc gtgatcgatg ccaccgggat gggcaccatt 480
gtccgggact ttgccagatg ctgccgtggc cctgatggcg gtcccctgga aatcagtctg 540
gagtctcagc aggactctag agagaagctg agacactccg gaggacctcc cgatccgcgg 600
tttgaccaca atggggagta ccctctcgtg gcgtctctgc ccgccgacct cgaagccatg 660
aagcaggtct tgatccagac ggcccgtctc atgtcaacgc agtatttccg catccctgcc 720
agcctagtta acacgctaaa ggaatcctgc aatcggatgc ttcgggaatc accagcgctc 780
agggacgaag gggagaatcc atggatttcg agcaacgatc tggtggtgtc gctgttgtgg 840
ctgtgtctga atcgcgttcg gtatcctgaa gataatacca acgtcattcc tccttccgat 900
tcctcggtct gcatggccgt gaatatccga gggcgtttgc agtcgcccat tgatccagga 960
tacgttggca acgccatcgt ccttctccgg gagagcgttg gcatgaatgc ttttctgcat 1020
aaaccgggcg acgatgatcc cctgggcgcc caatgttacg aaacagcgaa acggctaggc 1080
cgagaagcgt gggaagcagc cctggtgcgc atcgccctgg ccatccgccg caagctcaac 1140
accataaacg cgagttacgt gcgcagtgtt atatcctatc tggaggacgt gcccgacctg 1200
tccactgtgg cgtttggcca gacggactac cacatcagca gctggcggga tattggcgtc 1260
tacgaggctg attttggtgg ccacatgggc catcccagcg aaatgcgagt accagatggg 1320
atggtcgatg gcatgtttta catcttacct cgaaggcagg gaacacaccc ttgctgggag 1380
atccatgtta ctatccacca ggacacaatg aagcgactca ttgcagaccc tgtgtgggca 1440
cgatatacag tgagaaagcc ttcatcactc tgccgggatg aatga 1485
<210> 132
<211> 1485
<212> DNA
<213> 人工_BAU61551_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1485)
<400> 132
atg tca aag cct ctg ttt gaa gca tac ccc cta act ggt cta gat cat 48
Met Ser Lys Pro Leu Phe Glu Ala Tyr Pro Leu Thr Gly Leu Asp His
1 5 10 15
act ata ccc cct tgt tac gtc agg ttc tta tta act ttc cca gtt cca 96
Thr Ile Pro Pro Cys Tyr Val Arg Phe Leu Leu Thr Phe Pro Val Pro
20 25 30
gat gta gcc ctt gct gtt aac caa tta cag aaa ggc gcc gaa aat tta 144
Asp Val Ala Leu Ala Val Asn Gln Leu Gln Lys Gly Ala Glu Asn Leu
35 40 45
ata gag aaa ttg cct ttt ctt gca ggg tac cta gct agc tgt gaa aca 192
Ile Glu Lys Leu Pro Phe Leu Ala Gly Tyr Leu Ala Ser Cys Glu Thr
50 55 60
cct ggc gta aga ccg ggg cag tta gaa atc agg cca cca gca ggt gaa 240
Pro Gly Val Arg Pro Gly Gln Leu Glu Ile Arg Pro Pro Ala Gly Glu
65 70 75 80
aga aga cca gtg tgt ttg gtc gca cat cac agt aac tcg tac ctt gcg 288
Arg Arg Pro Val Cys Leu Val Ala His His Ser Asn Ser Tyr Leu Ala
85 90 95
gat tct tct gca aca agt acg acg gag caa ttg gga aca gct aac gaa 336
Asp Ser Ser Ala Thr Ser Thr Thr Glu Gln Leu Gly Thr Ala Asn Glu
100 105 110
aat tac ctt ccc gta cca ttc ttt cca gag ttg gac aaa cct gtt cca 384
Asn Tyr Leu Pro Val Pro Phe Phe Pro Glu Leu Asp Lys Pro Val Pro
115 120 125
ata ttc cgt gtc aaa gtt aac gct atg aca gat ggg att atc ttg ggg 432
Ile Phe Arg Val Lys Val Asn Ala Met Thr Asp Gly Ile Ile Leu Gly
130 135 140
ttc gct ttc cac cat agt gta ata gat gct acg gga atg ggt aca ata 480
Phe Ala Phe His His Ser Val Ile Asp Ala Thr Gly Met Gly Thr Ile
145 150 155 160
gtt agg gac ttc gcc agg tgc tgc aga ggt cct gat ggt ggg ccc tta 528
Val Arg Asp Phe Ala Arg Cys Cys Arg Gly Pro Asp Gly Gly Pro Leu
165 170 175
gag ata agt cta gaa agc caa caa gat tcg aga gaa aag ctg agg cac 576
Glu Ile Ser Leu Glu Ser Gln Gln Asp Ser Arg Glu Lys Leu Arg His
180 185 190
tca ggc ggt ccg cca gat ccc aga ttc gat cat aac gga gaa tac cca 624
Ser Gly Gly Pro Pro Asp Pro Arg Phe Asp His Asn Gly Glu Tyr Pro
195 200 205
ttg gtg gcc tca ctg cca gcg gac tta gaa gct atg aaa caa gtt tta 672
Leu Val Ala Ser Leu Pro Ala Asp Leu Glu Ala Met Lys Gln Val Leu
210 215 220
atc caa aca gcg agg ctg atg agt aca caa tac ttt aga ata cct gct 720
Ile Gln Thr Ala Arg Leu Met Ser Thr Gln Tyr Phe Arg Ile Pro Ala
225 230 235 240
agc ctt gtg aac act tta aaa gag tca tgt aat aga atg ctt cgt gaa 768
Ser Leu Val Asn Thr Leu Lys Glu Ser Cys Asn Arg Met Leu Arg Glu
245 250 255
tcc cct gca ctg agg gat gaa ggt gaa aac ccg tgg att agt tct aac 816
Ser Pro Ala Leu Arg Asp Glu Gly Glu Asn Pro Trp Ile Ser Ser Asn
260 265 270
gat tta gta gtg agt cta ctg tgg ctt tgt ttg aac agg gtg agg tac 864
Asp Leu Val Val Ser Leu Leu Trp Leu Cys Leu Asn Arg Val Arg Tyr
275 280 285
ccc gaa gac aat aca aat gtg att cca ccc tct gac agt tct gtt tgc 912
Pro Glu Asp Asn Thr Asn Val Ile Pro Pro Ser Asp Ser Ser Val Cys
290 295 300
atg gct gta aat ata aga ggg aga tta cag tcg ccg atc gat cca ggt 960
Met Ala Val Asn Ile Arg Gly Arg Leu Gln Ser Pro Ile Asp Pro Gly
305 310 315 320
tat gtt ggt aat gct att gta tta tta aga gaa tct gtt gga atg aat 1008
