工程化棒状杆菌属菌株
文献发布时间:2023-06-19 11:26:00
在本公开内容中引用的每项参考文献、专利和公布的专利申请通过引用以其整体并入本文。
本申请通过引用并入于2019年9月6日创建的且名称为“00820000060sequencelisting.txt”的195kb文本文件,其为本申请的序列表。
技术领域
本公开内容总体上涉及赖氨酸产生。
附图简述
图1A-1B是SEQ ID NO:3与三种棒状杆菌核糖核酸酶J(rnaJ)蛋白的氨基酸序列的多重比对,所述三种棒状杆菌核糖核酸酶J(rnaJ)蛋白被美国国家生物技术信息中心(NCBI)标识为参考号WP_011014791.1(SEQ ID NO:78)、WP_096456687.1(SEQ ID NO:79)和WP_023635875.1(SEQ ID NO:80)。方框标识了每个序列中与SEQ ID NO:3的氨基酸448对齐的氨基酸。图1A,与SEQ ID NO:3的氨基酸1-560比对。图1B,与SEQ ID NO:3的氨基酸561-718比对。
图2是SEQ ID NO:6与三种棒状杆菌accDA蛋白的氨基酸序列的多重比对,所述三种棒状杆菌accDA蛋白被NCBI标识为参考号WP_075348128(SEQ ID NO:81)、AGT04828.1(SEQ ID NO:82)和WP_066564978.1(SEQ ID NO:83)。方框标识了每个序列中与SEQ ID NO:6的氨基酸310对齐的氨基酸。
图3是SEQ ID NO:9与三种棒状杆菌cg1144蛋白的氨基酸序列的多重比对,所述三种棒状杆菌cg1144蛋白被NCBI标识为参考号WP_011897001(SEQ ID NO:84)、WP_066565124(SEQ ID NO:85)和EEW49979(SEQ ID NO:86)。方框标识了每个序列中与SEQ ID NO:9的氨基酸66对齐的氨基酸。
详细描述
本公开内容提供了用于赖氨酸的节省成本的产生的棒状杆菌属(Corynebacteria)的工程化菌株、用于产生该工程化菌株的工具和方法以及使用该工程化菌株产生赖氨酸的方法。
结构改变
本公开内容提供了以下四种可以被工程化到棒状杆菌属中以改进赖氨酸产生的结构改变:(a)在cg1383起始密码子前面插入启动子;(b)对天然phoU启动子的替代;(c)对天然cg3210启动子的替代;和(d)对天然cg0800启动子的替代。
a.在cg1383起始密码子前面插入启动子
cg1383也称为NCg11179,并且编码ABC型钼转运系统的ATP酶组分(例如,WP_011014216.1)。在一些实施方案中,启动子被插入在cg1383起始密码子前面。插入的启动子可以是来自被工程化的棒状杆菌属(Corynebacterium)的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
b.对天然phoU启动子的替代
phoU编码磷酸摄取调控因子(phosphate uptake regulator)(例如,BAB99964.1),并且也称为cgl2571和NCgl2482。在一些实施方案中,天然phoU启动子被phoU替代启动子替代。phoU替代启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
c.对天然cg3210启动子的替代
cg3210也称为NCg12802,并且编码细胞被膜相关的转录调控因子(cellenvelope-related transcriptional regulator)(例如,WP_011266029.1)。在一些实施方案中,天然cg3210启动子被cg3210替代启动子替代。cg3210替代启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
d.对天然cg0800启动子的替代
cg0800也称为NCg10668或prpR,并且编码MerR家族的转录调控因子(例如,WP_011013825.1)。在一些实施方案中,天然cg0800启动子被cg0800替代启动子替代。cg0800替代启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
另外的结构改变
在一些实施方案中,包含上述结构改变中的一种或更多种的工程化棒状杆菌属菌株还可以包含下述另外的结构改变(e)-(o)中的一种或更多种。
e.改变的棒状杆菌核糖核酸酶J蛋白
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含改变的棒状杆菌核糖核酸酶(rnaJ)蛋白。改变的棒状杆菌rnaJ蛋白包含G448S取代。这样的蛋白的一种实例在SEQ ID NO:3中示出。如本文使用的“包含G448S取代的棒状杆菌rnaJ蛋白”意指当使用NCBI的
表1.棒状杆菌核糖核酸蛋白的NCBI参考号
f.改变的棒状杆菌accDA蛋白
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含改变的棒状杆菌accDA蛋白。改变的棒状杆菌accDA蛋白包含G310E取代。这样的蛋白的一种实例在SEQID NO:6中示出。如本文使用的“包含G310E取代的棒状杆菌accDA蛋白”意指当使用美国国家生物技术信息中心的
表2.棒状杆菌accDA蛋白的NCBI参考号
g.改变的棒状杆菌cg1144蛋白
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含改变的棒状杆菌cg1144蛋白。改变的棒状杆菌cg1144蛋白包含P66S取代。这样的蛋白的一种实例在SEQID NO:9中示出。如本文使用的“包含P66S取代的棒状杆菌cg1144蛋白”意指当使用国家生物技术信息中心的
表3.棒状杆菌cg1144蛋白的NCBI参考号
h.在cg2766起始密码子前面插入启动子
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含插入在cg2766起始密码子前面的启动子。插入的启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
i.对天然actA启动子的替代
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含代替天然actA启动子的替代actA启动子。actA替代启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
j.对天然opcA编码序列的替代
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含代替天然opcA编码序列的不同棒状杆菌属的种或菌株的opcA蛋白的编码序列。合适的opcA蛋白列于表4A和表4B中。使用NCBI的
在一些实施方案中,替代opcA编码序列在驱动zwf和opcA两者的表达的启动子的控制下。在一些实施方案中,替代opcA编码序列处于插入的opcA启动子的控制下。插入的opcA启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
表4A.棒状杆菌opcA蛋白的NCBI参考号
表4B.棒状杆菌OpcA蛋白的NCBI参考号
k.插入至少一个另外的lysA、ask、asd、ddh和/或dapB的拷贝
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含至少一个另外的选自由lysA、ask、asd、ddh和dapB组成的组的至少一个基因的拷贝。另外的拷贝可以使用一个或更多个载体来提供。实施例1描述了两个可以用于插入ask、asd、ddh和dapB的拷贝的整合载体。
在一些实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ask、asd、ddh和dapB的拷贝。
在一些实施方案中,工程化棒状杆菌包含至少一个另外的ask、asd、ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、asd、ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ask、ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ask、asd和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ask、asd和ddh的拷贝。
在一些实施方案中,工程化棒状杆菌包含至少一个另外的asd、ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的ask、ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的ask、asd和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的ask、asd和ddh的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、asd和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、asd和ddh的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ask和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA、ask和ddh的拷贝。
在一些实施方案中,工程化棒状杆菌包含至少一个另外的ddh和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的asd和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的asd和ddh的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA和dapB的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA和ddh的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA和ask的拷贝。
在其他实施方案中,工程化棒状杆菌包含至少一个另外的lysA的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的ask的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的asd的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的ddh的拷贝。在其他实施方案中,工程化棒状杆菌包含至少一个另外的dapB的拷贝。
在任何上述其中工程化棒状杆菌包含至少一个另外的lysA的拷贝的实施方案中,另外的lysA的拷贝是下述密码子优化的形式。
l.对aceE的起始密码子的替代
在一些实施方案中,上述工程化棒状杆菌菌株中的任一种的基因组中的天然aceE基因的起始密码子被替代。在这些实施方案的一些中,起始密码子被GTG、CTG或TTG替代。在一些实施方案中,起始密码子被TTG替代。
m.对天然pyc基因启动子的替代
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含代替天然pyc基因启动子的pyc替代启动子。pyc替代启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
n.在zwf开放阅读框前面插入启动子
包含上述结构改变中的任一种的工程化棒状杆菌属菌株还可以包含插入在zwf开放阅读框前面的启动子。插入的启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
o.密码子优化的lysA编码序列的插入
包含上述结构改变中的任一种的工程化棒状杆菌属菌株可以包含处于启动子控制下并且包含终止子序列的密码子优化的lysA编码序列。启动子可以是来自被工程化的棒状杆菌属的种或菌株的不同基因的启动子,或者可以是异源启动子(即,另一种棒状杆菌属的种或菌株的启动子或者人工构建的启动子)。这些启动子包括,但不限于,
在一些实施方案中,启动子是Pcg0007_39(SEQ ID NO:20)。在这些实施方案的一些中,密码子优化的序列是SEQ ID NO:16。在这些实施方案的一些中,终止子是sod终止子(SEQ ID NO:17的核苷酸1436-1516)。在一些实施方案中,工程化棒状杆菌属的基因组包含核苷酸序列SEQ ID NO:17,该核苷酸序列SEQ ID NO:17是处于启动子Pcg0007_39(SEQ IDNO:20)的控制下并具有sod终止子(SEQ ID NO:17的核苷酸1436-1516)的密码子优化的lysA序列SEQ ID NO:16。
核酸和载体
可以使用载体将棒状杆菌属工程化以具有上述结构改变中的一种或更多种,导致与对应的天然棒状杆菌属(即未被工程化为包含所述结构改变的棒状杆菌属)相比改进的赖氨酸产生。这样的棒状杆菌属包括但不限于保藏为NRRL B-11474的棒状杆菌属、保藏为NRRL B-67439的棒状杆菌属以及表1、表2、表3、表4A和表4B中的棒状杆菌的种和菌株。将载体递送至棒状杆菌属的方法是熟知的,并且包括例如磷酸钙转染、DEAE-右旋糖酐介导的转染、显微注射、脂质介导的转染、电穿孔、接合和感染。
可以用于工程化上述改变的核酸是熟知的,并且在例如美国专利7,368,276、美国专利6,927,046、US 2017/0159045和WO 2017/00376以及下文实施例中进行了描述。编码上述改变的棒状杆菌rnaJ、accDA和cg1144蛋白的核酸可以被包含在载体中,在该载体中编码序列被可操作地连接至合适的调控序列,以用于在棒状杆菌属中表达。SEQ ID NO:2、SEQID NO:5和SEQ ID NO:8分别是编码SEQ ID NO:3、SEQ ID NO:6和SEQ ID NO:9的核苷酸序列的实例,但是可以使用编码改变的棒状杆菌蛋白的任何核苷酸序列。如本领域熟知的,核苷酸序列可以被优化以在各种棒状杆菌属的种或菌株中表达。
调控序列包括合适的mRNA核糖体结合位点和用于调控转录和翻译终止的序列,并且可以包括其他元件,诸如启动子或操作子。在被转化到宿主棒状杆菌属中之后,载体可以独立于宿主基因组复制或发挥作用,或者可以整合到基因组本身中。所使用的载体没有特别限制,并且可以是本领域已知的任何载体,条件是它能够在棒状杆菌属宿主中复制。参见,例如,Lee,2014;Knoppová等人,2007年;和Pátek&
载体可以包含至少一个选择性标志物,诸如抗生素抗性基因。合适的抗生素包括,例如,阿米卡星、氨苄青霉素、奥格门汀(阿莫西林加克拉维酸)、头孢唑啉、头孢西丁、头孢他啶、头孢噻呋、头孢菌素、氯霉素、恩诺沙星、氟苯尼考、庆大霉素、亚胺培南、卡那霉素、青霉素、沙氟沙星(sarafloxicin)、壮观霉素、链霉素、四环素、替卡西林和替米考星。
工程化棒状杆菌属菌株
本公开内容提供了工程化棒状杆菌属菌株,所述工程化棒状杆菌属菌株包含上述四种改变中的至少一种(即(a)在cg1383起始密码子前面插入替代启动子;(b)对天然phoU启动子的替代;(c)对天然cg3210启动子的替代;和(d)对天然cg0800启动子的替代)以及以下改变中的至少一种:
(e)rnaJ编码序列的改变,以编码包含G448S取代的核糖核酸酶J蛋白;
(f)天然accDA编码序列的改变,以编码包含G310E取代的乙酰辅酶a羧化酶羧基转移酶亚基α/β蛋白;
(g)天然cg1144编码序列的改变,以编码包含P66S取代的cg1144蛋白;
(h)在cg2766起始密码子前面插入启动子;
(i)对天然actA启动子的替代;
(j1)天然opcA编码序列被不同棒状杆菌属的种或菌株的opcA蛋白的编码序列替代;
(j2)天然opcA编码序列被处于插入的opcA启动子的控制下的不同棒状杆菌属的种或菌株的opcA蛋白的编码序列替代;
(k)插入至少一个另外的选自由ask、asd、ddh和dapB组成的组的至少一个基因的拷贝;
(l)对天然aceE基因起始密码子的替代;
(m)对天然pyc基因启动子的替代;
(n)在zwf开放阅读框前面插入另外的启动子;和
(o)插入密码子优化的lysA编码序列。
以下工程化棒状杆菌属的非限制性实施方案落入以上描述的范围内。
实施方案1.一种棒状杆菌属,所述棒状杆菌属根据布达佩斯条约(BudapestTreaty)的规定于2017年12月22日保藏在位于伊利诺伊州皮奥里亚大学街1815N号邮编61604(1815N.University Street,Peoria,IL 61604)的农业研究机构培养物保藏中心(NRRL)国际保藏单位(International Depository Authority),并且分配了登录号NRRLB-67535。
实施方案2.一种棒状杆菌属,所述棒状杆菌属具有主要由被保藏为NRRL B-67439的细菌的基因组组成但具有至多4种选自由以下组成的组的结构改变的细菌基因组:
(a)在cg1383起始密码子前面插入启动子;
(b)对天然phoU启动子的替代;
(c)对天然cg3210启动子的替代;和
(d)对天然cg0800启动子的替代。
实施方案3.一种棒状杆菌属,所述棒状杆菌属具有主要由被保藏为NRRL B-11474的细菌的基因组组成但具有以下(i)以及(ii)的细菌基因组:
(i)至少一种选自由以下组成的组的结构改变:
(a)在cg1383起始密码子前面插入启动子;
(b)对天然phoU启动子的替代;
(c)对天然cg3210启动子的替代;和
(d)对天然cg0800启动子的替代;以及
(ii)至少一种另外的选自由以下组成的组的结构改变:
(e)rnaJ编码序列的改变,以编码包含G448S取代的棒状杆菌rnaJ蛋白;
(f)天然accDA编码序列的改变,以编码包含G310E取代的棒状杆菌accDA蛋白;
(g)天然cg1144编码序列的改变,以编码包含P66S取代的棒状杆菌cg1144蛋白;
(h)在cg2766起始密码子前面插入启动子;
(i)对天然actA启动子的替代;
(j1)天然opcA编码序列被不同棒状杆菌属的opcA蛋白的替代opcA编码序列替代;
(j2)天然opcA编码序列被处于插入的opcA启动子的控制下的不同棒状杆菌属的opcA蛋白的替代opcA编码序列替代;
(k)插入至少一个另外的选自由lysA、ask、asd、ddh和dapB组成的组的至少一个基因的拷贝;
(1)天然aceE的起始密码子被GTG、CTG或TTG替代;
(m)天然pyc基因启动子被启动子Pcg1860(SEQ ID NO:11)替代;
(n)在zwf开放阅读框前面插入启动子Pcg0007_39(SEQ ID NO:20);和
(o)插入密码子优化的lysA编码序列。
实施方案4.根据实施方案2或3所述的棒状杆菌属,其中在所述cg1383起始密码子前面的启动子是启动子Pcg0007_39(SEQ ID NO:20)。
实施方案5.根据实施方案2-4中任一项所述的棒状杆菌属,其中所述天然phoU启动子被启动子Pcg0007_39(SEQ ID NO:20)替代。
实施方案6.根据实施方案2-5中任一项所述的棒状杆菌属,其中所述天然cg3210启动子被启动子Pcg0007_39(SEQ ID NO:20)替代。
实施方案7.根据实施方案2-6中任一项所述的棒状杆菌属,其中所述天然cg0800启动子被启动子Pcg0007_39(SEQ ID NO:20)替代。
实施方案8.根据实施方案3-7中任一项所述的棒状杆菌属,其中(e)所述天然rnaJ编码序列被改变以编码包含G448S取代的棒状杆菌rnaJ蛋白。
实施方案9.根据实施方案3-8中任一项所述的棒状杆菌属,其中(e)所述天然rnaJ编码序列被改变以编码氨基酸序列SEQ ID NO:3。
实施方案10.根据实施方案3-9中任一项所述的棒状杆菌属,其中(f)所述天然accDA编码序列被改变以编码包含所述G310E取代的棒状杆菌accDA蛋白。
实施方案11.根据实施方案3-10中任一项所述的棒状杆菌属细菌,其中(f)所述天然accDA编码序列被改变以编码氨基酸序列SEQ ID NO:6。
实施方案12.根据实施方案3-11中任一项所述的棒状杆菌属,其中(g)所述天然cg1144编码序列被改变以编码包含所述P66S取代的棒状杆菌cg1144蛋白。
实施方案13.根据实施方案3-12中任一项所述的棒状杆菌属细菌,其中(g)所述天然cg1144编码序列被改变以编码氨基酸序列SEQ ID NO:9。
实施方案14.根据实施方案3-13中任一项所述的棒状杆菌属细菌,其中(h)所述启动子被插入在所述cg2766起始密码子前面。
实施方案15.根据实施方案3-14中任一项所述的棒状杆菌属,其中(h)被插入在所述cg2766起始密码子前面的启动子是启动子Pcg0007_39(SEQ ID NO:20)。
实施方案16.根据实施方案3-15中任一项所述的棒状杆菌属细菌,其中(i)所述天然actA启动子被替代。
实施方案17.根据实施方案3-16中任一项所述的棒状杆菌属,其中(i)所述天然actA启动子被启动子Pcg0007_39(SEQ ID NO:20)替代。
实施方案18.根据实施方案3-17中任一项所述的棒状杆菌属细菌,其中(j)所述天然opcA编码序列被不同棒状杆菌属的opcA蛋白的替代opcA编码序列替代。
实施方案19.根据实施方案3-18中任一项所述的棒状杆菌属,其中所述天然opcA编码序列被选自由以下组成的组的棒状杆菌属的opcA蛋白的替代opcA编码序列替代:Corynebacterium vitaeruminis DSM 20294、马氏棒状杆菌(Corynebacteriummatruchotii)ATCC 33806、Corynebacterium falsenii DSM 44353、耐盐棒状杆菌(Corynebacterium halotolerans)YIM70093、产丙酮酸棒状杆菌(Corynebacteriumpyruviciproducens)ATCC BAA-1742、干酪棒状杆菌(Corynebacterium casei)UCMA 3821、解萜烯棒状杆菌(Corynebacterium terpenotabidum)Y-11和C.humireducensNBRC106098。
实施方案20.根据实施方案3-19中任一项所述的棒状杆菌属,其中所述天然opcA编码序列被C.humireducens NBRC 106098的opcA蛋白的替代opcA编码序列替代。
实施方案21.根据实施方案3-20中任一项所述的棒状杆菌属,其中所述替代opcA编码序列处于插入的opcA启动子的控制下。
实施方案22.根据实施方案21所述的棒状杆菌属,其中所述插入的opcA启动子是启动子Pcg0007_39(SEQ ID NO:20)。
实施方案23.根据实施方案3-22中任一项所述的棒状杆菌属细菌,其中(k)所述细菌的基因组包含至少一个另外的选自由lysA、ask、asd、ddh和dapB组成的组的至少一个基因的拷贝。
实施方案24.根据实施方案23所述的棒状杆菌属细菌,其中所述细菌的基因组包含(a)至少一个另外的lysA、ask、asd、ddh和dapB的拷贝。
实施方案24.根据实施方案23所述的棒状杆菌属细菌,其中所述细菌的基因组包含:
(b)至少一个另外的ask、asd、ddh和dapB的拷贝;
(c)至少一个另外的lysA、asd、ddh和dapB的拷贝;
(d)至少一个另外的lysA、ask、ddh和dapB的拷贝;
(e)至少一个另外的lysA、ask、asd和dapB的拷贝;或者
(f)至少一个另外的lysA、ask、asd和ddh的拷贝。
实施方案25.根据实施方案23所述的棒状杆菌属细菌,其中所述细菌的基因组包含:
(g)至少一个另外的asd、ddh和dapB的拷贝;
(h)至少一个另外的ask、ddh和dapB的拷贝;
(i)至少一个另外的ask、asd和dapB的拷贝;
(j)至少一个另外的ask、asd和ddh的拷贝;
(k)至少一个另外的lysA、ddh和dapB的拷贝;
(l)至少一个另外的lysA、asd和dapB的拷贝;
(m)至少一个另外的lysA、asd和ddh的拷贝;
(n)至少一个另外的lysA、ask和dapB的拷贝;或者
(o)至少一个另外的lysA、ask和ddh的拷贝。
实施方案26.根据实施方案23所述的棒状杆菌属细菌,其中所述细菌的基因组包含:
(p)至少一个另外的ddh和dapB的拷贝;
(q)至少一个另外的asd和dapB的拷贝;
(r)至少一个另外的asd和ddh的拷贝;
(s)至少一个另外的lysA和dapB的拷贝;
(t)至少一个另外的lysA和ddh的拷贝;
(u)至少一个另外的lysA和ask的拷贝。
实施方案27.根据实施方案23所述的棒状杆菌属细菌,其中所述细菌的基因组包含:
(v)至少一个另外的lysA的拷贝;
(w)至少一个另外的ask的拷贝;
(x)至少一个另外的asd的拷贝;
(y)至少一个另外的ddh的拷贝;或者
(z)至少一个另外的dapB的拷贝。
实施方案28.根据实施方案3-27中任一项所述的棒状杆菌属细菌,其中aceE的起始密码子被TTG、GTG或CTG替代。
实施方案29.根据实施方案3-28中任一项所述的棒状杆菌属细菌,其中aceE的起始密码子被TTG替代。
实施方案30.根据实施方案3-29中任一项所述的棒状杆菌属细菌,其中所述天然pyc基因启动子被替代。
实施方案31.根据实施方案3-30中任一项所述的棒状杆菌属细菌,其中所述天然pyc基因启动子被启动子Pcg1860(SEQ ID NO:11)替代。
实施方案32.根据实施方案3-31中任一项所述的棒状杆菌属细菌,其中启动子被插入在所述zwf开放阅读框前面。
实施方案33.根据实施方案3-32中任一项所述的棒状杆菌属细菌,其中启动子Pcg0007_39(SEQ ID NO:20)被插入在所述zwf开放阅读框前面。
实施方案34.根据实施方案3-33中任一项所述的棒状杆菌属,所述棒状杆菌属包含(o)密码子优化的lysA编码序列。
实施方案35.根据实施方案34所述的棒状杆菌属,其中所述密码子优化的lysA编码序列处于Pcg0007_39(SEQ ID NO:20)的控制下。
实施方案36.根据实施方案34或35所述的棒状杆菌属,所述棒状杆菌属包含所述密码子优化的lysA编码序列的异源终止子。
实施方案37.根据实施方案34-36中任一项所述的棒状杆菌属细菌,所述棒状杆菌属细菌包含核苷酸序列SEQ ID NO:17。
赖氨酸的产生
使用棒状杆菌属产生赖氨酸的方法是本领域熟知的,并且本公开内容中提供的工程化棒状杆菌属可以用于这些方法中的任一种。参见,例如,美国专利8,048,649、美国专利7,635,579、美国专利7,504,242、美国专利7,300,777、美国专利7,267,967、美国专利7,160,711、美国专利7,141,388、美国专利7,122,369、美国专利6,927,046、美国专利6,830,903、US2002/0086371、US 2004/0043458、US 2009/0325244、EP 1246921和EP1619252。
本领域技术人员将理解,存在落入所附权利要求书的范围内的上述实施方案的许多变化和组合。
实施例1.L-赖氨酸途径四基因构建体的制备
实施例1A.在bioD区中包含赖氨酸生物合成基因ask-asd-dapB-ddh的整合载体的构建。
pBKMS载体是pBR322衍生物,不能在谷氨酸棒状杆菌中复制,并且pBKMS载体包含卡那霉素抗性基因标志物和用于蔗糖反选择的处于强的合成启动子控制下的来自枯草芽孢杆菌(Bacillus subtilis)的果聚糖蔗糖酶sacB基因。用PmeI和XmaI将5.4kb ask-asd-ddh-dapB-orf2′盒(4Go)从pFC3-ask-asd-dapB-ddh质粒(美国专利7,368,276)中消化下来,并且连接到用HincII和XmaI线性化的pDElia11(美国专利6,927,046)中,以构建pD11-KBDH。
为了产生包含SpeI位点的9.4kb同源区,使用引物184f7(SEQ ID NO:70)和184F8(SEQ ID NO:71)从NRRL B-11474基因组DNA中扩增DNA片段,并且通过
实施例1B.包含赖氨酸生物合成基因ask-asd-dapB-ddh和用于靶向整合的FarI同源区的载体的构建。
NRRL B-11474衍生菌株的3′FarI同源区使用引物FarI 3P InfusF(SEQ ID NO:30)和FarI 3P InfusR AvrII(SEQ ID NO:31)通过PCR来扩增。将纯化的PCR片段通过
用PmeI和XmaI将5.4kb ask-asd-ddh-dapB-orf2′盒(4Go)从pFC3-ask-asd-dapB-ddh质粒(US7368276)中消化下来,并且连接到用HincII和XmaI线性化的pDElia11(美国专利6,927,046)中。将所得载体pD11-KBDH用NruI和SwaI消化,以消除ddh-dapB 2.9-kb片段,以产生pD11KD。然后将pD11KD用SmaI消化,并且将使用引物685(SEQ ID NO:74)和686(SEQID NO:75)从pD11KDBH扩增的1.36kb PCR片段使用
实施例2.谷氨酸棒状杆菌的等位基因替代
本实施例描述了下文实施例4中用于等位基因替代的方法。将菌株在培养基B(表5)或BHI肉汤(BD Biosciences)中在30℃培养,直到OD 660nm达到0.5。将细胞在4℃通过离心收获,在冰冷的去离子水中洗涤两次,并且重悬于冰冷的10%甘油中以产生电感受态(electrocompetent)谷氨酸棒状杆菌细胞。将质粒和细胞混合在一起,通过电穿孔进行转化,铺于含有10μg/ml卡那霉素的BHI琼脂板上,并且孵育直到已经整合了质粒的转化体(卡那霉素抗性、蔗糖敏感的)出现。将转化体在补充有另外的5%蔗糖(最终浓度为10%蔗糖)的培养基B(表5)中在30℃进一步培养过夜,并且铺于培养基B上,以选择通过第二次重组事件切除质粒并因此具有蔗糖抗性且卡那霉素敏感的菌株。通过PCR和测序进一步筛选克隆,以验证所期望突变的存在。
表5.实施例2和3中使用的培养基。
实施例3.基因组DNA提取
为了提取基因组DNA,使选择的菌株在培养基B(表5)中在30℃生长过夜。将培养物通过以5000×g(4℃)离心10分钟沉淀。将沉淀物悬浮于10ml包含25mM Tris-HCl pH 8.0、10mM EDTA、50mM葡萄糖和20mg/ml溶菌酶的溶液中,并且在37℃孵育2小时。在添加1.3ml10%SDS、67μl 10mg/ml RNA酶A和167μl蛋白酶K(20mg/ml储备液)后延长孵育另外2小时。将基因组DNA通过苯酚-氯仿提取进一步纯化,并且通过添加两倍体积的冰冷乙醇和0.1体积的3M乙酸钠(pH 5.2)沉淀。在-80℃孵育1小时后,将DNA沉淀物通过以14,000rpm(4℃)离心1小时分离,用70%乙醇洗涤,风干,并且溶解于无核酸酶的水中。
实施例4.将插入片段组装到pK18mobsacB衍生的载体中并且转化到谷氨酸棒状杆菌菌株中
实施例4A.用于整合和等位基因替代的质粒的组装
用于产生上游或下游同源重组区的所有DNA片段以及插入片段(inserts)使用聚合酶链式反应(PCR)从提取自菌株NRRL B-11474的纯化的基因组DNA或者从包含衍生自如实施例2中描述的该菌株的序列的质粒扩增或者化学合成(DNA 2.0)。将衍生自pK18mobsacB(Schaffer等人,Gene145:69-73,1994;登录号FJ437239)的pZ载体用作将这些变化引入谷氨酸棒状杆菌的载体骨架,所述pZ载体包含用于在酵母中进行选择的URA3基因。在酵母中通过同源重组将同源臂和插入片段组装到载体骨架中(Ma等人,Gene(58):201-16,1987)。每个DNA片段在每一侧都包含50-bp的重叠,以确保与其相邻部分的正确组装。为了组装,将线性化载体骨架、两条同源臂和任选地插入的DNA片段同时转化到酿酒酵母(Saccharomyces cerevisiae)CEN.PK中(Entian&Kotter,Methods in Microbiology36:629-66,2007),并且铺于不含尿嘧啶的合成完全琼脂板(Sigma)上。使用ZYMOPREP
实施例4B.用于引入rnaJ
两条同源臂使用PCR从如实施例3中描述获得的NRRL B-11474基因组DNA扩增。
约2.1kb的下游同源臂(SEQ ID NO:1的核苷酸2026至4101)使用引物SNP_084_P1(SEQ ID NO:38)和SNP_084_P2(SEQ ID NO:39)扩增。引物SNP_084_P1包含与实施例4A中描述的pZ骨架重叠的50个核苷酸的5′延伸。引物SNP_084_P2在位置25处包含G到A的取代,以引入G448S取代。类似地,约2.1kb的上游同源臂(SEQ ID NO:1的核苷酸1至2075)使用引物SNP_084_P3(SEQ ID NO:40)和SNP_084_P4(SEQ ID NO:41)扩增。引物SNP_084_P3在位置20处包含C到T的取代,以引入G448S取代。SNP_084_P4包含与实施例4A中描述的pZ重叠的50个碱基的5′延伸。两条同源臂的对齐产生由引物SNP_084_P2和SNP_084_P3限定的50-bp重叠。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带改变的rnaJ编码序列(SEQ ID NO:2),编码具有G448S取代的rnaJ氨基酸序列(SEQ ID NO:3)。
实施例4C.引入accDA
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2070bp的下游同源臂(SEQID NO:4的核苷酸2032至4101)使用引物SNP_033_P1(SEQ ID NO:46)和SNP_033_P2(SEQ IDNO:47)扩增。引物SNP_033_P1包含与实施例4A中描述的pZ载体骨架重叠的50个核苷酸的5′延伸。引物SNP_033_P2在位置20处包含G到A的取代,以引入G310E突变。类似地,约2.1kb的上游同源臂(SEQ ID NO:4的核苷酸1至2071)使用引物SNP_084_P3(SEQ ID NO:48)和SNP_084_P4(SEQ ID NO:49)扩增。引物SNP_033_P3在位置21处包含C到T的取代,以引入G310E突变。SNP_033_P4包含与实施例4A中描述的pZ骨架重叠的50个碱基的5′延伸。两条同源臂的对齐产生由引物SNP_033_P2和SNP_033_P3限定的40-bp重叠。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带改变的accDA编码序列(SEQ ID NO:5),编码具有G310E取代的accDA氨基酸序列(SEQ ID NO:6)。
实施例4D.用于引入cg1144
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2037bp的上游同源臂(SEQID NO:7的核苷酸1至2037)使用引物SNP_316_P1(SEQ ID NO:50)和SNP_316_P2(SEQ IDNO:51)扩增。引物SNP_316_P1包含与实施例4A中描述的pZ载体骨架重叠的50个核苷酸的5′延伸。引物SNP_316_P2在位置21处包含G到A的取代,以引入P66S突变。类似地,2070bp的下游同源臂(SEQ ID NO:7的核苷酸1998至4067)使用引物SNP_316_P3(SEQ ID NO:52)和SNP_316_P4(SEQ ID NO:53)扩增。引物SNP_316_P3在位置20处包含C到T的取代,以引入P66S突变。SNP_316_P4包含与实施例4A中描述的pZ骨架重叠的50个碱基的5′延伸。两条同源臂的对齐产生由引物SNP_316_P2和SNP_316_P3限定的40-bp重叠。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带改变的cg1144编码序列(SEQ ID NO:8),编码具有P66S取代的cg1144氨基酸序列(SEQ ID NO:9)。
实施例4E.用于天然pyc启动子被启动子Pcg1860替代的转化载体的构建
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2043bp的上游同源臂(SEQID NO:10的核苷酸1至2043)使用引物Pcg1860_pyc_P1(SEQ ID NO:42)和Pcg1860_pyc_P2(SEQ ID NO:43)扩增。类似地,2050bp的下游同源臂(SEQ ID NO:10的核苷酸2161至4210)使用引物Pcg1860_pyc_P3(SEQ ID NO:44)和Pcg1860_pyc_P4(SEQ ID NO:45)扩增。93-bpPcg1860核苷酸序列(SEQ ID NO:11)通过引物Pcg1860_pyc_P2和Pcg1860-pyc_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带启动子Pcg1860(SEQID NO:11)而不是pyc启动子(SEQ ID NO:12)。
实施例4F.用于引入aceE
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2072bp的上游同源臂(SEQID NO:13的核苷酸1至2072)使用引物SNP_aceE_P1(SEQ ID NO:54)和SNP_aceE_P2(SEQ IDNO:55)扩增。引物SNP_aceE_P1包含与实施例4A中描述的pZ载体骨架重叠的50个核苷酸的5′延伸。引物SNP_aceE_P2在位置22处包含T到A的取代,以引入A突变。类似地,2073bp的下游同源臂(SEQ ID NO:13的核苷酸2029至4101)使用引物SNP_aceE_P3(SEQ ID NO:56)和SNP_aceE_P4(SEQ ID NO:57)扩增。引物SNP_aceE_P3在位置23处包含A到T的取代,以引入T突变。SNP_aceE_P4包含与实施例4中描述的pZ骨架重叠的50个碱基的5′延伸。两条同源臂的对齐产生由引物SNP_aceE_P2和SNP_aceE_P3限定的44-bp重叠。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带位于aceE开放阅读框中的TTG起始密码子(SEQ ID NO:14)。
实施例4G.密码子优化的lysA编码序列的插入
将包含密码子优化的lysA片段SEQ ID NO:16的盒插入SEQ ID NO:15的核苷酸2048与2049之间,并且合成组装(DNA 2.0)到实施例4A中描述的pZ载体中。最终的质粒包含位于Pcg0007_39-lysA
实施例4H.在zwf编码序列上游插入启动子
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2050bp的上游同源臂(SEQID NO:19的核苷酸1至2050)使用引物Pcg0007_39-zwf_P1(SEQ ID NO:34)和Pcg0007_39-zwf_P2(SEQ ID NO:35)扩增。类似地,2050bp的下游同源臂(SEQ ID NO:19的核苷酸2052至4101)使用引物Pcg0007_39-zwf_P3(SEQ ID NO:36)和Pcg0007_39-zwf_P4(SEQ ID NO:37)扩增。93-bp Pcg0007_39核苷酸序列通过引物Pcg0007_39-zwf_P2和Pcg0007_39-pyc_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株结果为SEQ ID NO:19的核苷酸2051被启动子Pcg0007_39(SEQ ID NO:20)替代。所得菌株携带位于zwf ORF上游的启动子Pcg0007_39(SEQ ID NO:21)。
实施例4I.天然opcA编码序列被C.humireducens NBRC 106098的opcA编码序列替代
两条同源臂通过PCR从NRRL B-11474衍生菌株BS2CZ基因组DNA(实施例3)扩增。2039bp的上游同源臂(SEQ ID NO:22的核苷酸1至2039)使用引物opcA_P1(SEQ ID NO:58)和opcA_P2(SEQ ID NO:59)扩增。类似地,2083bp的下游同源臂(SEQ ID NO:22的核苷酸3000至5082)使用引物opcA_P3(SEQ ID NO:60)和opcA_P4(SEQ ID NO:61)扩增。由93-bp的Pcg0007_39核苷酸序列(SEQ ID NO:20)和C.humireducens opcA(SEQ ID NO:23)组成的基因盒携带引物opcA_P2和opcA_P3的5′延伸的序列重叠。然后如实施例4A中描述的,将两条同源臂和基因盒组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株结果为SEQ ID NO:22的核苷酸2040至2999被Pcg0007_39-opcA(SEQ ID NO:24)替代。
实施例4J.用于在cg2766起始密码子前面插入启动子Pcg0007_39的转化载体的构建
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2050bp的下游同源臂(SEQID NO:26的核苷酸2051至4100)使用引物Pcg0007_39-cg2766_P1(SEQ ID NO:62)和Pcg0007_39-cg2766_P2(SEQ ID NO:63)扩增。类似地,2050bp的上游同源臂(SEQ ID NO:26的核苷酸1至2050)使用引物Pcg0007_39-cg2766_P3(SEQ ID NO:64)和Pcg0007_39-cg2766_P4(SEQ ID NO:65)扩增。93-bp Pcg0007_39核苷酸序列(SEQ ID NO:20)通过引物Pcg0007_39-cg2766_P2和Pcg0007_39-cg2766_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带插入在cg2766前面的启动子Pcg0007_39(SEQ ID NO:27)。
实施例4K.用于actA启动子被启动子Pcg0007_39替代的转化载体的构建。
两条同源臂通过PCR从NRRL B-11474基因组DNA(实施例3)扩增。
2021bp的下游同源臂(SEQ ID NO:28的核苷酸2263至4283)使用引物Pcg0007_39-actA_P1(SEQ ID NO:66)和Pcg0007_39-actA_P2(SEQ ID NO:67)扩增。类似地,2050bp的上游同源臂(SEQ ID NO:28的核苷酸1至2050)使用引物Pcg0007_39-actA_P3(SEQ ID NO:68)和Pcg0007_39-actA_P4(SEQ ID NO:69)扩增。93-bp Pcg0007_39核苷酸序列(SEQ ID NO:20)通过引物Pcg0007_39-actA_P2和Pcg0007_39-actA_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带替代SEQ ID NO:28的核苷酸2051至2262并插入在actA前面的启动子Pcg0007_39(SEQ ID NO:29)。
实施例4L.用于在cg1383起始密码子前面插入启动子Pcg0007_39的转化载体的构建
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2043bp的上游同源臂(SEQID NO:98的核苷酸1至2043)使用引物Pcg0007_39-cg1383_P1(SEQ ID NO:99)和Pcg0007_39-cg1383_P2(SEQ ID NO:100)扩增。类似地,2050bp的下游同源臂(SEQ ID NO:98的核苷酸2044至4093)使用引物Pcg0007_39-cg1383_P3(SEQ ID NO:101)和Pcg0007_39-cg1383_P4(SEQ ID NO:102)扩增。93-bp Pcg0007_39核苷酸序列通过引物Pcg0007_39-cg1383_P2和Pcg0007_39-cg1383_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带插入在cg1383起始密码子前面的Pcg0007_39启动子(SEQ ID NO:103)。
实施例4M.用于天然phoU启动子被启动子Pcg0007_39替代的转化载体的构建
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。1935bp的上游同源臂(SEQID NO:104的核苷酸1至1935)使用引物Pcg0007_39-phoU_P1(SEQ ID NO:105)和Pcg0007_39-phoU_P2(SEQ ID NO:106)扩增。类似地,2050bp的下游同源臂(SEQ ID NO:104的核苷酸1995至4044)使用引物Pcg0007_39-phoU_P3(SEQ ID NO:107)和Pcg0007_39-phoU_P4(SEQID NO:108)扩增。93-bp Pcg0007_39核苷酸序列通过引物Pcg0007_39-phoU_P2和Pcg0007_39-phoU_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带位于phoU起始密码子前面并替代SEQ ID NO:104的核苷酸1936-1994的Pcg0007_39启动子(SEQ ID NO:109)。
实施例4N.用于天然cg3210启动子被启动子Pcg0007_39替代的转化载体的构建
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。1935bp的上游同源臂(SEQID NO:110的核苷酸1至2030)使用引物Pcg0007_39-cg3210_P1(SEQ ID NO:111)和Pcg0007_39-cg3210_P2(SEQ ID NO:112)扩增。类似地,2047bp的下游同源臂(SEQ ID NO:110的核苷酸2090至4136)使用引物Pcg0007_39-cg3210_P3(SEQ ID NO:113)和Pcg0007_39-3210_P4(SEQ ID NO:114)扩增。93-bp Pcg0007_39核苷酸序列通过引物Pcg0007_39-cg3210_P2和Pcg0007_39-cg3210_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带替代SEQ ID NO:110的核苷酸2031至2089并插入在cg3210起始密码子前面的Pcg0007_39(SEQ ID NO:115)。
实施例4O.用于天然cg0800启动子被启动子Pcg0007_39替代的转化载体的构建
两条同源臂通过PCR从NRRL B-11474基因组DNA扩增。2043bp的上游同源臂(SEQID NO:116的核苷酸1至2043)使用引物Pcg0007_39-cg0800_P3(SEQ ID NO:117)和Pcg0007_39-cg0800_P4(SEQ ID NO:118)扩增。类似地,2050bp的下游同源臂(SEQ ID NO:116的核苷酸2184至4233)使用引物Pcg0007_39-cg0800_P1(SEQ ID NO:119)和Pcg0007_39-0800_P2(SEQ ID NO:120)扩增。93-bp Pcg0007_39核苷酸序列通过引物Pcg0007_39-cg0800_P2和Pcg0007_39-cg0800_P3的5′延伸的重叠获得。然后如实施例4A中描述的,将两条同源臂组装到pZ中,并且如实施例2中描述转化到谷氨酸棒状杆菌NRRL B-11474及其衍生菌株中。所得菌株携带插入在cg0800起始密码子前面并替代SEQ ID NO:116的核苷酸2044-2183的Pcg0007_39启动子(SEQ ID NO:121)。
实施例5.在微量滴定板中评价改变的谷氨酸棒状杆菌菌株的赖氨酸产生
实施例5A.改变的谷氨酸棒状杆菌菌株的赖氨酸产生
先前实施例中描述的每种改变的菌株以及由组合不同改变产生的菌株的赖氨酸产生通过在微量滴定板中的三阶段发酵进行评价。使细胞在培养基C(表5)中生长。在48小时后,将10%的接种物转移至培养基D(表5)。在19小时后,将10%的细胞培养物转移至培养基E(表5),并且生长另外的24小时或96小时。培养条件为32℃,1000rpm(Infors HT)。
产生的赖氨酸的量使用偶联的赖氨酸氧化酶测定来确定。将培养物上清液添加至赖氨酸氧化酶反应溶液(250mM磷酸钾缓冲液、pH 7.5、824mg/ml苯酚、76mg/ml 4-氨基安替比林(4-amino antipyrene)(Sigma)、0.03mg/ml过氧化物酶(Sigma)、0.015单位/ml赖氨酸氧化酶(Sigma))中。将样品在25℃孵育40min,并且在490nm处读取吸光度。微量滴定板中右旋糖的量通过偶联葡萄糖氧化酶测定来确定。将培养物上清液添加至葡萄糖氧化酶反应溶液(275mM马来酸钠缓冲液pH 5.5、730mg/ml苯酚、680mg/ml 4-氨基安替比林(Sigma)、0.027mg/ml过氧化物酶(Sigma)、56mg/L葡萄糖氧化酶(Sigma))中。将样品在25℃孵育40min,并且在490nm处读取吸光度。
每种重组菌株及其直接亲本菌株产生的赖氨酸量(mM)示于表6中。表7A是基因组改变的列表,并且表7B鉴定了相对于保藏为NRRL B-11474的细菌的天然基因组,在测试菌株中存在何种改变。
表7A
表7B.修饰菌株的基因型
表8.序列表
参考文献
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Buchholz et al.,“Platform Engineering of Corynebacterium glutamicumwith Reduced Pyruvate Dehydrogenase Complex Activity for Improved Productionof 1-Lysine,1-Valine,and 2-Ketoisovalerate,”Applied Env.Microbiol.79,5566-75,2013
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序列表
<110> 阿彻丹尼尔斯米德兰德公司
<120> 工程化棒状杆菌属菌株
<130> 008200.00060
<140>
<141>
<150> US 62/728,384
<151> 2018-09-07
<160> 121
<170> FastSEQ for Windows Version 4.0
<210> 1
<211> 4101
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 1
aaaaactaga actgctcaag gccgttcgtg aggaagttgg ggatcgggcg aagctcatcg 60
ccggtgtcgg aaccaacaac acgcggacat ctgtggaact tgcggaagct gctgcttctg 120
ctggcgcaga cggcctttta gttgtaactc cttattactc caagccgagc caagagggat 180
tgctggcgca cttcggtgca attgctgcag caacagaggt tccaatttgt ctctatgaca 240
ttcctggtcg gtcaggtatt ccaattgaat ctgataccat gagacgcctg agtgaattac 300
