CN101010423A - 生产l-谷氨酸的微生物和生产l-谷氨酸的方法 - Google Patents
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Abstract
本发明涉及具有L-谷氨酸生产能力的棒状杆菌型细菌,其至少以与非突变菌株或野生型菌株相同的生长速率生长,而且其具有的胞内α-酮戊二酸脱氢酶活性小于非突变或野生型菌株的一半,并通过将突变引入到染色体odhA基因的编码区或表达控制区而获得,所述染色体odhA基因编码α-酮戊二酸脱氢酶复合物的Elo亚基。
Description
技术领域
本发明涉及生产L-谷氨酸的微生物和利用所述微生物生产L-谷氨酸的方法。L-谷氨酸广泛用于食品工业,例如,作为生产调味品的原料。
相关技术简述
已通过发酵方法利用生产L-谷氨酸的棒状杆菌型细菌如属于短杆菌属(Brevibacterium)和棒杆菌属(Corynebacterium)的细菌以工业规模常规生产L-谷氨酸。为了提高棒状杆菌型细菌的L-谷氨酸生产能力,将从自然界分离的菌株或这些菌株的人工突变体用作生产L-谷氨酸的棒状杆菌型细菌。
已经报导了许多使用重组DNA技术,通过增强L-谷氨酸生物合成酶活性而提高L-谷氨酸生产能力的技术。例如,已报导了具有扩增的柠檬酸合酶基因的棒状杆菌型细菌(JP07-121228B),和具有扩增的谷氨酸脱氢酶基因的棒状杆菌型细菌(EP955368)。
在物质的发酵生产中,维持细菌细胞的充分生长以充分生产目标物质是有必要的。相应地,培育目标物质的生物合成增强的菌株而不引起细菌生长下降是有必要的。在L-谷氨酸的生产中,已报导降低α-酮戊二酸脱氢酶活性是有利的(JP06-237779A和WO95/34672)。然而,还没有降低α-酮戊二酸脱氢酶活性同时维持足够的L-谷氨酸生产水平的报导。
发明内容
本发明的目的是提供提高棒状杆菌型细菌的L-谷氨酸生产能力的技术,所述棒状杆菌型细菌用于L-谷氨酸的发酵生产。当α-酮戊二酸脱氢酶基因被破坏时,通过TCA循环产生的能量减少,导致细菌生长的阻滞。结果,可能无法获得足够的L-谷氨酸产率。
因此,降解L-谷氨酸的能力降低同时维持正常的生长速率的突变菌株可用于有效地生产L-谷氨酸。通过将突变引入到编码棒状杆菌型细菌α-酮戊二酸脱氢酶复合物的E1o亚基的染色体odhA基因而获得了突变菌株,所述突变菌株具有降低的α-酮戊二酸脱氢酶活性,并且保持了几乎与野生型菌株相同的生长速率。发现利用这样的突变菌株有效地生产L-谷氨酸。
本发明的目的是提供生产L-谷氨酸的棒状杆菌型细菌,其包含
a)胞内α-酮戊二酸脱氢酶活性,该活性小于非突变或野生型菌株的一半,和
b)在染色体odhA基因的编码区或表达控制区中的突变,所述染色体odhA基因编码α-酮戊二酸脱氢酶复合物的E1o亚基,
其中所述细菌几乎以与非突变或野生型菌株相同的生长速率生长。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述odhA基因编码选自下组的蛋白质:
(A)包含SEQ ID NO:10的氨基酸序列的蛋白质,
(B)包含SEQ ID NO:10的氨基酸序列的蛋白质,其中在所述蛋白质中取代、缺失、插入、或添加了一个或几个氨基酸,并且其中所述蛋白质具有α-酮戊二酸脱氢酶复合物的E1o亚基的活性,
(C)包含SEQ ID NO:10的氨基酸37至1257的蛋白质,和
(D)包含SEQ ID NO:10的氨基酸37至1257的蛋白质,其中在所述蛋白质中取代、缺失、插入、或添加了一个或几个氨基酸,并且其中所述蛋白质具有α-酮戊二酸脱氢酶复合物的E1o亚基的活性。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述几个氨基酸是2至20个氨基酸。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述odhA基因选自下组:
(a)包含SEQ ID NO:9的核苷酸443至4213的基因,
(b)能够在严紧条件下与以下物质杂交的基因:包含SEQ ID NO:9的核苷酸443至4213的多核苷酸或由包含SEQ ID NO:9的核苷酸443至4213的多核苷酸制备的探针,并且其中所述基因编码蛋白质,该蛋白质具有α-酮戊二酸脱氢酶复合物的E1o亚基的活性,
(c)包含SEQ ID NO:9的核苷酸551至4213的基因,和
(d)能够在严紧条件下与以下物质杂交的基因:包含SEQ ID NO:9的核苷酸551至4213的多核苷酸或由包含SEQ ID NO:9的核苷酸551至4213的多核苷酸制备的探针,并且其中所述基因编码蛋白质,该蛋白质具有α-酮戊二酸脱氢酶复合物的E1o亚基的活性。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变被引入到编码焦磷酸硫胺素结合区的区域。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变包括氨基酸的缺失,所述氨基酸从下组选择:SEQ ID NO:10所示氨基酸序列中686位的Gly、687位的Leu、688位的Gly、713位的Asn、714位的Asn及其组合。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变被引入到包含SEQ ID NO:9的核苷酸2534至2548的区域。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变包括在包含SEQ ID NO:10的氨基酸698至702的区域中缺失一个或多个氨基酸。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变包括氨基酸的替换,所述氨基酸从下组选择:SEQ ID NO:10所示氨基酸序列中698位的Lys、699位的Leu、700位的Arg、702位的Tyr、及其组合。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变被引入到包含SEQ ID NO:9的核苷酸1094至1111的区域。