Tyr Val Gly Asn Ala Ile Val Leu Leu Arg Glu Ser Val Gly Met Asn
325 330 335
gcc ttt cta cat aaa cct ggt gat gac gac ccg ctt ggt gcc caa tgt 1056
Ala Phe Leu His Lys Pro Gly Asp Asp Asp Pro Leu Gly Ala Gln Cys
340 345 350
tac gag aca gct aaa aga ctt gga aga gaa gca tgg gaa gca gca tta 1104
Tyr Glu Thr Ala Lys Arg Leu Gly Arg Glu Ala Trp Glu Ala Ala Leu
355 360 365
gtc agg att gct ttg gca att agg cgt aag ttg aac act att aat gct 1152
Val Arg Ile Ala Leu Ala Ile Arg Arg Lys Leu Asn Thr Ile Asn Ala
370 375 380
tcc tat gtc aga tca gtt att agc tac tta gag gat gtg ccc gat cta 1200
Ser Tyr Val Arg Ser Val Ile Ser Tyr Leu Glu Asp Val Pro Asp Leu
385 390 395 400
tca acc gtt gcc ttc ggg caa act gat tat cat atc tcc agt tgg aga 1248
Ser Thr Val Ala Phe Gly Gln Thr Asp Tyr His Ile Ser Ser Trp Arg
405 410 415
gac att gga gtt tac gag gct gac ttt ggt ggt cat atg ggc cat cca 1296
Asp Ile Gly Val Tyr Glu Ala Asp Phe Gly Gly His Met Gly His Pro
420 425 430
tct gag atg aga gtc cct gac ggg atg gtc gat ggt atg ttt tac ata 1344
Ser Glu Met Arg Val Pro Asp Gly Met Val Asp Gly Met Phe Tyr Ile
435 440 445
cta cct aga aga caa ggt act cac cca tgt tgg gaa att cat gtg act 1392
Leu Pro Arg Arg Gln Gly Thr His Pro Cys Trp Glu Ile His Val Thr
450 455 460
ata cac cag gat acc atg aaa aga ctg atc gct gat ccg gtt tgg gca 1440
Ile His Gln Asp Thr Met Lys Arg Leu Ile Ala Asp Pro Val Trp Ala
465 470 475 480
aga tat acc gtt aga aag cct agt tct ttg tgc agg gac gag taa 1485
Arg Tyr Thr Val Arg Lys Pro Ser Ser Leu Cys Arg Asp Glu
485 490
<210> 133
<211> 494
<212> PRT
<213> 人工_BAU61551_针对其在酿酒酵母中的表达经密码子优化
<400> 133
Met Ser Lys Pro Leu Phe Glu Ala Tyr Pro Leu Thr Gly Leu Asp His
1 5 10 15
Thr Ile Pro Pro Cys Tyr Val Arg Phe Leu Leu Thr Phe Pro Val Pro
20 25 30
Asp Val Ala Leu Ala Val Asn Gln Leu Gln Lys Gly Ala Glu Asn Leu
35 40 45
Ile Glu Lys Leu Pro Phe Leu Ala Gly Tyr Leu Ala Ser Cys Glu Thr
50 55 60
Pro Gly Val Arg Pro Gly Gln Leu Glu Ile Arg Pro Pro Ala Gly Glu
65 70 75 80
Arg Arg Pro Val Cys Leu Val Ala His His Ser Asn Ser Tyr Leu Ala
85 90 95
Asp Ser Ser Ala Thr Ser Thr Thr Glu Gln Leu Gly Thr Ala Asn Glu
100 105 110
Asn Tyr Leu Pro Val Pro Phe Phe Pro Glu Leu Asp Lys Pro Val Pro
115 120 125
Ile Phe Arg Val Lys Val Asn Ala Met Thr Asp Gly Ile Ile Leu Gly
130 135 140
Phe Ala Phe His His Ser Val Ile Asp Ala Thr Gly Met Gly Thr Ile
145 150 155 160
Val Arg Asp Phe Ala Arg Cys Cys Arg Gly Pro Asp Gly Gly Pro Leu
165 170 175
Glu Ile Ser Leu Glu Ser Gln Gln Asp Ser Arg Glu Lys Leu Arg His
180 185 190
Ser Gly Gly Pro Pro Asp Pro Arg Phe Asp His Asn Gly Glu Tyr Pro
195 200 205
Leu Val Ala Ser Leu Pro Ala Asp Leu Glu Ala Met Lys Gln Val Leu
210 215 220
Ile Gln Thr Ala Arg Leu Met Ser Thr Gln Tyr Phe Arg Ile Pro Ala
225 230 235 240
Ser Leu Val Asn Thr Leu Lys Glu Ser Cys Asn Arg Met Leu Arg Glu
245 250 255
Ser Pro Ala Leu Arg Asp Glu Gly Glu Asn Pro Trp Ile Ser Ser Asn
260 265 270
Asp Leu Val Val Ser Leu Leu Trp Leu Cys Leu Asn Arg Val Arg Tyr
275 280 285
Pro Glu Asp Asn Thr Asn Val Ile Pro Pro Ser Asp Ser Ser Val Cys
290 295 300
Met Ala Val Asn Ile Arg Gly Arg Leu Gln Ser Pro Ile Asp Pro Gly
305 310 315 320
Tyr Val Gly Asn Ala Ile Val Leu Leu Arg Glu Ser Val Gly Met Asn
325 330 335
Ala Phe Leu His Lys Pro Gly Asp Asp Asp Pro Leu Gly Ala Gln Cys
340 345 350
Tyr Glu Thr Ala Lys Arg Leu Gly Arg Glu Ala Trp Glu Ala Ala Leu
355 360 365
Val Arg Ile Ala Leu Ala Ile Arg Arg Lys Leu Asn Thr Ile Asn Ala
370 375 380
Ser Tyr Val Arg Ser Val Ile Ser Tyr Leu Glu Asp Val Pro Asp Leu
385 390 395 400
Ser Thr Val Ala Phe Gly Gln Thr Asp Tyr His Ile Ser Ser Trp Arg
405 410 415
Asp Ile Gly Val Tyr Glu Ala Asp Phe Gly Gly His Met Gly His Pro
420 425 430
Ser Glu Met Arg Val Pro Asp Gly Met Val Asp Gly Met Phe Tyr Ile
435 440 445
Leu Pro Arg Arg Gln Gly Thr His Pro Cys Trp Glu Ile His Val Thr
450 455 460
Ile His Gln Asp Thr Met Lys Arg Leu Ile Ala Asp Pro Val Trp Ala
465 470 475 480
Arg Tyr Thr Val Arg Lys Pro Ser Ser Leu Cys Arg Asp Glu
485 490
<210> 134
<211> 1425
<212> DNA
<213> 罂粟(Papaver somniferum)_PsSalAT天然核苷酸序列
<400> 134
atggcaacaa tgtatagtgc tgctgttgaa gtgatctcta aggaaaccat taaacccaca 60
actccaaccc catctcaact taaaaacttc aatctgtcac ttctcgatca atgttttcct 120
ttatattatt atgttccaat cattcttttc tacccagcca ccgccgctaa tagtaccggt 180