ctacgatttt ggcggtcaag gacgccaagg gtgacctcgt tgcagccacg tcattgatca 360
aagaaacggg acttgcctgg tattcaggcg atgacccact aaaccttgtt tggcttgctt 420
tgggcggatc aggtttcatt tccgtaattg gacatgcagc ccccacagca ttacgtgagt 480
tgtacacaag cttcgaggaa ggcgacctcg tccgtgcgcg ggaaatcaac gccaaactat 540
caccgctggt agctgcccaa ggtcgcttgg gtggagtcag cttggcaaaa gctgctctgc 600
gtctgcaggg catcaacgta ggagatcctc gacttccaat tatggctcca aatgagcagg 660
aacttgaggc tctccgagaa gacatgaaaa aagctggagt tctataaata tgaatgattc 720
ccgaaatcgc ggccggaagg ttacccgcaa ggcgggccca ccagaagctg gtcaggaaaa 780
ccatctggat acccctgtct ttcaggcacc agatgcttcc tctaaccaga gcgctgtaaa 840
agctgagacc gccggaaacg acaatcggga tgctgcgcaa ggtgctcaag gatcccaaga 900
ttctcagggt tcccagaacg ctcaaggttc ccagaaccgc gagtccggaa ataacaaccg 960
caaccgttcc aacaacaacc gtcgcggtgg tcgtggacgt cgtggatccg gaaacgccaa 1020
tgagggcgcg aacaacaaca gtggtaacca gaaccgtcag ggcggaaacc gtggcaaccg 1080
cggtggcgga cgccgaaacg ttgttaagtc gatgcagggt gcggatctga cccagcgcct 1140
gccagagcca ccaaaggcac cggcaaacgg tctgcgtatt tacgcacttg gtggcatttc 1200
cgaaatcggt cgcaacatga ccgtgtttga gtacaacaac cgtctgctca tcgtggactg 1260
tggtgtgctc ttcccatctt caggtgagcc aggcgttgac ctgattcttc ctgacttcgg 1320
cccaattgag gatcacctgc accgcgtcga tgcattggtg gttactcacg gacacgaaga 1380
ccacattggt gctattccct ggctgctgaa gctgcgcaac gatatcccaa ttttggcatc 1440
ccgtttcacc ttggctctga ttgcagctaa gtgtaaggaa caccgtcagc gtccgaagct 1500
gatcgaggtc aacgagcagt ccaatgagga ccgcggaccg ttcaacattc gcttctgggc 1560
tgttaaccac tccatcccag actgccttgg tcttgctatc aagactcctg ctggtttggt 1620
catccacacc ggtgacatca agctggatca gactcctcct gatggacgcc caactgacct 1680
gcctgcattg tcccgtttcg gtgacgaggg cgtggacttg atgctgtgtg actccaccaa 1740
cgccaccacc cctggtgttt ctggatctga agctgatgtt gctccaaccc tgaagcgttt 1800
ggtcggcgat gctaagcagc gcgtcatttt ggcgtcgttt gcttccaacg tgtaccgcgt 1860
tcaggcagct gtggatgctg ctgttgcgtc caaccgtaag gttgcattca acggtcgttc 1920
catgattcgc aacatggaga tcgcggagaa gcttggttac ctgaaggcac ctcgcggaac 1980
cattatttcc atggatgatg cttctcgtat ggctcctcac aaggtcatgc tgattaccac 2040
tggtactcag ggtgagccta tggctgcgct gtctcgcatg gcgcgtcgtg agcaccgaca 2100
gatcactgtc cgtgatggag acttgattat cctttcttcc tccctggttc caggtaacga 2160
agaagcagtg ttcggtgtca tcaacatgct ggctcagatc ggtgcaactg ttgttaccgg 2220
tcgcgacgcc aaggtgcaca cctcgggcca cggctactcc ggagagctgt tgttcttgta 2280
caacgccgct cgtccgaaga acgctatgcc tgtccacggc gagtggcgcc acctgcgcgc 2340
caacaaggaa ctggctatct ccactggtgt taaccgcgac aacgttgtgc ttgcacaaaa 2400
cggtgttgtg gttgatatgg tcaacggtcg cgcacaggtt gttggtcaga ttccagttgg 2460
taacctttac gtcgacggtg tcaccatggg tgatattgac gcggacatcc ttgcagaccg 2520
tacctcgctt ggtgagggtg gcttgatctc gatcactgct gtcatcgaca accgcactgg 2580
ccgtctgttg gagcgcccaa ccgttcagac cagcggtttc tctgaggatg caaagtccat 2640
gatgggtgag gtcactgagc tgtccgaaac caccatgaat gatcttgcag ctgaaggtga 2700
aaacgatcct taccgcatgg ttcagcagct gcgccgcaag ctctctcgct tcgtcgagca 2760
gaagtggaag cgccagccgg tcatcatgcc aaccgtcatt ccgatgactg cggaaaccac 2820
gcacatcggt gacgatgagg ttcgcgcttc acgcgagtcc ctgtaaaagc atttcgcttt 2880
tcgacgtcgc gagcccgcag cacccaacct gggtgccgcg ggctcaacgc gtttctacac 2940
attgatctag tcttggttgc atgaatctcg cacaaacaga acgagcagca cttcaagcat 3000
cactgttgga aaaggggcca gacgcaccaa ccttgtgcgg cgaatggacc accaaagatc 3060
ttgcggcaca cctttatgtc cgtgagcaga agccaggagc agcgctgcgc agtatgcttc 3120
ccggcaagca ggatcctgca aaagaagagt ttgaagaagc gcttacccgc ccttacgtgg 3180
acgttgtcaa cgcgtgggct gaaggcccac aggggctgaa tccatggcga gcccttgact 3240
ctattggaaa tggcatggag catttcattc accatgaaga tgttctgcgt ggcgcgttgc 3300
agccaggcga tgaagtagaa gttcgtccca tggaccatgc gcacggcaaa gagctgcacc 3360
gcattttaaa acttctggct ccacgaatga tcaaatcacc gcgacccgtg gttctagctc 3420
cggtgggttt tccgaggatt gttcttcatg aaggtcgggg agtgtcggcc gatggtgaaa 3480
acgtctgtcg gattagcggt ggggtagggg agctgctgtt gtggctgtct ggccgtgatg 3540
ttgtgaaact aacatttgaa ggcaaccaag agggcgttgt tcggggaagt ttatagaaaa 3600
cttggacgaa tcgcgtggtg cttctgtgaa tagagttgtt tgagcgacta gagttaaggc 3660
catgactgtc agaaacggcg cacccaagcg gggaagctca cgatcatcgg gaaagtcgac 3720
ggggccaaaa accaccgcga cgaccaggcg aagcaaacca acaaccagca tctctcgggg 3780
tggccggtca gagactgctg cgtttacaac atcgatgacc gctcgacgcg ctgctgatac 3840
ttccgaagaa cggaccggca gcgcatttcg cgctgtggga tccggtatcg gaacgctatt 3900
tggtgccacc gcgagtggct tgggtaagat gacccgttcc attggtgaac ttggtcgtcg 3960
tcgaaacgaa gatgtactcg acgattttga tgatttcgag gaggagatcg caacgaaacc 4020
cgccacacga aaatcgcgga gcaaggctgt tgaacctgag ccggaattcg atgaggattt 4080
tgatgagcaa tcaggaaacc g 4101
<210> 2
<211> 2157
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的RnaJ蛋白的编码序列
<400> 2
atgaatgatt cccgaaatcg cggccggaag gttacccgca aggcgggccc accagaagct 60
ggtcaggaaa accatctgga tacccctgtc tttcaggcac cagatgcttc ctctaaccag 120
agcgctgtaa aagctgagac cgccggaaac gacaatcggg atgctgcgca aggtgctcaa 180
ggatcccaag attctcaggg ttcccagaac gctcaaggtt cccagaaccg cgagtccgga 240
aataacaacc gcaaccgttc caacaacaac cgtcgcggtg gtcgtggacg tcgtggatcc 300
ggaaacgcca atgagggcgc gaacaacaac agtggtaacc agaaccgtca gggcggaaac 360
cgtggcaacc gcggtggcgg acgccgaaac gttgttaagt cgatgcaggg tgcggatctg 420
acccagcgcc tgccagagcc accaaaggca ccggcaaacg gtctgcgtat ttacgcactt 480
ggtggcattt ccgaaatcgg tcgcaacatg accgtgtttg agtacaacaa ccgtctgctc 540
atcgtggact gtggtgtgct cttcccatct tcaggtgagc caggcgttga cctgattctt 600
cctgacttcg gcccaattga ggatcacctg caccgcgtcg atgcattggt ggttactcac 660
ggacacgaag accacattgg tgctattccc tggctgctga agctgcgcaa cgatatccca 720
attttggcat cccgtttcac cttggctctg attgcagcta agtgtaagga acaccgtcag 780
cgtccgaagc tgatcgaggt caacgagcag tccaatgagg accgcggacc gttcaacatt 840
cgcttctggg ctgttaacca ctccatccca gactgccttg gtcttgctat caagactcct 900
gctggtttgg tcatccacac cggtgacatc aagctggatc agactcctcc tgatggacgc 960
ccaactgacc tgcctgcatt gtcccgtttc ggtgacgagg gcgtggactt gatgctgtgt 1020
gactccacca acgccaccac ccctggtgtt tctggatctg aagctgatgt tgctccaacc 1080
ctgaagcgtt tggtcggcga tgctaagcag cgcgtcattt tggcgtcgtt tgcttccaac 1140
gtgtaccgcg ttcaggcagc tgtggatgct gctgttgcgt ccaaccgtaa ggttgcattc 1200
aacggtcgtt ccatgattcg caacatggag atcgcggaga agcttggtta cctgaaggca 1260
cctcgcggaa ccattatttc catggatgat gcttctcgta tggctcctca caaggtcatg 1320
ctgattacca ctggtactca gagtgagcct atggctgcgc tgtctcgcat ggcgcgtcgt 1380
gagcaccgac agatcactgt ccgtgatgga gacttgatta tcctttcttc ctccctggtt 1440
ccaggtaacg aagaagcagt gttcggtgtc atcaacatgc tggctcagat cggtgcaact 1500
gttgttaccg gtcgcgacgc caaggtgcac acctcgggcc acggctactc cggagagctg 1560
ttgttcttgt acaacgccgc tcgtccgaag aacgctatgc ctgtccacgg cgagtggcgc 1620
cacctgcgcg ccaacaagga actggctatc tccactggtg ttaaccgcga caacgttgtg 1680
cttgcacaaa acggtgttgt ggttgatatg gtcaacggtc gcgcacaggt tgttggtcag 1740
attccagttg gtaaccttta cgtcgacggt gtcaccatgg gtgatattga cgcggacatc 1800
cttgcagacc gtacctcgct tggtgagggt ggcttgatct cgatcactgc tgtcatcgac 1860
aaccgcactg gccgtctgtt ggagcgccca accgttcaga ccagcggttt ctctgaggat 1920
gcaaagtcca tgatgggtga ggtcactgag ctgtccgaaa ccaccatgaa tgatcttgca 1980
gctgaaggtg aaaacgatcc ttaccgcatg gttcagcagc tgcgccgcaa gctctctcgc 2040
ttcgtcgagc agaagtggaa gcgccagccg gtcatcatgc caaccgtcat tccgatgact 2100
gcggaaacca cgcacatcgg tgacgatgag gttcgcgctt cacgcgagtc cctgtaa 2157
<210> 3
<211> 718
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的RnaJ蛋白
<400> 3
Met Asn Asp Ser Arg Asn Arg Gly Arg Lys Val Thr Arg Lys Ala Gly
1 5 10 15
Pro Pro Glu Ala Gly Gln Glu Asn His Leu Asp Thr Pro Val Phe Gln
20 25 30
Ala Pro Asp Ala Ser Ser Asn Gln Ser Ala Val Lys Ala Glu Thr Ala
35 40 45
Gly Asn Asp Asn Arg Asp Ala Ala Gln Gly Ala Gln Gly Ser Gln Asp
50 55 60
Ser Gln Gly Ser Gln Asn Ala Gln Gly Ser Gln Asn Arg Glu Ser Gly
65 70 75 80
Asn Asn Asn Arg Asn Arg Ser Asn Asn Asn Arg Arg Gly Gly Arg Gly
85 90 95
Arg Arg Gly Ser Gly Asn Ala Asn Glu Gly Ala Asn Asn Asn Ser Gly
100 105 110
Asn Gln Asn Arg Gln Gly Gly Asn Arg Gly Asn Arg Gly Gly Gly Arg
115 120 125
Arg Asn Val Val Lys Ser Met Gln Gly Ala Asp Leu Thr Gln Arg Leu
130 135 140
Pro Glu Pro Pro Lys Ala Pro Ala Asn Gly Leu Arg Ile Tyr Ala Leu
145 150 155 160
Gly Gly Ile Ser Glu Ile Gly Arg Asn Met Thr Val Phe Glu Tyr Asn
165 170 175
Asn Arg Leu Leu Ile Val Asp Cys Gly Val Leu Phe Pro Ser Ser Gly
180 185 190
Glu Pro Gly Val Asp Leu Ile Leu Pro Asp Phe Gly Pro Ile Glu Asp
195 200 205
His Leu His Arg Val Asp Ala Leu Val Val Thr His Gly His Glu Asp
210 215 220
His Ile Gly Ala Ile Pro Trp Leu Leu Lys Leu Arg Asn Asp Ile Pro
225 230 235 240
Ile Leu Ala Ser Arg Phe Thr Leu Ala Leu Ile Ala Ala Lys Cys Lys
245 250 255
Glu His Arg Gln Arg Pro Lys Leu Ile Glu Val Asn Glu Gln Ser Asn
260 265 270
Glu Asp Arg Gly Pro Phe Asn Ile Arg Phe Trp Ala Val Asn His Ser
275 280 285
Ile Pro Asp Cys Leu Gly Leu Ala Ile Lys Thr Pro Ala Gly Leu Val
290 295 300
Ile His Thr Gly Asp Ile Lys Leu Asp Gln Thr Pro Pro Asp Gly Arg
305 310 315 320
Pro Thr Asp Leu Pro Ala Leu Ser Arg Phe Gly Asp Glu Gly Val Asp
325 330 335
Leu Met Leu Cys Asp Ser Thr Asn Ala Thr Thr Pro Gly Val Ser Gly
340 345 350
Ser Glu Ala Asp Val Ala Pro Thr Leu Lys Arg Leu Val Gly Asp Ala
355 360 365
Lys Gln Arg Val Ile Leu Ala Ser Phe Ala Ser Asn Val Tyr Arg Val
370 375 380
Gln Ala Ala Val Asp Ala Ala Val Ala Ser Asn Arg Lys Val Ala Phe
385 390 395 400
Asn Gly Arg Ser Met Ile Arg Asn Met Glu Ile Ala Glu Lys Leu Gly
405 410 415
Tyr Leu Lys Ala Pro Arg Gly Thr Ile Ile Ser Met Asp Asp Ala Ser
420 425 430
Arg Met Ala Pro His Lys Val Met Leu Ile Thr Thr Gly Thr Gln Ser
435 440 445
Glu Pro Met Ala Ala Leu Ser Arg Met Ala Arg Arg Glu His Arg Gln
450 455 460
Ile Thr Val Arg Asp Gly Asp Leu Ile Ile Leu Ser Ser Ser Leu Val
465 470 475 480
Pro Gly Asn Glu Glu Ala Val Phe Gly Val Ile Asn Met Leu Ala Gln
485 490 495
Ile Gly Ala Thr Val Val Thr Gly Arg Asp Ala Lys Val His Thr Ser
500 505 510
Gly His Gly Tyr Ser Gly Glu Leu Leu Phe Leu Tyr Asn Ala Ala Arg
515 520 525
Pro Lys Asn Ala Met Pro Val His Gly Glu Trp Arg His Leu Arg Ala
530 535 540
Asn Lys Glu Leu Ala Ile Ser Thr Gly Val Asn Arg Asp Asn Val Val
545 550 555 560
Leu Ala Gln Asn Gly Val Val Val Asp Met Val Asn Gly Arg Ala Gln
565 570 575
Val Val Gly Gln Ile Pro Val Gly Asn Leu Tyr Val Asp Gly Val Thr
580 585 590
Met Gly Asp Ile Asp Ala Asp Ile Leu Ala Asp Arg Thr Ser Leu Gly
595 600 605
Glu Gly Gly Leu Ile Ser Ile Thr Ala Val Ile Asp Asn Arg Thr Gly
610 615 620
Arg Leu Leu Glu Arg Pro Thr Val Gln Thr Ser Gly Phe Ser Glu Asp
625 630 635 640
Ala Lys Ser Met Met Gly Glu Val Thr Glu Leu Ser Glu Thr Thr Met
645 650 655
Asn Asp Leu Ala Ala Glu Gly Glu Asn Asp Pro Tyr Arg Met Val Gln
660 665 670
Gln Leu Arg Arg Lys Leu Ser Arg Phe Val Glu Gln Lys Trp Lys Arg
675 680 685
Gln Pro Val Ile Met Pro Thr Val Ile Pro Met Thr Ala Glu Thr Thr
690 695 700
His Ile Gly Asp Asp Glu Val Arg Ala Ser Arg Glu Ser Leu
705 710 715
<210> 4
<211> 4101
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 4
gctgcagaca ggtagtcacc tgcgatagcc agaccgttct gggttccgga gaaggaagca 60
ccaccggtgt agaagtcggt ggattcgctg gtgctcttgc ccacgcgaag caccacggtc 120
atcgtcacaa taatgaatac gacgaagaca ctgatgttaa gaatcggatt accgacgccc 180
tcagaaacag cgtcttgtgc aaggagaata gtggaattca tatctgatta accttccatc 240
ttctggcgaa tagcagcctg acgaggctcc aagttcctgt tcgcaaacat gacgtagacg 300
taagtaataa tgaatgtggt tacgaactga aggaagccga agattaggcc gatgttgatt 360
gcgccgaaaa ctggggttgc catccactct gatgcaaagg atgcaaccag aacgtagacg 420
acgtaccaca agaagaaggc aacggtcatt gggaaagcaa aggaacggaa cttgctgcgg 480
agctctccaa attctgggct cgcctgcatg tcccgaaact cttgagccgt cggctggcga 540
cgcccctggg tgatttgttc tgcattcact ggaaacttat ctcgctttca tatccctcga 600
gcgggagtcg gtgatcggcc actctttaag caatgccggc tttaaaataa agcaacttat 660
atgtttctca ccacatctgg ccgacgaccg ctaagtatgt tgtcgatcac agctaaacgt 720
gtgaatgtga agttacctaa ctcacattgc aatgcgatag cgatttggaa aactcactcc 780
ccccaatacc ttaacttgaa cttaaaaata gtggtttacc tgcatttata aaagttcccg 840
atctaccccc tctttacccc gaaatacccc ttttgcaaag attgcaaaca caacagtgca 900
atagttaacg ggcttcacac gtcacctttc tgtccggttt taggctatgt tcgggacgtc 960
taggcaaaag tagttttgtg agatgaaacg cataatccgt cattttttac gcaatcgata 1020
gcctaaattg ggcttagatc ttccgcctct aaataggtat gcaaagacat tcgaattaat 1080
taacaaagcc atttttcggc cgtggagaag cgttttccga ctatggtgtg gggcatggaa 1140
cacacttcag cattgacgct catagactcg gttttggacc ctgacagctt catttcttgg 1200
gatgaaactc cccaatatga caacctcaat caaggctatg cagagacctt ggagcgggct 1260
cgaagcaagg ccaaatgcga tgaatcggtg attactggag aaggcaccgt ggagggcatt 1320
ccggtagccg ttattttgtc cgatttttcc ttcctcggcg gttctttggg cacggtcgcg 1380
tcggtgcgca tcatgaaggc gattcaccgc gccacagagc tgaaactccc actgctggtc 1440
tcccctgctt ccggtggtgc gcgcatgcag gaagacaatc gagcctttgt catgatggtg 1500
tccataaccg cggctgtgca acgccaccgc gaagcacatt tgccgttcct ggtgtatttg 1560
cgcaatccca cgatgggtgg cgccatggcc tcgtggggtt catctgggca tctcactttt 1620
gcggaacccg gcgcgcagat cggtttcctg ggccctcgcg tggtggagtt gaccaccggg 1680
cgtgcgcttc cagacggtgt gcagcaggcg gagaatttgg tgaaaactgg tgtgattgat 1740
ggcattgtgt cgccgctcca attgcgtgca gcggtggcaa aaacgctcaa gattattcag 1800
cctgcagagg caaccgatcg tttttctcca acaactcctg gcgcagcgct tccggtgatg 1860
gaggcgattg cgcgttctcg tgacccgcac aggcctggaa tcggggagat tatggaaacg 1920
ttgggggcag acgtcgtcag gctatctggt gcgcgtgctg gcgcgttgag ctcggctgtg 1980
cgcgttgccc tagcgcgcat cgggggccgg cctgtggtgc tgattggaca ggatcggcgc 2040
ttcacgcttg ggccgcagga gctgcgtttt gcgcgccgtg gcatttcgtt ggcgcgcgag 2100
ctgaacctgc cgatcgtgtc catcatcgac acctccggcg ccgaattgtc acaggcggct 2160
gaggagctcg gcatcgcaag ctcgattgcg cgcaccttgt ccaagcttat cgacgctccc 2220
ctccccaccg tttcggtcat tattggtcag ggcgttggcg gtggcgcgct ggccatgctg 2280
cccgccgatc tggtctacgc ggccgaaaac gcgtggctgt ccgcattgcc tccagagggc 2340
gcctcggcca tcctcttccg cgacaccaac cacgccgcgg aaatcataga gcgacaaggc 2400
gtgcaggcgc acgcactttt aagccaaggg cttatcgacg ggatcgtcgc cgaaaccgag 2460
cactttgttg aagaaattct cggcacaatc agcaacgccc tctccgaatt ggataacaat 2520
ccggagaggg caggacgcga cagtcgcttc acacgatttg agcgtttagc gcagtaaaga 2580
aaattatgcg ctgatcaaat cgatgatgaa caccagggta cggccagaca gtgggtggcc 2640
ggaaccctca gggccgtaag cagcctctgg cggaatggtc agctgacgac gtccgccgac 2700
cttcatgcct ggaattcctt cttgccaacc tgcaatgagg ccgttgagtg ggaactggct 2760
ggtctgtcca cgatcccagg aagagtcaaa ctcctcgccg gtttcaaggt caacgcccac 2820
atagtggacc tcaacttctc cgcctgggcg ggcttctgcg ccttcgccaa cgatgatgtc 2880
agagattacg agatcttccg gtgctggacc gaccggtagc tcaatctgtg gcttttccat 2940
gtgctgtaca tgctccttga aaatcgtatt ttttggtttc atgcaaaaac ggccgaaacc 3000
atccctgtca ttatgacaga cgaagtggct tcggccgaag ttgaacgaga atcatccgct 3060
aaatttagcg ctcctcgcga ggaaccaact tgcgggattc cttgccggtg tagacctggc 3120
gtgggcggtt gatcttgttg cctgctgcac cgagctgctc gcggtagtga gcgatccatc 3180
ctggcagacg accgattgcg aacaatacgg tgaagaagtc agttgggaag cccattgcgc 3240
ggtagatcag gccggtgtag aagtctacgt tcgggtagag cttgcgggag atgaagtaat 3300
catcagccag tgcaatttct tccagcttga ttgccagatc cagaagatcg tcgccaccga 3360
ggtgctcgag gatctcgtgt gcggtctcct tgacgattgc tgcacgtgga tcgtagttct 3420
tgtaaacgcg gtgtccgaag cccatgaggc ggacgccgtc ttccttgttc ttgaccttgt 3480
tcatgaacgc ggttgcgtcg ccaccgtggt tgttcttgat gtcttcgagc atctccagaa 3540
cagcctggtt tgcgccaccg tgcagtgggc cggacagagc gttgatgcca ccagcgatgg 3600
agacaaacat gttggcctgt gcggaaccga tcatacgaac ggtggaggtg gagcagttct 3660
gctcgtggtc agcgtgcagg atgagcagct tgtccagagc cttgaccatg attgggtcga 3720
tctcgtatgg ctcggttggg taaccgaaca tcatgcgcag gaagttctca cgcgcgttga 3780
gggagttgtc tgggtacatg taaggagcac ccttgcgtgc gcggtgtgcg tacgcagcca 3840
gcattggaac ctttgccatg aggcgaacgg ttgccttatc aagctgtgcc tcatcgagtg 3900
ggttcagctg atcctggtag taggtagaca aaatgttaac cgaggaagcc aaggttgcca 3960
ttgggtgagc gtcgcgtggg aacacgttga actgggactt gaagtcctcg tccagaaggg 4020
tgtggtggcg aatctcgtcg ttaaacttgt gaagctcatc tggggttggt agctcaccgt 4080
tgataagtag gtaagaaacc t 4101
<210> 5
<211> 1455
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的accDA蛋白的编码序列
<400> 5
atggtgtggg gcatggaaca cacttcagca ttgacgctca tagactcggt tttggaccct 60
gacagcttca tttcttggga tgaaactccc caatatgaca acctcaatca aggctatgca 120
gagaccttgg agcgggctcg aagcaaggcc aaatgcgatg aatcggtgat tactggagaa 180
ggcaccgtgg agggcattcc ggtagccgtt attttgtccg atttttcctt cctcggcggt 240
tctttgggca cggtcgcgtc ggtgcgcatc atgaaggcga ttcaccgcgc cacagagctg 300
aaactcccac tgctggtctc ccctgcttcc ggtggtgcgc gcatgcagga agacaatcga 360
gcctttgtca tgatggtgtc cataaccgcg gctgtgcaac gccaccgcga agcacatttg 420
ccgttcctgg tgtatttgcg caatcccacg atgggtggcg ccatggcctc gtggggttca 480
tctgggcatc tcacttttgc ggaacccggc gcgcagatcg gtttcctggg ccctcgcgtg 540
gtggagttga ccaccgggcg tgcgcttcca gacggtgtgc agcaggcgga gaatttggtg 600
aaaactggtg tgattgatgg cattgtgtcg ccgctccaat tgcgtgcagc ggtggcaaaa 660
acgctcaaga ttattcagcc tgcagaggca accgatcgtt tttctccaac aactcctggc 720
gcagcgcttc cggtgatgga ggcgattgcg cgttctcgtg acccgcacag gcctggaatc 780
ggggagatta tggaaacgtt gggggcagac gtcgtcaggc tatctggtgc gcgtgctggc 840
gcgttgagct cggctgtgcg cgttgcccta gcgcgcatcg ggggccggcc tgtggtgctg 900
attggacagg atcggcgctt cacgcttgag ccgcaggagc tgcgttttgc gcgccgtggc 960
atttcgttgg cgcgcgagct gaacctgccg atcgtgtcca tcatcgacac ctccggcgcc 1020
gaattgtcac aggcggctga ggagctcggc atcgcaagct cgattgcgcg caccttgtcc 1080
aagcttatcg acgctcccct ccccaccgtt tcggtcatta ttggtcaggg cgttggcggt 1140
ggcgcgctgg ccatgctgcc cgccgatctg gtctacgcgg ccgaaaacgc gtggctgtcc 1200
gcattgcctc cagagggcgc ctcggccatc ctcttccgcg acaccaacca cgccgcggaa 1260
atcatagagc gacaaggcgt gcaggcgcac gcacttttaa gccaagggct tatcgacggg 1320
atcgtcgccg aaaccgagca ctttgttgaa gaaattctcg gcacaatcag caacgccctc 1380
tccgaattgg ataacaatcc ggagagggca ggacgcgaca gtcgcttcac acgatttgag 1440
cgtttagcgc agtaa 1455
<210> 6
<211> 484
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的accDA蛋白
<400> 6
Met Val Trp Gly Met Glu His Thr Ser Ala Leu Thr Leu Ile Asp Ser
1 5 10 15
Val Leu Asp Pro Asp Ser Phe Ile Ser Trp Asp Glu Thr Pro Gln Tyr
20 25 30
Asp Asn Leu Asn Gln Gly Tyr Ala Glu Thr Leu Glu Arg Ala Arg Ser
35 40 45
Lys Ala Lys Cys Asp Glu Ser Val Ile Thr Gly Glu Gly Thr Val Glu
50 55 60
Gly Ile Pro Val Ala Val Ile Leu Ser Asp Phe Ser Phe Leu Gly Gly
65 70 75 80
Ser Leu Gly Thr Val Ala Ser Val Arg Ile Met Lys Ala Ile His Arg
85 90 95
Ala Thr Glu Leu Lys Leu Pro Leu Leu Val Ser Pro Ala Ser Gly Gly
100 105 110
Ala Arg Met Gln Glu Asp Asn Arg Ala Phe Val Met Met Val Ser Ile
115 120 125
Thr Ala Ala Val Gln Arg His Arg Glu Ala His Leu Pro Phe Leu Val
130 135 140
Tyr Leu Arg Asn Pro Thr Met Gly Gly Ala Met Ala Ser Trp Gly Ser
145 150 155 160
Ser Gly His Leu Thr Phe Ala Glu Pro Gly Ala Gln Ile Gly Phe Leu
165 170 175
Gly Pro Arg Val Val Glu Leu Thr Thr Gly Arg Ala Leu Pro Asp Gly
180 185 190
Val Gln Gln Ala Glu Asn Leu Val Lys Thr Gly Val Ile Asp Gly Ile
195 200 205
Val Ser Pro Leu Gln Leu Arg Ala Ala Val Ala Lys Thr Leu Lys Ile
210 215 220
Ile Gln Pro Ala Glu Ala Thr Asp Arg Phe Ser Pro Thr Thr Pro Gly
225 230 235 240
Ala Ala Leu Pro Val Met Glu Ala Ile Ala Arg Ser Arg Asp Pro His
245 250 255
Arg Pro Gly Ile Gly Glu Ile Met Glu Thr Leu Gly Ala Asp Val Val
260 265 270
Arg Leu Ser Gly Ala Arg Ala Gly Ala Leu Ser Ser Ala Val Arg Val
275 280 285
Ala Leu Ala Arg Ile Gly Gly Arg Pro Val Val Leu Ile Gly Gln Asp
290 295 300
Arg Arg Phe Thr Leu Glu Pro Gln Glu Leu Arg Phe Ala Arg Arg Gly
305 310 315 320
Ile Ser Leu Ala Arg Glu Leu Asn Leu Pro Ile Val Ser Ile Ile Asp
325 330 335
Thr Ser Gly Ala Glu Leu Ser Gln Ala Ala Glu Glu Leu Gly Ile Ala
340 345 350
Ser Ser Ile Ala Arg Thr Leu Ser Lys Leu Ile Asp Ala Pro Leu Pro
355 360 365
Thr Val Ser Val Ile Ile Gly Gln Gly Val Gly Gly Gly Ala Leu Ala
370 375 380
Met Leu Pro Ala Asp Leu Val Tyr Ala Ala Glu Asn Ala Trp Leu Ser
385 390 395 400
Ala Leu Pro Pro Glu Gly Ala Ser Ala Ile Leu Phe Arg Asp Thr Asn
405 410 415
His Ala Ala Glu Ile Ile Glu Arg Gln Gly Val Gln Ala His Ala Leu
420 425 430
Leu Ser Gln Gly Leu Ile Asp Gly Ile Val Ala Glu Thr Glu His Phe
435 440 445
Val Glu Glu Ile Leu Gly Thr Ile Ser Asn Ala Leu Ser Glu Leu Asp
450 455 460
Asn Asn Pro Glu Arg Ala Gly Arg Asp Ser Arg Phe Thr Arg Phe Glu
465 470 475 480
Arg Leu Ala Gln
<210> 7
<211> 4067
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 7
cacctgcgac aacaccgaga accgcgagga agagcaccac gcgcatgagg ccagtgatca 60
ggatgccggt gatagaggaa ttggcaacag ctttgacgct gttggggccg gttcgtccgg 120
agtccagcat gcggtgtgcg ccagcgtaag tgatgtatcc accgacggtt ccacccacaa 180
gtgtggtgat gacaagccag tcgatggtgt caggaagtac tgcattcttc agcgccgagc 240
caactggagg ttgggagacg aaagccacgt agacggtgag caggatcatc acgacgccga 300
ggaccacgag gaatttgtcc agagcagcac caagtcgctt gaacaagaag atcgcaatcg 360
cgatggccgc ggtgatcacg ccaccaactt tgacgtccca gccaagcaga gcgttaagcc 420
caaggccacc accggcgatg ttgccgatgt tgaatactat gccgccaata cagaccagta 480
cggccagcac ccaaccaaaa cctgggataa cagtattgcc gagttcttgg gcgcgcattt 540
cagagatgcc gatgatgcgc cacacattca gctgcaccgc gatgtcaatg aggatcgaca 600
ccaggatcgc aaatgcgaaa gctgcgccga gctggttggt gaagacagcg gtttgggtga 660
ggaaccctgg gccgatggca gaagttgcca tgaggaagat ggcacccatc agagcgcttc 720
ggccagaggc ggttactgca gcttgcttga gggatggcgc attggctttt gcaatgtcgc 780
caggcccagg ggttccagtg ctggctgaga ctagctggct gtcatcgaca tttttgttgt 840
cggccattgt cgcgctcctt tcgtgggggg ctttcaaaat gattggggta gcttgcacac 900
caatcgcaga ttgttgaaca atcttggtga gttgagagtg agtttagacc caatgtgtcc 960
tgtggcacaa tcccccatag ggattaatag tcattgcata cggccagacc acccccgcaa 1020
gtgcctaggt gtgtccccag tcctttaaga tcagtcacat gacgcaggca atagcagcat 1080
cccttgattt agcggctcga atcaccgcca aaattgatca aggagtgctc actccaggta 1140
ctcgactacc cgaggttgct ttggcagaag aacttggcgt ttcacggaac acgctgcgtg 1200
aagcttttcg ggtactcatg caagacggac tggtggatca tattcccaac cgtggggttt 1260
tcgtgcacac gttcaccaag tcggatgtgg aagatatcta tgcttaccgc acatttatcg 1320
aggttgctgc gattaggtcg gcgcggaaaa atcctcagtt gctggagcag tctttggggg 1380
taatgcgaga ggcctacgaa aggggtgctg cagccaatgc cgtgggtgat tggcaaactg 1440
tcggttctgc caacagtgct tttcacttgg cgattgtgga cctagcagga gtggctaggt 1500
tgtcagcaga cgctcgaaaa gtgttggcgc tggctcgcat cggattcatg gccacctaca 1560
acgtggagac attccatagc atttacgtgg aaaagaacca tcaaatcttg aagtatttgg 1620
ctgccggtga attcgaagag gcggaacaat acctccagaa atacttcgaa gattcgcgcg 1680
atgatttgtc tgcgcaccta ccggaatttt gaaattgcga tagcatagat gacttgacac 1740
tgttgcagca ggcggttttc acctggcctc ttaggatttg gtcaaatggt tgcaacgacg 1800
ccagggaagg aggcgcaccc catggcattt ggatttttta gtagacgtaa gaaaaagaac 1860
aaagacaaaa acccgaatga aaattcagca gtgcccgcac actctgaaga ttcacctcag 1920
gaggtttttg agggtaatgg tcgtcaggta ggcgacccca ttgaacagca ggttgatcga 1980
gatgctaaag gtcgtctcac agcggcggat ttcttgccgg acgctgatct gccacagctg 2040
aatcgttcgc gtgcaaatat gctgcgccgt gaattggagt accgtttttc actccagaat 2100
gcccacatta atatcgatgg aaacacggcc atgattcagc gttcagatgg cggggcagca 2160
catgtctcgt tgcgcaccct cgcgatgaat gcagctggcc ttgataactt tgatcaactc 2220
cctgaactgg tggaaagctt cgttcacggc acgctggccg atgcaacatt aaacgatctt 2280
tctactgctg acctgtataa agcactgcgc cttcgcctgc tgccaacacc tggtgaaggc 2340
gacgatctag ttgagcatgg actcgaccgg gaaagccaga tccgcgacga ttcaatcctg 2400
