工程化棒状杆菌属菌株

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

Becker et al.，“Systems level engineering of Corynebacteriumglutamicum-Reprogramming translational efficiency for superior production，”Engineering in Life Sciences 10，430-38，2010

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

Knoppová et al.，Curr.Microbiol.55，234-39，2007

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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