agcagtaacc atcatgatga tcttgacttg cttaagagtt ctctttccaa aacactagtt 240
cacttttatc caatggctgg taggatgata gacaatattc tggtcgactg tcatgaccaa 300
gggattaact tttacaaagt taaaattaga ggtaaaatgt gtgagttcat gtcgcaaccg 360
gatgtgccac taagccagct tcttccctct gaagttgttt ccgcgagtgt ccctaaggaa 420
gcactggtga tcgttcaagt gaacatgttt gactgtggtg gaacagccat ttgttcgagt 480
gtatcacata agattgccga tgcagctaca atgagtacgt tcattcgtag ttgggcaagc 540
accactaaaa catctcgtag tgggggttca actgctgccg ttacagatca gaaattgatt 600
ccttctttcg actcggcatc tctattccca cctagtgaac gattgacatc tccatcaggg 660
atgtcagaga taccattttc cagtacccca gaggatacag aagatgataa aactgtcagc 720
aagagatttg tgttcgattt tgcaaagata acatctgtac gtgaaaagtt gcaagtattg 780
atgcatgata actacaaaag ccgcaggcaa acaagggttg aggtggttac ttctctaata 840
tggaagtccg tgatgaaatc cactccagcc ggttttttac cagtggtaca tcatgccgtg 900
aaccttagaa agaaaatgga cccaccatta caagatgttt cattcggaaa tctatctgta 960
actgtttcgg cgttcttacc agcaacaaca acgacaacaa caaatgcggt caacaagaca 1020
atcaatagta cgagtagtga atcacaagtg gtacttcatg agttacatga ttttatagct 1080
cagatgagga gtgaaataga taaggtcaag ggtgataaag gtagcttgga gaaagtcatt 1140
caaaattttg cttctggtca tgatgcttca ataaagaaaa tcaatgatgt tgaagtgata 1200
aacttttgga taagtagctg gtgcaggatg ggattatacg agattgattt tggttgggga 1260
aagccaattt gggtaacagt tgatccaaat atcaagccga acaagaattg ttttttcatg 1320
aatgatacga aatgtggtga aggaatagaa gtttgggcga gctttcttga ggatgatatg 1380
gctaagttcg agcttcacct aagtgaaatc cttgaattga tttga 1425
<210> 135
<211> 1425
<212> DNA
<213> 人工_PsSalAT_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1425)
<400> 135
atg gca aca atg tac tca gct gca gtt gag gtt ata tct aag gaa acg 48
Met Ala Thr Met Tyr Ser Ala Ala Val Glu Val Ile Ser Lys Glu Thr
1 5 10 15
ata aaa cca acc act cca acc cca agc caa ttg aag aat ttc aat tta 96
Ile Lys Pro Thr Thr Pro Thr Pro Ser Gln Leu Lys Asn Phe Asn Leu
20 25 30
tct tta tta gac cag tgc ttt ccc ttg tac tac tat gtc ccc atc atc 144
Ser Leu Leu Asp Gln Cys Phe Pro Leu Tyr Tyr Tyr Val Pro Ile Ile
35 40 45
ttg ttc tac cct gcg act gct gca aac tcc act ggt tcc tcg aac cac 192
Leu Phe Tyr Pro Ala Thr Ala Ala Asn Ser Thr Gly Ser Ser Asn His
50 55 60
cat gat gat cta gat ctt ctg aag agc tcc ctt agc aag aca ctt gtt 240
His Asp Asp Leu Asp Leu Leu Lys Ser Ser Leu Ser Lys Thr Leu Val
65 70 75 80
cac ttc tac cct atg gcc ggt agg atg atc gat aac ata ttg gtt gac 288
His Phe Tyr Pro Met Ala Gly Arg Met Ile Asp Asn Ile Leu Val Asp
85 90 95
tgc cac gat cag ggt atc aat ttc tat aaa gtt aaa atc agg ggc aag 336
Cys His Asp Gln Gly Ile Asn Phe Tyr Lys Val Lys Ile Arg Gly Lys
100 105 110
atg tgt gaa ttc atg tct cag cct gat gtg cca ctg tcg cag ctg cta 384
Met Cys Glu Phe Met Ser Gln Pro Asp Val Pro Leu Ser Gln Leu Leu
115 120 125
cct agt gaa gtg gta tcc gca tct gtc cca aaa gag gcc ttg gtc ata 432
Pro Ser Glu Val Val Ser Ala Ser Val Pro Lys Glu Ala Leu Val Ile
130 135 140
gtc caa gtt aat atg ttc gat tgc ggt ggg acg gcc atc tgc tcg tcg 480
Val Gln Val Asn Met Phe Asp Cys Gly Gly Thr Ala Ile Cys Ser Ser
145 150 155 160
gtc agt cat aag atc gca gac gcc gca acc atg tca aca ttt att aga 528
Val Ser His Lys Ile Ala Asp Ala Ala Thr Met Ser Thr Phe Ile Arg
165 170 175
tct tgg gcg agt acc acc aaa act tca agg tct ggc ggg tca acc gcc 576
Ser Trp Ala Ser Thr Thr Lys Thr Ser Arg Ser Gly Gly Ser Thr Ala
180 185 190
gct gtt act gac cag aag ttg att cct agc ttt gat tcg gca agc tta 624
Ala Val Thr Asp Gln Lys Leu Ile Pro Ser Phe Asp Ser Ala Ser Leu
195 200 205
ttc cca cct tcc gaa agg ttg act tca cca agc ggg atg tct gag ata 672
Phe Pro Pro Ser Glu Arg Leu Thr Ser Pro Ser Gly Met Ser Glu Ile
210 215 220
cca ttt tct tca acc cct gaa gat acc gag gac gac aaa aca gtt agc 720
Pro Phe Ser Ser Thr Pro Glu Asp Thr Glu Asp Asp Lys Thr Val Ser
225 230 235 240
aaa aga ttc gtg ttt gac ttt gca aag ata aca tct gtt aga gaa aag 768
Lys Arg Phe Val Phe Asp Phe Ala Lys Ile Thr Ser Val Arg Glu Lys
245 250 255
ctt cag gta tta atg cac gac aac tac aaa agc agg cgt cag acc agg 816
Leu Gln Val Leu Met His Asp Asn Tyr Lys Ser Arg Arg Gln Thr Arg
260 265 270
gtt gag gtc gta acg agc ctg atc tgg aag agt gtc atg aag tca aca 864
Val Glu Val Val Thr Ser Leu Ile Trp Lys Ser Val Met Lys Ser Thr
275 280 285
cca gct ggg ttc ctt ccc gtc gtg cat cat gcg gta aat ttg agg aag 912