cgcaccttca cctctgacat gtcgatcgcg ctggtgctcg ataccgagca tgccatccgc 2460
atccagccac tcaaagagct cgaggagttc gatgacctca gcgccctaga gcgggctgcg 2520
gaccgcaata cctggcaaga gctttacgac gcaaacgttg acgcttcctt cgtcgacgct 2580
gaatcagaca gcgaagggtc atcattttgg gctttcgaat ctaactcgta ctacctgggt 2640
agtgcaccac tgttcctcaa cgatttgttg gcaaagtggg cacctgacct ggaccaaagt 2700
gatggcgtca tctttgctgt ccctgatcgt gatctgttga ttgcgcgtcc tgtgaccacc 2760
ggcgaagatc tgatgaacgg aatcaccgcg atggtgagga tcgcgatgcg ctttggcctc 2820
gggaacccga cgtcgataag cccgcgcctg cacctgctgc gcgacaacca ggtgaccacc 2880
ttcaccgact tccgcgtcgt ctctcctgaa atggaagctg aatgggaaga cagcgcgttt 2940
gacgcgccac cggccggcgc gatcggcatt gaggtgcgcc cagatccgta tctgatggag 3000
cgcctccaac agggcggctt tggtgatttc ggagatttcg gcaagccccg cgatctagat 3060
atgtagcgaa aaaaggacct tcacttctta agaagtgaag gtccttattt atcgggttct 3120
agtgttagaa cttgttctcg cgctcggtgt gagccatagc aagaacgtcg aggcgcttgt 3180
ccagctcttc ctcggtgagc ttctcgccat caaccaagcc caaatcgatg acagtctggc 3240
ggatggtctt gccctcagcc aaagcagtct tagccacctt tgcagcagct tcgtagccga 3300
ttgcagagtt cagtggggta acgatggaag gtgaagactc agcgagctcc ttcatgtgtg 3360
cctcgtttgg ctcaatgcca tcaacgagac gggttgcgaa cacgcgggaa gtgttagcca 3420
gcaggcgagc ggactcaagc acgttgcgag ccatcactgg gatgaacacg ttgagctcga 3480
actggccctg ggtgccggag aacgcaacag ctgcgtcatt gccgataacc tgagcggaaa 3540
cctgggtagc ggtctcacag agaactgggt tgaccttgcc tggcatgatg gaggaacccg 3600
gctgcaggtc tgggagacgg atctcgccaa gaccggtcag tgggccggag cccatgaggc 3660
ggatatcgtt agcgatcttg tacaaggaga cagcgataac gcgcattgcg ccggagaact 3720
caacaagagc gtcgcgtgca gcctgagcct cgaagtggtt ctcagcttcc ttgagctcct 3780
tgacgtcggt caagttgatc agttcagcaa caaccttgcc gccgaaatca gcggaggtgt 3840
tgataccggt accagcagcg gtgccaccaa tagccagctc accaaggcga ggaagagtag 3900
cctcaacgcg ctcgatgccg agctggatct ggcgagcgta gccaccgaac tcctggccca 3960
gggttactgg aacagcgtcc atcaggtggg tgcggccgga cttaacaacc tcagaccact 4020
cgttagcctt cttcgccaaa gactcgtgca gaactttcag gcctggg 4067
<210> 8
<211> 1245
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的cg1144蛋白的编码序列
<400> 8
atggcatttg gattttttag tagacgtaag aaaaagaaca aagacaaaaa cccgaatgaa 60
aattcagcag tgcccgcaca ctctgaagat tcacctcagg aggtttttga gggtaatggt 120
cgtcaggtag gcgaccccat tgaacagcag gttgatcgag atgctaaagg tcgtctcaca 180
gcggcggatt tcttgtcgga cgctgatctg ccacagctga atcgttcgcg tgcaaatatg 240
ctgcgccgtg aattggagta ccgtttttca ctccagaatg cccacattaa tatcgatgga 300
aacacggcca tgattcagcg ttcagatggc ggggcagcac atgtctcgtt gcgcaccctc 360
gcgatgaatg cagctggcct tgataacttt gatcaactcc ctgaactggt ggaaagcttc 420
gttcacggca cgctggccga tgcaacatta aacgatcttt ctactgctga cctgtataaa 480
gcactgcgcc ttcgcctgct gccaacacct ggtgaaggcg acgatctagt tgagcatgga 540
ctcgaccggg aaagccagat ccgcgacgat tcaatcctgc gcaccttcac ctctgacatg 600
tcgatcgcgc tggtgctcga taccgagcat gccatccgca tccagccact caaagagctc 660
gaggagttcg atgacctcag cgccctagag cgggctgcgg accgcaatac ctggcaagag 720
ctttacgacg caaacgttga cgcttccttc gtcgacgctg aatcagacag cgaagggtca 780
tcattttggg ctttcgaatc taactcgtac tacctgggta gtgcaccact gttcctcaac 840
gatttgttgg caaagtgggc acctgacctg gaccaaagtg atggcgtcat ctttgctgtc 900
cctgatcgtg atctgttgat tgcgcgtcct gtgaccaccg gcgaagatct gatgaacgga 960
atcaccgcga tggtgaggat cgcgatgcgc tttggcctcg ggaacccgac gtcgataagc 1020
ccgcgcctgc acctgctgcg cgacaaccag gtgaccacct tcaccgactt ccgcgtcgtc 1080
tctcctgaaa tggaagctga atgggaagac agcgcgtttg acgcgccacc ggccggcgcg 1140
atcggcattg aggtgcgccc agatccgtat ctgatggagc gcctccaaca gggcggcttt 1200
ggtgatttcg gagatttcgg caagccccgc gatctagata tgtag 1245
<210> 9
<211> 414
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的cg1144蛋白
<400> 9
Met Ala Phe Gly Phe Phe Ser Arg Arg Lys Lys Lys Asn Lys Asp Lys
1 5 10 15
Asn Pro Asn Glu Asn Ser Ala Val Pro Ala His Ser Glu Asp Ser Pro
20 25 30
Gln Glu Val Phe Glu Gly Asn Gly Arg Gln Val Gly Asp Pro Ile Glu
35 40 45
Gln Gln Val Asp Arg Asp Ala Lys Gly Arg Leu Thr Ala Ala Asp Phe
50 55 60
Leu Ser Asp Ala Asp Leu Pro Gln Leu Asn Arg Ser Arg Ala Asn Met
65 70 75 80
Leu Arg Arg Glu Leu Glu Tyr Arg Phe Ser Leu Gln Asn Ala His Ile
85 90 95
Asn Ile Asp Gly Asn Thr Ala Met Ile Gln Arg Ser Asp Gly Gly Ala
100 105 110
Ala His Val Ser Leu Arg Thr Leu Ala Met Asn Ala Ala Gly Leu Asp
115 120 125
Asn Phe Asp Gln Leu Pro Glu Leu Val Glu Ser Phe Val His Gly Thr
130 135 140
Leu Ala Asp Ala Thr Leu Asn Asp Leu Ser Thr Ala Asp Leu Tyr Lys
145 150 155 160
Ala Leu Arg Leu Arg Leu Leu Pro Thr Pro Gly Glu Gly Asp Asp Leu
165 170 175
Val Glu His Gly Leu Asp Arg Glu Ser Gln Ile Arg Asp Asp Ser Ile
180 185 190
Leu Arg Thr Phe Thr Ser Asp Met Ser Ile Ala Leu Val Leu Asp Thr
195 200 205
Glu His Ala Ile Arg Ile Gln Pro Leu Lys Glu Leu Glu Glu Phe Asp
210 215 220
Asp Leu Ser Ala Leu Glu Arg Ala Ala Asp Arg Asn Thr Trp Gln Glu
225 230 235 240
Leu Tyr Asp Ala Asn Val Asp Ala Ser Phe Val Asp Ala Glu Ser Asp
245 250 255
Ser Glu Gly Ser Ser Phe Trp Ala Phe Glu Ser Asn Ser Tyr Tyr Leu
260 265 270
Gly Ser Ala Pro Leu Phe Leu Asn Asp Leu Leu Ala Lys Trp Ala Pro
275 280 285
Asp Leu Asp Gln Ser Asp Gly Val Ile Phe Ala Val Pro Asp Arg Asp
290 295 300
Leu Leu Ile Ala Arg Pro Val Thr Thr Gly Glu Asp Leu Met Asn Gly
305 310 315 320
Ile Thr Ala Met Val Arg Ile Ala Met Arg Phe Gly Leu Gly Asn Pro
325 330 335
Thr Ser Ile Ser Pro Arg Leu His Leu Leu Arg Asp Asn Gln Val Thr
340 345 350
Thr Phe Thr Asp Phe Arg Val Val Ser Pro Glu Met Glu Ala Glu Trp
355 360 365
Glu Asp Ser Ala Phe Asp Ala Pro Pro Ala Gly Ala Ile Gly Ile Glu
370 375 380
Val Arg Pro Asp Pro Tyr Leu Met Glu Arg Leu Gln Gln Gly Gly Phe
385 390 395 400
Gly Asp Phe Gly Asp Phe Gly Lys Pro Arg Asp Leu Asp Met
405 410
<210> 10
<211> 4210
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 10
cgcagtcatc cggtggagaa gacttctcgt ggtacctcga acacgtccca ggatcaatgg 60
cccggttggg ttgctggccg gggcacggac ccaagcaaga cctccatcaa agtgacctgg 120
ttgtggatga gcgagccatc ggagttggcg tcaggctctt tggctccctt gtgcagcagt 180
acagtagccg atctgaagct ttcttaaatt cctaatgggg gtagtgtgta gggctggctc 240
taaattgctg acacgtcacc caggctaatt cagcagtaat catttagact tggaatcgct 300
taccagtggt ttcaacaatg cattcaccca gctcacacgt gtggaggtgc cttaatggca 360
aagaggatcg taattatcgg cggtggacct gcaggctatg aagccgcact cgcaggcgct 420
aaatacggtg cagaagttac cgttattgaa gatgtcggag ttggcggatc cgcagtcacc 480
atggactgtg taccttcaaa gtccttcatc gctggtaccg gtatcaaaac cgacctccga 540
cgtgctgatg acatgggact taaccgtggg cttggaaaag cacacctaga aatcgatgca 600
ctgaacatcc gtgtgaagga ccttgcgaaa gcacagtccg aagatatctt gggccagctg 660
cagcgctcag atgtccgcat gattaacggt gtgggccgct ttgatgatta caacaccaag 720
caaaccaccc actacattaa agtcacccac agcgatggct ccgaagaaac cattgagtgc 780
gatctggtgc tggttgcaac tggtgcaacc ccccgcattc ttaaaggtgc agagccagac 840
ggcgagcgca tcttgacctg gcgtcaggtc tatgacattg aagaactccc cacccacctt 900
atcgtggttg gttccggtgt gaccggtgcg gaatttgtct ctgcgtttgc tgaactcggc 960
gtcaaagtca ccatggtggc atcccgtgac cgcattttgc ctcacgatga cgcagatgcc 1020
gcagacgtgc tggaaaccgt tctggctgag cgcggagtat ccctggaaaa gcatgcccgc 1080
gtggagtctg tcacccgcac cgaagacggt ggcgtgtgtg ttcgcactgc tgacggacgt 1140
gaaatctacg gttctcacgc gttgatgact gttggttcca ttccaaacac ggcagatctt 1200
ggcctggaga acatcggtgt tgagctggca ccatccggcc atatcaaggt tgaccgcgtc 1260
tcccgcacca acatccccgg tgtgtacgca gcaggtgact gtactgacct attcccactg 1320
gcgtccgttg cagcgatgca gggccgtatc gccatgtatc acgcactcgg tgaaggcgtg 1380
agccccatcc gtttgaagac tgttgccacc gcagtgttta cccgcccaga gatcgcagca 1440
gtaggtatca cccatgcaca ggttgattcc ggcgaagtgt ctgctcgcgt gattgtgctt 1500
cctttggcta ctaacccacg cgccaagatg cgttccctgc gccacggttt tgtgaagctg 1560
ttctgccgcc gtaactctgg cctgatcatc ggtggtgtcg tggtggcacc gaccgcgtct 1620
gagctgatcc taccgatcgc tgtggcagtg accaaccgtc tgacagttgc tgatctggct 1680
gataccttcg cggtgtaccc atcattgtca ggttcgatta ctgaagcagc acgtcagctg 1740
gttcaacatg atgatctagg ctaatttttc tgagtctcag attttaagaa aacccaggat 1800
tgctttgtgc actcctgggt tttcactttg ttaagcagtt ttggggaaaa gtgcaaagtt 1860
tgcaaagttt agaaatattt taagaggtaa gatgtctgca ggtggaagcg tttaaatgcg 1920
ttaaacttgg ccaaatgtgg caacctttgc aaggtgaaaa actggggcgg ggttagatcc 1980
tggggggttt atttcattca ctttggcttg aagtcgtgca ggtcagggga gtgttgcccg 2040
aaaacattga gaggaaaaca aaaaccgatg tttgattggg ggaatcgggg gttacgatac 2100
taggacgaag tgactgctat cacccttggc ggtctcttgt tgaaaggaat aattactcta 2160
gtgtcgactc acacatcttc aacgcttcca gcattcaaaa agatcttggt agcaaaccgc 2220
ggcgaaatcg cggtccgtgc tttccgtgca gcactcgaaa ccggtgcagc cacggtagct 2280
atttaccccc gtgaagatcg gggatcattc caccgctctt ttgcttctga agctgtccgc 2340
attggtactg aaggctcacc agtcaaggcg tacctggaca tcgatgaaat tatcggtgca 2400
gctaaaaaag ttaaagcaga tgctatttac ccgggatatg gcttcctgtc tgaaaatgcc 2460
cagcttgccc gcgagtgcgc ggaaaacggc attactttta ttggcccaac cccagaggtt 2520
cttgatctca ccggtgataa gtctcgtgcg gtaaccgccg cgaagaaggc tggtctgcca 2580
gttttggcgg aatccacccc gagcaaaaac atcgatgaca tcgttaaaag cgctgaaggc 2640
cagacttacc ccatctttgt aaaggcagtt gccggtggtg gcggacgcgg tatgcgcttt 2700
gtttcttcac ctgatgagct ccgcaaattg gcaacagaag catctcgtga agctgaagcg 2760
gcattcggcg acggttcggt atatgtcgaa cgtgctgtga ttaaccccca gcacattgaa 2820
gtgcagatcc ttggcgatcg cactggagaa gttgtacacc tttatgaacg tgactgctca 2880
ctgcagcgtc gtcaccaaaa agttgtcgaa attgcgccag cacagcattt ggatccagaa 2940
ctgcgtgatc gcatttgtgc ggatgcagta aagttctgcc gctccattgg ttaccagggc 3000
gcgggaaccg tggaattctt ggtcgatgaa aagggcaacc acgttttcat cgaaatgaac 3060
ccacgtatcc aggttgagca caccgtgact gaagaagtca ccgaggtgga cctggtgaag 3120
gcgcagatgc gcttggctgc tggtgcaacc ttgaaggaat tgggtctgac ccaagataag 3180
atcaagaccc acggtgcagc actgcagtgc cgcatcacca cggaagatcc aaacaacggc 3240
ttccgcccag ataccggaac tatcaccgcg taccgctcac caggcggagc tggcgttcgt 3300
cttgacggtg cagctcagct cggtggcgaa atcaccgcac actttgactc catgctggtg 3360
aaaatgacct gccgtggttc cgactttgaa actgctgttg ctcgtgcaca gcgcgcgttg 3420
gctgagttca ccgtgtctgg tgttgcaacc aacattggtt tcttgcgtgc gttgctgcgg 3480
gaagaggact tcacttccaa gcgcatcgcc accggattta tcggcgatca cccacacctc 3540
cttcaggctc cacctgcgga tgatgagcag ggacgcatcc tggattactt ggcagatgtc 3600
accgtgaaca agcctcatgg tgtgcgtcca aaggatgttg cagcaccaat cgataagctg 3660
cccaacatca aggatctgcc actgccacgc ggttcccgtg accgcctgaa gcagcttggc 3720
ccagccgcgt ttgctcgtga tctccgtgag caggacgcac tggcagttac tgataccacc 3780
ttccgcgatg cacaccagtc tttgcttgcg acccgagtcc gctcattcgc actgaagcct 3840
gcggcagagg ccgtcgcaaa gctgactcct gagcttttgt ccgtggaggc ctggggcggc 3900
gcgacctacg atgtggcgat gcgtttcctc tttgaggatc cgtgggacag gctcgacgag 3960
ctgcgcgagg cgatgccgaa tgtaaacatt cagatgctgc ttcgcggccg caacaccgtg 4020
ggatacaccc cgtacccaga ctccgtctgc cgcgcgtttg ttaaggaagc tgccacctcc 4080
ggcgtggaca tcttccgcat cttcgacgcg cttaacgacg tctcccagat gcgtccagca 4140
atcgacgcag tcctggagac caacaccgcg gtagccgagg tggctatggc ttattctggt 4200
gatctctctg 4210
<210> 11
<211> 93
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子1860
<400> 11
cttagctttg acctgcacaa atagttgcaa attgtcccac atacacataa agtagcttgc 60
gtatttaaaa ttatgaacct aaggggttta gca 93
<210> 12
<211> 3516
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子1860和pyc编码序列
<400> 12
cttagctttg acctgcacaa atagttgcaa attgtcccac atacacataa agtagcttgc 60
gtatttaaaa ttatgaacct aaggggttta gcagtgtcga ctcacacatc ttcaacgctt 120
ccagcattca aaaagatctt ggtagcaaac cgcggcgaaa tcgcggtccg tgctttccgt 180
gcagcactcg aaaccggtgc agccacggta gctatttacc cccgtgaaga tcggggatca 240
ttccaccgct cttttgcttc tgaagctgtc cgcattggta ctgaaggctc accagtcaag 300
gcgtacctgg acatcgatga aattatcggt gcagctaaaa aagttaaagc agatgctatt 360
tacccgggat atggcttcct gtctgaaaat gcccagcttg cccgcgagtg cgcggaaaac 420
ggcattactt ttattggccc aaccccagag gttcttgatc tcaccggtga taagtctcgt 480
gcggtaaccg ccgcgaagaa ggctggtctg ccagttttgg cggaatccac cccgagcaaa 540
aacatcgatg acatcgttaa aagcgctgaa ggccagactt accccatctt tgtaaaggca 600
gttgccggtg gtggcggacg cggtatgcgc tttgtttctt cacctgatga gctccgcaaa 660
ttggcaacag aagcatctcg tgaagctgaa gcggcattcg gcgacggttc ggtatatgtc 720
gaacgtgctg tgattaaccc ccagcacatt gaagtgcaga tccttggcga tcgcactgga 780
gaagttgtac acctttatga acgtgactgc tcactgcagc gtcgtcacca aaaagttgtc 840
gaaattgcgc cagcacagca tttggatcca gaactgcgtg atcgcatttg tgcggatgca 900
gtaaagttct gccgctccat tggttaccag ggcgcgggaa ccgtggaatt cttggtcgat 960
gaaaagggca accacgtttt catcgaaatg aacccacgta tccaggttga gcacaccgtg 1020
actgaagaag tcaccgaggt ggacctggtg aaggcgcaga tgcgcttggc tgctggtgca 1080
accttgaagg aattgggtct gacccaagat aagatcaaga cccacggtgc agcactgcag 1140
tgccgcatca ccacggaaga tccaaacaac ggcttccgcc cagataccgg aactatcacc 1200
gcgtaccgct caccaggcgg agctggcgtt cgtcttgacg gtgcagctca gctcggtggc 1260
gaaatcaccg cacactttga ctccatgctg gtgaaaatga cctgccgtgg ttccgacttt 1320
gaaactgctg ttgctcgtgc acagcgcgcg ttggctgagt tcaccgtgtc tggtgttgca 1380
accaacattg gtttcttgcg tgcgttgctg cgggaagagg acttcacttc caagcgcatc 1440
gccaccggat ttatcggcga tcacccacac ctccttcagg ctccacctgc ggatgatgag 1500
cagggacgca tcctggatta cttggcagat gtcaccgtga acaagcctca tggtgtgcgt 1560
ccaaaggatg ttgcagcacc aatcgataag ctgcccaaca tcaaggatct gccactgcca 1620
cgcggttccc gtgaccgcct gaagcagctt ggcccagccg cgtttgctcg tgatctccgt 1680
gagcaggacg cactggcagt tactgatacc accttccgcg atgcacacca gtctttgctt 1740
gcgacccgag tccgctcatt cgcactgaag cctgcggcag aggccgtcgc aaagctgact 1800
cctgagcttt tgtccgtgga ggcctggggc ggcgcgacct acgatgtggc gatgcgtttc 1860
ctctttgagg atccgtggga caggctcgac gagctgcgcg aggcgatgcc gaatgtaaac 1920
attcagatgc tgcttcgcgg ccgcaacacc gtgggataca ccccgtaccc agactccgtc 1980
tgccgcgcgt ttgttaagga agctgccacc tccggcgtgg acatcttccg catcttcgac 2040
gcgcttaacg acgtctccca gatgcgtcca gcaatcgacg cagtcctgga gaccaacacc 2100
gcggtagccg aggtggctat ggcttattct ggtgatctct ctgatccaaa tgaaaagctc 2160
tacaccctgg attactacct aaagatggca gaggagatcg tcaagtctgg cgctcacatt 2220
ctggccatta aggatatggc tggtctgctt cgcccagctg cggtaaccaa gctggtcacc 2280
gcactgcgcc gtgaattcga tctgccagtg cacgtgcaca cccacgacac tgcgggtggc 2340
cagttggcta cctactttgc tgcagctcaa gctggtgcag atgctgttga cggtgcttcc 2400
gcaccactgt ctggcaccac ctcccagcca tccctgtctg ccattgttgc tgcattcgcg 2460
cacacccgtc gcgataccgg tttgagcctc gaggctgttt ctgacctcga gccgtactgg 2520
gaagctgtgc gcggactgta cctgccattt gagtctggaa ccccaggccc aaccggtcgc 2580
gtctaccgcc acgaaatccc aggcggacag ttgtccaacc tgcgtgcaca ggccaccgca 2640
ctgggccttg ctgatcgctt cgagctcatc gaagacaact acgcagccgt taatgagatg 2700
ctgggacgcc caaccaaggt caccccatcc tccaaggttg ttggcgacct cgcactccac 2760
ctggttggtg cgggtgtaga tccagcagac tttgctgcag acccacaaaa gtacgacatc 2820
ccagactctg tcatcgcgtt cctgcgcggc gagcttggta accctccagg tggctggcca 2880
gaaccactgc gcacccgcgc actggaaggc cgctccgaag gcaaggcacc tctgacggaa 2940
gttcctgagg aagagcaggc gcacctcgac gctgatgatt ccaaggaacg tcgcaacagc 3000
ctcaaccgcc tgctgttccc gaagccaacc gaagagttcc tcgagcaccg tcgccgcttc 3060
ggcaacacct ctgcgctgga tgatcgtgaa ttcttctacg gactggtcga gggccgcgag 3120
actttgatcc gcctgccaga tgtgcgcacc ccactgcttg ttcgcctgga tgcgatctct 3180
gagccagacg ataagggtat gcgcaatgtt gtggccaacg tcaacggcca gatccgccca 3240
atgcgtgtgc gtgaccgctc cgttgagtct gtcaccgcaa ccgcagaaaa ggcagattcc 3300
tccaacaagg gccatgttgc tgcaccattc gctggtgttg tcactgtgac tgttgctgaa 3360
ggtgatgagg tcaaggctgg agatgcagtc gcaatcatcg aggctatgaa gatggaagca 3420
acaatcactg cttctgttga cggcaagatt gaacgcgttg tggttcctgc tgcaacgaag 3480
gtggaaggtg gcgacttgat cgtcgtcgtt tcctaa 3516
<210> 13
<211> 4101
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 13
catcgtctaa aaggcctggg cgaccacgta aagacgggca cgacgagaag atcatcgacg 60
caactttacg gctcatcgac agcaatcgtc ccgtcacggt caatgcagtt gtcaaagaaa 120
gcggagtggc acgtgcagcg gtttatcgac gctggcccag gctagtggat ctagtagcgg 180
aagctttaga tgccgggcga gctccagttg aaatagatac cccaggggac atcaaagaga 240
ccttgattga tgggctgttt acaaatcagg cgaaaaccac tggagtctcc tatcctcgtc 300
agcgatttcg caaacggctc gagttggtga tgtcagatca agaattacag ctcgcctaat 360
ggaattcaca tgtgaagaga cgtcgagaag caaatattcg cgcgctgcaa gtcgcgcaag 420
aaaaaggcca aatccgggcg gatctagaca tcgaggcgtg cctcgatgca atccttgggg 480
tgttttatta ccaatcggtc gcgcgtggag taaatttcac cgaccaaggt acaacgcaac 540
gatgcagaga agccttggag gtgatctggc atggaatgga accttaaatt caggttctga 600
cgaggtgcga agcaagttgt cgcgcgccgc acctcagtat ccggatcaac ttaatttcga 660
agtgctgggt tttctcgcgc atacccaatg cgtaccgatg tgctcatgag cgaaaaacag 720
gccacgataa gtttcttaaa acttatcgtg gcctgcttct atatttgtgc gccctgacgg 780
gctcgaaccg ccgacctgct gggtgtaaac cagctgctct tccagctgag ctaaaggcgc 840
gcacgtgctt ttctagaacc accttggtgg cctcgaaagc aacgagtgaa atactaacac 900
acaatctcca cagacctaaa atcgctgctc aggccgtgga aattagcgat tgttaaggct 960
tcttgtttcc acgctggacg aggcaagaac cttgccaatt accgagacgt tccgccttgg 1020
tctgcacgag acctgccagt tgtgctgatt cagagataac tccaggagcc agggctcctt 1080
ctttaccaat gccaggagtc aacacccaga tacgaccatt ctcagcgagg gagcggatgg 1140
aatccacaag tccgtcgacg agatcgccgt catcctcgcg ccaccagagc agcacgacat 1200
cgcacagctc gtcggtttct tcatcgagta gttcctcacc gattgcatct tcgatggact 1260
cgctgatcag cgtgtcggaa tcttcatccc atccaatttc ttgaacgata tgacccgatt 1320
gaatgccgag tagttgagca taatcctggg caccttgctt gactgcgccc ggagcgtcgg 1380
ccactttaat aatcctcctc gtgtgggccc cgatgtgttt ttcgattaca tggattcaac 1440
atgaaaccgc ggggctattg atatatccga attgcacatt accgtccaac cggtactttg 1500
aaccaccttt ccctggaatt ttttcctttt cctccccctt tacgctcaag aatcaatgaa 1560
ttcaatcact ggccagcgat taacttttcg agttttcagt cttggatttc cacaattctc 1620
ttcaaaataa tggtggctag atttttcatc aaaccctcac caaaaggaca tcagacctgt 1680
agttttatgc gattcgcgtc aaacgtgaga gaaacatcac atctcacggg aaactacccg 1740
ataattcttt gcaaaacttt gcaaagggta atgaacatgc agctagtttc cgtagaaatg 1800
ttctttaaaa aatccacaac aattgccagg aagcacaccg attgatggat acctgaaatc 1860
ccagtgagcg caccactccc cttacgtcac agtctgtaaa acaaatcttc ggtgttgcgt 1920
atccttgtta ataacttatg cgttgaccca ttcgtgcact tcggtgtgcc acaattaggt 1980
acgaccaaga atgggaccgg gaaaccggga cgtataaacg aaataaaaca ttccaacagg 2040
aggtgtggaa atggccgatc aagcaaaact tggtggtaag ccctcggatg actctaactt 2100
cgcgatgatc cgcgatggcg tggcatctta tttgaacgac tcagatccgg aggagaccaa 2160
cgagtggatg gattcactcg acggattact ccaggagtct tctccagaac gtgctcgtta 2220
cctcatgctt cgtttgcttg agcgtgcatc tgcaaagcgc gtatctcttc ccccaatgac 2280
gtcaaccgac tacgtcaaca ccattccaac ctctatggaa cctgaattcc caggcgatga 2340
ggaaatggag aagcgttacc gtcgttggat tcgctggaac gcagccatca tggttcaccg 2400
cgctcagcga ccaggcatcg gcgtcggcgg acacatttcc acttacgcag gcgcagcccc 2460
tctgtacgaa gttggcttca accacttctt ccgcggcaag gatcacccag gcggcggcga 2520
ccagatcttc ttccagggcc acgcatcacc aggtatgtac gcacgtgcat tcatggaggg 2580
tcgcctttct gaagacgatc tcgatggctt ccgtcaggaa gtttcccgtg agcagggtgg 2640
cattccgtcc taccctcacc cacacggtat gaaggacttc tgggagttcc caactgtgtc 2700
catgggtctt ggcccaatgg atgccattta ccaggcacgt ttcaaccgct acctcgaaaa 2760
ccgtggcatc aaggacacct ctgaccagca cgtctgggcc ttccttggcg acggcgaaat 2820
ggacgagcca gaatcacgtg gtctcatcca gcaggctgca ctgaacaacc tggacaacct 2880
gaccttcgtg gttaactgca acctgcagcg tctcgacgga cctgtccgcg gtaacaccaa 2940
gatcatccag gaactcgagt ccttcttccg tggcgcaggc tggtctgtga tcaaggttgt 3000
ttggggtcgc gagtgggatg aacttctgga gaaggaccag gatggtgcac ttgttgagat 3060
catgaacaac acctccgatg gtgactacca gaccttcaag gctaacgacg gcgcatatgt 3120
tcgtgagcac ttcttcggac gtgacccacg caccgcaaag ctcgttgaga acatgaccga 3180
cgaagaaatc tggaagctgc cacgtggcgg ccacgattac cgcaaggttt acgcagccta 3240
caagcgagct cttgagacca aggatcgccc aaccgtcatc cttgctcaca ccattaaggg 3300
ctacggactc ggccacaact tcgaaggccg taacgcaacc caccagatga agaagctgac 3360
gcttgatgat ctgaagttgt tccgcgacaa gcagggcatc ccaatcaccg atgagcagct 3420
ggagaaggat ccttaccttc ctccttacta ccacccaggt gaagacgctc ctgaaatcaa 3480
gtacatgaag gaacgtcgcg cagcgctcgg tggctacctg ccagagcgtc gtgagaacta 3540
cgatccaatt caggttccac cactggataa gcttcgctct gtccgtaagg gctccggcaa 3600
gcagcagatc gctaccacta tggcgactgt tcgtaccttc aaggaactga tgcgcgataa 3660
gggcttggct gatcgccttg tcccaatcat tcctgatgag gcacgtacct tcggtcttga 3720
ctcttggttc ccaaccttga agatctacaa cccgcacggt cagaactacg tgcctgttga 3780
ccacgacctg atgctctcct accgtgaggc acctgaagga cagatcctgc acgaaggcat 3840
caacgaggct ggttccgtgg catcgttcat cgctgcgggt acctcctacg ccacccacgg 3900
caaggccatg attccgctgt acatcttcta ctcgatgttc ggattccagc gcaccggtga 3960
ctccatctgg gcagcagccg atcagatggc acgtggcttc ctcttgggcg ctaccgcagg 4020
tcgcaccacc ctgaccggtg aaggcctcca gcacatggat ggacactccc ctgtcttggc 4080
ttccaccaac gagggtgtcg a 4101
<210> 14
<211> 2051
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 改变的aceE编码序列
<400> 14
ttggccgatc aagcaaaact tggtggtaag ccctcggatg actctaactt cgcgatgatc 60
cgcgatggcg tggcatctta tttgaacgac tcagatccgg aggagaccaa cgagtggatg 120
gattcactcg acggattact ccaggagtct tctccagaac gtgctcgtta cctcatgctt 180
cgtttgcttg agcgtgcatc tgcaaagcgc gtatctcttc ccccaatgac gtcaaccgac 240
tacgtcaaca ccattccaac ctctatggaa cctgaattcc caggcgatga ggaaatggag 300
aagcgttacc gtcgttggat tcgctggaac gcagccatca tggttcaccg cgctcagcga 360
ccaggcatcg gcgtcggcgg acacatttcc acttacgcag gcgcagcccc tctgtacgaa 420
gttggcttca accacttctt ccgcggcaag gatcacccag gcggcggcga ccagatcttc 480
ttccagggcc acgcatcacc aggtatgtac gcacgtgcat tcatggaggg tcgcctttct 540
gaagacgatc tcgatggctt ccgtcaggaa gtttcccgtg agcagggtgg cattccgtcc 600
taccctcacc cacacggtat gaaggacttc tgggagttcc caactgtgtc catgggtctt 660
ggcccaatgg atgccattta ccaggcacgt ttcaaccgct acctcgaaaa ccgtggcatc 720
aaggacacct ctgaccagca cgtctgggcc ttccttggcg acggcgaaat ggacgagcca 780
gaatcacgtg gtctcatcca gcaggctgca ctgaacaacc tggacaacct gaccttcgtg 840
gttaactgca acctgcagcg tctcgacgga cctgtccgcg gtaacaccaa gatcatccag 900
gaactcgagt ccttcttccg tggcgcaggc tggtctgtga tcaaggttgt ttggggtcgc 960
gagtgggatg aacttctgga gaaggaccag gatggtgcac ttgttgagat catgaacaac 1020
acctccgatg gtgactacca gaccttcaag gctaacgacg gcgcatatgt tcgtgagcac 1080
ttcttcggac gtgacccacg caccgcaaag ctcgttgaga acatgaccga cgaagaaatc 1140
tggaagctgc cacgtggcgg ccacgattac cgcaaggttt acgcagccta caagcgagct 1200
cttgagacca aggatcgccc aaccgtcatc cttgctcaca ccattaaggg ctacggactc 1260
ggccacaact tcgaaggccg taacgcaacc caccagatga agaagctgac gcttgatgat 1320
ctgaagttgt tccgcgacaa gcagggcatc ccaatcaccg atgagcagct ggagaaggat 1380
ccttaccttc ctccttacta ccacccaggt gaagacgctc ctgaaatcaa gtacatgaag 1440
gaacgtcgcg cagcgctcgg tggctacctg ccagagcgtc gtgagaacta cgatccaatt 1500
caggttccac cactggataa gcttcgctct gtccgtaagg gctccggcaa gcagcagatc 1560
gctaccacta tggcgactgt tcgtaccttc aaggaactga tgcgcgataa gggcttggct 1620
gatcgccttg tcccaatcat tcctgatgag gcacgtacct tcggtcttga ctcttggttc 1680
ccaaccttga agatctacaa cccgcacggt cagaactacg tgcctgttga ccacgacctg 1740
atgctctcct accgtgaggc acctgaagga cagatcctgc acgaaggcat caacgaggct 1800
ggttccgtgg catcgttcat cgctgcgggt acctcctacg ccacccacgg caaggccatg 1860
attccgctgt acatcttcta ctcgatgttc ggattccagc gcaccggtga ctccatctgg 1920
gcagcagccg atcagatggc acgtggcttc ctcttgggcg ctaccgcagg tcgcaccacc 1980
ctgaccggtg aaggcctcca gcacatggat ggacactccc ctgtcttggc ttccaccaac 2040
gagggtgtcg a 2051
<210> 15
<211> 4099
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 15
gaatcttatc ccatggctaa gaaagtagac acctcgaacg ctacccccgc tctagccctt 60
cttacggaga ggcaaattcc ttttgagctg gatgttcatg atgtagatcc aaaatcatca 120