Pro Ala Gly Phe Leu Pro Val Val His His Ala Val Asn Leu Arg Lys
290 295 300
aag atg gac cca cca ttg cag gat gtc tcc ttc ggc aac ctg agt gtt 960
Lys Met Asp Pro Pro Leu Gln Asp Val Ser Phe Gly Asn Leu Ser Val
305 310 315 320
act gtc tca gca ttc ttg ccc gcc aca act acc acc acc aca aat gcc 1008
Thr Val Ser Ala Phe Leu Pro Ala Thr Thr Thr Thr Thr Thr Asn Ala
325 330 335
gtt aat aag aca ata aat tca act tca tcg gag agt cag gtg gtg cta 1056
Val Asn Lys Thr Ile Asn Ser Thr Ser Ser Glu Ser Gln Val Val Leu
340 345 350
cat gaa ttg cac gat ttt atc gca caa atg aga agt gag ata gac aaa 1104
His Glu Leu His Asp Phe Ile Ala Gln Met Arg Ser Glu Ile Asp Lys
355 360 365
gtt aaa ggc gat aag ggt agc cta gaa aag gtg att cag aac ttt gcc 1152
Val Lys Gly Asp Lys Gly Ser Leu Glu Lys Val Ile Gln Asn Phe Ala
370 375 380
tct ggt cat gac gct tca ata aag aaa ata aat gac gta gag gtc ata 1200
Ser Gly His Asp Ala Ser Ile Lys Lys Ile Asn Asp Val Glu Val Ile
385 390 395 400
aat ttc tgg ata tct tca tgg tgc aga atg ggc ttg tat gag atc gac 1248
Asn Phe Trp Ile Ser Ser Trp Cys Arg Met Gly Leu Tyr Glu Ile Asp
405 410 415
ttc ggc tgg ggc aag ccg att tgg gtt aca gtt gac cct aat atc aag 1296
Phe Gly Trp Gly Lys Pro Ile Trp Val Thr Val Asp Pro Asn Ile Lys
420 425 430
cca aat aag aat tgc ttc ttt atg aac gac aca aaa tgc gga gaa gga 1344
Pro Asn Lys Asn Cys Phe Phe Met Asn Asp Thr Lys Cys Gly Glu Gly
435 440 445
ata gag gta tgg gca agc ttc ctg gaa gac gac atg gcc aag ttt gaa 1392
Ile Glu Val Trp Ala Ser Phe Leu Glu Asp Asp Met Ala Lys Phe Glu
450 455 460
ttg cac tta tcg gag ata ttg gaa ttg atc taa 1425
Leu His Leu Ser Glu Ile Leu Glu Leu Ile
465 470
<210> 136
<211> 474
<212> PRT
<213> 人工_PsSalAT_针对其在酿酒酵母中的表达经密码子优化
<400> 136
Met Ala Thr Met Tyr Ser Ala Ala Val Glu Val Ile Ser Lys Glu Thr
1 5 10 15
Ile Lys Pro Thr Thr Pro Thr Pro Ser Gln Leu Lys Asn Phe Asn Leu
20 25 30
Ser Leu Leu Asp Gln Cys Phe Pro Leu Tyr Tyr Tyr Val Pro Ile Ile
35 40 45
Leu Phe Tyr Pro Ala Thr Ala Ala Asn Ser Thr Gly Ser Ser Asn His
50 55 60
His Asp Asp Leu Asp Leu Leu Lys Ser Ser Leu Ser Lys Thr Leu Val
65 70 75 80
His Phe Tyr Pro Met Ala Gly Arg Met Ile Asp Asn Ile Leu Val Asp
85 90 95
Cys His Asp Gln Gly Ile Asn Phe Tyr Lys Val Lys Ile Arg Gly Lys
100 105 110
Met Cys Glu Phe Met Ser Gln Pro Asp Val Pro Leu Ser Gln Leu Leu
115 120 125
Pro Ser Glu Val Val Ser Ala Ser Val Pro Lys Glu Ala Leu Val Ile
130 135 140
Val Gln Val Asn Met Phe Asp Cys Gly Gly Thr Ala Ile Cys Ser Ser
145 150 155 160
Val Ser His Lys Ile Ala Asp Ala Ala Thr Met Ser Thr Phe Ile Arg
165 170 175
Ser Trp Ala Ser Thr Thr Lys Thr Ser Arg Ser Gly Gly Ser Thr Ala
180 185 190
Ala Val Thr Asp Gln Lys Leu Ile Pro Ser Phe Asp Ser Ala Ser Leu
195 200 205
Phe Pro Pro Ser Glu Arg Leu Thr Ser Pro Ser Gly Met Ser Glu Ile
210 215 220
Pro Phe Ser Ser Thr Pro Glu Asp Thr Glu Asp Asp Lys Thr Val Ser
225 230 235 240
Lys Arg Phe Val Phe Asp Phe Ala Lys Ile Thr Ser Val Arg Glu Lys
245 250 255
Leu Gln Val Leu Met His Asp Asn Tyr Lys Ser Arg Arg Gln Thr Arg
260 265 270
Val Glu Val Val Thr Ser Leu Ile Trp Lys Ser Val Met Lys Ser Thr
275 280 285
Pro Ala Gly Phe Leu Pro Val Val His His Ala Val Asn Leu Arg Lys
290 295 300
Lys Met Asp Pro Pro Leu Gln Asp Val Ser Phe Gly Asn Leu Ser Val
305 310 315 320
Thr Val Ser Ala Phe Leu Pro Ala Thr Thr Thr Thr Thr Thr Asn Ala
325 330 335
Val Asn Lys Thr Ile Asn Ser Thr Ser Ser Glu Ser Gln Val Val Leu
340 345 350
His Glu Leu His Asp Phe Ile Ala Gln Met Arg Ser Glu Ile Asp Lys
355 360 365
Val Lys Gly Asp Lys Gly Ser Leu Glu Lys Val Ile Gln Asn Phe Ala
370 375 380
Ser Gly His Asp Ala Ser Ile Lys Lys Ile Asn Asp Val Glu Val Ile
385 390 395 400
Asn Phe Trp Ile Ser Ser Trp Cys Arg Met Gly Leu Tyr Glu Ile Asp
405 410 415
Phe Gly Trp Gly Lys Pro Ile Trp Val Thr Val Asp Pro Asn Ile Lys
420 425 430
Pro Asn Lys Asn Cys Phe Phe Met Asn Asp Thr Lys Cys Gly Glu Gly
435 440 445
Ile Glu Val Trp Ala Ser Phe Leu Glu Asp Asp Met Ala Lys Phe Glu
450 455 460
Leu His Leu Ser Glu Ile Leu Glu Leu Ile
465 470
<210> 137
<211> 1443
<212> DNA
<213> AstC_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(1443)
<400> 137
atg act aag atc aac cca tac aag ggt atc ttg gtt gaa ttg aag gac 48
Met Thr Lys Ile Asn Pro Tyr Lys Gly Ile Leu Val Glu Leu Lys Asp
1 5 10 15
atc gtt ttc act tct tct tct gac caa atc aag ttg cca atc aac act 96
Ile Val Phe Thr Ser Ser Ser Asp Gln Ile Lys Leu Pro Ile Asn Thr
20 25 30
ttc aag tct atc ttg tgt tgt ggt gct act gct caa tac caa tgt ggt 144
Phe Lys Ser Ile Leu Cys Cys Gly Ala Thr Ala Gln Tyr Gln Cys Gly
35 40 45
aag atc aac aga gct caa tac tac tct aga ttg gct aga gac ttc gct 192
Lys Ile Asn Arg Ala Gln Tyr Tyr Ser Arg Leu Ala Arg Asp Phe Ala
50 55 60
ttg tct ttg gct gac gtt act gct ttg ttc gac act gtt caa gct act 240
Leu Ser Leu Ala Asp Val Thr Ala Leu Phe Asp Thr Val Gln Ala Thr
65 70 75 80
atc aga cca gaa gaa tct ttc ttg gct ttc ttg gct gaa ttg aag tct 288
Ile Arg Pro Glu Glu Ser Phe Leu Ala Phe Leu Ala Glu Leu Lys Ser
85 90 95
aga ttc ggt gaa caa ttg aag ttg tac gct gtt gct aac atg tct aga 336
Arg Phe Gly Glu Gln Leu Lys Leu Tyr Ala Val Ala Asn Met Ser Arg
100 105 110
gaa gac tac gct atg ttg aag tct ttg cca atc gac tgg tct ttg ttc 384
Glu Asp Tyr Ala Met Leu Lys Ser Leu Pro Ile Asp Trp Ser Leu Phe
115 120 125
gac ggt gtt ttc ttg tct gct gac ttg ggt atg aga aag cca gaa ttg 432
Asp Gly Val Phe Leu Ser Ala Asp Leu Gly Met Arg Lys Pro Glu Leu
130 135 140
aga ttc ttc aga cac gtt ttg gaa tct atc tct atg aag cca gaa gac 480
Arg Phe Phe Arg His Val Leu Glu Ser Ile Ser Met Lys Pro Glu Asp
145 150 155 160
act atc ttg gtt gac aac gac act gac aac atc ttg tgt gct ttg tct 528
Thr Ile Leu Val Asp Asn Asp Thr Asp Asn Ile Leu Cys Ala Leu Ser
165 170 175
atg ggt ttg aag ggt atc ttg ttc ggt tct act tct gtt cca caa gct 576
Met Gly Leu Lys Gly Ile Leu Phe Gly Ser Thr Ser Val Pro Gln Ala
180 185 190
ttg act aac ttg ttg gaa tac gac cac atc tct aga gct gaa caa ttc 624
Leu Thr Asn Leu Leu Glu Tyr Asp His Ile Ser Arg Ala Glu Gln Phe
195 200 205
ttg aga tct cac gct aag tct ttg cac tct gtt act cac act ggt gtt 672
Leu Arg Ser His Ala Lys Ser Leu His Ser Val Thr His Thr Gly Val
210 215 220
act atc aga gaa aac ttc gct caa ttg ttg atc ttg gaa gct act ggt 720
Thr Ile Arg Glu Asn Phe Ala Gln Leu Leu Ile Leu Glu Ala Thr Gly
225 230 235 240
gac atc gac ttg gtt gaa ttg gaa tac cac cca act act tgg aac tac 768
Asp Ile Asp Leu Val Glu Leu Glu Tyr His Pro Thr Thr Trp Asn Tyr
245 250 255
ttc atc ggt act cca gtt ttg act caa act gaa ttc cca cac gac ttg 816
Phe Ile Gly Thr Pro Val Leu Thr Gln Thr Glu Phe Pro His Asp Leu
260 265 270
gac act act tct ttg gct act act gtt ttg gac aga cca aag gac atc 864
Asp Thr Thr Ser Leu Ala Thr Thr Val Leu Asp Arg Pro Lys Asp Ile
275 280 285
gct aac gaa atc atg gac gaa atg ttg aag tac aga tct gac gac gac 912
Ala Asn Glu Ile Met Asp Glu Met Leu Lys Tyr Arg Ser Asp Asp Asp
290 295 300
ttg atg ttg act ttc ttc act gac ttc aag aac aga gtt gac cca gtt 960
Leu Met Leu Thr Phe Phe Thr Asp Phe Lys Asn Arg Val Asp Pro Val
305 310 315 320
gtt tgt tgt aac gtt ttg tct ttg ttc tac aag tac ggt aga ggt cac 1008
Val Cys Cys Asn Val Leu Ser Leu Phe Tyr Lys Tyr Gly Arg Gly His
325 330 335
gaa ttg cac cac act ttg gct tgg gtt aga caa gtt ttg atc aga aga 1056
Glu Leu His His Thr Leu Ala Trp Val Arg Gln Val Leu Ile Arg Arg
340 345 350
gct tac atc aac ggt act gct ttc tac cca atg cca gaa gct ttc ttg 1104
Ala Tyr Ile Asn Gly Thr Ala Phe Tyr Pro Met Pro Glu Ala Phe Leu
355 360 365
tac ttc ttc ttc aga ttc ttg caa cac atc act cac ttg cca caa ttg 1152
Tyr Phe Phe Phe Arg Phe Leu Gln His Ile Thr His Leu Pro Gln Leu
370 375 380
tac gac ggt ttg aag gtt ttg ttg aag gaa aga ttg caa gaa aga gtt 1200
Tyr Asp Gly Leu Lys Val Leu Leu Lys Glu Arg Leu Gln Glu Arg Val
385 390 395 400
ggt gtt cca gtt gac cca atc tct ttg tct atg aga ttg atc gct tgt 1248
Gly Val Pro Val Asp Pro Ile Ser Leu Ser Met Arg Leu Ile Ala Cys
405 410 415
aac ggt gtt ggt atc cac gac aga atg ggt ttg aac gct ttg ttg tct 1296
Asn Gly Val Gly Ile His Asp Arg Met Gly Leu Asn Ala Leu Leu Ser
420 425 430
atg caa aac cca gac ggt tct tgg gac ttg ggt act atg tac cac tac 1344
Met Gln Asn Pro Asp Gly Ser Trp Asp Leu Gly Thr Met Tyr His Tyr
435 440 445
gct tct aag aga ttg cca atc ggt aac caa ggt gtt tct act gct atg 1392
Ala Ser Lys Arg Leu Pro Ile Gly Asn Gln Gly Val Ser Thr Ala Met
450 455 460
gct atc aag gct atc aag caa tgt caa gct aac caa tgt gct ggt atc 1440
Ala Ile Lys Ala Ile Lys Gln Cys Gln Ala Asn Gln Cys Ala Gly Ile
465 470 475 480
taa 1443
<210> 138
<211> 480
<212> PRT
<213> AstC_针对其在酿酒酵母中的表达经密码子优化
<400> 138
Met Thr Lys Ile Asn Pro Tyr Lys Gly Ile Leu Val Glu Leu Lys Asp
1 5 10 15