aagggcttcg cattggatgc ctctgaagta atgggtgtgg agccggaagt ggtgtttaaa 180
acgctcatgg cagatattga tggtgaacac gtggtcgcga ttgttccagc cagcagcacg 240
ttgaatctca agcagttggc taaggctgga aaaggtaagc atgcaaacat gatggatcgc 300
agccgtgcac aggtagtcac ggggtatgtc cctggtggaa tctcaccgat agggcagaag 360
aataagcacc gcgtgttttt ggatgagtct gcaattctcc aggagcgaat ctacgtcagc 420
gcaggacgcc gaggctggtc actgattatc gccccggatg atgttcttct ggctaccgat 480
ggggtttacg cggatattgc tgatcattca taaaaggtaa aacccacccc cgcaggggtg 540
cgtcgtaaag caagctcctt ttcttagtac ggcgcgatag tggactcgtg cgcgtcgatg 600
ttgatgcggc cgtgcacgga cgggaatgcg cgctgcgcgc agttctcgcg ggtgcacact 660
cggcagccgg acccgatggg ggtggcggtg gataggtcct ggaggttgaa gccgcgggag 720
tacacggtgc ggtcggcgtg gcgcgcttcg cagccgagcc cgatggcgaa cattttgtcc 780
acttcgccga accgggcttc gtggtgtcgc acggtgcgtg agatccacag gtagttgcgt 840
ccgtcgggca tttgcgcgaa ttggcggagc acttggccgg ggttggtgaa ggtttcaaac 900
acgttccaca gggggcaggt gccgccgtag tgggtgaagt ggaagccggt ggcggattgg 960
cgtttggaca tgttgccggc gcggtcgacg cgcacgaagg taaacgggat gccgcgcagg 1020
ttgggacgct gcagggtgga caggcggtgg gcggtcgtct cgtagcccac gccgaaaagc 1080
tgacctaggt attcgatgtc gtagccggat ttttcggcct cggagtggaa gattttgtag 1140
ggcagcatta cggcggcggc gaagtaggag gcgacgccgc ggatggctag ggtgcgggct 1200
tcgggggtgg accagatgcc gtcgtcgacg atgccttcga tgaggtcgtt ggcttctagg 1260
tagccgagtt cggtggccat gcggaaggcg cgttgtccgg ggttgaggcg tgcgtggatt 1320
gtcagtaggc gcgtctcggg gtcaaagtgg tgcagcgtgc cggattcctc tttggaggag 1380
gtgatggtga catcgtgatc catttgcagg cgcctggcga tcgaatcttc catggcgcgg 1440
gaatcgtacg gctgccagcc cagttgcgcg gcgatggctt cggcgcggcg gtcgagcgca 1500
tcgaagtagt tttggcgggc gtaaatgaaa tcgcgcacct cttcgtgcgg catgctcacg 1560
gcctccgcga tggggcgacg ttcctcaggc gtgttggtgc gattgtccac tgcgatggag 1620
aacttgtcgc gtacgtttcg gtagcgctgg tgcatttcca ccatggcgcg cgccagctgc 1680
gggtggttgt acaccatctc cgatagctct tggagctcca cgttcgcggg gttgatctcc 1740
cggtccaaca tgacatcctg aacttcagct aaaaggcggg aatcatcgtc gcgggagaaa 1800
aacgttgcgt ctacgccgaa cgcctcggtg atgcgcaata acaccggtac ggtgagcggg 1860
cgtacgtcgt gctcaatctg atttacataa ctggcagata agccaagggt tgctgccaac 1920
gacgcctggc tcaggtctct ttcgcggcgc agttggcgca gcctggaccc cacatatgtc 1980
tttcccatgg cgcaactata gcgtgatcac catcacctta accactcttg gcattgaggt 2040
gttgcaaagt tgttgatttt cgcttttcga cgcagcccgc cgccatcggg tgcccggcgt 2100
ggtcaggcca catgcgcccc gggaactttt tgggcaccta cggtgcaaca gttgcgaaaa 2160
ttgtgtcacc tgcgcaaagc cttgcttcta ttcgggaaat tcgggtgtct aaactttttg 2220
gttgatacca aacggggtta gaaactgttc agatcggtat cctgtgagga agctcacctt 2280
ggttttagaa tgttgaaaaa gcctcacgtt tccgcaggta gagcacactc aattaaatga 2340
gcgtcaaacg acaataaagt aaggctaccc taataactgg ggttttatgc ctctaaacag 2400
tcagttgggg gcggtagggg agcgtcccat gactggttaa tgcctcgatc tgggacgtac 2460
agtaacagcg acactggagg tgccatgact gttagaaatc ccgaccgtga ggcaatccgt 2520
cacggaaaaa ttacgacgga ggcgctgcgt gagcgtcccg catacccgac ctgggcaatg 2580
aagctgacca tggccatcac tggcctaatc ttcggtggct tcgttcttgt tcacatgatc 2640
ggaaacctga aaatcttcat gccggactac gcagccgatt ctgcgcatcc gggtgaagca 2700
caagtagatg tctacggcga gttcctgcgt gagatcggat ccccgatcct cccacacggc 2760
tcagtcctct ggatcctacg tattatcctg ctggtcgcat tggttctgca catctactgt 2820
gcattcgcat tgaccggccg ttctcaccag tcccgcggaa agttccgccg taccaacctc 2880
gttggcggct tcaactcctt cgcgacccgc tccatgctgg tgaccggaat cgttctcctt 2940
gcgttcatta tcttccacat cctcgacctg accatgggtg ttgctccagc agccccaact 3000
tcattcgagc acggcgaagt atacgcaaat atggtggctt cctttagccg ctggcctgta 3060
gcaatttggt acatcattgc caacctggtc ctgttcgtcc acctgtctca cggcatctgg 3120
cttgcagtct ctgacctggg aatcaccgga cgtcgctgga gggcaatcct cctcgcagtt 3180
gcgtacatcg ttcctgcact ggtcctgatc ggcaacatca ccattccgtt cgccatcgct 3240
gttggctgga ttgcgtaaag gttaggaaga atttatgagc actcactctg aaaccacccg 3300
cccagagttc atccacccag tctcagtcct cccagaggtc tcagctggta cggtccttga 3360
cgctgctgag ccagcaggcg ttcccaccaa agatatgtgg gaataccaaa aagaccacat 3420
gaacctggtc tccccactga accgacgcaa gttccgcgtc ctcgtcgttg gcaccggcct 3480
gtccggtggc gctgcagcag cagccctcgg cgaactcgga tacgacgtca aggcgttcac 3540
ctaccacgac gcacctcgcc gtgcgcactc cattgctgca cagggtggcg ttaactccgc 3600
ccgcggcaag aaggtagaca acgacggcgc ataccgccac gtcaaggaca ccgtcaaggg 3660
cggcgactac cgtggccgcg agtccgactg ctggcgtctc gccgtcgagt ccgtccgcgt 3720
catcgaccac atgaacgcca tcggtgcacc attcgcccgc gaatacggtg gcgccttggc 3780
aacccgttcc ttcggtggtg tgcaggtctc ccgtacctac tacacccgtg gacaaaccgg 3840
acagcagctg cagctctcca ccgcatccgc actacagcgc cagatccacc tcggctccgt 3900
agagatcttc acccataacg aaatggttga cgtaattgtc accgaacgta atggtgaaaa 3960
gcgctgcgaa ggcctgatca tgcgcaacct gatcaccggc gagctcaccg cacacaccgg 4020
ccatgccgtt atcctggcaa ccggtggcta cggcaacgtg taccacatgt ccaccctggc 4080
gaagaactcc aacgcctcg 4099
<210> 16
<211> 1338
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 密码子优化的lysA编码序列
<400> 16
atggctaccg tagaaaactt caatgaacta cccgcgcatg tctggcctcg taatgcagtt 60
cgccaggaag atggcgttgt tacggtagcg ggagtgccgc ttcctgatct tgcagaagag 120
tacggtacac ctctctttgt tgttgacgaa gatgatttcc gttctcgatg tcgtgatatg 180
gctaccgcat ttgggggacc aggcaatgtg cattacgctt ccaaagcgtt ccttactaaa 240
actattgcac gctgggtaga cgaagaggga ctagcgttgg atatcgcgtc tattaatgaa 300
ctgggaattg cgttggcggc tggattcccg gcttcacgaa tcacggcaca tggtaataat 360
aaaggtgtcg agtttctgcg cgctcttgta cagaatggtg tgggtcatgt agtgcttgat 420
tctgctcaag aactagaact tcttgactat gttgcagcgg gcgaaggcaa gattcaagat 480
gttttgattc gtgttaagcc aggcattgaa gctcatactc atgaattcat cgcgacgagt 540
catgaagatc agaaatttgg attcagtctg gcgtcaggtt ccgctttcga agcggctaaa 600
gctgctaata acgctgagaa tcttaatctg gtagggctac attgtcatgt cggatcccag 660
gtttttgatg ctgagggttt caaacttgcg gctgaacgcg ttttgggttt gtatagtcaa 720
attcatagtg agcttggtgt agctctacca gaacttgatc ttggtggggg atatggtatt 780
gcgtatactg cggctgaaga accgcttaat gtagctgaag ttgcgtccga tttgcttacg 840
gctgtcggta aaatggcagc agaattgggt atcgatgcgc caactgttct tgtggaaccg 900
ggacgggcta ttgcgggtcc atctacagtt actatttacg aggtcggaac gactaaagat 960
gtgcatgtcg atgatgataa gactcgtcgc tatattgctg tcgacggtgg aatgtccgat 1020
aacattcgtc ctgcgctcta tggatctgag tatgatgcac gcgtcgtatc tcgttttgct 1080
gagggtgacc cggtttcaac tcgcatcgtg ggatctcatt gtgaatctgg agacattctg 1140
atcaatgacg aaatttatcc gtctgatatt acgagtggtg attttcttgc tttggctgcg 1200
actggcgcat attgttacgc aatgtctagt cgttataacg cttttactcg tccagctgtt 1260
gtatccgtgc gggcgggatc atctcgcctt atgcttagac gtgaaacctt ggacgacatt 1320
ctttcccttg aggcgtag 1338
<210> 17
<211> 1516
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子cg0007_39-密码子优化的lysA编码序列-sod终止子
<400> 17
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg gctaccgtag aaaacttcaa 120
tgaactaccc gcgcatgtct ggcctcgtaa tgcagttcgc caggaagatg gcgttgttac 180
ggtagcggga gtgccgcttc ctgatcttgc agaagagtac ggtacacctc tctttgttgt 240
tgacgaagat gatttccgtt ctcgatgtcg tgatatggct accgcatttg ggggaccagg 300
caatgtgcat tacgcttcca aagcgttcct tactaaaact attgcacgct gggtagacga 360
agagggacta gcgttggata tcgcgtctat taatgaactg ggaattgcgt tggcggctgg 420
attcccggct tcacgaatca cggcacatgg taataataaa ggtgtcgagt ttctgcgcgc 480
tcttgtacag aatggtgtgg gtcatgtagt gcttgattct gctcaagaac tagaacttct 540
tgactatgtt gcagcgggcg aaggcaagat tcaagatgtt ttgattcgtg ttaagccagg 600
cattgaagct catactcatg aattcatcgc gacgagtcat gaagatcaga aatttggatt 660
cagtctggcg tcaggttccg ctttcgaagc ggctaaagct gctaataacg ctgagaatct 720
taatctggta gggctacatt gtcatgtcgg atcccaggtt tttgatgctg agggtttcaa 780
acttgcggct gaacgcgttt tgggtttgta tagtcaaatt catagtgagc ttggtgtagc 840
tctaccagaa cttgatcttg gtgggggata tggtattgcg tatactgcgg ctgaagaacc 900
gcttaatgta gctgaagttg cgtccgattt gcttacggct gtcggtaaaa tggcagcaga 960
attgggtatc gatgcgccaa ctgttcttgt ggaaccggga cgggctattg cgggtccatc 1020
tacagttact atttacgagg tcggaacgac taaagatgtg catgtcgatg atgataagac 1080
tcgtcgctat attgctgtcg acggtggaat gtccgataac attcgtcctg cgctctatgg 1140
atctgagtat gatgcacgcg tcgtatctcg ttttgctgag ggtgacccgg tttcaactcg 1200
catcgtggga tctcattgtg aatctggaga cattctgatc aatgacgaaa tttatccgtc 1260
tgatattacg agtggtgatt ttcttgcttt ggctgcgact ggcgcatatt gttacgcaat 1320
gtctagtcgt tataacgctt ttactcgtcc agctgttgta tccgtgcggg cgggatcatc 1380
tcgccttatg cttagacgtg aaaccttgga cgacattctt tcccttgagg cgtagtaggc 1440
atttttagta cgtgcaataa ccactctggt ttttccaggg tggttttttg atgccctttt 1500
tggagtcttc aactgg 1516
<210> 18
<211> 445
<212> PRT
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 18
Met Ala Thr Val Glu Asn Phe Asn Glu Leu Pro Ala His Val Trp Pro
1 5 10 15
Arg Asn Ala Val Arg Gln Glu Asp Gly Val Val Thr Val Ala Gly Val
20 25 30
Pro Leu Pro Asp Leu Ala Glu Glu Tyr Gly Thr Pro Leu Phe Val Val
35 40 45
Asp Glu Asp Asp Phe Arg Ser Arg Cys Arg Asp Met Ala Thr Ala Phe
50 55 60
Gly Gly Pro Gly Asn Val His Tyr Ala Ser Lys Ala Phe Leu Thr Lys
65 70 75 80
Thr Ile Ala Arg Trp Val Asp Glu Glu Gly Leu Ala Leu Asp Ile Ala
85 90 95
Ser Ile Asn Glu Leu Gly Ile Ala Leu Ala Ala Gly Phe Pro Ala Ser
100 105 110
Arg Ile Thr Ala His Gly Asn Asn Lys Gly Val Glu Phe Leu Arg Ala
115 120 125
Leu Val Gln Asn Gly Val Gly His Val Val Leu Asp Ser Ala Gln Glu
130 135 140
Leu Glu Leu Leu Asp Tyr Val Ala Ala Gly Glu Gly Lys Ile Gln Asp
145 150 155 160
Val Leu Ile Arg Val Lys Pro Gly Ile Glu Ala His Thr His Glu Phe
165 170 175
Ile Ala Thr Ser His Glu Asp Gln Lys Phe Gly Phe Ser Leu Ala Ser
180 185 190
Gly Ser Ala Phe Glu Ala Ala Lys Ala Ala Asn Asn Ala Glu Asn Leu
195 200 205
Asn Leu Val Gly Leu His Cys His Val Gly Ser Gln Val Phe Asp Ala
210 215 220
Glu Gly Phe Lys Leu Ala Ala Glu Arg Val Leu Gly Leu Tyr Ser Gln
225 230 235 240
Ile His Ser Glu Leu Gly Val Ala Leu Pro Glu Leu Asp Leu Gly Gly
245 250 255
Gly Tyr Gly Ile Ala Tyr Thr Ala Ala Glu Glu Pro Leu Asn Val Ala
260 265 270
Glu Val Ala Ser Asp Leu Leu Thr Ala Val Gly Lys Met Ala Ala Glu
275 280 285
Leu Gly Ile Asp Ala Pro Thr Val Leu Val Glu Pro Gly Arg Ala Ile
290 295 300
Ala Gly Pro Ser Thr Val Thr Ile Tyr Glu Val Gly Thr Thr Lys Asp
305 310 315 320
Val His Val Asp Asp Asp Lys Thr Arg Arg Tyr Ile Ala Val Asp Gly
325 330 335
Gly Met Ser Asp Asn Ile Arg Pro Ala Leu Tyr Gly Ser Glu Tyr Asp
340 345 350
Ala Arg Val Val Ser Arg Phe Ala Glu Gly Asp Pro Val Ser Thr Arg
355 360 365
Ile Val Gly Ser His Cys Glu Ser Gly Asp Ile Leu Ile Asn Asp Glu
370 375 380
Ile Tyr Pro Ser Asp Ile Thr Ser Gly Asp Phe Leu Ala Leu Ala Ala
385 390 395 400
Thr Gly Ala Tyr Cys Tyr Ala Met Ser Ser Arg Tyr Asn Ala Phe Thr
405 410 415
Arg Pro Ala Val Val Ser Val Arg Ala Gly Ser Ser Arg Leu Met Leu
420 425 430
Arg Arg Glu Thr Leu Asp Asp Ile Leu Ser Leu Glu Ala
435 440 445
<210> 19
<211> 4508
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 19
cttctccgac tacatgcgtc ctgcagttcg tcttgcagct ctcatggaga ccgacgctta 60
ctacgtctgg acccacgact ccatcggtct gggcgaagat ggcccaaccc accagcctgt 120
tgaaaccttg gctgcactgc gcgccatccc aggtctgtcc gtcctgcgtc ctgcagatgc 180
gaacgagacc gcccaggctt gggctgcagc acttgagtac aaggaaggcc ctaagggtct 240
tgcactaacc cgccagaaca ttcctgttct ggaaggcacc aaggagaagg ctgctgaagg 300
cgttcgccgc ggtggctacg tcctggttga gggttccaag gaaaccccag atgtgatcct 360
catgggctcc ggctccgagg ttcagcttgc agttaacgct gcgaaggctc tggaagctga 420
gggcgttgca gctcgcgttg tttccgttcc ttgcatggat tggttccagg agcaggacgc 480
agagtacatc gagtccgttc tgcctgcagc tgtgaccgct cgtgtgtctg ttgaagctgg 540
catcgcaatg ccttggtacc gcttcttggg cacccagggc cgtgctgtct cccttgagca 600
cttcggtgct tctgcggatt accagaccct gtttgagaag ttcggcatca ccaccgatgc 660
agtcgtggca gcggccaagg actccattaa cggttaattg ccctgctgtt tttagcttca 720
acccggggca atatgattct ccggaatttt attgccccgg gttgttgttg ttaatcggta 780
caaagggtct taagcacatc ccttacttgc ctgctctcct tgagcacagt tcaagaacaa 840
ttcttttaag gaaaatttag tttcatgtct cacattgatg atcttgcaca gctcggcact 900
tccacttggc tcgacgacct ctcccgcgag cgcattactt ccggcaatct cagccaggtt 960
attgaggaaa agtctgtagt cggtgtcacc accaacccag ctattttcgc agcagcaatg 1020
tccaagggcg attcctacga cgctcagatc gcagagctca aggccgctgg cgcatctgtt 1080
gaccaggctg tttacgccat gagcatcgac gatgttcgca atgcttgtga tctgttcacc 1140
ggcatcttcg agtcctccaa cggctacgac ggccgcgtgt ccatcgaggt tgacccacgt 1200
atctctgctg accgcgacgc aaccctggct caggccaagg agctgtgggc aaaggttgat 1260
cgtccaaacg tcatgatcaa gatccctgca accccaggtt ctttgccagc aatcaccgac 1320
gctttggctg agggcatcag cgttaacgtc accctgatct tctccgttgc tcgctaccgc 1380
gaagtcatcg ctgcgttcat cgagggcatc aagcaggctg ctgcaaacgg ccacgacgta 1440
tccaagattc actctgtggc ttccttcttc gtctcccgcg tcgacgttga gatcgacaag 1500
cgcctcgagg caatcggatc cgatgaggct ttggctctgc gcggcaaggc aggcgttgcc 1560
aacgctcagc gcgcttacgc tgtgtacatg gagcttttcg acgccgccga gctgcctgaa 1620
ggtgccaaca ctcagcgccc tctgtgggca tccaccggcg tgaagaaccc tgcgtacgct 1680
gcaactcttt acgtttccga gctggctggt ccaaacaccg tcaacaccat gccagaaggc 1740
accatcgacg cggttctgga gcagggcaac ctgcacggcg acaccctgtc caactccgcg 1800
gcagaagctg acgctgtgtt ctcccagctt gaggctctgg gcgttgactt ggcagatgtc 1860
ttccaggtcc tggagaccga gggtgtggac aagttcgttg cttcttggag cgaactgctt 1920
gagtccatgg aagctcgcct gaagtagaat cggcacgctg catcagtaac ggcgacataa 1980
aatcgaatca gttcgatctt gtgtggccgt tacacatctt tcattaaaga aaggatcgtg 2040
acgctaccat cgtgagcaca aacacgaccc ccaccagctg gacaaaccca ctgcgcgacc 2100
cgcaggataa acgactcccc cgcatcgctg gcccttccgg catggtgatc ttcggtgtca 2160
ctggcgactt ggctcgaaag aagctgcttc ccgccattta tgatctagca aaccgcggat 2220
tgctgccccc aggattctcg ttggtaggtt acggccgccg cgaatggtcc aaagaagact 2280
ttgaaaaata cgtacgcgat gccgcaagtg ctggtgctcg tacggaattt cgtgaaaatg 2340
tttgggagcg cctcgccgag ggtatggaat ttgttcgcgg caactttgat gatgatgcag 2400
ctttcgacaa cctcgctgca acactcaagc gcatcgacaa aacccgcggc accgccggca 2460
actgggctta ctacctgtcc attccaccag attccttcac agcggtctgc caccagctgg 2520
agcgttccgg catggctgaa tccaccgaag aagcatggcg ccgcgtgatc atcgagaagc 2580
ctttcggcca caacctcgaa tccgcacacg agctcaacca gctggtcaac gcagtcttcc 2640
cagaatcttc tgtgttccgc atcgaccact atttgggcaa ggaaacagtt caaaacatcc 2700
tggctctgcg ttttgctaac cagctgtttg agccactgtg gaactccaac tacgttgacc 2760
acgtccagat caccatggct gaagatatcg gcttgggtgg acgtgctggt tactacgacg 2820
gcatcggtgc agcccgcgac gtcatccaga accacctgat ccagctcttg gctctggttg 2880
ccatggaaga accaatttct ttcgtgccag cgcagctgca ggcagaaaag atcaaggtgc 2940
tctctgcgac aaagccatgc tacccattgg ataaaacctc cgctcgtggt cagtacgctg 3000
ccggttggca gggctctgag ttagtcaagg gacttcgcga agaagatggc ttcaaccctg 3060
agtccaccac tgagactttt gcggcttgta ccttagagat cacgtctcgt cgctgggctg 3120
gtgtgccgtt ctacctgcgc accggtaagc gtcttggtcg ccgtgttact gagattgccg 3180
tggtgtttaa agacgcacca caccagcctt tcgacggcga catgactgta tcccttggcc 3240
aaaacgccat cgtgattcgc gtgcagcctg atgaaggtgt gctcatccgc ttcggttcca 3300
aggttccagg ttctgccatg gaagtccgtg acgtcaacat ggacttctcc tactcagaat 3360
ccttcactga agaatcacct gaagcatacg agcgcctcat tttggatgcg ctgttggatg 3420
aatccagcct cttccccacc aacgaggaag tggaactgag ctggaagatt ctggatccaa 3480
ttcttgaagc atgggacgcc gatggagaac cagaggatta cccagcaggt acgtggggtc 3540
caaagagcgc tgatgaaatg ctttcccgca acggtcacac ctggcgcagg ccataattta 3600
ggggcaaaaa atgatctttg aacttccgga taccaccacc cagcaaattt ccaagaccct 3660
aactcgactg cgtgaatcgg gcacccaggt caccaccggc cgagtgctca ccctcatcgt 3720
ggtcactgac tccgaaagcg atgtcgctgc agttaccgag tccaccaatg aagcctcgcg 3780
cgagcaccca tctcgcgtga tcattttggt ggttggcgat aaaactgcag aaaacaaagt 3840
tgacgcagaa gtccgtatcg gtggcgacgc tggtgcttcc gagatgatca tcatgcatct 3900
caacggacct gtcgctgaca agctccagca tgtcgtcaca ccactgttgc ttcctgacac 3960
ccccatcgtt gcttggtggc caggtgaatc accaaagaat ccttcccagg atccaattgg 4020
acgcatcgca caacgacgca tcactgatgc tttgtacgac cgtgatgacg cactagaaga 4080
tcgtgttgag aactatcacc cccctgcgta cgctgcaact ctttacgttt ccgagctggc 4140
tggtccaaac accgtcaaca ccatgccaga aggcaccatc gacgcggttc tggagcaggg 4200
caacctgcac ggcgacaccc tgtccaactc cgcggcagaa gctgacgctg tgttctccca 4260
gcttgaggct ctgggcgttg acttggcaga tgtcttccag gtcctggaga ccgagggtgt 4320
ggacaagttc gttgcttctt ggagcgaact gcttgagtcc atggaagctc gcctgaagta 4380
gaatcggcac gctgcatcag taacggcgac ataaaatcga atcagttcga tcttgtgtgg 4440
ccgttacaca tctttcatta aagaaaggat cgtgacgcta ccattgccgt ttctcgcgtt 4500
gtgtgtgg 4508
<210> 20
<211> 97
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子cg0007_39
<400> 20
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaa 97
<210> 21
<211> 1643
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子cg0007_39插入和zwf编码区
<400> 21
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaagtg agcacaaaca cgacccccac 120
cagctggaca aacccactgc gcgacccgca ggataaacga ctcccccgca tcgctggccc 180
ttccggcatg gtgatcttcg gtgtcactgg cgacttggct cgaaagaagc tgcttcccgc 240
catttatgat ctagcaaacc gcggattgct gcccccagga ttctcgttgg taggttacgg 300
ccgccgcgaa tggtccaaag aagactttga aaaatacgta cgcgatgccg caagtgctgg 360
tgctcgtacg gaatttcgtg aaaatgtttg ggagcgcctc gccgagggta tggaatttgt 420
tcgcggcaac tttgatgatg atgcagcttt cgacaacctc gctgcaacac tcaagcgcat 480
cgacaaaacc cgcggcaccg ccggcaactg ggcttactac ctgtccattc caccagattc 540
cttcacagcg gtctgccacc agctggagcg ttccggcatg gctgaatcca ccgaagaagc 600
atggcgccgc gtgatcatcg agaagccttt cggccacaac ctcgaatccg cacacgagct 660
caaccagctg gtcaacgcag tcttcccaga atcttctgtg ttccgcatcg accactattt 720
gggcaaggaa acagttcaaa acatcctggc tctgcgtttt gctaaccagc tgtttgagcc 780
actgtggaac tccaactacg ttgaccacgt ccagatcacc atggctgaag atatcggctt 840
gggtggacgt gctggttact acgacggcat cggtgcagcc cgcgacgtca tccagaacca 900
cctgatccag ctcttggctc tggttgccat ggaagaacca atttctttcg tgccagcgca 960
gctgcaggca gaaaagatca aggtgctctc tgcgacaaag ccatgctacc cattggataa 1020
aacctccgct cgtggtcagt acgctgccgg ttggcagggc tctgagttag tcaagggact 1080
tcgcgaagaa gatggcttca accctgagtc caccactgag acttttgcgg cttgtacctt 1140
agagatcacg tctcgtcgct gggctggtgt gccgttctac ctgcgcaccg gtaagcgtct 1200
tggtcgccgt gttactgaga ttgccgtggt gtttaaagac gcaccacacc agcctttcga 1260
cggcgacatg actgtatccc ttggccaaaa cgccatcgtg attcgcgtgc agcctgatga 1320
aggtgtgctc atccgcttcg gttccaaggt tccaggttct gccatggaag tccgtgacgt 1380
caacatggac ttctcctact cagaatcctt cactgaagaa tcacctgaag catacgagcg 1440
cctcattttg gatgcgctgt tggatgaatc cagcctcttc cccaccaacg aggaagtgga 1500
actgagctgg aagattctgg atccaattct tgaagcatgg gacgccgatg gagaaccaga 1560
ggattaccca gcaggtacgt ggggtccaaa gagcgctgat gaaatgcttt cccgcaacgg 1620
tcacacctgg cgcaggccat aat 1643
<210> 22
<211> 5082
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 22
ccctgcgtac gctgcaactc tttacgtttc cgagctggct ggtccaaaca ccgtcaacac 60
catgccagaa ggcaccatcg acgcggttct ggagcagggc aacctgcacg gcgacaccct 120
gtccaactcc gcggcagaag ctgacgctgt gttctcccag cttgaggctc tgggcgttga 180
cttggcagat gtcttccagg tcctggagac cgagggtgtg gacaagttcg ttgcttcttg 240
gagcgaactg cttgagtcca tggaagctcg cctgaagtag aatcggcacg ctgcatcagt 300
aacggcgaca taaaatcgaa tcagttcgat cttgtgtggc cgttacacat ctttcattaa 360
agaaaggatc gtgacgctac cattgccgtt tctcgcgttg tgtgtggtac tacgtgggga 420
cctaagcgtg tattatggaa acgtctgtat cggataagta gcgaggagtg ttcgttaaaa 480
gtgagcacaa acacgacccc caccagctgg acaaacccac tgcgcgaccc gcaggataaa 540
cgactccccc gcatcgctgg cccttccggc atggtgatct tcggtgtcac tggcgacttg 600
gctcgaaaga agctgcttcc cgccatttat gatctagcaa accgcggatt gctgccccca 660
ggattctcgt tggtaggtta cggccgccgc gaatggtcca aagaagactt tgaaaaatac 720
gtacgcgatg ccgcaagtgc tggtgctcgt acggaatttc gtgaaaatgt ttgggagcgc 780
ctcgccgagg gtatggaatt tgttcgcggc aactttgatg atgatgcagc tttcgacaac 840
ctcgctgcaa cactcaagcg catcgacaaa acccgcggca ccgccggcaa ctgggcttac 900
tacctgtcca ttccaccaga ttccttcaca gcggtctgcc accagctgga gcgttccggc 960
atggctgaat ccaccgaaga agcatggcgc cgcgtgatca tcgagaagcc tttcggccac 1020
aacctcgaat ccgcacacga gctcaaccag ctggtcaacg cagtcttccc agaatcttct 1080
gtgttccgca tcgaccacta tttgggcaag gaaacagttc aaaacatcct ggctctgcgt 1140
tttgctaacc agctgtttga gccactgtgg aactccaact acgttgacca cgtccagatc 1200
accatggctg aagatatcgg cttgggtgga cgtgctggtt actacgacgg catcggtgca 1260
gcccgcgacg tcatccagaa ccacctgatc cagctcttgg ctctggttgc catggaagaa 1320
ccaatttctt tcgtgccagc gcagctgcag gcagaaaaga tcaaggtgct ctctgcgaca 1380
aagccatgct acccattgga taaaacctcc gctcgtggtc agtacgctgc cggttggcag 1440
ggctctgagt tagtcaaggg acttcgcgaa gaagatggct tcaaccctga gtccaccact 1500
gagacttttg cggcttgtac cttagagatc acgtctcgtc gctgggctgg tgtgccgttc 1560
tacctgcgca ccggtaagcg tcttggtcgc cgtgttactg agattgccgt ggtgtttaaa 1620
gacgcaccac accagccttt cgacggcgac atgactgtat cccttggcca aaacgccatc 1680
gtgattcgcg tgcagcctga tgaaggtgtg ctcatccgct tcggttccaa ggttccaggt 1740
tctgccatgg aagtccgtga cgtcaacatg gacttctcct actcagaatc cttcactgaa 1800
gaatcacctg aagcatacga gcgcctcatt ttggatgcgc tgttggatga atccagcctc 1860
ttccccacca acgaggaagt ggaactgagc tggaagattc tggatccaat tcttgaagca 1920
tgggacgccg atggagaacc agaggattac ccagcaggta cgtggggtcc aaagagcgct 1980
gatgaaatgc tttcccgcaa cggtcacacc tggcgcaggc cataatttag gggcaaaaaa 2040
tgatctttga acttccggat accaccaccc agcaaatttc caagacccta actcgactgc 2100
gtgaatcggg cacccaggtc accaccggcc gagtgctcac cctcatcgtg gtcactgact 2160
ccgaaagcga tgtcgctgca gttaccgagt ccaccaatga agcctcgcgc gagcacccat 2220
ctcgcgtgat cattttggtg gttggcgata aaactgcaga aaacaaagtt gacgcagaag 2280
tccgtatcgg tggcgacgct ggtgcttccg agatgatcat catgcatctc aacggacctg 2340
tcgctgacaa gctccagcat gtcgtcacac cactgttgct tcctgacacc cccatcgttg 2400
cttggtggcc aggtgaatca ccaaagaatc cttcccagga tccaattgga cgcatcgcac 2460
aacgacgcat cactgatgct ttgtacgacc gtgatgacgc actagaagat cgtgttgaga 2520
actatcaccc aggtgatacc gacatgacgt gggcgcgcct tacccagtgg cggggacttg 2580
ttgcctcctc attggatcac ccaccacaca gcgaaatcac ttccgtgagg ctgaccggcg 2640
caagcggcag tacctcggtg gatttggctg caggctggtt ggcgcgcagg ctgaacgtgc 2700
ctgtgatccg cgaggtgaca gatgctccca ccgtgtcaac cgatgagttt ggtactccac 2760
tgctggctat ccagcgcctg gagatcgttc gcaccaccgg ctcgatcatc atcaccatcc 2820
atgacgctca tacccttcag gtagagatgc cggaatccgg caatgcccca tcgctggtgg 2880
ctattggtcg tcgaagtgag tccgactgct tgtctgagga gcttcgccac atggatccag 2940
atttgggcta ccagcacgca ctatccggct tgtccagcgt caagctggaa accgtctaag 3000
gagaaataca acactatggt tgatgtagta cgcgcacgcg atactgaaga tttggttgca 3060
caggctgcct ccaaattcat tgaggttgtt gaagcagcaa ctgccaataa tagcaccgca 3120
caggtagtgc tcaccggtgg tggcgccggc atcaagttgc tggaaaagct cagcgttgat 3180
gcggctgacc ttgcctggga tcgcattcat gtgttcttcg gcgatgagcg caatgtccct 3240
gtcagtgatt ctgagtccaa tgagggccag gctcgtgagg cactgttgtc caaggtttct 3300
atccctgaag ccaacattca cggatatggt ctcggcgacg tagatcttgc agaggcagcc 3360
cgcgcttacg aagctgtgtt ggatgaattc gcaccaaacg gctttgatct tcacctgctc 3420
ggcatgggtg gcgaaggcca tatcaactcc ctgttccctc acaccgatgc agtcaaggaa 3480
tcctccgcaa aggtcatcgc ggtgtttgat tcccctaagc ctccttcaga gcgtgcaact 3540
ctaacccttc ctgcggttca ctccgcaaag cgggtgtggt tgctggtttc tggtgcggag 3600
aaggctgagg cagctgcggc gatcgtcaac ggtgagcctg ctgttgagtg gcctgctgct 3660
ggagctaccg gatctgagga aacggtattg ttcttggctg atgatgctgc aggaaatctc 3720
taagtagcgc cagctctaac aagaagctct aacaagaagc ttcaacaaga agctctaacg 3780
aaaagcacta acaaactaat ccgggtgcga accttcatct gaatcgatgg aggttcgcac 3840
ccggattttt ggctcagcgt tagattctca ccctaaaacc gcgcacttaa aaaggttagc 3900
tttctatacg tgcaccgcct ctttgattgg ttcatgcgag gcgatccgaa agtcttcgct 3960