Ile Val Phe Thr Ser Ser Ser Asp Gln Ile Lys Leu Pro Ile Asn Thr
20 25 30
Phe Lys Ser Ile Leu Cys Cys Gly Ala Thr Ala Gln Tyr Gln Cys Gly
35 40 45
Lys Ile Asn Arg Ala Gln Tyr Tyr Ser Arg Leu Ala Arg Asp Phe Ala
50 55 60
Leu Ser Leu Ala Asp Val Thr Ala Leu Phe Asp Thr Val Gln Ala Thr
65 70 75 80
Ile Arg Pro Glu Glu Ser Phe Leu Ala Phe Leu Ala Glu Leu Lys Ser
85 90 95
Arg Phe Gly Glu Gln Leu Lys Leu Tyr Ala Val Ala Asn Met Ser Arg
100 105 110
Glu Asp Tyr Ala Met Leu Lys Ser Leu Pro Ile Asp Trp Ser Leu Phe
115 120 125
Asp Gly Val Phe Leu Ser Ala Asp Leu Gly Met Arg Lys Pro Glu Leu
130 135 140
Arg Phe Phe Arg His Val Leu Glu Ser Ile Ser Met Lys Pro Glu Asp
145 150 155 160
Thr Ile Leu Val Asp Asn Asp Thr Asp Asn Ile Leu Cys Ala Leu Ser
165 170 175
Met Gly Leu Lys Gly Ile Leu Phe Gly Ser Thr Ser Val Pro Gln Ala
180 185 190
Leu Thr Asn Leu Leu Glu Tyr Asp His Ile Ser Arg Ala Glu Gln Phe
195 200 205
Leu Arg Ser His Ala Lys Ser Leu His Ser Val Thr His Thr Gly Val
210 215 220
Thr Ile Arg Glu Asn Phe Ala Gln Leu Leu Ile Leu Glu Ala Thr Gly
225 230 235 240
Asp Ile Asp Leu Val Glu Leu Glu Tyr His Pro Thr Thr Trp Asn Tyr
245 250 255
Phe Ile Gly Thr Pro Val Leu Thr Gln Thr Glu Phe Pro His Asp Leu
260 265 270
Asp Thr Thr Ser Leu Ala Thr Thr Val Leu Asp Arg Pro Lys Asp Ile
275 280 285
Ala Asn Glu Ile Met Asp Glu Met Leu Lys Tyr Arg Ser Asp Asp Asp
290 295 300
Leu Met Leu Thr Phe Phe Thr Asp Phe Lys Asn Arg Val Asp Pro Val
305 310 315 320
Val Cys Cys Asn Val Leu Ser Leu Phe Tyr Lys Tyr Gly Arg Gly His
325 330 335
Glu Leu His His Thr Leu Ala Trp Val Arg Gln Val Leu Ile Arg Arg
340 345 350
Ala Tyr Ile Asn Gly Thr Ala Phe Tyr Pro Met Pro Glu Ala Phe Leu
355 360 365
Tyr Phe Phe Phe Arg Phe Leu Gln His Ile Thr His Leu Pro Gln Leu
370 375 380
Tyr Asp Gly Leu Lys Val Leu Leu Lys Glu Arg Leu Gln Glu Arg Val
385 390 395 400
Gly Val Pro Val Asp Pro Ile Ser Leu Ser Met Arg Leu Ile Ala Cys
405 410 415
Asn Gly Val Gly Ile His Asp Arg Met Gly Leu Asn Ala Leu Leu Ser
420 425 430
Met Gln Asn Pro Asp Gly Ser Trp Asp Leu Gly Thr Met Tyr His Tyr
435 440 445
Ala Ser Lys Arg Leu Pro Ile Gly Asn Gln Gly Val Ser Thr Ala Met
450 455 460
Ala Ile Lys Ala Ile Lys Gln Cys Gln Ala Asn Gln Cys Ala Gly Ile
465 470 475 480
<210> 139
<211> 606
<212> DNA
<213> AstI_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(606)
<400> 139
atg act aga caa tct cac tac caa gct atc atc ttg gac ttg ggt aac 48
Met Thr Arg Gln Ser His Tyr Gln Ala Ile Ile Leu Asp Leu Gly Asn
1 5 10 15
gtt gtt ttc gaa tgg gac act tct caa aac cca cca act gct gct cca 96
Val Val Phe Glu Trp Asp Thr Ser Gln Asn Pro Pro Thr Ala Ala Pro
20 25 30
aac caa atc tct ttg ttg aga act tct atg aag tct cca gtt tac cac 144
Asn Gln Ile Ser Leu Leu Arg Thr Ser Met Lys Ser Pro Val Tyr His
35 40 45
tct tac gaa aga ggt caa ttg tct act gaa gaa tgt cac aga ttg ttg 192
Ser Tyr Glu Arg Gly Gln Leu Ser Thr Glu Glu Cys His Arg Leu Leu
50 55 60
ggt gaa tct ttg cac gtt gac cca ggt caa atc aag gaa gct ttc gac 240
Gly Glu Ser Leu His Val Asp Pro Gly Gln Ile Lys Glu Ala Phe Asp
65 70 75 80
ttg gct aga caa tct ttg aga tct aac cca gct ttg ttg gac ttc atc 288
Leu Ala Arg Gln Ser Leu Arg Ser Asn Pro Ala Leu Leu Asp Phe Ile
85 90 95
aga caa ttg aag caa act aga ggt gtt gct gtt tac gct atg tct aac 336
Arg Gln Leu Lys Gln Thr Arg Gly Val Ala Val Tyr Ala Met Ser Asn
100 105 110
atc cca caa gct gaa atc gaa tac ttg aag gaa tct aga gct ggt gac 384
Ile Pro Gln Ala Glu Ile Glu Tyr Leu Lys Glu Ser Arg Ala Gly Asp
115 120 125
atg gaa gtt ttc gac gaa gtt ttc gct tct ggt tac gtt ggt tct aga 432
Met Glu Val Phe Asp Glu Val Phe Ala Ser Gly Tyr Val Gly Ser Arg
130 135 140
aag cca gaa act gaa ttc tac aga aga gtt atg ggt gaa atc ggt ttg 480
Lys Pro Glu Thr Glu Phe Tyr Arg Arg Val Met Gly Glu Ile Gly Leu
145 150 155 160
aag gct gaa aga gtt gtt ttc gtt gac gac aag gaa gaa aac gtt gac 528
Lys Ala Glu Arg Val Val Phe Val Asp Asp Lys Glu Glu Asn Val Asp
165 170 175
gtt gct aga ggt ttg ggt ttg tac ggt gtt tgt ttc ggt ggt gtt gaa 576
Val Ala Arg Gly Leu Gly Leu Tyr Gly Val Cys Phe Gly Gly Val Glu
180 185 190
gaa ttg aga ggt cac ttg ttg ggt atc taa 606
Glu Leu Arg Gly His Leu Leu Gly Ile
195 200
<210> 140
<211> 201
<212> PRT
<213> AstI_针对其在酿酒酵母中的表达经密码子优化
<400> 140
Met Thr Arg Gln Ser His Tyr Gln Ala Ile Ile Leu Asp Leu Gly Asn