gtacagcggc gaggtgtttc cgaccgctag ttgtgctgcg agttcgccat aagctggaga 4020
gagcttgaag gcgtgtccgg atcctccggt gagcacaatg acattgtcca ccgcgtcgat 4080
aattggagat ttgtctgcag tgtaggtgtc atagtggacg ctgaaacggt tgggctctgg 4140
gttgacccct gggaagagtt cgtgggtttt ccgaccaaat tcagagactg cgtcccggtc 4200
gaggcggaga tcttcatctt ctacatgagc gcttaatgga acgccccact catccaatcc 4260
ggcaattttg atgctgtacc catcgacgca tggtgctcca aatacgtgga agccatcacg 4320
gtcacggatg aagcatggca gattttccgg ctggaagtcc actgggttgt tgggcaggaa 4380
ccaagtgagc actaggcgtc gcacttcaag cagtggcgcg atggagggca cgagctcgct 4440
tgtccagctg ccggtggtga cgataacgcg gtccacgatt gtggtttctt cgcctgcttg 4500
gatgactacg tgatcgccgt tgtcttcgat gctggtgatt ttttggtgat cgcgtacttg 4560
ggcaccattt gccttggctg tttcgattgc actgaacact gctagttcgg ggcgcagggc 4620
tccaccttga agatcaacaa ttccagcttc atcatcgcgg aagtctagac ctgggtaacg 4680
cttgcgcatc tgcgcggcgg taagtcgttc atgtggcagc tcataacgtt ccactgattc 4740
caccaggcgt tggaagggtg cttcgtcttc cttgccggtg cttaagacac cgaagttgtg 4800
gaagagttcg cgtccggaaa tctcgctcag tgatgaccat agtgctcgtg cgcgtttgag 4860
caacggaacg taggtgctgc cttcgtggta ggccatgcga aacagtcggg actcccctgt 4920
gaatgcgccg tagccatggg agatgccgaa ttgttcaaag ccgatggcct ctacacctgg 4980
gatgttactt aagtgccaca gtgccatgga gccggttgat ccaaggccga ttaccgcaat 5040
tttcattgtt gtttcttctc tcaagccagt gcgggtgatt cc 5082
<210> 23
<211> 951
<212> DNA
<213> C. humireducens
<400> 23
atgaagatcg aattgcctga caccgatacc cgcaacattg cagcctccct gttggaagcc 60
cgcgaacaca actcccagac gactggtcgt gtactcaccc tgatcgtagt ggccgcattg 120
gatgatgacg tcgaccatat catctccgtc acccgtgacg cgagccatga gcacccagct 180
cgcgttctcg tgctccttac aggccgtgaa gacgcggact ctcgcctcga tgctgaagtt 240
cgcgttggag gtcacgccgg cgcgtcagaa atggtaatta tgcgcatcac cggagaaatg 300
gtgaaccact tggcggccgt cgtcacccca cttctgctgc ctgacactcc tgtagtggcg 360
tggtggccta ccactgcccc ggaaaaccca tcaggacacc ctctgggaca gctcgcccag 420
cgacgtatca caaacgcaga tcatccggat tccgaatctg tactccgtct tcgtcgagcg 480
acttacgctc cgggtgattc tgacatggtg tggtccaaga tcactgcctg gcgtggtatc 540
gtcgcttctt ctctcgatcg ccatccacat gaagacgtgg aggcggccgt gctcactgga 600
cctgctgatg aaccacaaat cgatatcgca gcgggttggc tggcagatcg cctgggtgtc 660
ccggtgcgcc gcgaggtctg tgagcccggc aacgatgccc gcctgttccc gattcgccgt 720
cttaagtttg agcgcgcttc aggtcctgtt cgtgtagagg tactcgacca tcgcactgtg 780
ttggtggccc ttccaggccg accagaatcg ctcgtggcca tgaacgaacg ctccgacgct 840
gattgcctca gcgaggaatt gcgccacctg gaccccgatc tggcatacgc caacgccttg 900
cgcggactca cccgcgtgaa gggcatcaat ggccacccag cgtcgtcctg a 951
<210> 24
<211> 1048
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子cg0007_39-C. humireducense opcA编码序列
<400> 24
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg aagatcgaat tgcctgacac 120
cgatacccgc aacattgcag cctccctgtt ggaagcccgc gaacacaact cccagacgac 180
tggtcgtgta ctcaccctga tcgtagtggc cgcattggat gatgacgtcg accatatcat 240
ctccgtcacc cgtgacgcga gccatgagca cccagctcgc gttctcgtgc tccttacagg 300
ccgtgaagac gcggactctc gcctcgatgc tgaagttcgc gttggaggtc acgccggcgc 360
gtcagaaatg gtaattatgc gcatcaccgg agaaatggtg aaccacttgg cggccgtcgt 420
caccccactt ctgctgcctg acactcctgt agtggcgtgg tggcctacca ctgccccgga 480
aaacccatca ggacaccctc tgggacagct cgcccagcga cgtatcacaa acgcagatca 540
tccggattcc gaatctgtac tccgtcttcg tcgagcgact tacgctccgg gtgattctga 600
catggtgtgg tccaagatca ctgcctggcg tggtatcgtc gcttcttctc tcgatcgcca 660
tccacatgaa gacgtggagg cggccgtgct cactggacct gctgatgaac cacaaatcga 720
tatcgcagcg ggttggctgg cagatcgcct gggtgtcccg gtgcgccgcg aggtctgtga 780
gcccggcaac gatgcccgcc tgttcccgat tcgccgtctt aagtttgagc gcgcttcagg 840
tcctgttcgt gtagaggtac tcgaccatcg cactgtgttg gtggcccttc caggccgacc 900
agaatcgctc gtggccatga acgaacgctc cgacgctgat tgcctcagcg aggaattgcg 960
ccacctggac cccgatctgg catacgccaa cgccttgcgc ggactcaccc gcgtgaaggg 1020
catcaatggc cacccagcgt cgtcctga 1048
<210> 25
<211> 316
<212> PRT
<213> C. humireducens
<400> 25
Met Lys Ile Glu Leu Pro Asp Thr Asp Thr Arg Asn Ile Ala Ala Ser
1 5 10 15
Leu Leu Glu Ala Arg Glu His Asn Ser Gln Thr Thr Gly Arg Val Leu
20 25 30
Thr Leu Ile Val Val Ala Ala Leu Asp Asp Asp Val Asp His Ile Ile
35 40 45
Ser Val Thr Arg Asp Ala Ser His Glu His Pro Ala Arg Val Leu Val
50 55 60
Leu Leu Thr Gly Arg Glu Asp Ala Asp Ser Arg Leu Asp Ala Glu Val
65 70 75 80
Arg Val Gly Gly His Ala Gly Ala Ser Glu Met Val Ile Met Arg Ile
85 90 95
Thr Gly Glu Met Val Asn His Leu Ala Ala Val Val Thr Pro Leu Leu
100 105 110
Leu Pro Asp Thr Pro Val Val Ala Trp Trp Pro Thr Thr Ala Pro Glu
115 120 125
Asn Pro Ser Gly His Pro Leu Gly Gln Leu Ala Gln Arg Arg Ile Thr
130 135 140
Asn Ala Asp His Pro Asp Ser Glu Ser Val Leu Arg Leu Arg Arg Ala
145 150 155 160
Thr Tyr Ala Pro Gly Asp Ser Asp Met Val Trp Ser Lys Ile Thr Ala
165 170 175
Trp Arg Gly Ile Val Ala Ser Ser Leu Asp Arg His Pro His Glu Asp
180 185 190
Val Glu Ala Ala Val Leu Thr Gly Pro Ala Asp Glu Pro Gln Ile Asp
195 200 205
Ile Ala Ala Gly Trp Leu Ala Asp Arg Leu Gly Val Pro Val Arg Arg
210 215 220
Glu Val Cys Glu Pro Gly Asn Asp Ala Arg Leu Phe Pro Ile Arg Arg
225 230 235 240
Leu Lys Phe Glu Arg Ala Ser Gly Pro Val Arg Val Glu Val Leu Asp
245 250 255
His Arg Thr Val Leu Val Ala Leu Pro Gly Arg Pro Glu Ser Leu Val
260 265 270
Ala Met Asn Glu Arg Ser Asp Ala Asp Cys Leu Ser Glu Glu Leu Arg
275 280 285
His Leu Asp Pro Asp Leu Ala Tyr Ala Asn Ala Leu Arg Gly Leu Thr
290 295 300
Arg Val Lys Gly Ile Asn Gly His Pro Ala Ser Ser
305 310 315
<210> 26
<211> 4100
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 26
acacaaagaa gcaccaaccg atccggcgct tgcgcgtttg ctccctgatt ttcaacacga 60
gggcgatgag gaatacgacg gcgataattc tttcctccgt tcacttcatg aaggcgatat 120
cactcgcgcg aagctggaga atctgcgtgt gatcaatgat gcgctgggcc ccgacggcaa 180
tgttgcggtt accgcctctg aggaggaagc acacgcatgg ttggccgcgc tcaatgatat 240
ccgcctgtac gttgcctccg gtgatgcacg cggcggagaa gccgccgagg aagaccgcga 300
aaacctcgtg cagtggctgg cctacaatca agagtccttg ctggaagcga tgatgaatta 360
atgcttatcg acgtcccggg cttcctttta ggccacgtca cgaaggggga tacgggttgc 420
tcagtggtca ttgcacctaa cggtgcattt gcgggcgtcg atgtccgtgg gggaggccca 480
ggcaccaggg aaaccgacct tctagaacca cacaattctg tgcagcaagc acatgccgtg 540
gtgttgtgtg gcggttcggc gttcgggttg gctgctgccg atggagtgat gacagcccta 600
gaaaaccgcg gtattggttt ccctgtccgt cccgaagggc ctatcgtgcc aatcgttcca 660
ggcgctgtga tttttgattt gttggtgggt gaccccaaaa ataggcccac cgcagctgat 720
ggggaacagg ctgtagaaaa cgctttcgct ggtacacaca acggttcggg cagtgtcggt 780
gcaggaaccg gcgctacggc aggtcggctg cgtggcggtt ttggccaaag ctcgcgccgg 840
gtcggaaagt acaccatcgc ggcaggtgtg gtggcgaatc ctgttgggga agtcgtggac 900
ctaaagactg gggctttgtt tggtaggccc gaagtgatgg gggtgggcgt cgataagcta 960
aaaagcgcgg cagagacgct gaacacgacc atcggcgtcg tggcaactga cgcgccggtg 1020
acaaaagccc aggcgaagcg cttggcgctg gtggcccatg acggattggc gagggccgtg 1080
cggccgtcgc attcgccgat ggacggtgac acattttttg ctatgtcatc gggtgatggt 1140
agtggcgtta ccccggttga gctgggggaa ttgtcggctc atgctgcaga ttgcgtacag 1200
gacgctatta tcgacgccat acttaccgcg agtccgggac tcgggctcaa aagcttcagg 1260
gaacttttac catgagttac aacagcccgt ataacaacac gaatttcagc accactggcg 1320
cgttccaacc tgctggtggg ccggtgaagc cgtggaataa gcctgatggc agcctgaatc 1380
agcagctgaa aaataaatcc cgtgtgcgca caggtcttac catcgccatc ggttatgttg 1440
tggtgatttg ggcggtgcat ttggcatcca tcgtgattgc gctgctcact ggcttcaacc 1500
tgaccaactt tggtattcat ccgctggata ccagtgcact gtggggtatt ttcgcctcac 1560
cgttgctgca tggaagcttc agtcacctca ttggaaatac cgttccaggc tttatattca 1620
gtttcctcat cggtatgagt ggcaagcgcg tgttctggga agtcacgatt atcgcaggtc 1680
tcatcggcgg tcttggtaca tggattttcg gtggaatcgg caccaaccac atcggtgcgt 1740
ccggcctgat ttatggctgg cttggctacc tgatcgtgcg tggaattttc aataaggata 1800
ttaaacagtt cctgcttggc atagttttgg cgttcattta ctccggtttg ttctggggtc 1860
tgctacctac tcaaatcggt gtgtcctggc agggccacct tttcggtgca cttggtggaa 1920
tcggtgcggg tgcttttatc gcctcggatg acccggcagc gttgaaagcg aagaagcagc 1980
agaagaaact agaaaagcaa caacgccaaa gaggcttgta gttttcacct agcgactaca 2040
cttaattgac atgacaagtg agaattccga atcccaggac atttggctaa ccgatgagca 2100
acaagatgtg tggctcgatg tgtggacaat gcgaatcggc ctgcctgctc gcttggatgc 2160
tcaactgaaa gaagctgcgg gtgtcagcca ctttgagtac ttcaccatgg cgcagatttc 2220
tatggccccg gaacatcggg tgcgcatgag tgagcttgct gagctgtccg atatgacgct 2280
atcgcatcta tctagagtgg ttactcgcct agaaaaggct ggctgggtga agcgtgttcc 2340
cgatcccgat gatggtcgcg ccaccgttgc tgtgctcacg gactctgggt gggagaaagt 2400
taaagcaaca gcccctggtc atgtgaagga agtgcgtcgt ttggtgtttg acgatctcac 2460
tccagaagaa ctcaaggtaa tgggcactgc aatgaagaag attgtgaacc gactcgatat 2520
gtccaacagg ctgccgcggg tgtagttaga agttttaaga aatgtagaca acggcgttgt 2580
caccacctgt gcaggtgaca tactcgccgt tgtcttcgca gttcatggtg tagtttccgc 2640
cagtgacagg gctggtggct gtcaccctgc cggatagttc attggtgtct aaatagtggt 2700
ttacaaaagc atctcggacg gcttgggcaa aacctgctga ggtggatgct gacccggtgt 2760
acacattgtt caaattgccg gcatcggtct gggtggcagc agcatcattg gctggtgatg 2820
ctcccgcagg aagtgatggt tgcgcagggc ggtttcgggg agtttccgtt gcactgcttg 2880
ggggctgatc ctctgtcgcg ctcggtgact caggggattc cgaaatagtt tgctccacag 2940
attcagaggt gcttggactt gccacagggc tcgaggaatt atttggtgca acgaggctgt 3000
tgaacaacaa gacaaacgca atcacgatga tggccaccaa gatcaagatc aacccaatga 3060
gactgccttt gggtgaacgt ttcgcgggcg ccacagctgt aggagagcca atagtctccg 3120
aagttgctcg caccgcagcg aatcgtctgg tttcttgatc aggtgaatca gccattgcca 3180
tattgtgaca catcttggac ggataaaaag ggaagcaacg cgaggtagct tatgatggca 3240
accgtgactg atttcagtgg atctatgatt gaacgccccg tgccaggtgc tgatgcgccg 3300
attggaattt ttgattctgg agttggcgga ttaaccgtag ctcgcacaat catcgatcaa 3360
ttgccacatg aatcagttat ttatatcggt gatactgcca atggccctta tggtccgttg 3420
cctatcgcta aggtccgtga gcacgccatc cgcattgccg atgagttggt ggaacgcgga 3480
tgcaagatga ttgtcattgc ctgtaacact gcgtccgctg cgtttctccg agatgcccgt 3540
gaacgataca gtgtgccagt cgtggaagtt attcttcccg cagtaaggcg tgcggtggca 3600
tccacccgca atggcaaagt gggcgtgatc ggcacagtgg gaaccattaa ctccggtgcg 3660
taccaggatc ttttctctgc aagcccctcc attgaggtca acgcagtggc atgcccacgg 3720
tttgtggatt tcgtggaacg cggaattacc agcggcaggc agatcctcaa cattgcgcag 3780
gattatttag agcctctgca agcagaaggg gtggacactc ttgtgcttgg atgcacccac 3840
tatccactgc tctccggtgt cattcagttg gcaatggggg accacgtaag tttggtctct 3900
agcgcggaag aaactgcgaa agacgtgctg agaatcttga gccagcaaga tcttttagcc 3960
gatccggaca tgcatcctga gccaagttat agctttgaat caacaggcga tccggaaatc 4020
tttgcacaat taagccgccg attccttgga ccaattgttt ccctagtgag acaaaacgag 4080
ggataacccc aggtgtgtgt 4100
<210> 27
<211> 595
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子cg0007_39-cg2766
<400> 27
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg acaagtgaga attccgaatc 120
ccaggacatt tggctaaccg atgagcaaca agatgtgtgg ctcgatgtgt ggacaatgcg 180
aatcggcctg cctgctcgct tggatgctca actgaaagaa gctgcgggtg tcagccactt 240
tgagtacttc accatggcgc agatttctat ggccccggaa catcgggtgc gcatgagtga 300
gcttgctgag ctgtccgata tgacgctatc gcatctatct agagtggtta ctcgcctaga 360
aaaggctggc tgggtgaagc gtgttcccga tcccgatgat ggtcgcgcca ccgttgctgt 420
gctcacggac tctgggtggg agaaagttaa agcaacagcc cctggtcatg tgaaggaagt 480
gcgtcgtttg gtgtttgacg atctcactcc agaagaactc aaggtaatgg gcactgcaat 540
gaagaagatt gtgaaccgac tcgatatgtc caacaggctg ccgcgggtgt agtta 595
<210> 28
<211> 4283
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 28
cgcatgatgt gctgatgcag atcatccacc gcgtcatcgc ttttagccat ctctagggca 60
agatctggtt caggatcaat gaggatctga cgaatatgat ctgtcatgtc agctgcgagg 120
cgggccatct ccttgaaata gccacgcatg tcctccggga tcaccggatc ggggtaacgg 180
cgacgcacgg aattagctac gtgcattgcc agagcaccca tacgggtaat ttcctcgacg 240
atgtagatgg aggagacaac ctggcgaaga tcacgggcta caggagtttc aagcgccaaa 300
agttcgacgg cttgagactc gcatccagta cgaagctctt caagcttttc tgcattgcct 360
aaagcgcttt ctgccgatgc caaagaagtg cgcaggaggg catcagtcgc cttatccatg 420
cactccttag ttagatcaca caaaatgatt agatcgtgtg cgaattcatc caattgctgt 480
ctatatgcag tacgcatggc aacactataa ggcgataatg gtatttctgc agacctaaaa 540
caccccttta cggttgaatc acctaataat gggggatagc caactattgg cgggggtaag 600
tattaaatta actaccctcc ggagttttcc atttctgcgc ctttaggaac agtggcatcc 660
tcaggatcgt ccaaccagcc atctggaagt gccacctttg caggagcgcc ctgtcgacct 720
cgtgcacctt ccgcgtcctc tgccttggcg gtggagtcag cccatggtgc aaggagatcc 780
tcaagctcca cataggtgga aaccttggcc agattggagc ggaattcgcc gccaacaggg 840
aaaccgcgca ggtaccaacc catgtgctta cgcagatcgc gcagcccctt ggtttcgcca 900
tcatgctgca tgaggagttc tgcgtggcgc aggatgattt gggtaacttc gccgaaggta 960
ggctcctctg ggatttcttc tccacgaaca gcagcagaca gctcagcaaa gagccaaggc 1020
ctgcccaggc aaccacgccc aaccacgacg ccatcgcagc cagtttgctc catcatgcgc 1080
gttgcatcgg atgccgcgaa aatatcgcca ttgcccaaaa ctgggatgcc ggtatctgcc 1140
aaatgctcct tcaggcgcgc gatctcgttc caatcagcct caccggaata gcgctgcgcc 1200
gcagtgcggg cgtgaagcgc tacggacttg gcgcctgcgt cgacagcaat gcgtccagca 1260
tccaagtgag tatggtgctc atcatcaata ccaacgcgga acttcaccgt caccggaatg 1320
tccgtgcctt ccgtagcctt cacagccgcg gaaacgatgt tttcaaacaa acggcgcttg 1380
taaggaatcg cagaaccgcc accacggcgc gtgacctttg gaaccgggca gccaaagttc 1440
atatcaatat gatccgccaa gttttcatca acgatcatct tcgccgcttc gtaggtgtac 1500
ttcgggtcaa ccgtgtacag ctgcaagctt cggggatttt catccggcgc gaaggtggtc 1560
atgtgcatgg ttttctcatt gcgctcaaca agagcacgcg cagtcaccat ttcacagacg 1620
tacagccccg agattgttcc cgtgcgttgc atttcctgtt cgcggcacag cgtgcggaaa 1680
gcaacgttgg ttacaccagc catgggggct agaaccacag gggaggcaag gtcaaagggg 1740
ccgattttta gagtcaccta actattgtcc cccgtgaatc aggttgggca aaatatttga 1800
agcaaattgt gagcagggcg caactaggaa agtggtgtgc tttcactttt tgggggctgg 1860
ggttgggtta agcttcgcgg gctctagggt tggtctgagc tttattcctg agctttattc 1920
ctggtctttg ggaggcttgc aaacaggggg catgcaaatt tgggggtaat gctgggcctt 1980
gaaatcccac tatcacagat agtattcggg catttcctgt cacgatggtt tatccttggg 2040
acacaacatc aaagtggggt acatcatatg cttccggttg aagtgaccta tctgaaaaga 2100
ttggtcgaac cttgaagcaa tggtgtgaac tgcgttaacg aattttgtcg gacgttaaaa 2160
tggtcgcatt ctgcttgctg aagtggcaca cctatgtgtt ctgcttgggt atagcagtgc 2220
gggaaaaatt tgaaaaagtc cgattacctg aggaggtatt caatgtctga tcgcattgct 2280
tcagaaaagc tgcgctccaa gctcatgtcc gccgacgagg cggcacagtt tgttaaccac 2340
ggtgacaagg ttggtttctc tggcttcacc ggcgctggct acccaaaggc actgcctacg 2400
gcaatcgcta accgggctaa agaagcacac ggtgcaggca acgactacgc aatcgacctg 2460
ttcactggcg catcgaccgc ccctgactgc gatggcgtac ttgcagaagc tgacgctatc 2520
cgctggcgca tgccatacgc atctgatcca atcatgcgta acaagatcaa ctccggctcc 2580
atgggatact ccgatatcca cctgtcccac tccggccagc aggttgaaga gggcttcttc 2640
ggccagctca acgtagctgt cattgaagtc acccgcatca ctgaagacgg acacctcatc 2700
ccttcttcct ccgtgggtaa caacgttgag tggctcaacg ctgcagagaa ggtcatcctc 2760
gaggttaact cttggcagtc cgagaacctc gagggcatgc acgacatctg gtctgtacca 2820
gcactgccaa accgcatcgc tgtgccaatc aacaaggccg gcgaccgcat cggtaagacc 2880
tacatcgaat tcgacaccga caaggttgtc gcaattgttg aaaccaacac cccagaccgc 2940
aacgcaccat ttaagcctgt cgatgacatc tctaagaaga tcgccggtaa cttcctcgac 3000
ttcctcgaag gtgaagttgc agctggccgt ctttcctacg atggttacgt catgcagtcc 3060
ggcgtgggca acgtgcctaa tgctgtgatg gcaggcctgc tggagtccaa gtttgagaac 3120
atccaggctt acaccgaggt tatccaggac ggcatggttg accttatcga cgccggcaag 3180
atgaccgttg cttctgcaac ctccttctcc ttgtctcctg agtacgcaga gaagatgaac 3240
aacgacgcaa agcgttaccg tgagtccatc atcctgcgcc cacagcagat ctctaaccac 3300
ccagaggtga tccgtcgcgt cggcctgatc gccaccaacg gtcttatcga agctgacatc 3360
tatggcaacg ttaactccac caacgtttct ggctcccgcg tcatgaacgg catcggcggc 3420
tccggcgact tcacccgtaa cggctacatc tccagcttca tcaccccttc agaggcaaag 3480
ggcggcgcaa tctctgcgat cgttcctttc gcatcccaca tcgaccacac cgagcacgat 3540
gtcatggttg ttatctctga gtacggttac gcagaccttc gtggcctggc tccacgtgag 3600
cgcgttgcca agatgatcgg cctggcacac cctgattacc gcccactgct cgaggagtac 3660
tacgctcgcg caacctccgg tgacaacaag tacatgcaga cccctcatga tcttgcaacc 3720
gcgtttgatt tccacatcaa cctggctaag aacggctcca tgaaggcata agttttttct 3780
tggtttagaa accgccgcct cgacaacatt tcgaggcggc ggtttctttt atcacctggg 3840
ttttgagcgt taaattagac caggtcaggc tagtgtttgg tagctaattg agggcgattt 3900
taataaggcc ggtgccatgt actaatatgg tctgagttgg gcctatagct cagttggtag 3960
agctacggac ttttaatccg caggtcttgg gttcgagtcc caatgggccc acatcttaag 4020
tacccctgtt ttggagaatg ctccgagcca ggggtacttt tcttttcctc acacacagta 4080
gctgctgaga aaaatgaaga ccttttgtta ggttgggagt atgaccaacc catacgaggc 4140
cttcataccg ctcaagcatc gtacggggat tgaacccgag cacacctttt gggaatggga 4200
aaacaaaagg gttcacattg caaggagacg tcgagaagcg cccgtccgcg ttatcgtggt 4260
gcatgggcta ggcacccata gtg 4283
<210> 29
<211> 1606
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子cg0007_39-actA
<400> 29
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg tctgatcgca ttgcttcaga 120
aaagctgcgc tccaagctca tgtccgccga cgaggcggca cagtttgtta accacggtga 180
caaggttggt ttctctggct tcaccggcgc tggctaccca aaggcactgc ctacggcaat 240
cgctaaccgg gctaaagaag cacacggtgc aggcaacgac tacgcaatcg acctgttcac 300
tggcgcatcg accgcccctg actgcgatgg cgtacttgca gaagctgacg ctatccgctg 360
gcgcatgcca tacgcatctg atccaatcat gcgtaacaag atcaactccg gctccatggg 420
atactccgat atccacctgt cccactccgg ccagcaggtt gaagagggct tcttcggcca 480
gctcaacgta gctgtcattg aagtcacccg catcactgaa gacggacacc tcatcccttc 540
ttcctccgtg ggtaacaacg ttgagtggct caacgctgca gagaaggtca tcctcgaggt 600
taactcttgg cagtccgaga acctcgaggg catgcacgac atctggtctg taccagcact 660
gccaaaccgc atcgctgtgc caatcaacaa ggccggcgac cgcatcggta agacctacat 720
cgaattcgac accgacaagg ttgtcgcaat tgttgaaacc aacaccccag accgcaacgc 780
accatttaag cctgtcgatg acatctctaa gaagatcgcc ggtaacttcc tcgacttcct 840
cgaaggtgaa gttgcagctg gccgtctttc ctacgatggt tacgtcatgc agtccggcgt 900
gggcaacgtg cctaatgctg tgatggcagg cctgctggag tccaagtttg agaacatcca 960
ggcttacacc gaggttatcc aggacggcat ggttgacctt atcgacgccg gcaagatgac 1020
cgttgcttct gcaacctcct tctccttgtc tcctgagtac gcagagaaga tgaacaacga 1080
cgcaaagcgt taccgtgagt ccatcatcct gcgcccacag cagatctcta accacccaga 1140
ggtgatccgt cgcgtcggcc tgatcgccac caacggtctt atcgaagctg acatctatgg 1200
caacgttaac tccaccaacg tttctggctc ccgcgtcatg aacggcatcg gcggctccgg 1260
cgacttcacc cgtaacggct acatctccag cttcatcacc ccttcagagg caaagggcgg 1320
cgcaatctct gcgatcgttc ctttcgcatc ccacatcgac cacaccgagc acgatgtcat 1380
ggttgttatc tctgagtacg gttacgcaga ccttcgtggc ctggctccac gtgagcgcgt 1440
tgccaagatg atcggcctgg cacaccctga ttaccgccca ctgctcgagg agtactacgc 1500
tcgcgcaacc tccggtgaca acaagtacat gcagacccct catgatcttg caaccgcgtt 1560
tgatttccac atcaacctgg ctaagaacgg ctccatgaag gcataa 1606
<210> 30
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物FarI 3p InfusF
<400> 30
atttcacacc gcatatgcgg catccttcag accgc 35
<210> 31
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物FarI 3p InfusR AvrII
<400> 31
tgggatccgt cgacctgcag cctaggatct ctggaccgga atc 43
<210> 32
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物FarI 5p InfusF
<400> 32
gtccagagat cctaggatcc cgccgagaaa accg 34
<210> 33
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物FarI 5p Infus R
<400> 33
aacagctcac tctagaagtg gaggtcatct gcac 34
<210> 34
<211> 71
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-zwf_P1
<400> 34
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt cttctccgac 60
tacatgcgtc c 71
<210> 35
<211> 95
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-zwf_P2
<400> 35
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcaatggtag cgtcacgatc ctttc 95
<210> 36
<211> 92
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-zwf_P3
<400> 36
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaagtgagca caaacacgac cc 92
<210> 37
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-zwf_P4
<400> 37
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt gggtgatagt 60
tctcaacacg atcttc 76
<210> 38
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_084_P1
<400> 38
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt cggtttcctg 60
attgctcatc aaaatc 76
<210> 39
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_084_P2
<400> 39
catgctgatt accactggta ctcagagtga gcctatggct gcgctgtctc 50
<210> 40
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_084_P3
<400> 40
gagacagcgc agccataggc tcactctgag taccagtggt aatcagcatg 50
<210> 41
<211> 73
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_084_P4
<400> 41
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt aaaaactaga 60
actgctcaag gcc 73
<210> 42
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg1860_pyc_P1
<400> 42
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt cgcagtcatc 60
cggtggagaa gacttc 76
<210> 43
<211> 91
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg1860_pyc_P2
<400> 43
attttaaata cgcaagctac tttatgtgta tgtgggacaa tttgcaacta tttgtgcagg 60
tcaaagctaa gtttcgggca acactcccct g 91
<210> 44
<211> 94
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg1860_pyc_P3
<400> 44
agttgcaaat tgtcccacat acacataaag tagcttgcgt atttaaaatt atgaacctaa 60
ggggtttagc agtgtcgact cacacatctt caac 94
<210> 45
<211> 77
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg1860_pyc_P4
<400> 45
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt cagagagatc 60
accagaataa gccatag 77
<210> 46
<211> 77
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_033_P1
<400> 46
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt aggtttctta 60
cctacttatc aacggtg 77
<210> 47
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_033_P2
<400> 47
gatcggcgct tcacgcttga gccgcaggag ctgcgttttg 40
<210> 48
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_033_P3
<400> 48
caaaacgcag ctcctgcggc tcaagcgtga agcgccgatc 40
<210> 49
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_033_P4
<400> 49
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt gctgcagaca 60
ggtagtcacc 70
<210> 50
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_316_P1
<400> 50
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt cacctgcgac 60
aacaccgaga ac 72
<210> 51
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_316_P2
<400> 51
ctgtggcaga tcagcgtccg acaagaaatc cgccgctgtg 40
<210> 52
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_316_P3
<400> 52
cacagcggcg gatttcttgt cggacgctga tctgccacag 40
<210> 53
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SNP_316_P4
<400> 53
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt cccaggcctg 60
aaagttctgc 70
<210> 54
<211> 71
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SCS_aceE_P1
<400> 54
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt catcgtctaa 60
aaggcctggg c 71
<210> 55
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SCS_aceE_P2
<400> 55
caagttttgc ttgatcggcc aattccacac ctcctgttgg aatg 44
<210> 56
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SCS_aceE_P3
<400> 56
cattccaaca ggaggtgtgg aattggccga tcaagcaaaa cttg 44
<210> 57
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物SCS_aceE_P4
<400> 57
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tcgacaccct 60
cgttggtgga ag 72
<210> 58
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物opcA_P1
<400> 58
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt ccctgcgtac 60
gctgcaactc tttacg 76
<210> 59
<211> 74