1 5 10 15
Val Val Phe Glu Trp Asp Thr Ser Gln Asn Pro Pro Thr Ala Ala Pro
20 25 30
Asn Gln Ile Ser Leu Leu Arg Thr Ser Met Lys Ser Pro Val Tyr His
35 40 45
Ser Tyr Glu Arg Gly Gln Leu Ser Thr Glu Glu Cys His Arg Leu Leu
50 55 60
Gly Glu Ser Leu His Val Asp Pro Gly Gln Ile Lys Glu Ala Phe Asp
65 70 75 80
Leu Ala Arg Gln Ser Leu Arg Ser Asn Pro Ala Leu Leu Asp Phe Ile
85 90 95
Arg Gln Leu Lys Gln Thr Arg Gly Val Ala Val Tyr Ala Met Ser Asn
100 105 110
Ile Pro Gln Ala Glu Ile Glu Tyr Leu Lys Glu Ser Arg Ala Gly Asp
115 120 125
Met Glu Val Phe Asp Glu Val Phe Ala Ser Gly Tyr Val Gly Ser Arg
130 135 140
Lys Pro Glu Thr Glu Phe Tyr Arg Arg Val Met Gly Glu Ile Gly Leu
145 150 155 160
Lys Ala Glu Arg Val Val Phe Val Asp Asp Lys Glu Glu Asn Val Asp
165 170 175
Val Ala Arg Gly Leu Gly Leu Tyr Gly Val Cys Phe Gly Gly Val Glu
180 185 190
Glu Leu Arg Gly His Leu Leu Gly Ile
195 200
<210> 141
<211> 591
<212> DNA
<213> AstK_针对其在酿酒酵母中的表达经密码子优化
<220>
<221> CDS
<222> (1)..(591)
<400> 141
atg tgt act act ttc aag gct gct atc ttc gac atg ggt ggt gtt ttg 48
Met Cys Thr Thr Phe Lys Ala Ala Ile Phe Asp Met Gly Gly Val Leu
1 5 10 15
ttc act tgg aac cca atc gtt gac act caa gtt tct ttg aag gac ttg 96
Phe Thr Trp Asn Pro Ile Val Asp Thr Gln Val Ser Leu Lys Asp Leu
20 25 30
ggt act atc atc aac tct gaa act tgg gaa caa ttc gaa aga ggt aag 144
Gly Thr Ile Ile Asn Ser Glu Thr Trp Glu Gln Phe Glu Arg Gly Lys
35 40 45
atc gaa cca gac gac tgt tac cac caa ttg ggt tct caa atc ggt ttg 192
Ile Glu Pro Asp Asp Cys Tyr His Gln Leu Gly Ser Gln Ile Gly Leu
50 55 60
cca ggt tct gaa atc gct gct act ttc aga caa act act ggt tgt ttg 240
Pro Gly Ser Glu Ile Ala Ala Thr Phe Arg Gln Thr Thr Gly Cys Leu
65 70 75 80
aga cca gac gct aga atg act tct ttg ttg aga gaa ttg aag ggt caa 288
Arg Pro Asp Ala Arg Met Thr Ser Leu Leu Arg Glu Leu Lys Gly Gln
85 90 95
ggt gtt gct gtt tac atg atg act aac atc cca gct cca gac ttc cac 336
Gly Val Ala Val Tyr Met Met Thr Asn Ile Pro Ala Pro Asp Phe His
100 105 110
caa ttg aga gaa atg cac tac gaa tgg gac ttg ttc gac ggt atc ttc 384
Gln Leu Arg Glu Met His Tyr Glu Trp Asp Leu Phe Asp Gly Ile Phe
115 120 125
gct tct gct ttg gaa ggt atg aga aag cca gac ttg gaa ttc tac gaa 432
Ala Ser Ala Leu Glu Gly Met Arg Lys Pro Asp Leu Glu Phe Tyr Glu
130 135 140
cac gtt ttg aag caa atc gac act tct gct gct gaa act atc ttc gtt 480
His Val Leu Lys Gln Ile Asp Thr Ser Ala Ala Glu Thr Ile Phe Val
145 150 155 160
gac gac aag ttg gaa aac gtt atc gct gct caa gct gtt ggt atg gtt 528
Asp Asp Lys Leu Glu Asn Val Ile Ala Ala Gln Ala Val Gly Met Val
165 170 175
ggt ttg cac ttg act gac tct ttg gct act tgt atg gaa ttg aga caa 576
Gly Leu His Leu Thr Asp Ser Leu Ala Thr Cys Met Glu Leu Arg Gln
180 185 190
ttg gtt ggt tgt taa 591
Leu Val Gly Cys
195
<210> 142
<211> 196
<212> PRT
<213> AstK_针对其在酿酒酵母中的表达经密码子优化
<400> 142
Met Cys Thr Thr Phe Lys Ala Ala Ile Phe Asp Met Gly Gly Val Leu
1 5 10 15
Phe Thr Trp Asn Pro Ile Val Asp Thr Gln Val Ser Leu Lys Asp Leu
20 25 30
Gly Thr Ile Ile Asn Ser Glu Thr Trp Glu Gln Phe Glu Arg Gly Lys
35 40 45
Ile Glu Pro Asp Asp Cys Tyr His Gln Leu Gly Ser Gln Ile Gly Leu
50 55 60
Pro Gly Ser Glu Ile Ala Ala Thr Phe Arg Gln Thr Thr Gly Cys Leu
65 70 75 80
Arg Pro Asp Ala Arg Met Thr Ser Leu Leu Arg Glu Leu Lys Gly Gln
85 90 95
Gly Val Ala Val Tyr Met Met Thr Asn Ile Pro Ala Pro Asp Phe His
100 105 110
Gln Leu Arg Glu Met His Tyr Glu Trp Asp Leu Phe Asp Gly Ile Phe
115 120 125
Ala Ser Ala Leu Glu Gly Met Arg Lys Pro Asp Leu Glu Phe Tyr Glu
130 135 140
His Val Leu Lys Gln Ile Asp Thr Ser Ala Ala Glu Thr Ile Phe Val
145 150 155 160
Asp Asp Lys Leu Glu Asn Val Ile Ala Ala Gln Ala Val Gly Met Val
165 170 175
Gly Leu His Leu Thr Asp Ser Leu Ala Thr Cys Met Glu Leu Arg Gln
180 185 190
Leu Val Gly Cys
195
<210> 143
<211> 479
<212> PRT
<213> 碳核曲霉(Aspergillus sclerotiicarbonarius)_PYI04555.1 蛋白质序列
<400> 143
Met Gly Ala Ser Val Ser Phe Gln Pro Phe Val Pro Thr Pro Leu Asp
1 5 10 15
His Ala Met Pro Pro Ile Tyr Val Ser Gln Phe Leu Cys Phe Pro Thr
20 25 30
Thr Thr Pro Gln Ser Ala Ile Gln Ser Leu Gln Val Gly Ile Glu Arg
35 40 45
Leu Phe Glu Arg Leu Pro Phe Leu Ala Gly Glu Ile Leu Ile Asn Glu
50 55 60
His Thr Gly Ala Ile Lys Val Gln Ala Pro Ser Ala Leu Ile Arg Glu
65 70 75 80
Ile Pro Tyr Met Ala Leu Arg Ala His Pro Asp Leu Tyr Leu Pro Ala