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物opcA_P2
<400> 59
atacacgctt aggtccccac gtagtaccac acacaacgcg agaaacggca tttttgcccc 60
taaattatgg cctg 74
<210> 60
<211> 79
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物opcA_P3
<400> 60
gcggactcac ccgcgtgaag ggcatcaatg gccacccagc gtcgtcctga ggagaaatac 60
aacactatgg ttgatgtag 79
<210> 61
<211> 75
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物opcA_P4
<400> 61
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt ggaatcaccc 60
gcactggctt gagag 75
<210> 62
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物pcg0007_39-cg2766_P1
<400> 62
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt acacacacct 60
ggggttatcc 70
<210> 63
<211> 98
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物pcg0007_39-cg2766_P2
<400> 63
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaaatgacaa gtgagaattc cgaatccc 98
<210> 64
<211> 103
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物pcg0007_39-cg2766_P3
<400> 64
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcagtcaatt aagtgtagtc gctaggtgaa aac 103
<210> 65
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物pcg0007_39-cg2766_P4
<400> 65
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt acacaaagaa 60
gcaccaaccg 70
<210> 66
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物184f7引物pcg0007_39-actA_P1
<400> 66
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt cactatgggt 60
gcctagccca tg 72
<210> 67
<211> 95
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物184f7引物pcg0007_39-actA_P2
<400> 67
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaaatgtctg atcgcattgc ttcag 95
<210> 68
<211> 97
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物184f7引物pcg0007_39-actA_P3
<400> 68
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcagatgttg tgtcccaagg ataaacc 97
<210> 69
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物184f7引物pcg0007_39-actA_P4
<400> 69
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt cgcatgatgt 60
gctgatgcag 70
<210> 70
<211> 48
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物184f7
<400> 70
cggtatttca caccgcatac tagtaccatc cagaatcgtt tctaagcg 48
<210> 71
<211> 42
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物184F8
<400> 71
gggatccgtc gacctgcatc cacgaagcca cgagataatc tg 42
<210> 72
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物244-4Go F2 Inf SpeI
<400> 72
ggatcttggt actagtcgct gagacagttg agagaag 37
<210> 73
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物253-4Go R3
<400> 73
ggatcttggt actagtagga attcgatgcg gatcc 35
<210> 74
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物685
<400> 74
gaattcctgc agcccggggt accacactgt ttccttg 37
<210> 75
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物686
<400> 75
tcgagctcgg tacccgggac tagtccaagt gcgtggcgag ttt 43
<210> 76
<211> 46
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物687
<400> 76
ccaagcttgc atgcctgcag gactagtaac gggcggtgaa gggcaa 46
<210> 77
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物693
<400> 77
cttcgaaatg tttgacctgc aggacagcct ttaaacgctc ctttt 45
<210> 78
<211> 718
<212> PRT
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 78
Met Asn Asp Ser Arg Asn Arg Gly Arg Lys Val Thr Arg Lys Ala Gly
1 5 10 15
Pro Pro Glu Ala Gly Gln Glu Asn His Leu Asp Thr Pro Val Phe Gln
20 25 30
Ala Pro Asp Ala Ser Ser Asn Gln Ser Ala Val Lys Ala Glu Thr Ala
35 40 45
Gly Asn Asp Asn Arg Asp Ala Ala Gln Gly Ala Gln Gly Ser Gln Asp
50 55 60
Ser Gln Gly Ser Gln Asn Ala Gln Gly Ser Gln Asn Arg Glu Ser Gly
65 70 75 80
Asn Asn Asn Arg Asn Arg Ser Asn Asn Asn Arg Arg Gly Gly Arg Gly
85 90 95
Arg Arg Gly Ser Gly Asn Ala Asn Glu Gly Ala Asn Asn Asn Ser Gly
100 105 110
Asn Gln Asn Arg Gln Gly Gly Asn Arg Gly Asn Arg Gly Gly Gly Arg
115 120 125
Arg Asn Val Val Lys Ser Met Gln Gly Ala Asp Leu Thr Gln Arg Leu
130 135 140
Pro Glu Pro Pro Lys Ala Pro Ala Asn Gly Leu Arg Ile Tyr Ala Leu
145 150 155 160
Gly Gly Ile Ser Glu Ile Gly Arg Asn Met Thr Val Phe Glu Tyr Asn
165 170 175
Asn Arg Leu Leu Ile Val Asp Cys Gly Val Leu Phe Pro Ser Ser Gly
180 185 190
Glu Pro Gly Val Asp Leu Ile Leu Pro Asp Phe Gly Pro Ile Glu Asp
195 200 205
His Leu His Arg Val Asp Ala Leu Val Val Thr His Gly His Glu Asp
210 215 220
His Ile Gly Ala Ile Pro Trp Leu Leu Lys Leu Arg Asn Asp Ile Pro
225 230 235 240
Ile Leu Ala Ser Arg Phe Thr Leu Ala Leu Ile Ala Ala Lys Cys Lys
245 250 255
Glu His Arg Gln Arg Pro Lys Leu Ile Glu Val Asn Glu Gln Ser Asn
260 265 270
Glu Asp Arg Gly Pro Phe Asn Ile Arg Phe Trp Ala Val Asn His Ser
275 280 285
Ile Pro Asp Cys Leu Gly Leu Ala Ile Lys Thr Pro Ala Gly Leu Val
290 295 300
Ile His Thr Gly Asp Ile Lys Leu Asp Gln Thr Pro Pro Asp Gly Arg
305 310 315 320
Pro Thr Asp Leu Pro Ala Leu Ser Arg Phe Gly Asp Glu Gly Val Asp
325 330 335
Leu Met Leu Cys Asp Ser Thr Asn Ala Thr Thr Pro Gly Val Ser Gly
340 345 350
Ser Glu Ala Asp Val Ala Pro Thr Leu Lys Arg Leu Val Gly Asp Ala
355 360 365
Lys Gln Arg Val Ile Leu Ala Ser Phe Ala Ser Asn Val Tyr Arg Val
370 375 380
Gln Ala Ala Val Asp Ala Ala Val Ala Ser Asn Arg Lys Val Ala Phe
385 390 395 400
Asn Gly Arg Ser Met Ile Arg Asn Met Glu Ile Ala Glu Lys Leu Gly
405 410 415
Tyr Leu Lys Ala Pro Arg Gly Thr Ile Ile Ser Met Asp Asp Ala Ser
420 425 430
Arg Met Ala Pro His Lys Val Met Leu Ile Thr Thr Gly Thr Gln Gly
435 440 445
Glu Pro Met Ala Ala Leu Ser Arg Met Ala Arg Arg Glu His Arg Gln
450 455 460
Ile Thr Val Arg Asp Gly Asp Leu Ile Ile Leu Ser Ser Ser Leu Val
465 470 475 480
Pro Gly Asn Glu Glu Ala Val Phe Gly Val Ile Asn Met Leu Ala Gln
485 490 495
Ile Gly Ala Thr Val Val Thr Gly Arg Asp Ala Lys Val His Thr Ser
500 505 510
Gly His Gly Tyr Ser Gly Glu Leu Leu Phe Leu Tyr Asn Ala Ala Arg
515 520 525
Pro Lys Asn Ala Met Pro Val His Gly Glu Trp Arg His Leu Arg Ala
530 535 540
Asn Lys Glu Leu Ala Ile Ser Thr Gly Val Asn Arg Asp Asn Val Val
545 550 555 560
Leu Ala Gln Asn Gly Val Val Val Asp Met Val Asn Gly Arg Ala Gln
565 570 575
Val Val Gly Gln Ile Pro Val Gly Asn Leu Tyr Val Asp Gly Val Thr
580 585 590
Met Gly Asp Ile Asp Ala Asp Ile Leu Ala Asp Arg Thr Ser Leu Gly
595 600 605
Glu Gly Gly Leu Ile Ser Ile Thr Ala Val Ile Asp Asn Arg Thr Gly
610 615 620
Arg Leu Leu Glu Arg Pro Thr Val Gln Thr Ser Gly Phe Ser Glu Asp
625 630 635 640
Ala Lys Ser Met Met Gly Glu Val Thr Glu Leu Ser Glu Thr Thr Met
645 650 655
Asn Asp Leu Ala Ala Glu Gly Glu Asn Asp Pro Tyr Arg Met Val Gln
660 665 670
Gln Leu Arg Arg Lys Leu Ser Arg Phe Val Glu Gln Lys Trp Lys Arg
675 680 685
Gln Pro Val Ile Met Pro Thr Val Ile Pro Met Thr Ala Glu Thr Thr
690 695 700
His Ile Gly Asp Asp Glu Val Arg Ala Ser Arg Glu Ser Leu
705 710 715
<210> 79
<211> 709
<212> PRT
<213>谷氨酸棒状杆菌(C. glutamicum)
<400> 79
Met Asn Asp Ser Arg Asn Arg Gly Arg Lys Val Thr Arg Lys Ala Gly
1 5 10 15
Pro Pro Glu Ala Gly Gln Glu Asn His Val Glu Thr Pro Val Phe Gln
20 25 30
Ala Pro Asp Ala Ser Ser Asn Gln Ser Thr Val Lys Ala Glu Thr Ala
35 40 45
Gly Asn Asp Asn Arg Asp Ser Ser Gln Gly Ser Gln Ala Gln Gly Ser
50 55 60
Gln Ala Lys Glu Ser Gly Asp Ala Lys Ser Gly Asn Asn Asn Arg Asn
65 70 75 80
Arg Ser Ser Asn Asn Arg Arg Gly Gly Arg Gly Arg Arg Gly Ser Gly
85 90 95
Asn Ala Asn Glu Gly Ser Asn Asn Asn Gly Gly Asn Arg Gly Asn Arg
100 105 110
Gly Asn Arg Gly Gly Gly Arg Arg Asn Val Val Lys Ser Met Gln Gly
115 120 125
Ala Asp Leu Thr Gln Arg Leu Pro Glu Pro Pro Lys Ala Pro Ser Asn
130 135 140
Gly Leu Arg Ile Tyr Ala Leu Gly Gly Ile Ser Glu Ile Gly Arg Asn
145 150 155 160
Met Thr Val Phe Glu Tyr Asn Asn Arg Leu Leu Ile Val Asp Cys Gly
165 170 175
Val Leu Phe Pro Ser Ser Gly Glu Pro Gly Val Asp Leu Ile Leu Pro
180 185 190
Asp Phe Gly Pro Ile Glu Asp His Leu His Arg Val Asp Ala Leu Val
195 200 205
Val Thr His Gly His Glu Asp His Ile Gly Ala Ile Pro Trp Leu Leu
210 215 220
Lys Leu Arg His Asp Ile Pro Ile Leu Ala Ser Arg Phe Thr Leu Ala
225 230 235 240
Leu Ile Ala Ala Lys Cys Lys Glu His Arg Gln Arg Pro Lys Leu Ile
245 250 255
Glu Val Asn Glu Lys Ser Ser Glu Asp Arg Gly Pro Phe Asn Ile Arg
260 265 270
Phe Trp Ala Val Asn His Ser Ile Pro Asp Cys Leu Gly Leu Ala Ile
275 280 285
Lys Thr Pro Ala Gly Leu Val Ile His Thr Gly Asp Ile Lys Leu Asp
290 295 300
Gln Thr Pro Pro Asp Gly Arg Pro Thr Asp Leu Pro Ala Leu Ser Arg
305 310 315 320
Phe Gly Asp Glu Gly Val Asp Leu Met Leu Cys Asp Ser Thr Asn Ala
325 330 335
Thr Thr Pro Gly Val Ser Gly Ser Glu Ala Asp Val Ala Pro Thr Leu
340 345 350
Lys Arg Leu Val Gly Asp Ala Lys Gln Arg Val Ile Leu Ala Ser Phe
355 360 365
Ala Ser Asn Val Tyr Arg Val Gln Ala Ala Val Asp Ala Ala Val Ala
370 375 380
Ala Asn Arg Lys Val Ala Phe Asn Gly Arg Ser Met Ile Arg Asn Met
385 390 395 400
Glu Ile Ala Glu Lys Leu Gly Tyr Leu Lys Ala Pro Arg Gly Thr Ile
405 410 415
Ile Ser Met Asp Asp Ala Ser Arg Met Ala Pro His Lys Val Met Leu
420 425 430
Ile Thr Thr Gly Thr Gln Gly Glu Pro Met Ala Ala Leu Ser Arg Met
435 440 445
Ala Arg Arg Glu His Arg Gln Ile Thr Val Arg Asp Gly Asp Leu Ile
450 455 460
Ile Leu Ser Ser Ser Leu Val Pro Gly Asn Glu Glu Ala Val Phe Gly
465 470 475 480
Val Ile Asn Met Leu Ala Gln Ile Gly Ala Thr Val Val Thr Gly Arg
485 490 495
Asp Ala Lys Val His Thr Ser Gly His Gly Tyr Ser Gly Glu Leu Leu
500 505 510
Phe Leu Tyr Asn Ala Ala Arg Pro Lys Asn Ala Met Pro Val His Gly
515 520 525
Glu Trp Arg His Leu Arg Ala Asn Lys Glu Leu Ala Ile Ser Thr Gly
530 535 540
Val Asn Arg Asp Asn Val Val Leu Ala Gln Asn Gly Val Val Val Asp
545 550 555 560
Leu Val Asn Gly Arg Ala Gln Val Val Gly Gln Ile Pro Val Gly Asn
565 570 575
Leu Tyr Val Asp Gly Val Thr Met Gly Asp Ile Asp Ala Asp Ile Leu
580 585 590
Ala Asp Arg Thr Ser Leu Gly Glu Gly Gly Leu Ile Ser Ile Thr Ala
595 600 605
Val Ile Asp Asn Arg Thr Gly Arg Leu Leu Glu Arg Pro Thr Val Gln
610 615 620
Thr Ser Gly Phe Ser Glu Asp Ala Lys Ser Met Met Gly Glu Val Thr
625 630 635 640
Glu Leu Ala Glu Thr Thr Met Asn Asp Leu Ala Ala Glu Gly Glu Asn
645 650 655
Asp Pro Tyr Arg Met Val Gln Gln Met Arg Arg Lys Leu Ser Arg Phe
660 665 670
Val Glu Gln Lys Trp Lys Arg Gln Pro Val Ile Met Pro Thr Val Ile
675 680 685
Pro Met Thr Ala Glu Thr Arg His Ile Asp Asp Asp Glu Val Arg Ala
690 695 700
Ser Arg Glu Ser Leu
705
<210> 80
<211> 683
<212> PRT
<213>溃疡棒状杆菌(C. ulcerans)
<400> 80
Met Thr Glu Pro Arg Asn Arg Ser Arg Lys Val Thr Arg Lys Ala Gly
1 5 10 15
Pro Pro Glu Thr Thr Glu Ala Pro Ala Phe Gln Ala Pro Asp Thr Ser
20 25 30
Val Glu Val Ser Ala Lys Asp Glu Ala Glu Lys Lys Thr Ser Gly Glu
35 40 45
Gly Ser Ser Asn Gln Glu Ala Ser Ser Lys Gly Asn Arg Gly Arg Ser
50 55 60
Asn Ser Gly Arg Ala Gly Asn Gly Thr Arg Asn Gly Arg Ser Arg Arg
65 70 75 80
Gly Gly Asn Pro Gln Asn Asn Lys Ser Asn Arg Gly Arg Arg Asn Val
85 90 95
Val Lys Ser Met Gln Gly Ala Asp Leu Thr Glu Arg Leu Pro Glu Pro
100 105 110
Pro Lys Ala Pro Lys Asn Gly Leu Arg Ile Tyr Ala Leu Gly Gly Ile
115 120 125
Ser Glu Ile Gly Arg Asn Met Thr Val Phe Glu Tyr Asn Asn Lys Met
130 135 140
Leu Leu Val Asp Cys Gly Val Leu Phe Pro Ser Ser Gly Glu Pro Gly
145 150 155 160
Val Asp Leu Ile Leu Pro Asp Phe Gly Pro Ile Glu Asp Lys Leu Asp
165 170 175
Lys Val Glu Ala Leu Val Val Thr His Gly His Glu Asp His Ile Gly
180 185 190
Ala Ile Pro Trp Leu Leu Lys Leu Arg Pro Asp Leu Pro Ile Tyr Ala
195 200 205
Ser Lys Phe Thr Leu Ala Leu Ile Ala Ala Lys Cys Arg Glu His Arg
210 215 220
Gln Arg Pro Lys Leu Ile Glu Val Asn Glu Lys Ser Asp Ile Asn Arg
225 230 235 240
Gly Pro Phe Asn Ile Arg Phe Trp Ala Val Asn His Ser Ile Pro Asp
245 250 255
Cys Leu Gly Leu Ala Ile Lys Thr Gly Ala Gly Leu Val Ile His Thr
260 265 270
Gly Asp Ile Lys Leu Asp Gln Thr Pro Thr Asp Gly Arg Pro Thr Asp
275 280 285
Leu Pro Ala Leu Ser Arg Phe Gly Asp Glu Gly Val Asp Leu Met Leu
290 295 300
Cys Asp Ser Thr Asn Ala Thr Thr Pro Gly Val Ser Gly Ser Glu Ala
305 310 315 320
Asp Ile Ala Pro Thr Leu Lys Arg Leu Val Gly Asp Ala Lys Gln Arg
325 330 335
Val Ile Leu Ala Ser Phe Ala Ser Asn Val Tyr Arg Val Gln Ala Ala
340 345 350
Val Asp Ala Ala Val Ala Ser Gly Arg Lys Val Ala Phe Asn Gly Arg
355 360 365
Ser Met Ile Arg Asn Met Glu Ile Ala Glu Lys Met Gly Tyr Leu Lys
370 375 380
Ala Pro Arg Gly Thr Ile Val Ser Met Asp Asp Ala Ala Lys Met Ala
385 390 395 400
Pro His Lys Val Met Leu Ile Thr Thr Gly Thr Gln Gly Glu Pro Met
405 410 415
Ala Ala Leu Ser Arg Met Ala Arg Arg Glu His Arg Gln Ile Thr Val
420 425 430
Arg Asp Gly Asp Leu Ile Ile Leu Ser Ser Ser Leu Val Pro Gly Asn
435 440 445
Glu Glu Ala Val Phe Gly Val Ile Asn Met Leu Ala Gln Ile Gly Ala
450 455 460
Thr Val Val Thr Ser Arg Asp Ala Lys Val His Thr Ser Gly His Gly
465 470 475 480
Tyr Ser Gly Glu Leu Leu Phe Leu Tyr Asn Ala Ala Arg Pro Val Asn
485 490 495
Ala Met Pro Val His Gly Glu Trp Arg His Leu Arg Ala Asn Lys Glu
500 505 510
Leu Ala Ile Ser Thr Gly Val Glu Arg Asp Arg Val Val Leu Ala Gln
515 520 525
Asn Gly Val Val Val Asp Leu Val Asp Gly Arg Ala Arg Val Val Gly
530 535 540
Gln Ile Gln Ile Gly Asn Leu Tyr Val Asp Gly Val Thr Met Gly Asp
545 550 555 560
Ile Asp Ala Gly Val Leu Glu Glu Arg Thr Ser Leu Gly Glu Gly Gly
565 570 575
Leu Ile Ala Ile Thr Ala Val Ile Asp Asn Arg Thr Gly Arg Leu Leu
580 585 590
Glu Arg Pro Thr Val Gln Ala Lys Gly Phe Ser Glu Asp Ala Val Ala
595 600 605
Met Met Pro Glu Val Thr Glu Leu Val Glu Asn Thr Met Thr Asp Leu
610 615 620
Ala Gly Glu Gly Glu Asn Asp Pro Tyr Arg Met Val Gln Gln Leu Arg
625 630 635 640
Arg Arg Val Ser Arg Phe Val Glu Gln Lys Trp Arg Arg Lys Pro Met
645 650 655
Ile Met Pro Thr Val Ile Pro Thr Thr Gln Thr Gln Val Glu Ala Asp
660 665 670
Glu Glu Glu Ile Arg Ala Thr Arg Glu Ser Leu
675 680
<210> 81
<211> 484
<212> PRT
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 81
Met Val Trp Gly Met Glu His Thr Ser Ala Leu Thr Leu Ile Asp Ser
1 5 10 15
Val Leu Asp Pro Asp Ser Phe Ile Ser Trp Asp Glu Thr Pro Gln Tyr
20 25 30
Asp Asn Leu Asn Gln Gly Tyr Ala Glu Thr Leu Glu Arg Ala Arg Ser
35 40 45
Lys Ala Lys Cys Asp Glu Ser Val Ile Thr Gly Glu Gly Thr Val Glu
50 55 60
Gly Ile Pro Val Ala Val Ile Leu Ser Asp Phe Ser Phe Leu Gly Gly
65 70 75 80
Ser Leu Gly Thr Val Ala Ser Val Arg Ile Met Lys Ala Ile His Arg
85 90 95
Ala Thr Glu Leu Lys Leu Pro Leu Leu Val Ser Pro Ala Ser Gly Gly
100 105 110
Ala Arg Met Gln Glu Asp Asn Arg Ala Phe Val Met Met Val Ser Ile
115 120 125
Thr Ala Ala Val Gln Arg His Arg Glu Ala His Leu Pro Phe Leu Val
130 135 140
Tyr Leu Arg Asn Pro Thr Met Gly Gly Ala Met Ala Ser Trp Gly Ser
145 150 155 160
Ser Gly His Leu Thr Phe Ala Glu Pro Gly Ala Gln Ile Gly Phe Leu
165 170 175
Gly Pro Arg Val Val Glu Leu Thr Thr Gly Arg Ala Leu Pro Asp Gly
180 185 190
Val Gln Gln Ala Glu Asn Leu Val Lys Thr Gly Val Ile Asp Gly Ile
195 200 205
Val Ser Pro Leu Gln Leu Arg Ala Ala Val Ala Lys Thr Leu Lys Ile
210 215 220
Ile Gln Pro Ala Glu Ala Thr Asp Arg Phe Ser Pro Thr Thr Pro Gly
225 230 235 240
Ala Ala Leu Pro Val Met Glu Ala Ile Ala Arg Ser Arg Asp Pro His
245 250 255
Arg Pro Gly Ile Gly Glu Ile Met Glu Thr Leu Gly Ala Asp Val Val
260 265 270
Arg Leu Ser Gly Ala Arg Ala Gly Ala Leu Ser Ser Ala Val Arg Val
275 280 285
Ala Leu Ala Arg Ile Gly Gly Arg Pro Val Val Leu Ile Gly Gln Asp
290 295 300
Arg Arg Phe Thr Leu Gly Pro Gln Glu Leu Arg Phe Ala Arg Arg Gly
305 310 315 320
Ile Ser Leu Ala Arg Glu Leu Asn Leu Pro Ile Val Ser Ile Ile Asp
325 330 335
Thr Ser Gly Ala Glu Leu Ser Gln Ala Ala Glu Glu Leu Gly Ile Ala
340 345 350
Ser Ser Ile Ala Arg Thr Leu Ser Lys Leu Ile Asp Ala Pro Leu Pro
355 360 365
Thr Val Ser Val Ile Ile Gly Gln Gly Val Gly Gly Gly Ala Leu Ala
370 375 380
Met Leu Pro Ala Asp Leu Val Tyr Ala Ala Glu Asn Ala Trp Leu Ser
385 390 395 400
Ala Leu Pro Pro Glu Gly Ala Ser Ala Ile Leu Phe Arg Asp Thr Asn
405 410 415
His Ala Ala Glu Ile Ile Glu Arg Gln Gly Val Gln Ala His Ala Leu
420 425 430
Leu Ser Gln Gly Leu Ile Asp Gly Ile Val Ala Glu Thr Glu His Phe
435 440 445
Val Glu Glu Ile Leu Gly Thr Ile Ser Asn Ala Leu Ser Glu Leu Asp
450 455 460
Asn Asn Pro Glu Arg Ala Gly Arg Asp Ser Arg Phe Thr Arg Phe Glu
465 470 475 480
Arg Leu Ala Gln
<210> 82
<211> 480
<212> PRT
<213> 谷氨酸棒状杆菌(C. glutamicum) MB001
<400> 82
Met Glu His Thr Ser Ala Leu Thr Leu Ile Asp Ser Val Leu Asp Pro
1 5 10 15
Asp Ser Phe Ile Ser Trp Asn Glu Thr Pro Gln Tyr Asp Asn Leu Asn
20 25 30
Gln Gly Tyr Ala Glu Thr Leu Glu Arg Ala Arg Ser Lys Ala Lys Cys
35 40 45
Asp Glu Ser Val Ile Thr Gly Glu Gly Thr Val Glu Gly Ile Pro Val
50 55 60
Ala Val Ile Leu Ser Asp Phe Ser Phe Leu Gly Gly Ser Leu Gly Thr
65 70 75 80
Val Ala Ser Val Arg Ile Met Lys Ala Ile His Arg Ala Thr Glu Leu
85 90 95
Lys Leu Pro Leu Leu Val Ser Pro Ala Ser Gly Gly Ala Arg Met Gln
100 105 110
Glu Asp Asn Arg Ala Phe Val Met Met Val Ser Ile Thr Ala Ala Val
115 120 125
Gln Arg His Arg Glu Ala His Leu Pro Phe Leu Val Tyr Leu Arg Asn
130 135 140
Pro Thr Met Gly Gly Ala Met Ala Ser Trp Gly Ser Ser Gly His Leu
145 150 155 160
Thr Phe Ala Glu Pro Gly Ala Gln Ile Gly Phe Leu Gly Pro Arg Val
165 170 175
Val Glu Leu Thr Thr Gly His Ala Leu Pro Asp Gly Val Gln Gln Ala
180 185 190
Glu Asn Leu Val Lys Thr Gly Val Ile Asp Gly Ile Val Ser Pro Leu
195 200 205
Gln Leu Arg Ala Ala Val Ala Lys Thr Leu Lys Val Ile Gln Pro Val
210 215 220
Glu Ala Thr Asp Arg Phe Ser Pro Thr Thr Pro Gly Val Ala Leu Pro
225 230 235 240
Val Met Glu Ala Ile Ala Arg Ser Arg Asp Pro Gln Arg Pro Gly Ile
245 250 255
Gly Glu Ile Met Glu Thr Leu Gly Ala Asp Val Val Lys Leu Ser Gly
260 265 270
Ala Arg Ala Gly Ala Leu Ser Pro Ala Val Arg Val Ala Leu Ala Arg
275 280 285
Ile Gly Gly Arg Pro Val Val Leu Ile Gly Gln Asp Arg Arg Phe Thr
290 295 300
Leu Gly Pro Gln Glu Leu Arg Phe Ala Arg Arg Gly Ile Ser Leu Ala
305 310 315 320
Arg Glu Leu Asn Leu Pro Ile Val Ser Ile Ile Asp Thr Ser Gly Ala
325 330 335
Glu Leu Ser Gln Ala Ala Glu Glu Leu Gly Ile Ala Ser Ser Ile Ala
340 345 350
Arg Thr Leu Ser Lys Leu Ile Asp Ala Pro Leu Pro Thr Val Ser Val
355 360 365
Ile Ile Gly Gln Gly Val Gly Gly Gly Ala Leu Ala Met Leu Pro Ala
370 375 380
Asp Leu Val Tyr Ala Ala Glu Asn Ala Trp Leu Ser Ala Leu Pro Pro
385 390 395 400
Glu Gly Ala Ser Ala Ile Leu Phe Arg Asp Thr Asn His Ala Ala Glu
405 410 415
Ile Ile Glu Arg Gln Gly Val Gln Ala His Ala Leu Leu Ser Gln Gly
420 425 430
Leu Ile Asp Gly Ile Val Ala Glu Thr Glu His Phe Val Glu Glu Ile
435 440 445
Leu Gly Thr Ile Ser Asn Ala Leu Ser Glu Leu Asp Asn Asn Pro Glu
450 455 460
Arg Ala Gly Arg Asp Ser Arg Phe Thr Arg Phe Glu Arg Leu Ala Gln
465 470 475 480
<210> 83
<211> 480
<212> PRT
<213>克氏棒状杆菌(C. crudilactis)
<400> 83
Met Ala Tyr Thr Ser Ala Leu Ser Leu Ile Asp Thr Val Leu Asp Ser
1 5 10 15
Asp Ser Phe Ala Ser Trp Asp Thr Pro Pro Arg Tyr Gly Asn Ile Asp
20 25 30
Pro Lys Tyr Ala Glu Thr Leu Ala Lys Ala Arg Ser Lys Ser Glu Cys
35 40 45
Asn Glu Ser Ile Ile Thr Gly Glu Gly Thr Val Asp Gly Ile Ala Val
50 55 60
Ala Val Ile Leu Ser Asp Phe Ser Phe Leu Gly Gly Ser Leu Gly Thr
65 70 75 80
Val Ala Ser Met Arg Ile Met Asn Ala Ile His Arg Ala Thr Glu Arg
85 90 95
Lys Met Pro Leu Leu Val Ser Pro Ala Ser Gly Gly Ala Arg Met Gln
100 105 110
Glu Asp Asn Arg Ala Phe Val Ile Met Val Ser Ile Thr Ala Ala Val
115 120 125
Gln Arg His Arg Glu Ala His Leu Pro Phe Leu Val Tyr Leu Arg Asn
130 135 140
Pro Thr Met Gly Gly Ala Met Ala Ser Trp Gly Ser Ser Gly His Ile
145 150 155 160
Thr Phe Ala Glu Pro Gly Ala Gln Ile Gly Phe Leu Gly Pro Arg Val
165 170 175
Val Glu Leu Ser Thr Gly Lys Pro Leu Pro Glu Gly Val Gln Gln Ala
180 185 190
Glu Asn Leu Val Ala Arg Gly Val Ile Asp Gly Ile Val Ser Pro Met
195 200 205
Gln Leu Arg Ala Ala Val Ser Lys Thr Val Lys Val Leu Gln Pro Ala
210 215 220
Val Leu Gly Ala His Phe Pro Thr Thr Ala Pro Gly Ser Gly Leu Pro
225 230 235 240
Val Met Glu Ala Ile Ala Arg Ser Arg Asp Pro Arg Arg Pro Gly Ile
245 250 255
Gly Glu Ile Ile Glu Thr Leu Gly Glu Asp Val Val Lys Leu Ser Gly
260 265 270
Ala Arg Arg Gly Thr Ala Met Arg Ala Val Arg Val Gly Leu Ala Arg
275 280 285
Leu Ala Gly Arg Pro Val Val Leu Ile Gly Gln Asp Arg His Phe Ala
290 295 300
Leu Gly Pro Gln Asp Leu Ser Phe Ala Arg Arg Gly Ile Ser Leu Ala
305 310 315 320
Arg Glu Leu Asn Leu Pro Ile Val Ser Ile Ile Asp Thr Ser Gly Ala
325 330 335
Glu Leu Ser Gln Asp Ala Glu Glu Leu Gly Ile Ala Ser Ser Ile Ala
340 345 350
Arg Thr Leu Ala Lys Leu Ile Asp Ala Pro Leu Pro Thr Val Ser Val
355 360 365
Ile Leu Gly Gln Gly Val Gly Gly Gly Ala Leu Ala Met Leu Pro Ala
370 375 380
Asp Lys Val Tyr Ala Ala Glu Asn Ala Trp Leu Ser Ala Leu Pro Pro
385 390 395 400
Glu Gly Ala Ser Ala Ile Leu Phe Arg Ser Thr Asp His Ala Ala Glu
405 410 415
Ile Met Glu Arg Gln Gly Val Gln Ala Gln Thr Leu Leu Ser Gln Gly
420 425 430
Leu Ile Asp Gly Ile Ile Ser Glu Ala Val Pro Phe Val Glu Glu Val
435 440 445
Leu Gly Thr Ile Ser Asn Ala Leu Ser Glu Leu Ala Ser Asn Pro Glu
450 455 460
Arg Ala Gly Arg Glu Ser Arg Phe Lys Arg Phe Glu Arg Leu Ala Arg
465 470 475 480
<210> 84
<211> 414
<212> PRT
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 84
Met Ala Phe Gly Phe Phe Ser Arg Arg Lys Lys Lys Asn Lys Asp Lys
1 5 10 15
Asn Pro Asn Glu Asn Ser Ala Val Pro Ala His Ser Glu Asp Ser Pro
20 25 30
Gln Glu Val Phe Glu Gly Asn Gly Arg Gln Val Gly Asp Pro Ile Glu
35 40 45
Gln Gln Val Asp Arg Asp Ala Arg Gly Arg Leu Thr Ala Ala Asp Phe
50 55 60
Leu Pro Asp Ala Asp Leu Pro Gln Leu Asn Arg Ser Arg Ala Asn Met
65 70 75 80
Leu Arg Arg Glu Leu Glu Tyr Arg Phe Ser Leu Gln Asn Ala His Ile
85 90 95
Asn Ile Asp Gly Asn Thr Ala Met Ile Gln Arg Ser Asp Gly Gly Ala
100 105 110
Ala His Val Ser Leu Arg Thr Leu Ala Met Asn Ala Ala Gly Leu Asp
115 120 125
Asn Phe Asp Gln Leu Pro Glu Leu Val Glu Ser Phe Val His Gly Thr
130 135 140
Leu Ala Asp Ala Thr Leu Asn Asp Leu Ser Thr Ala Asp Leu Tyr Lys
145 150 155 160
Ala Leu Arg Leu Arg Leu Leu Pro Thr Pro Gly Glu Gly Asp Asp Leu
165 170 175
Val Glu His Gly Leu Asp Arg Glu Ser Gln Ile Arg Asp Asp Ser Ile
180 185 190
Leu Arg Thr Phe Thr Ser Asp Met Ser Ile Ala Leu Val Leu Asp Thr
195 200 205
Glu His Ala Ile Arg Ile Gln Pro Leu Lys Glu Leu Glu Glu Phe Asp
210 215 220
Asp Leu Ser Ala Leu Glu Arg Ala Ala Asp Arg Asn Thr Trp Gln Glu
225 230 235 240
Leu Tyr Asp Ala Asn Val Asp Ala Ser Phe Val Asp Ala Glu Ser Asp
245 250 255
Ser Glu Gly Ser Ser Phe Trp Ala Phe Glu Ser Asn Ser Tyr Tyr Leu
260 265 270
Gly Ser Ala Pro Leu Phe Leu Asn Asp Leu Leu Ala Lys Trp Ala Pro
275 280 285
Asp Leu Asp Gln Ser Asp Gly Val Ile Phe Ala Val Pro Asp Arg Asp
290 295 300
Leu Leu Ile Ala Arg Pro Val Thr Thr Gly Glu Asp Leu Met Asn Gly
305 310 315 320
Ile Thr Ala Met Val Arg Ile Ala Met Arg Phe Gly Leu Gly Asn Pro
325 330 335
Thr Ser Ile Ser Pro Arg Leu His Leu Leu Arg Asp Asn Gln Val Thr
340 345 350
Thr Phe Thr Asp Phe Arg Val Val Ser Pro Glu Met Glu Ala Glu Trp
355 360 365
Glu Asp Ser Ala Phe Asp Ala Pro Pro Ala Gly Ala Ile Gly Ile Glu
370 375 380
Val Arg Pro Asp Pro Tyr Leu Met Glu Arg Leu Gln Gln Gly Gly Phe
385 390 395 400
Gly Asp Phe Gly Asp Phe Gly Lys Pro Arg Asp Leu Asp Met
405 410
<210> 85
<211> 383
<212> PRT
<213>克氏棒状杆菌(C. crudilactis)
<400> 85
Met Ala Phe Gly Phe Phe Gly Arg Lys Lys Glu Arg Lys Asp Arg His
1 5 10 15
Ser Gly Asp Pro Glu Arg Asp Ser Lys Ser Arg Leu Thr Ser Ala Asp