85 90 95
Lys Gln Cys Ala Thr Thr Pro Ile Glu Arg Gln Leu Lys Thr Asn Ser
100 105 110
Leu Asp Glu Ser Tyr His Pro Leu Pro Ala Ala Leu Pro Leu Ser Gln
115 120 125
Pro Gln Pro Val Ile Arg Phe Gln Ala Asn Thr Leu Ala Asp Gly Ile
130 135 140
Leu Phe Ala Val Ser Tyr His His Cys Ile Phe Asp Gly Thr Gly Cys
145 150 155 160
Gly Gln Ile Leu Glu Met Leu Ala Gln Cys Cys Ser Ala Ser Asp Asp
165 170 175
Lys Ile Ser Leu Pro Thr Asp Cys His Thr Asp Val Leu Leu Arg Glu
180 185 190
Tyr Ile Ser Asn Leu Ser Pro Thr Thr Asn Ile Pro His Asp Tyr Thr
195 200 205
Gln Ala Tyr Ser Thr Thr Val Gln Pro Asp Pro Asp Ala Ser Asp Pro
210 215 220
Asp Thr Ser Pro Ala Ile Pro Ser Ser Leu Tyr Thr Glu Ala Phe Thr
225 230 235 240
Phe Pro Ser Gln Gln Ile Thr Thr Leu Arg Asp Ala Cys Asn His Leu
245 250 255
Leu Pro Lys Leu Pro Ser Thr Ser Asn Ala His Pro His Lys Pro Thr
260 265 270
Pro Asn Pro Leu Ser Ser Asn Asp Val Leu Thr Ala Leu Ile Ala Leu
275 280 285
Cys Ile Thr Arg Ala Thr Asn Thr Thr Thr Pro Pro Leu Gln Pro Asn
290 295 300
Asn His Ser Leu Ser Met Ala Val Asn Leu Arg Thr Arg Ile Gln Pro
305 310 315 320
Gln Val Pro Asp His Phe Leu Gly Asn Phe Ala Thr Leu Leu Pro Ile
325 330 335
His Phe Thr Ser Pro Val His Thr Gln Gln Ser Asp Leu Leu Leu Thr
340 345 350
Thr Glu Pro Pro Asp Pro Ala Leu Ile His Leu Thr Thr Leu Ala Ser
355 360 365
Gln Ile Arg Ser Ser Leu Ser Thr Val Asn Thr Asp Tyr Ile Cys Gly
370 375 380
Leu Met Thr Asp Leu Arg Thr Arg Arg Asn Ala Gly Glu Asn Ser Ser
385 390 395 400
Leu Leu Ile Glu Gly Ile Lys Ile Ser Ser Trp Arg His Leu Ser Val
405 410 415
Tyr Lys Pro Asp Phe Gly Pro Gly Leu Gly Lys Ile Ala Gly Phe Glu
420 425 430
Phe Gln Ala Gly Leu Met Asp Asn Leu Val Val Ile Leu Pro Trp Arg
435 440 445
Asn Gly Asp Trp Asp Val Arg Val Thr Leu Leu Glu Arg Asp Met Arg
450 455 460
Gly Phe Arg Glu Asp Arg Leu Val Arg Trp Ala Leu Gly Ser Gly
465 470 475
<210> 144
<211> 474
<212> PRT
<213> 鞭苔(Bazzania trilobata)_ERR364415-1_contig_8546 蛋白质序列
<400> 144
Met Ala Arg Ala Pro Pro Pro Pro Pro Pro Gly Leu Arg Met Arg Asp
1 5 10 15
Thr Val Leu Ser Ile Val Lys Pro Ile Arg Lys Thr Gln His Leu Glu
20 25 30
Thr Ile Asp Ala Thr Phe Val Asp Leu Met Arg Met Asp Ser Phe Ile
35 40 45
Pro Val Ile Phe Ala Tyr Arg Pro Ala Asp Lys Ser Glu Ala Ala Tyr
50 55 60
Ser Arg Leu Val Asn Arg Ile Lys Glu Ser Leu Gln Lys Val Leu Val
65 70 75 80
Pro Phe Phe Gly Phe Ala Gly Arg Trp Val Pro Ser Ser Gly Gly Ser
85 90 95
Arg Arg Leu Leu Cys Asn Asp Glu Gly Val Pro Phe Ile Glu Ala Phe
100 105 110
Val Asp Glu Glu Leu Asp Ser Val Val Lys Ala Ser Ala Ala Phe Gln
115 120 125
Pro Val Thr Glu Leu Asn Gly Leu Gly Val Leu Gly Met Asp Met Thr
130 135 140
Ser Tyr Asp Gln Arg Met Pro Pro Glu Gly Gly Gln Pro Cys Val Val
145 150 155 160
Ala Gln Val Thr Arg Phe Lys Cys Gly Gly Val Val Leu Gly Val Ala
165 170 175
Phe Asn His Thr His Thr Asp Gly Gln Gly Phe Tyr Thr Phe Met Arg
180 185 190
Ala Trp Ser Asp Phe Ser Arg Thr Asn Gly Thr Ala Ile Lys Val Asp
195 200 205
His Asn Arg Ala Leu Pro Glu Leu Ala Ser Leu Ser Gln Phe Phe Ile
210 215 220
Lys Gln His Asp Arg Ile Gly Gly Lys Thr Ser Thr Asp Arg Val Asn
225 230 235 240
Asp His Cys Ser Lys Val Pro Glu Arg Leu Ala Leu Lys Ala Phe Glu
245 250 255
Val Arg Ala Ser Lys Ile Lys Ala Ala Lys Leu Ala Ala Glu Asp Gly
260 265 270
Gly Val Gly Tyr Val Ser Thr Val Asp Cys Ile Val Ala His Leu Trp
275 280 285
Lys Thr Leu Ala Arg Leu Pro Pro Val Val Leu Asp Gly Arg Glu Ile
290 295 300
Thr Val Phe Ser Pro Val Glu Gly Arg Asn Arg Phe Leu Asp Pro Pro
305 310 315 320
Arg Pro Asn Met Cys Gly Asn Cys Phe Ala Ala Met Val Thr Pro Lys
325 330 335
Ile Pro Thr Gln Glu Leu Leu Glu Met Pro Leu Ala Ala Ile Ala Gly
340 345 350
Lys Gln Arg Glu Lys Leu Ser Thr Thr Arg Arg Glu Glu Trp Phe Gly
355 360 365
Gln Gln Ser Phe Arg Glu Leu Ala Ser Ala Met Asn Thr Ser Lys Ser
370 375 380
Ala Leu Leu Ile Val Thr Ser Trp Phe Asn Phe Pro Met Tyr Glu Ile
385 390 395 400
Asp Phe Gly Ala Gly Lys Pro Phe Phe Ala Ser Thr Thr Asn Met Ile
405 410 415
Ser Pro Ile Asn Gly Val Cys Cys Gly Val Ile Ala Pro Pro Thr Pro
420 425 430
Gly Ser Cys Ser Ser Ile Ala Thr Leu Tyr Ile Leu Cys Leu Pro Ala
435 440 445
Val Leu Glu Ala Leu Glu Asn Val Pro Asp Phe Leu Ser Phe Phe Val
450 455 460
Pro His Pro Asn His Lys Asp Asn Ser Gln
465 470
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