20 25 30
Leu Leu Pro Asp Thr Asp Leu Pro Gln Leu Asn Arg Ser Arg Ala Asn
35 40 45
Met Leu Arg Gln Glu Leu Glu Tyr Arg Phe Ala Leu Gln Asp Ala His
50 55 60
Ile Asn Ile Asp Gly His Ser Ala Met Val Gln Arg Ala Asp Gly Gly
65 70 75 80
Ala Ala His Val Ser Leu Arg Thr Leu Ala Met Asn Ala Ala Gly Leu
85 90 95
Glu Asn Phe Asp Gln Leu Pro Glu Leu Val Glu Asn Phe Val His Gly
100 105 110
Thr Leu Ala Asp Ala Thr Leu Lys Asp Leu Ser Thr Ala Asp Leu Tyr
115 120 125
Lys Ala Leu Arg Leu Arg Leu Leu Pro Thr Pro Ser Glu Asn Asp Glu
130 135 140
Leu Ala Glu Tyr Gly Leu Thr Ile Glu Ser Lys Ile Arg Asp Asp Ser
145 150 155 160
Ile Leu Arg Thr Phe Thr Ser Asp Met Ser Ile Ala Leu Val Leu Asp
165 170 175
Thr Glu His Ala Ile Arg Ile Gln Pro Leu Lys Glu Leu Glu Arg Phe
180 185 190
Asp Asp Leu Gly Ala Leu Glu Arg Ala Ala Asp Arg Asn Thr Trp Gln
195 200 205
Glu Leu Tyr Asp Ala Ser Val Asp Val Thr Phe Val Asp Ala Glu Ser
210 215 220
Asp Ser Glu Gly Ser Ser Phe Trp Thr Phe Glu Ser Asn Ser Tyr Tyr
225 230 235 240
Leu Gly Ser Ala Pro Leu Phe Leu Gly Asp Leu Leu Ala Lys Trp Ala
245 250 255
Pro Asp Leu Asp Gln Ser Asp Gly Val Ile Phe Ala Val Pro Asp Arg
260 265 270
Asp Leu Leu Ile Ala Arg Pro Ile Ser Thr Gly Glu Asn Leu Met Asn
275 280 285
Gly Ile Thr Ala Met Val Arg Thr Ala Met Arg Phe Gly Leu Gly Asn
290 295 300
Pro Thr Ser Ile Ser Pro Arg Leu His Met Leu His Asp Asn Gln Val
305 310 315 320
Ile Thr Phe Thr Asp Tyr Arg Ile Val Ser Pro Glu Met Glu Ala Glu
325 330 335
Trp Glu Glu Asn Ser Phe Asp Ala Pro Pro Ala Gly Ala Ile Gly Ile
340 345 350
Glu Val Arg Pro Asp Pro Tyr Leu Met Glu Arg Leu Gln Gln Gly Gly
355 360 365
Phe Gly Asp Phe Gly Asp Phe Gly Lys Pro Arg Asp Leu Gly Met
370 375 380
<210> 86
<211> 373
<212> PRT
<213>有效棒状杆菌(C. efficiens) YS-314
<400> 86
Met Glu Gln Lys Val Glu Arg Asp Gly Asn Gly Ile Leu Thr Pro Ala
1 5 10 15
Asp Phe Leu Pro Asp Thr Asp Leu Pro Gln Leu Asn Arg Ser Arg Ala
20 25 30
Asn Met Leu Arg Arg Glu Leu Glu Tyr Arg Phe Ala Leu Lys Asp Ala
35 40 45
Gln Ile Thr Ile Asp Gly Ala Ser Ala Met Val His Arg Pro Asp Gly
50 55 60
Gly Ala Ala His Val Ser Leu Arg Thr Leu Ala Met Asn Ala Ala Gly
65 70 75 80
Leu Asp Asp Leu Gly Gln Leu Pro Glu Leu Val Asp Asn Phe Val His
85 90 95
Gly Thr Leu Ala Asp Ala Thr Leu Ser Thr Leu Ser Thr Ala Asp Leu
100 105 110
Tyr Lys Ser Leu Arg Leu Arg Leu Leu Pro Asn Pro Glu Gln Gly Asp
115 120 125
Ser Ile Leu Glu Thr Ser Asp Gly Val Glu Arg Gln Ile Arg Glu Thr
130 135 140
Ser Val Leu Arg Asp Phe Thr Ser Asp Thr Ser Ile Ala Leu Val Leu
145 150 155 160
Asp Thr Glu His Ala Ile Arg Ile Gln Pro Leu His Glu Leu Glu Glu
165 170 175
Phe Asp Ser Leu Asp Ala Leu Glu Arg Ala Ala Asp Arg Asn Thr Trp
180 185 190
Gln Glu Leu Leu Asp Ala Asp Val Asp Val Thr Tyr Phe Asn Asn Asp
195 200 205
Glu Gly Gly Glu Gly Ser Asp Phe Trp Ala Phe Glu Ser Ser Ser Tyr
210 215 220
Tyr Leu Gly Ser Ser Pro Leu Phe Leu Thr Asp Leu Leu His Arg Trp
225 230 235 240
Ala Pro Asp Leu Asp Gln Ser Arg Gly Val Ile Phe Ala Val Pro Asp
245 250 255
Arg Asp Leu Leu Ile Ala Arg Pro Val Thr Thr Gly Glu Asn Leu Met
260 265 270
Asn Gly Ile Thr Ser Met Val Arg Ile Ala Met Arg Phe Gly Leu Gly
275 280 285
Asn Pro Thr Ser Ile Ser Pro Arg Leu His Leu Leu Tyr Asp Asn Gln
290 295 300
Ile Asn Thr Phe Thr Asp Tyr Arg Ile Ile Glu Pro Thr Glu Asp Glu
305 310 315 320
Glu Trp Ala Val Pro Thr Ala Asp Ala Pro Gln Pro Gly Ser Ile Gly
325 330 335
Ile Glu Val Arg Pro Asp Ala Tyr Leu Met Glu Met Leu Gln Gln Asp
340 345 350
Gly Phe Gly Asp Ser Phe Asp Asp Phe Gly Pro Arg Asp Leu Gly Pro
355 360 365
Asp Asp Phe Arg Tyr
370
<210> 87
<211> 316
<212> PRT
<213> C. vitaeruminis DSM 20294
<400> 87
Met Ile Phe Glu Leu Pro Asp Ser Ser Thr Arg Asp Ile Ala Lys Thr
1 5 10 15
Leu Val Arg Ile Arg Asp Thr Gly Gly Gln Val Thr Thr Ser Arg Val
20 25 30
Leu Ser Leu Ile Val Ile Ala Arg Lys Glu Asp Asp Val Glu Ala Val
35 40 45
Ile Arg Ala Thr Asn Glu Ala Ser Arg Glu His Pro Ser Arg Val Ile
50 55 60
Val Leu Ile Gln Gly Asp Pro Thr Val Pro Ser Ser Arg Leu Asp Ala
65 70 75 80
Glu Val Arg Ile Gly Gly Asp Ala Gly Ala Ser Glu Leu Ile Leu Ile
85 90 95
Asn Leu Asp Gly Glu Val Ala Asn Asn Leu Val His Val Val Thr Pro
100 105 110
Leu Leu Leu Pro Asp Thr Pro Ile Val Thr Trp Trp Ala Tyr Ala Ala
115 120 125
Pro Val Asp Pro Ser Ser Asp Pro Met Gly Ile Ile Ala Gln Arg Arg
130 135 140
Ile Thr Asp Ser Leu Phe Asp Pro Pro Gln Asp Ala Leu Tyr Asn Arg
145 150 155 160
Arg Asn His Tyr Val Pro Gly Asp Ser Asp Ile Ala Trp Ala Arg Leu
165 170 175
Thr Pro Trp Arg Gly Val Leu Ala Ser Thr Leu Asp Gln Pro Pro His
180 185 190
Glu Met Val Gln Glu Val Arg Val Val Gly Pro Gln Asp Ser Pro Ser
195 200 205
Val Asp Leu Ala Ala Gly Trp Leu Ile Asp Arg Leu Gly Val Pro Val
210 215 220
Ser Arg Leu Val Asp Glu His Ala Gln Ala Cys Asp Ala Asp Thr Ile
225 230 235 240
Pro Val Asn Arg Val Glu Leu Asp Arg Pro Ser Gly Thr Val Val Leu
245 250 255
Glu Val Leu Pro Asp Gln Asp Thr Leu Ser Val Thr Phe Pro Gly Lys
260 265 270
Glu Val Ala Leu Val Ala Ile Ser Arg Arg Ser Thr Ser Asp Cys Leu
275 280 285
Ala Glu Glu Leu Arg His Leu Asp Pro Asp Val Ala Tyr Gln Arg Ala
290 295 300
Leu Ser Gly Leu Ala His Val Asp Tyr Pro Gln Gly
305 310 315
<210> 88
<211> 317
<212> PRT
<213> 马氏棒状杆菌(C. matruchotii) ATCC 33806
<400> 88
Met Ile Phe Thr Leu Ser Asp Thr Asp Thr His Glu Ile Ala Arg Thr
1 5 10 15
Leu Val Glu Ile Arg Asp Asp Gly Ser Gln Gly Thr Thr Ser Arg Val
20 25 30
Leu Thr Leu Ile Ala Val Ala Lys Asp Thr Asp Asn Ile Arg Ser Val
35 40 45
Ile Ser Ala Ser Lys Asp Ala Ser Arg Glu His Pro Cys Arg Val Ile
50 55 60
Val Leu Val Thr Gly Asp Pro Asp Arg Asn Thr Arg Ile Asp Ala Glu
65 70 75 80
Val Arg Ile Gly Gly Asp Ala Gly Ala Ser Glu Ile Ile Val Ile Asp
85 90 95
Leu Glu Gly Glu Val Thr Arg His Leu Val His Val Val Thr Pro Leu
100 105 110
Leu Leu Pro Asp Thr Pro Ile Val Val Trp Trp Pro Tyr Thr Ala Pro
115 120 125
Val Asn Pro Ala Glu Asp Pro Leu Gly Arg Ile Ala Gln Arg Arg Ile
130 135 140
Thr Asp Ser Gln Lys Asp Pro Leu Val Asp Ala Leu Tyr Asn Arg Arg
145 150 155 160
Asn Ser Tyr Thr Asp Gly Asp Ser Asp Leu Ala Trp Ala Arg Ile Thr
165 170 175
Pro Trp Arg Gly Val Val Ala Ser Ala Leu Asp Gln Pro Pro Phe Glu
180 185 190
Pro Val Ile Glu Ala Lys Val Tyr Gly Ala Thr Asn Cys Pro Ser Val
195 200 205
Asp Leu Ala Ala Gly Trp Leu Ala Ala Arg Leu Gly Val Pro Val Thr
210 215 220
Arg Ile Ala Val Asp Asp Pro Ser Val Lys Lys Gln Ala Ser Leu Asn
225 230 235 240
Val Thr Pro Val Glu Arg Val Glu Leu Val Arg Pro Ser Gly Ala Thr
245 250 255
Val Leu Glu Ala Leu Asp Ser Gln Thr Ile Gly Val Ser Val Pro Asn
260 265 270
Arg Glu Pro Ala Arg Val Ala Val Asn Leu Arg Thr Asp Ala Asp Cys
275 280 285
Leu Ala Glu Glu Leu Arg His Leu Asp Pro Asp His Ala Tyr Asp Glu
290 295 300
Val Leu Arg Gly Leu Asn Gln Val Ser Tyr Pro Val Val
305 310 315
<210> 89
<211> 353
<212> PRT
<213> C. falsenii DSM 44353
<400> 89
Met Ile Ile Asp Leu Pro Asn Thr Ser Thr His Ile Ile Ser Gln Lys
1 5 10 15
Leu Ser Ala Leu Arg Glu Glu Arg Gly Glu Val Ala Thr Gly Arg Val
20 25 30
Leu Thr Leu Ile Val Val Ala Ser Ser Glu Asp Asp Leu Pro Arg Ile
35 40 45
Ile Asp Ala Thr His Glu Ala Ser Arg Glu His Pro Ala Arg Val Leu
50 55 60
Ile Leu Val Asn His Arg Lys Ser Asp Glu Val His Leu Asp Ala Glu
65 70 75 80
Ile Arg Leu Gly Gly Asp Ala Gly Ala Ser Glu Ile Ile Val Met His
85 90 95
Asn His Gly Glu Leu Val Ala His Arg Ala Ser Val Val Thr Pro Leu
100 105 110
Leu Leu Pro Asp Thr Pro Ile Val Ala Trp Trp Pro Thr Asn Gly Pro
115 120 125
Arg Asn Pro Ala Ala Asp Ala Ile Gly Met Leu Ala Thr Arg Arg Ile
130 135 140
Thr Asp Ser Leu His Asp Asp Asp Thr Asp Ala Leu Phe Arg Arg Arg
145 150 155 160
Met Thr Tyr Thr Arg Gly Asp Ser Asp Met Ala Trp Ser Arg Ile Thr
165 170 175
Leu Trp Arg Gly Leu Leu Ala Ser Ser Leu Asp Gln Pro Pro Tyr Glu
180 185 190
Pro Ile Asp Ser Val Glu Leu Thr Gly Pro Ala Asp Asp Pro Ser Thr
195 200 205
Asp Ile Ala Ala Gly Trp Leu Ala Glu Lys Leu Gln Val Pro Val Thr
210 215 220
Arg His Asn Ser Gly Thr Asp Ala Ile Pro Arg Asp Asp Lys Gly Asn
225 230 235 240
Lys Thr Ile Ser Ile Glu Lys Ala Val Leu His Arg Glu Ser Ala Thr
245 250 255
Val Glu Val Thr Val Val Asp Ser Glu Thr Val Ser Met Lys Val Gly
260 265 270
Asp Lys Glu Ser Leu Val Thr Leu Gly Arg Arg Pro Leu Ala Asp Cys
275 280 285
Leu Ala Glu Glu Leu Arg His Leu Asp Pro Asp Tyr Ala Phe Gly His
290 295 300
Ala Leu Arg Gly Leu Val Arg Val Asn Arg Pro Glu Arg Thr Asn Arg
305 310 315 320
Leu Pro Asn Tyr Arg Gly Ala Ser Ile Pro Asp Glu Ala Pro Asp Arg
325 330 335
Trp Ser Ala Ser Val Ala Asp Asp His Phe Ala Glu Gln Glu Ser Arg
340 345 350
Arg
<210> 90
<211> 310
<212> PRT
<213> 耐盐棒状杆菌(C. halotolerans) YIM 70093
<400> 90
Met Ile Ile Thr Leu Pro Asp Thr Asp Ser Arg Glu Val Ala Ala Ser
1 5 10 15
Leu Leu Glu Ala Arg Glu Gln Asn Ser Arg Thr Thr Gly Arg Val Leu
20 25 30
Thr Leu Ile Val Ala Ala Asn Leu Asn Asp Asn Val Asp Ala Ile Ile
35 40 45
Ser Ala Ala Arg Gly Ala Ser His Glu His Pro Ala Arg Val Leu Val
50 55 60
Phe Ile Thr Gly Glu Thr Ser Gly Lys Ala Gln Leu Asp Ala Glu Val
65 70 75 80
Arg Val Gly Gly Asp Ala Gly Ala Ser Glu Met Val Ile Met Arg Val
85 90 95
Ser Gly Glu Leu Ala Gly His Leu Glu Ser Val Val Thr Pro Leu Leu
100 105 110
Leu Pro Asp Thr Pro Ile Val Thr Trp Trp Pro Thr Glu Ala Pro Glu
115 120 125
Asn Pro Ser Ala His Pro Leu Gly Arg Leu Ser Gln Arg Arg Ile Thr
130 135 140
Asn Ala Gln Arg Asp Gly Gly Glu Asp Ile Leu Asp Arg Leu Arg Ala
145 150 155 160
Ala Tyr Thr Pro Gly Asp Ser Asp Met Met Trp Ser Arg Ile Thr Ala
165 170 175
Trp Arg Gly Ile Val Ala Ser Ser Leu Asp Gln His Pro His Gly Ala
180 185 190
Val Glu Ser Ala Thr Leu Val Gly Pro Pro Glu Asp Pro Ser Val Asp
195 200 205
Ile Ala Ala Gly Trp Leu Ala Asp Arg Leu Gly Val Glu Ile Ile Arg
210 215 220
Glu Asp Cys Glu Pro Ala Glu Asp Pro Thr Leu Phe Pro Ile Arg Ser
225 230 235 240
Leu Arg Phe Asn Arg Ala Pro Gly Pro Leu Lys Val Asp Val Ile Asp
245 250 255
His Arg Thr Val Gln Val Thr Leu Pro Gly Arg Pro Glu Ser Leu Val
260 265 270
Ala Met Ser Val Arg Ser Asp Ser Glu Cys Leu Ser Glu Glu Leu Arg
275 280 285
His Leu Asp Pro Asp Thr Ala Tyr Ala Gln Ala Leu Arg Ala Leu Asn
290 295 300
Arg Val Thr His Arg Ser
305 310
<210> 91
<211> 325
<212> PRT
<213> 产丙酮酸棒状杆菌(C. pyruviciproducens) ATCC BAA-1742
<400> 91
Met His Gln Leu Arg Asn Thr Ser Thr Lys Glu Ile Ala Ser Lys Leu
1 5 10 15
Glu Gln Ile Glu Thr Ser Gly Met Gln Pro Arg Thr Asn His Val Leu
20 25 30
Thr Leu Ile Val Val Ile Asn Gly Lys Thr Asn Lys Asp Arg Leu Phe
35 40 45
His Thr Ile Ser Ser Ser Ser Lys Glu His Pro Ser Arg Val Leu Val
50 55 60
Leu Glu Pro Leu Ala Glu Glu His Ser Leu Gly Phe His Ala Ser Ser
65 70 75 80
Val Ile Asp Ala Glu Leu His Val Gly Gly Asp Ala Gly Ala Ser Glu
85 90 95
Ile Ile Trp Met Gln Leu Arg Gly Glu Val Thr Glu His Leu Asp Ser
100 105 110
Val Val Thr Pro Leu Leu Leu Pro Asp Thr Pro Ile Val Val Trp Trp
115 120 125
Pro Ala Ser Ala Pro Val Asn Pro Ser His Asp Pro Leu Gly Lys Leu
130 135 140
Ala Met Arg Arg Ile Thr Asp Ser Phe Phe Asp Pro Ala Glu Asp Ala
145 150 155 160
Ile Tyr Arg Arg Arg Ser Asn Tyr Thr Pro Gly Asp Ser Asp Leu Cys
165 170 175
Trp Ser Arg Leu Thr Pro Trp Arg Gly Ile Leu Ala Ser Ala Leu Asp
180 185 190
Gln Pro Pro Phe Glu Pro Val Arg Ser Ala Lys Val Phe Gly Leu Ala
195 200 205
Val Ser Pro Thr Val Asp Leu Ala Ala Gly Trp Leu Ala Asp Val Leu
210 215 220
Gly Val Pro Val Tyr Arg Glu Val Arg Gly Thr Ala Asp Arg Pro Val
225 230 235 240
Asp His Asn Gly Asn Pro Ala Thr Pro Val Ala Glu Val Ile Leu Glu
245 250 255
Arg Asp Thr Gly Ser Val Arg Leu Val Val Ala Asp Ser Glu Thr Ile
260 265 270
Glu Leu Thr Ile Gly Asp Gly Pro Ala Thr Lys Ile Ala Gln Gly Arg
275 280 285
Arg Pro Asp Val Asp Cys Leu Ser Glu Glu Met Arg His Leu Val Pro
290 295 300
Asp Gln Ala Tyr Ala Gly Ala Leu Asp Gly Leu Ser Lys Val Thr Tyr
305 310 315 320
Leu Glu Ala Asp Gln
325
<210> 92
<211> 303
<212> PRT
<213> 干酪棒状杆菌(C. casei) UCMA 3821
<400> 92
Met Ile Thr Pro Met Pro Asn Thr Thr Thr Ser Lys Ile Ala Arg Gly
1 5 10 15
Leu Val Glu Ala Gln Glu His Tyr Thr Leu Thr Thr Ser Arg Val Leu
20 25 30
Thr Leu Ile Val Val Val Asn Val Asp Asp Glu Val Asp Ala Val Leu
35 40 45
Asp Ser Val Arg Asp Ala Ser His Glu His Pro Ser Arg Val Leu Val
50 55 60
Val Ile Thr Gly Ser Arg Asp Ala Glu Pro Leu Leu Asn Ala Glu Leu
65 70 75 80
Arg Val Gly Gly Glu Ala Gly Ala Ser Glu Ile Val Val Met His Leu
85 90 95
Phe Gly Pro Leu Ala Asp Asn Ala Glu Ser Val Val Thr Pro Leu Leu
100 105 110
Leu Pro Asp Thr Pro Ile Val Val Trp Trp Pro Thr Ala Ala Pro Glu
115 120 125
Gly Pro Ser Lys Thr Pro Ile Gly Lys Leu Gly Gln Arg Arg Ile Thr
130 135 140
Tyr Ala Ala Ala Lys Gly Asp Leu Arg Arg Leu Ser Ser Gly Tyr Ala
145 150 155 160
Pro Gly Asp Thr Asp Met Ala Trp Gly Glu Ile Thr Gln Trp Arg Gly
165 170 175
Ile Val Ala Ser Ser Leu Asp Arg Leu Pro His Glu Pro Val Lys Arg
180 185 190
Val Glu Ile Ser Gly Pro Ser Asp Ser Leu Ala Val Asp Leu Ala Ala
195 200 205
Ala Trp Met Leu Asp Arg Leu Ala Val Pro Val Lys Arg Val Val Ser
210 215 220
Asp Ala Pro Gly Glu Lys Phe Thr Ile His Ser Val Arg Phe Phe Arg
225 230 235 240
Glu Thr Ser Asp Ile Ile Ile Asp Ala Ser Glu Glu Gly Ser Val Arg
245 250 255
Val Asn Val Pro Gly Ser Pro Glu Ser Met Val Ala Leu Asn Thr Arg
260 265 270
Ser Arg Ala Glu Ile Leu Ser Glu Glu Leu Arg His Leu Asp Ala Asp
275 280 285
Glu Ala Tyr Ala His Thr Leu Arg Gly Leu Ala Gln Val Asp Tyr
290 295 300
<210> 93
<211> 350
<212> PRT
<213> 解萜烯棒状杆菌(C. terpenotabidum) Y-11
<400> 93
Met Thr Thr Pro Glu Ser Thr Gly Val Gly Ser Ser Gly Val Gly Gln
1 5 10 15
Leu Asp Gly Arg Met Asn Leu Gly Arg Thr Thr Thr His Arg Ile Thr
20 25 30
Arg Ala Leu Arg Ser Leu Arg Glu Lys Arg Gly Glu Val Thr Thr Gly
35 40 45
Arg Val Leu Thr Leu Ile Val Ser Ala Arg Ser Glu Glu Asp Leu Asp
50 55 60
Ser Val Val Arg Gly Ala Asn Glu Ala Ser Arg Glu His Pro Ala Arg
65 70 75 80
Val Ile Val Val Ile His Asp Arg Ser Thr Asp Glu Val Ser Leu Asp
85 90 95
Ala Glu Leu Ile Val Gly Gly Gln Ala Gly Ala Ala Glu Ile Ile Phe
100 105 110
Met Arg Leu His Gly Glu Leu Ala Asp His Ala Asp Ser Val Val Met
115 120 125
Pro Leu Leu Leu Pro Asp Thr Pro Val Val Val Trp Trp Pro Phe Thr
130 135 140
Ser Pro Arg Asn Pro Ala Gly Asp Ser Leu Gly Arg Leu Ala Ser Arg
145 150 155 160
Arg Ile Thr Asp Ser Phe Ser Asp Ala Ala Ala Gly Lys Gly Ala Lys
165 170 175
Ala Ile Phe Arg Arg Arg Ile Gly Tyr Thr Pro Gly Asp Ser Asp Leu
180 185 190
Val Trp Ser Arg Ile Thr Pro Trp Arg Gly Leu Leu Ser Ser Ala Val
195 200 205
Glu Gln His Ala Gly Asp Thr Val Thr Gly Val Asp Ile Thr Gly Pro
210 215 220
Ala Glu Asp Pro Ala Val Asp Ile Ala Ala Gly Trp Leu Ala Asp Arg
225 230 235 240
Phe Ser Val Pro Val Thr Arg Tyr Ala Gly Thr Ser Pro Ala Val Pro
245 250 255
Ile Asp Asp Glu Gly Leu Ala Thr Ala Pro Val Glu Arg Val Val Leu
260 265 270
His Phe Thr Gly Gly Asp Leu Val Met Glu Val Leu Asp His Arg Ser
275 280 285
Val Arg Ile Leu Asp Gly Glu Thr Asp Asn Ile Val Thr Leu His Arg
290 295 300
Arg Thr Leu Gly Glu Cys Leu Ala Glu Glu Leu Arg His Leu Glu Pro
305 310 315 320
Asp Ala Ala Phe Gly Asp Ala Leu His Gly Leu Pro Arg Val Arg Val
325 330 335
Ala Arg Glu Pro Ala Pro Ala Pro Val Glu Glu Ser His Arg
340 345 350
<210> 94
<211> 319
<212> PRT
<213> 谷氨酸棒状杆菌(C. glutamicum) NRRL B-11474
<400> 94
Met Ile Phe Glu Leu Pro Asp Thr Thr Thr Gln Gln Ile Ser Lys Thr
1 5 10 15
Leu Thr Arg Leu Arg Glu Ser Gly Thr Gln Val Thr Thr Gly Arg Val
20 25 30
Leu Thr Leu Ile Val Val Thr Asp Ser Glu Ser Asp Val Ala Ala Val
35 40 45
Thr Glu Ser Thr Asn Glu Ala Ser Arg Glu His Pro Ser Arg Val Ile
50 55 60
Ile Leu Val Val Gly Asp Lys Thr Ala Glu Asn Lys Val Asp Ala Glu
65 70 75 80
Val Arg Ile Gly Gly Asp Ala Gly Ala Ser Glu Met Ile Ile Met His
85 90 95
Leu Asn Gly Pro Val Ala Asp Lys Leu Gln His Val Val Thr Pro Leu
100 105 110
Leu Leu Pro Asp Thr Pro Ile Val Ala Trp Trp Pro Gly Glu Ser Pro
115 120 125
Lys Asn Pro Ser Gln Asp Pro Ile Gly Arg Ile Ala Gln Arg Arg Ile
130 135 140
Thr Asp Ala Leu Tyr Asp Arg Asp Asp Ala Leu Glu Asp Arg Val Glu
145 150 155 160
Asn Tyr His Pro Gly Asp Thr Asp Met Thr Trp Ala Arg Leu Thr Gln
165 170 175
Trp Arg Gly Leu Val Ala Ser Ser Leu Asp His Pro Pro His Ser Glu
180 185 190
Ile Thr Ser Val Arg Leu Thr Gly Ala Ser Gly Ser Thr Ser Val Asp
195 200 205
Leu Ala Ala Gly Trp Leu Ala Arg Arg Leu Asn Val Pro Val Ile Arg
210 215 220
Glu Val Thr Asp Ala Pro Thr Val Ser Thr Asp Glu Phe Gly Thr Pro
225 230 235 240
Leu Leu Ala Ile Gln Arg Leu Glu Ile Val Arg Thr Thr Gly Ser Ile
245 250 255
Ile Ile Thr Ile His Asp Ala His Thr Leu Gln Val Glu Met Pro Glu
260 265 270
Ser Gly Asn Ala Pro Ser Leu Val Ala Ile Gly Arg Arg Ser Glu Ser
275 280 285
Asp Cys Leu Ser Glu Glu Leu Arg His Met Asp Pro Asp Leu Gly Tyr
290 295 300
Gln His Ala Leu Ser Gly Leu Ser Ser Val Lys Leu Glu Thr Val
305 310 315
<210> 95
<211> 87
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子3121
<400> 95
gtggctaaaa cttttggaaa cttaagttac ctttaatcgg aaacttattg aattcgggtg 60
aggcaactgc aactctggac ttaaagc 87
<210> 96
<211> 98
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子0755
<400> 96
aataaattta taccacacag tctattgcaa tagaccaagc tgttcagtag ggtgcatggg 60
agaagaattt cctaataaaa actcttaagg acctccaa 98
<210> 97
<211> 86
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 启动子3381
<400> 97
cgccggataa atgaattgat tattttaggc tcccagggat taagtctagg gtggaatgca 60
gaaatatttc ctacggaagg tccgtt 86
<210> 98
<211> 4093
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 98
gtgatggcat cgcgcacctc aaactgtgca cccgcttcta agccaagcgc tccaagatca 60
aggcgaacag tagcctcacg agcactacgt ggatccaagt tgaccacaat caacacggta 120
tttccggtca aagcatcaac cttggagtag gcgatgatct gatcattgtc tgcttcgtgg 180
aagtggatgt tgcgtagctg ctgcaaggca gggttcgcgc gacggatctg gttgagcaga 240
gcgatgtaat cctcgaggga atcgccacgc tccagagcgc cctcgaaatc gcggggacgc 300
agctcgtact tctcagaatc caagtactct tccgaaccag gcttgacggc ctcgtgctca 360
aagagctcat atccggaata tacgccccac acaggagaca tcgtggcggc caatgcggcg 420
cggatagcga acattgcgcg tccaccatgc tgcagagacg catgcaaaat gtcgggagtg 480
ttcacaaaca ggttcggacg agaaatatcc gccatgcggg cgatctcagt agcgaactcg 540
gtgagctcct ccttggtgac cttccacgtg aagtaggtgt aagactggga gaaaccaatc 600
ttggccaagc catacagacg tgccgggcga gtaaacgcct ccgctaggaa aatgacctca 660
gggtttgatt tatggatggc agaaataagc cactgccaga aattagcggg cttagtgtgc 720
gggttgtcca cgcggaatgt ggtcacaccc aaatccaccc agaacttcac cacacgatag 780
acctcttcat agatcttcgg agcagcatta tcaaagttga tgggataaat atcctggtac 840
ttcttcggtg ggttttctgc atacgcaatg gtgccatcag ccaacaccgt gaaaaactcg 900
cggtgttcct gcgcccatgg atgatcaggg gcagcctgca gagctagatc gagtgcaact 960
tccaaattga gttcccgtgc gcgagccaac aacgcctgga aatcttcaat ggtgcccaac 1020
cgcggatgcg ttgcatcatg cccaccatct ttagaaccaa tagcccacgg cgaacccaca 1080
tcatgaggtt ccggggtcag cgtattattg cggcccttgc ggttgacctc gccaatcgga 1140
tggatcggcg ggaagtaaac agtatcgaag cccatcttcg cgacacgctc caacgcctga 1200
gcagtggtag cgaaagtgcc atgaacgggg gtgccggact catcccaacc acctgtggaa 1260
cgagggaaaa gctcatacca agagttgacc aaagcggcac gacgctcgac acgaacctgc 1320
aggttttcac ccatcgtgac caactcacgc aacgggttaa gttctaaaag atgagtgaca 1380
ctcgcggtga gcgctggggc gagacgtgcg cgtacatcgc cgccacgccg cagagaggag 1440
gcgacgtcga aaagcgcagt cctatcctct ttagacaaat tctccgcagc ccgttcaaac 1500
agctgagccc cgtgctcaaa gtcgttatac aactcatctg aaccctggcc tgcctcaatt 1560
ttagtggtaa tcgcatgacg ccacgtagac ataggatccg accacgcatc cacacgaaat 1620
gtccaattac cagggacatc aggaacgaaa aatgcattcg actgatcctg attatcagga 1680
gtggggcgca tatgaatctg aataggctca gccgccactg aagaatcctc cggaccggac 1740
acattcaacg tcgcggcaat tgcatcatgg ccttcacgcc acacaacagc cgagacagga 1800
acaatctcac ccactactgc tttcgctgga tttccatcca aaatccgggg acgaacatca 1860
tcgataccga gacggccagt cacagttttc tcctcaacat gcagtgtgaa gtgcgaacaa 1920
acgcacacaa gcttaaaagt tcgatcaagc agacacaccg gcaattagcc gggggtatgt 1980
ctccattatg aaagcttaac ggagctgtgt cacttccaca cggaaccgga gaaatcagtc 2040
aagatgggag atgtgactaa ctctataaaa aactctgcag ctgcatcttt tccctctgat 2100
tcgagtgaac tcaatgacga attcaatgaa gacctcctca tcgatttcca aggagtcttc 2160
tttaagaggg atggtcgaaa cctcgtcggc cccgtcgact ggcaagtaga actagatgaa 2220
cgctgggtaa ttatcggacc aaacggagcc ggcaaaacca cactcatccg catggcatcc 2280
gccgaagaat tcccctccgg aggaatggca cgcctcatgg gagaacgcat cggcaaaact 2340
gacatgcgcg acctccgctc catgatcggc gtcagctcct ccgcattagg caaccgaatc 2400
ccctccgaag aaaaagtctc tgacctagtc atctccgcag gctacgcaat cctcggccgc 2460
tggcgcgaag actacgacga aatggacttc ggacaagcca ccgaaatcct cgaacaagtc 2520
ggagccatgc acctagccga ccgcacctgg ggaaccctct ccgaaggcga acgcaaacga 2580
gtcctggtcg cacgcgcact catgaccaac ccggaactcc tcatccttga cgaaccaacc 2640
gcaggaatgg acctcggcgg acgcgaagac ctcgtcggct acctcggaga actcgccatg 2700
gacccagacg cacctgccat cgtcatgatc acccaccacg tcgaagaaat ccccgccgga 2760
ttcacccacg caatgctcct cgacgaaggt gaaatcgtag cccaaggctt gatcaacacc 2820
gtcatgacaa acgagaacct atccaaagca ttccaccagc caatccaagt agaccgcatc 2880
ggggaacgct actttgcccg ccgtgtgaga accgccagga gtcatagggc tcagtaggtt 2940
ttttggagtt gtgggccggt gttggagggg tggttgttta ataggcgact tcacgtatgc 3000
ggacttcatg tgcactgact tcttgcgctc aacttctcat gtgctgactt cactccggat 3060
tgctggcatt tggtagtgat gtcgtgcgat gagaagttga gtatgcgaag tcggtaccga 3120
gaagtcgacc tttaggacat ttttagagga gtccacgcag agactctgtc tccgtaagtt 3180
cgatttcacg gttacgattt cgaactgtcg atatcccggg tgcggtttct caattacaat 3240
ttcgcacttc caatttctca agagcgatac cactaggaaa agtgccattt tccgattgaa 3300
atcgccgttg agaaattgag cgcaaggtat cgagtttgcg aaatcggtaa taagaaatcg 3360
gtaatgagaa atcgtaaccg tgaaatcgaa ctttaaaccc aaatgaccga aaaagttcaa 3420
accaacaagc atcaagtgac atattctcgt gcggactgcg cacggaatcc atagactggt 3480
ctgcagaagt tttacgtgtg ggtttgcttc ggatggggct ttaacaagct tcacagatgt 3540
tggtttttca taggcaagct atccagaagc aggctttaca gaaagtcagg gtgtggcaat 3600
ggtgatgcag ggcattggtg gtaggaagct ggcggcgacg gtgctgttgg ttcgggatgg 3660
gatcatcaat gggcgtcctg atgtggaggt ttacattcag gagcgtgtgt ctactatggc 3720
taattttcct cgggcgacgg tgtttccggg tgggggtgtt gattctcggg attttgcgga 3780
tggtcacggt aaggaagtgt ggaggggacc tagcgcggag gagtggggtg tgcgtttagg 3840
cgtggagcct catgttgcgt atgcgttggt ttttgctgcg gttcgtgagt tgtttgaaga 3900
ggccggtacg ttgcttgcgg agcatacgga tggttctggt ttggtgaaga atgctggtca 3960
gtatcacgga tatcgggagt tgttggagac tcatgagatg tcgctgacgg atatgttgca 4020
gagtgagaat ttggcgattc gtagtgattt gattgtgcct tttgccaggt gggcgagccc 4080
tgagggtaat agg 4093
<210> 99
<211> 74
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-cg1383_P1
<400> 99
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt gtgatggcat 60
cgcgcacctc aaac 74
<210> 100
<211> 96
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-cg1383_P2
<400> 100
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcacttgact gatttctccg gttccg 96
<210> 101
<211> 103
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-cg1383_P3
<400> 101
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaaatgggag atgtgactaa ctctataaaa aac 103
<210> 102
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-cg1383_P4
<400> 102
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt cctattaccc 60
tcagggctcg 70
<210> 103
<211> 991
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 插入在cg1383起始密码子前面的Pcg0007_39启动子
<400> 103
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg ggagatgtga ctaactctat 120
aaaaaactct gcagctgcat cttttccctc tgattcgagt gaactcaatg acgaattcaa 180
tgaagacctc ctcatcgatt tccaaggagt cttctttaag agggatggtc gaaacctcgt 240
cggccccgtc gactggcaag tagaactaga tgaacgctgg gtaattatcg gaccaaacgg 300
agccggcaaa accacactca tccgcatggc atccgccgaa gaattcccct ccggaggaat 360
ggcacgcctc atgggagaac gcatcggcaa aactgacatg cgcgacctcc gctccatgat 420
cggcgtcagc tcctccgcat taggcaaccg aatcccctcc gaagaaaaag tctctgacct 480
agtcatctcc gcaggctacg caatcctcgg ccgctggcgc gaagactacg acgaaatgga 540
cttcggacaa gccaccgaaa tcctcgaaca agtcggagcc atgcacctag ccgaccgcac 600
ctggggaacc ctctccgaag gcgaacgcaa acgagtcctg gtcgcacgcg cactcatgac 660
caacccggaa ctcctcatcc ttgacgaacc aaccgcagga atggacctcg gcggacgcga 720
agacctcgtc ggctacctcg gagaactcgc catggaccca gacgcacctg ccatcgtcat 780
gatcacccac cacgtcgaag aaatccccgc cggattcacc cacgcaatgc tcctcgacga 840
aggtgaaatc gtagcccaag gcttgatcaa caccgtcatg acaaacgaga acctatccaa 900
agcattccac cagccaatcc aagtagaccg catcggggaa cgctactttg cccgccgtgt 960
gagaaccgcc aggagtcata gggctcagta g 991
<210> 104
<211> 4044
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 104
gacaggtgga tatcggagta tcccatggag ccggagttga tcttgttacg catgattgga 60
tcagatgcgt atggcatgcg ccagcggata gcgtcagctt ctgcaagtac gccatcgcag 120
tcaggggcgg tcgatgcgcc agtgaacagg tcgattgcgt agtcgttgcc tgcaccgtgt 180
gcttctttag cccggttagc gattgccgta ggcagtgcct ttgggtagcc agcgccggtg 240
aagccagaga aaccaacctt gtcaccgtgg ttaacaaact gtgccgcctc gtcggcggac 300
atgagcttgg agcgcagctt ttctgaagca atgcgatcag acatttttaa cgaacactcc 360
tcgctactta tccgatacag acgtttccat aatacacgct taggtcccca cgtagtacca 420
cacacaacgc gagaaacggc agatgttgtg tcccaaggat aaaccatcgt gacaggaaat 480
gcccgaatac tatctgtgat agtgggattt caaggcccag cattaccccc aaatttgcat 540
gccccctgtt tgcaagcctc ccaaagacca ggaataaagc tcaggaataa agctcagacc 600
aaccctagag cccgcgaagc ttaacccaac cccagccccc aaaaagtgaa agcacaccac 660
tttcctagtt gcgccctgct cacaatttgc ttcaaatatt ttgcccaacc tgattcacgg 720
gggacaatag ttaggtgact ctaaaaatcg gcccctttga ccttgcctcc cctgtggttc 780
tagcccccat ggctggtgta accaacgttg ctttccgcac gctgtgccgc gaacaggaaa 840
tgcaacgcac gggaacaatc tcggggctgt acgtctgtga aatggtgact gcgcgtgctc 900
ttgttgagcg caatgagaaa accatgcaca tgaccacctt cgcgccggat gaaaatcccc 960
gaagcttgca gctgtacacg gttgacccga agtacaccta cgaagcggcg aagatgatcg 1020
ttgatgaaaa cttggcggat catattgata tgaactttgg ctgcccggtt ccaaaggtca 1080
cgcgccgtgg tggcggttct gcgattcctt acaagcgccg tttgtttgaa aacatcgttt 1140
ccgcggctgt gaaggctacg gaaggcacgg acattccggt gacggtgaag ttccgcgttg 1200
gtattgatga tgagcaccat actcacttgg atgctggacg cattgctgtc gacgcaggcg 1260
ccaagtccgt agcgcttcac gcccgcactg cggcgcagcg ctattccggt gaggctgatt 1320
ggaacgagat cgcgcgcctg aaggagcatt tggcagatac cggcatccca gttttgggca 1380
atggcgatat tttcgcggca tccgatgcaa cgcgcatgat ggagcaaact ggctgcgatg 1440
gcgtcgtggt tgggcgtggt tgcctgggca ggccttggct ctttgctgag ctgtctgctg 1500
ctgttcgtgg agaagaaatc ccagaggagc ctaccttcgg cgaagttacc caaatcatcc 1560
tgcgccacgc agaactcctc atgcagcatg atggcgaaac caaggggctg cgcgatctgc 1620
gtaagcacat gggttggtac ctgcgcggtt tccctgttgg cggcgaattc cgctccaatc 1680
tggccaaggt ttccacctat gtggagcttg aggatctcct tgcaccatgg gctgactcca 1740
ccgccaaggc agaggacgcg gaaggtgcac gaggtcgaca gggcgctcct gcaaaggtgg 1800
cacttccaga tggctggttg gacgatcctg aggatgccac tgttcctaaa ggcgcagaaa 1860
tggaaaactc cggagggtag ttaatttaat acttaccccc gccaatagtt ggctatcccc 1920
cattattagg tgattcaacc gtaaaggggt gttttaggtc tgcagaaata ccattatcgc 1980
cttatagtgt tgccatgcgt actgcatata gacagcaatt ggatgaattc gcacacgatc 2040
taatcatttt gtgtgatcta actaaggagt gcatggataa ggcgactgat gccctcctgc 2100
gcacttcttt ggcatcggca gaaagcgctt taggcaatgc agaaaagctt gaagagcttc 2160
gtactggatg cgagtctcaa gccgtcgaac ttttggcgct tgaaactcct gtagcccgtg 2220
atcttcgcca ggttgtctcc tccatctaca tcgtcgagga aattacccgt atgggtgctc 2280
tggcaatgca cgtagctaat tccgtgcgtc gccgttaccc cgatccggtg atcccggagg 2340
acatgcgtgg ctatttcaag gagatggccc gcctcgcagc tgacatgaca gatcatattc 2400
gtcagatcct cattgatcct gaaccagatc ttgccctaga gatggctaaa agcgatgacg 2460
cggtggatga tctgcatcag cacatcatgc gtattctcac gctgcgtcct tggcctcacg 2520
acaccaagag cgcggttgat ctgacgctgc tttcccgctt ctacgagcgt tacgccgatc 2580
acacggtaaa cgtggccgcc cgtatcattt acctgtccac cgggctgcac ccggaggagt 2640
acatggaaaa gcgcgagcaa caaagggccg atgccgacat ggagaagcgc tgggccgagc 2700
tggagcggca gttccgcacc agcgagtaaa aagctgcttc tcgacgctaa aagccactac 2760
ccaccatctg aaattaatca ggtggtgggt agtggcttta attcttcgat ttatccgaag 2820
cggccggaga tgtaatcttc ggtttccttc tgatctgggt tttcgaagat cttcttggta 2880
ggtccgattt caaccagacg acctggccta ccggtcgcct ccagggagta gaacgcggtc 2940
tgatcggaca cacgtgcagc ctgctgcatg ttgtgggtca cgatgacgat ggtgaactct 3000
tccttcagct cgtggataag gtcctccaca gccagggttg agattgggtc aagcgcggag 3060
caaggctcgt ccatgaggag gatttctggc tcaaccgcga tcgcgcgagc gatgcacaga 3120
cgctgctgct gaccaccgga gaggccgccg cctggcttgt ccagacgatc cttaacctct 3180
tcccacaggt ttgcgccacg aagagacttc tcagcaactt ccttgagctt cttcttgttc 3240
ttctcgccgg aaagcttcag acctgcaacc acgttgtcct caatggacat ggttgggaat 3300
gggttagcct tctggaagac catgccgatg gtgttacgga ctgcaactgg gtcgatcttg 3360
gagccgtaga tgttctcgcc gtcgagaagg atctcaccct tgacgtatgc acctggggtg 3420
acctcgtgca tacggttgat ggagcgaaga actgtggact tgccacagcc ggatggtccg 3480
atgaatgcgg tgacagagcg tgcgggaact tcgaggttca cgttctgcac tgcgtggaaa 3540
tcaccgtagt agatgttgac atcattgagc ttgagcttcg acatcgcttt agatactcct 3600
gttgtatttt ctgagattgt tattgcttga cagagaactt ggcggagatg attcgtgcgc 3660
caatgttcag gacagcgatg atgagcacca gggtgagggc tgcgccccac agcttgtcca 3720
gcgttgctgg tgcggtgccg gccttgtaca tatcaagcat catcagtgga agggaagcct 3780
gcggaccgct gaatgggttc cagttgatgg cctgggagga accgaccaag accagaactg 3840
gtgcggactc acccatgaca cgtgcgactg cgagcatgac gccggtgacg ataccggaca 3900
gtgcggttgg gagaacgatc tttgcgatgg tcttccactt tggcacgccc agtgcgtagg 3960
acgcttcacg cagatcctga ggaacaacgc ggagcatttc ttcggtgttt cggatgatca 4020
ctggaaccat cagaatcacc agtg 4044
<210> 105
<211> 73
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-phoU_P1
<400> 105
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt gacaggtgga 60
tatcggagta tcc 73
<210> 106
<211> 98
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-phoU_P2
<400> 106
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcaaatcacc taataatggg ggatagcc 98
<210> 107
<211> 100
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-phoU_P3
<400> 107
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaaatgcgta ctgcatatag acagcaattg 100
<210> 108
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物Pcg0007_39-phoU_P4
<400> 108
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt cactggtgat 60
tctgatggtt cc 72
<210> 109
<211> 832
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 在phoU起始密码子前面的Pcg0007_39启动子
<400> 109
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg cgtactgcat atagacagca 120
attggatgaa ttcgcacacg atctaatcat tttgtgtgat ctaactaagg agtgcatgga 180
taaggcgact gatgccctcc tgcgcacttc tttggcatcg gcagaaagcg ctttaggcaa 240
tgcagaaaag cttgaagagc ttcgtactgg atgcgagtct caagccgtcg aacttttggc 300
gcttgaaact cctgtagccc gtgatcttcg ccaggttgtc tcctccatct acatcgtcga 360
ggaaattacc cgtatgggtg ctctggcaat gcacgtagct aattccgtgc gtcgccgtta 420
ccccgatccg gtgatcccgg aggacatgcg tggctatttc aaggagatgg cccgcctcgc 480
agctgacatg acagatcata ttcgtcagat cctcattgat cctgaaccag atcttgccct 540
agagatggct aaaagcgatg acgcggtgga tgatctgcat cagcacatca tgcgtattct 600
cacgctgcgt ccttggcctc acgacaccaa gagcgcggtt gatctgacgc tgctttcccg 660
cttctacgag cgttacgccg atcacacggt aaacgtggcc gcccgtatca tttacctgtc 720
caccgggctg cacccggagg agtacatgga aaagcgcgag caacaaaggg ccgatgccga 780
catggagaag cgctgggccg agctggagcg gcagttccgc accagcgagt aa 832
<210> 110
<211> 4137
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 110
cggcgtcggc aaatttgtag agggcttctt cggtgaaggt tccggcaggc cccaaatagg 60
ccacaacaat tggtgcgtcg ctcatggtta cacagcttaa cccgccgaac taaggtgggt 120
gtccatgtct gttgctcaat tagcgaaccg cttggcccaa ctctcccccg ccgagcatgg 180
ttttgcatgg ttcgaccctg aaatcaccgc tggccatggc gttggcccgt tgcatggcat 240
ggtgattcca gccaaggacc tcaacgatgt cgtgggcatg cccaccgcgt tcggaaatgc 300
atctcggcgc aaggtagcaa cagacaccga tccgttcatc caaaatctca tcgaccgcgg 360
cgcgatcatc gctggcaaaa cccaaaccag cgagctcggc atgacggcgt attgcgagcc 420
catcggcatg gacgcaccca gcaacccggt tttacctggt cacacgcctg gaggttcgtc 480
cggtggtgcg gccgttgctg ttgccaggtc gcttgtcgac gccgcccacg cctcagacgg 540
tggcggctcg atccgggttc cagccgccgc gtgtggcctg gtcgggttta aacccgccca 600
cgattcccga ggtggaaacc cctccacgca ggggtttatc acccgcgacg tggccaccca 660
agtgcgtttg cacgcacttc aaccacgcac cagacgcctg cgcatcggcg ttctcgctga 720
gcccatccat gctaattcgc ttgtcgacgc ccccttcctg agcatcctcg aatccaccgc 780
ccacctcctg gagaaagctg gccacgaaat agtgtccgta cccctccctt atgacgcttg 840
ggcttttgac gcttatacag aagttttcat gatgaaatcc gctggactaa ccaacctggg 900
tagccccatt acaagatggt tgagtgaaca aggccgtagt ctttctcctt ctgatagaca 960
atcatgtgtc aaggcttttg attccgtggc tgagactgta cacggagcat gggacatcga 1020
tgttttatta acccctacct tggcttatgc acctcccaag attgggtact tttcatccat 1080
gccacctgaa gaagacttcc ttgcacaaac caaatggacg ccgtgggcaa cgctgttcaa 1140
catgaccggt ggtgcagcca tcagcgtgcc tgttgaaggt gtcggcattc atcttggtgg 1200
gatacgcgta cgagatgaag acctcttagg attagcagca tttcttgaag gagctgtggc 1260
atgagtagtt cagtaatgtc accgcgtcga ataaaggcgg aaatacttat cgttttagcc 1320
atcacctttg gcatgagcgg tctgcgatca atcctgcgcc tcatcgacga tctcctcgca 1380
ccggttgcgc tcaacgacca gcaagttgca ctgaatgcct caatggcaaa ctccgcatgg 1440
ctggatctcg ccttgcaatt gtgttccgcc ggtgtgctgt tttcctgggg agcgctagcg 1500
atctatctgc ttggcgaacg attccgctgg ataaaaccca aagattggac atggggtgca 1560
ggccttgcag cgctcatcgg cattcccggg ctgatcttct atgcgagtgc cgttcatctg 1620
ggtctttcca aacaggtcgt gcccaccact ttggaaacct ggtgggaaat cccagtgctg 1680
cttatatggt cagccgccaa cgcatttggt gaagagatcg tggtggtcat gtggttcttc 1740
accagactgc gccaactgaa gtggagcgtg ccagcagtca ttgtggcatc ttcagttttg 1800
cgcggttcct accacctcta ccagggaatc tctgcgggtt ttggcaacat catcatggga 1860
gtagcgttcg catatttctt tcaccgcacg ggcaaaatct ggccactggt aatcgctcac 1920
ttcttgattg atgcggtggc ttttgtgggc tactccgcaa ttggcgggaa tttaagctgg 1980
ttagggcttt aaactgaagt tcctatttta ttgggattag aaaaacacga gagatttaag 2040
gcaaatccgc taaatacatc tgtcccgcct aggcgctact ctattaacca tggattcacc 2100
aggacagggc gaaatagccc gcgatagtca aggacgcccg atactggacc ggtatggtcg 2160
gccggttcgg gttcgtccgc aaccgcggca aactcccccg actcctcgaa cccctcccgt 2220
gaatgaaaca agggtctatc agccaaggca aacgccaccg cgccagactc ctccaaggca 2280
gacacctcca cgacaaatgc ctcctaggca aaccccaccg aggcaggttc ccccacaaca 2340
gcagtaccag cagcctggtc agatcggaca ccttcgcccg cagccgccag tgattacaaa 2400
tggtggaggg agacgtcgaa aagcaatatc ttttaagccc cgtggctgcc ttggctcgct 2460
cgcgggcctg gttgctgtgg tgctggtgct cgcttttgtg gtcgcgctgt gggcggatgc 2520
caagctgaat cgcgtggatg cgacgcccgc gacgcaggtg gcgaataccg caggcacgaa 2580
ctggctgctg gtgggttcgg actcgcggca gggtttaagt gatgaggata ttgagcggct 2640
gggtaccggc ggagatatcg gtgtgggtcg tacggacacg atcatggtgt tgcatatgcc 2700
gcgtactggc gagccgacgc tgttgtcgat tccgcgtgat tcttatgtca atgtccctgg 2760
ctggggcatg gataaggcaa acgcggcgtt cactgtgggt ggcccgcagc tgctcacgca 2820
gaccgtggag gaggcgactg gtctgcgaat tgatcactat gcggaaatcg gcatgggtgg 2880
tttggcgaac atggttgatg ccgtgggcgg cgtggaaatg tgtcctgctg agccgatgta 2940
tgatccgctg gccaacctgg atattcaagc cggctgccag gaatttgatg gggcgaccgc 3000
gctgggatac gtgcgcactc gtgccacatc cctgggtgat ctggaccgcg tggtgcgcca 3060
gcgggagttt ttctccgcgc tgctgagtac ggctacgtcc ccgggcacgt tgctgaatcc 3120
gttccgcacc ttcccgatga tctccaacgc ggtgggaaca ttcaccgtcg gcgagggcga 3180
tcacgtgtgg cacctggccc gattggcgct ggcgatgcgc ggaggaatcg tgacggagac 3240
cgtgccgatt gcctcattcg cagattacga tgtgggaaat gttgcgatct gggacgaagc 3300
tggagccgaa gcactattta gctccatgcg ctaaaacccc aggtaatcat tcacaggaag 3360
tgtccagtca actttttggt taattgatta tcttacctag agacttggat acttccctcc 3420
acggattgct aacgatccag ttttatcgtt ggactcatga aaaagatcca catgattcca 3480
gcggccctcc tcatcggttc cctcgcactt gtcagctgcg cgccttcaca acccacccag 3540
aacgcgcaga cggtttcttc tgcaatgatg gaaactccta cttatacttc atcatctagg 3600
tcggtggaat ctaccgcggg gacgatcgcc gccaccgagg aagccgcgat gacgctgctg 3660
gcgaatagtt cgggggatcc gggatctgat tctgggggtg cgcgctatag cctcaattcg 3720
ctcaaggttt ccgagcaagc cgcagcgaac gccgtattaa aggccgtgct taacgacgcc 3780
tcctaccaag agttcgcaga ctcctcctac ttagaaatca caggaacgcc ctccgccgac 3840
ggaacatggg gcatctcatt tggcggacct tccgaatcgg cctccgtgga attctccgac 3900
ggcagcgtca gcttctcccc cgtcgacatg acagtgcccg caacacgatt gcctcagatg 3960
ggcgcatttt atgaaaccct gaccgaagaa caattagaca tgctggagac tggcctcgcg 4020
gtttccacct tggattcgag ccaacaggaa atgcttctgg acttggtctc caactcgatc 4080
ggcctggcag atactgaaac cactgcgact gcaattagtg aaatccgagc aaaactt 4137
<210> 111
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg3210-p1
<400> 111
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt cggcgtcggc 60
aaatttgtag 70
<210> 112
<211> 103
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg3210-p2
<400> 112
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcatcgtgtt tttctaatcc caataaaata gga 103
<210> 113
<211> 93
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg3210-p3
<400> 113
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaaatggatt caccaggaca ggg 93
<210> 114
<211> 80
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg3210-p4
<400> 114
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt agttttgctc 60
ggatttcact aattgcagtc 80
<210> 115
<211> 1342
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 在cg3210前面的Pcg0007_39
<400> 115
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg gattcaccag gacagggcga 120
aatagcccgc gatagtcaag gacgcccgat actggaccgg tatggtcggc cggttcgggt 180
tcgtccgcaa ccgcggcaaa ctcccccgac tcctcgaacc cctcccgtga atgaaacaag 240
ggtctatcag ccaaggcaaa cgccaccgcg ccagactcct ccaaggcaga cacctccacg 300
acaaatgcct cctaggcaaa ccccaccgag gcaggttccc ccacaacagc agtaccagca 360
gcctggtcag atcggacacc ttcgcccgca gccgccagtg attacaaatg gtggagggag 420
acgtcgaaaa gcaatatctt ttaagccccg tggctgcctt ggctcgctcg cgggcctggt 480
tgctgtggtg ctggtgctcg cttttgtggt cgcgctgtgg gcggatgcca agctgaatcg 540
cgtggatgcg acgcccgcga cgcaggtggc gaataccgca ggcacgaact ggctgctggt 600
gggttcggac tcgcggcagg gtttaagtga tgaggatatt gagcggctgg gtaccggcgg 660
agatatcggt gtgggtcgta cggacacgat catggtgttg catatgccgc gtactggcga 720
gccgacgctg ttgtcgattc cgcgtgattc ttatgtcaat gtccctggct ggggcatgga 780
taaggcaaac gcggcgttca ctgtgggtgg cccgcagctg ctcacgcaga ccgtggagga 840
ggcgactggt ctgcgaattg atcactatgc ggaaatcggc atgggtggtt tggcgaacat 900
ggttgatgcc gtgggcggcg tggaaatgtg tcctgctgag ccgatgtatg atccgctggc 960
caacctggat attcaagccg gctgccagga atttgatggg gcgaccgcgc tgggatacgt 1020
gcgcactcgt gccacatccc tgggtgatct ggaccgcgtg gtgcgccagc gggagttttt 1080
ctccgcgctg ctgagtacgg ctacgtcccc gggcacgttg ctgaatccgt tccgcacctt 1140
cccgatgatc tccaacgcgg tgggaacatt caccgtcggc gagggcgatc acgtgtggca 1200
cctggcccga ttggcgctgg cgatgcgcgg aggaatcgtg acggagaccg tgccgattgc 1260
ctcattcgca gattacgatg tgggaaatgt tgcgatctgg gacgaagctg gagccgaagc 1320
actatttagc tccatgcgct aa 1342
<210> 116
<211> 4233
<212> DNA
<213> 谷氨酸棒状杆菌(C. glutamicum)
<400> 116
cttttgccct cggtggccaa acatccgctg agtcctacct tgtcattgac aagatcatcg 60
atgcggcccg caagtccggc gccgacgcca tccaccctgg ctacggcttc ctcgcagaaa 120
acgctgactt cgcagaagca gtcatcaacg aaggcctgat ctggattgga ccttcacctg 180
agtccatccg cgccctcggc gacaaggtca ccgctcgcca catcgcagat accgccaagg 240
ctccaatggc tcctggcacc aaggaaccag taaaagacgc agcagaagtt gtggctttcg 300
ctgaagaatt cggtctccca atcgccatca aggcagcttt cggtggcggc ggacgtggca 360
tgaaggttgc ctacaagatg gaagaagtcg ctgacctctt cgagtccgca acccgtgaag 420
caaccgcagc gttcggccgc ggcgagtgct tcgtggagcg ctacctggac aaggcacgcc 480
acgttgaggc tcaggtcatc gccgataagc acggcaacgt tgttgtcgcc ggaacccgtg 540
actgctccct gcagcgccgt ttccagaagc tcgtcgaaga agcaccagca ccattcctca 600
ccgatgacca gcgcgagcgt ctccactcct ccgcgaaggc tatctgtaag gaagctggct 660
actacggtgc aggcaccgtt gagtacctcg ttggctccga cggactgatc tccttcctcg 720
aggtcaacac ccgcctccag gtggaacacc cagtcaccga agagaccacc ggcatcgacc 780
tggtccgcga aatgttccgc atcgcagaag gccacgagct ctccatcaag gaagatccag 840
ctccacgcgg ccacgcattc gagttccgca tcaacggcga agacgctggc tccaacttca 900
tgcctgcacc aggcaagatc accagctacc gcgagccaca gggcccaggc gtccgcatgg 960
actccggtgt cgttgaaggt tccgaaatct ccggacagtt cgactccatg ctggcaaagc 1020
tgatcgtttg gggcgacacc cgcgagcagg ctctccagcg ctcccgccgt gcacttgcag 1080
agtacgttgt cgagggcatg ccaaccgtta tcccattcca ccagcacatc gtggaaaacc 1140
cagcattcgt gggcaacgac gaaggcttcg agatctacac caagtggatc gaagaggttt 1200
gggataaccc aatcgcacct tacgttgacg cttccgagct cgacgaagat gaggacaaga 1260
ccccagcaca gaaggttatt gtggagatca acggccgtcg cgttgaggtt gcactcccag 1320
gcgatctggc actcggtggc accgctggtc ctaagaagaa ggccaagaag cgtcgcgcag 1380
gtggcgcaaa ggctggcgta tccggcgatg cagtggcagc tccaatgcag ggcactgtca 1440
tcaaggtcaa cgtcgaagaa ggcgctgaag tcaacgaagg cgacaccgtt gttgtcctcg 1500
aggctatgaa gatggaaaac cctgtgaagg ctcataagtc cggaaccgta accggcctta 1560
ctgtcgctgc aggcgagggt gtcaacaagg gcgttgttct cctcgagatc aagtaagttc 1620
ttggattcct gttagtcaaa aaccgggtcg catctgatgt tcagatgtgg cccggtttta 1680
ctatttgtga atacactctg cccctgttgt acattcgaca acaagatttc ggtaaaaagg 1740
tggtcaaaat caggaaatct tgttgtcgaa tgtacaccgc attgtgggct ttcacggttc 1800
gctgcgttga ttgttattta gccagcccgc tccccacact taaccggtga tcaacgagcc 1860
cacatacccg gcgcggtgct ggaacagccc gagaaattca agatcgtttg gggttggctg 1920
ctcgcccgct gatacttctc gttcaatgga gtggatgact gcttcgcgga ttgcggttcc 1980
atcggcgtag ccttgccagt tgatctcggt ggtgtctttc atggtgtgct ccccttgtgg 2040
ttttttattt tctttaacgc taatcggctg acggctttca atgcacaagg taaaatttgt 2100
gtaattgtgt cgtgtggatt tgaaggtttt tgtaaatcta gttaaatcat gaggtcataa 2160
gcttttcggg aggtggtcaa aagatgtcga agctttacgc gggggcaagg atcaatgcac 2220
tgcgccgaac ccaccaactg acccaatcag cattagccga caagcttgat ctctccacga 2280
gctatctcaa ccagttggaa aatgatggcc ggccgcttac tgccacggtg cttttgcaac 2340
tgatgaaagt ctttgatgtt gaggccagtt atttctcccc tgaccggggc acagccacgg 2400
ctacccgact ggcagaaacc ttggctatga atcaggggcc gacgatgtcg atggatgatc 2460
tgttagattt cgcggatcgt ttccctcagt tggcacagca tattatccag cctgctgagg 2520
tcgatcccgc acatagttct gcgcatgatt ttgttcggga ttattttgcc acccacaaga 2580
actacattga ttctcttgat cgccttggtg aggagttggc aaccgccatt ggtcagccgg 2640
ggcttcgggt taccaggctc gcgcagttgc ttgatgcgga gtacaacatc acggtgcgtt 2700
tccgggcgcc agatattact ggccggaggc actttgatcc ccaatcgcgt cagattctgc 2760
tgcggcaaga tctcagcgag gcgcagcagt gttttcagtt ggcggaggaa ttgacgtttc 2820
ttgctcatgc agagctcttg gataccctga ccacagatca accggatctc ccttatgagg 2880
cagctatccg cctggctaag gtgggtctct cccaatactt cgcggctgct gttgtcatgc 2940
cgtacacccg ctttttggaa ttcgcccagg acaagcacta tgacatcgag ttgatctctg 3000
aggcgtttgg agtgtctttc gagtccgcgt gccaccgctt gtctacgctg cagcgttcgg 3060
gggcgtcagg ggtgccgttt ttctttgtgc gctcggaccg ggcaggaaat atttccaaga 3120
ggcaatctgc agctacgttc catttctcgc gaacagacgg cacttgtcct ttgtgggcgc 3180
tgcatcgagc ttttgaacgt caaggaaaca tcacccgcca ggttgctcgc atgccggatg 3240
gccggaccta tttgtggctc gcacgcgcgg tgaaaggtcg aactcatggt ttcgggcatc 3300
ctgctgcgga attcgccatc ggcctgggct gcgatatcag cgaggcacca ggcttggtgt 3360
attcccaagg cctcaatttg gatccagagt ccgccgcaga gatcggccct ggttgtcgga 3420
tctgtcctcg ggagaactgt gtgcagcgtg cattcccacc atcgggtcaa gaatctatcc 3480
gcccagcccc tgtccaactc ctcaactaaa aaggcagcac ttaaaaaaca ctatggaacc 3540
catggaacta aacggcggtc gtttctacgc ccgcaagctc acctcagata atcggattga 3600
tgatgtcccg gcgctctcac tgatccttcc cgatgcggca cagttcatcg cggacgccac 3660
cacagcctgg cacgaggaca ccgaatacac ctgggctatc tgcgagcaga ccagtgttga 3720
actactggct ttggcacgcc ttgatcccaa gcagcgcaca ctcgaggtcg tgcccgcagg 3780
tgatcccgcc acccaattgc ccaatgatcc agtgctcgct ccgaaaacca tcggcgatgc 3840
tgtggcggag ggccgcgaca ccatcaccag atgggcacaa ggctatttag aaatcagcct 3900
cactgattag gttctctccg aaatgggcac cacctccctt tcttccactc cgttgtaggc 3960
agacaatggt cggatcagcg cattgttttc aaattgctcc acgatgtgtg ccgtccaccc 4020
tgacactcgg gccagcacaa acagcggtgt gaaaaagtcg acggggaatc ccagcatgta 4080
ataggcaggg ccggccggga agtcgaggtt gggtttaatg ccagtgcgag cctccatgac 4140
ttcctgcatc gactcataca tgtgcaccca tttttgacca cggtgacgag cagcaaggga 4200
gcgcatggat ttctccatgg aggggaccct gga 4233
<210> 117
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg0800-p1
<400> 117
tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tccagggtcc 60
cctccatgga gaaatc 76
<210> 118
<211> 92
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg0800-p2
<400> 118
tactacgtgg ggacctaagc gtgtattatg gaaacgtctg tatcggataa gtagcgagga 60
gtgttcgtta aaaatgtcga agctttacgc gg 92
<210> 119
<211> 93
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg0800-p3
<400> 119
tatccgatac agacgtttcc ataatacacg cttaggtccc cacgtagtac cacacacaac 60
gcgagaaacg gcaaaaccac aaggggagca cac 93
<210> 120
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> Pcg0007_39-cg0800-p4
<400> 120
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt cttttgccct 60
cggtggccaa ac 72
<210> 121
<211> 1423
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 在cg0800前面的Pcg0007_39
<400> 121
tgccgtttct cgcgttgtgt gtggtactac gtggggacct aagcgtgtat tatggaaacg 60
tctgtatcgg ataagtagcg aggagtgttc gttaaaaatg tcgaagcttt acgcgggggc 120
aaggatcaat gcactgcgcc gaacccacca actgacccaa tcagcattag ccgacaagct 180
tgatctctcc acgagctatc tcaaccagtt ggaaaatgat ggccggccgc ttactgccac 240
ggtgcttttg caactgatga aagtctttga tgttgaggcc agttatttct cccctgaccg 300
gggcacagcc acggctaccc gactggcaga aaccttggct atgaatcagg ggccgacgat 360
gtcgatggat gatctgttag atttcgcgga tcgtttccct cagttggcac agcatattat 420
ccagcctgct gaggtcgatc ccgcacatag ttctgcgcat gattttgttc gggattattt 480
tgccacccac aagaactaca ttgattctct tgatcgcctt ggtgaggagt tggcaaccgc 540
cattggtcag ccggggcttc gggttaccag gctcgcgcag ttgcttgatg cggagtacaa 600
catcacggtg cgtttccggg cgccagatat tactggccgg aggcactttg atccccaatc 660
gcgtcagatt ctgctgcggc aagatctcag cgaggcgcag cagtgttttc agttggcgga 720
ggaattgacg tttcttgctc atgcagagct cttggatacc ctgaccacag atcaaccgga 780
tctcccttat gaggcagcta tccgcctggc taaggtgggt ctctcccaat acttcgcggc 840
tgctgttgtc atgccgtaca cccgcttttt ggaattcgcc caggacaagc actatgacat 900
cgagttgatc tctgaggcgt ttggagtgtc tttcgagtcc gcgtgccacc gcttgtctac 960
gctgcagcgt tcgggggcgt caggggtgcc gtttttcttt gtgcgctcgg accgggcagg 1020
aaatatttcc aagaggcaat ctgcagctac gttccatttc tcgcgaacag acggcacttg 1080
tcctttgtgg gcgctgcatc gagcttttga acgtcaagga aacatcaccc gccaggttgc 1140
tcgcatgccg gatggccgga cctatttgtg gctcgcacgc gcggtgaaag gtcgaactca 1200
tggtttcggg catcctgctg cggaattcgc catcggcctg ggctgcgata tcagcgaggc 1260
accaggcttg gtgtattccc aaggcctcaa tttggatcca gagtccgccg cagagatcgg 1320
ccctggttgt cggatctgtc ctcgggagaa ctgtgtgcag cgtgcattcc caccatcggg 1380
tcaagaatct atccgcccag cccctgtcca actcctcaac taa 1423
- 工程化棒状杆菌属菌株
- 含有棒状杆菌属菌株及其培养物的用于预防、治疗或改善胃肠道疾病的组合物