本发明的又一目的是提供如上所述的棒状杆菌型细菌,其中所述突变包括在包含SEQ ID NO:10的氨基酸218至224的区域中缺失一个或多个氨基酸。
本发明的又一目的是提供生产L-谷氨酸的方法,包括:
a)在培养基中培养根据权利要求1的棒状杆菌型细菌和
b)从培养基和/或所述细菌收集L-谷氨酸。
本发明的又一目的是提供编码突变的α-酮戊二酸脱氢酶的基因,所述基因选自下组:
(a)包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸443至4213的基因,或者包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸443至4210的基因,
(b)能够在严紧条件下与以下物质杂交的基因:包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸443至4213的多核苷酸、包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸443至4210的多核苷酸、由从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸443至4213制备的探针、或由从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸443至4210制备的探针,并且其中所述基因编码具有α-酮戊二酸脱氢酶活性的蛋白质,通过与E2o和E3亚基形成复合物,该蛋白质的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半,
(c)包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸551至4213的基因,或者包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸551至4210的基因,和
(d)能够在严紧条件下与以下物质杂交的基因:包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸551至4213的多核苷酸、包含从SEQ JD NO:44、46和48组成的组中选择的多核苷酸的核苷酸551至4210的多核苷酸、由从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸551至4213制备的探针、或由从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸551至4210制备的探针,并且其中所述基因编码具有α-酮戊二酸脱氢酶活性的蛋白质,通过与E2o和E3亚基形成复合物,该蛋白质的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半。
本发明的又一目的是提供突变的α-酮戊二酸脱氢酶,其选自下组:
(a)从SEQ ID NO:12、14、16、45、47和49组成的组中选择的蛋白质,
(b)从SEQ ID NO:12、14、16、45、47和49组成的组中选择的蛋白质,其中在所述蛋白质中取代、缺失或添加了一个或几个氨基酸,并且其中通过与E2o和E3亚基形成复合物,所述蛋白质显示的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半,
(c)包含从SEQ ID NO:12、45、47和49组成的组中选择的氨基酸序列的氨基酸37至1256的蛋白质,包含氨基酸序列SEQ ID NO:14的氨基酸37至1255的蛋白质,或者包含氨基酸序列SEQ ID NO:16的氨基酸37至1254的蛋白质,和
(d)包含从SEQ ID NO:12、45、47和49组成的组中选择的氨基酸序列的氨基酸37至1256的蛋白质,包含氨基酸序列SEQ ID NO:14的氨基酸37至1255的蛋白质,或者包含氨基酸序列SEQ ID NO:16的氨基酸37至1254的蛋白质,其中在所述蛋白质中取代、缺失、或添加了一个或几个氨基酸,并且其中通过与E2o和E3亚基形成复合物,所述蛋白质显示的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半。
附图说明
图1显示质粒pBS3的构建。
图2显示质粒pBS4S的构建。
图3显示质粒pBSOAGN的构建。
图4显示质粒pBSOA2-2的构建。
图5显示培养各菌株时L-谷氨酸浓度的变化。
图6显示各菌株的生长速率。
图7显示培养各菌株时残糖(residual sugar)量的变化。
优选实施方案的说明
以下将详细描述本发明。
<1>本发明的棒状杆菌型细菌
本发明的棒状杆菌型细菌具有生产L-谷氨酸的能力,并且几乎以与非突变菌株或野生型菌株相同的生长速率生长。通过将突变引入到odhA基因的编码区或表达控制区,本发明的棒状杆菌型细菌所具有的胞内α-酮戊二酸脱氢酶(以下也称为″α-KGDH″)活性小于非突变菌株或野生型菌株的活性的一半,所述odhA基因编码α-KGDH复合物的E1o亚基。
α-KGDH也称为酮戊二酸脱氢酶或2-酮戊二酸脱氢酶。
在本发明中,棒状杆菌型细菌的例子包括常规的棒状杆菌型细菌,也包括曾经分类为短杆菌属但目前分类为棒杆菌属的细菌(Int.J.Syst.Bacteriol.,41,255(1991)),以及非常接近棒杆菌属细菌的短杆菌属细菌。这样的棒状杆菌型细菌的例子包括下述的细菌。
嗜乙酰乙酸棒杆菌(Corynebacterium acetoacidophilum)
醋谷棒杆菌(Corynebacterium acetoglutamicum)
烷醇棒杆菌(Corynebacterium alkanolyticum)
美棒杆菌( Corynebacterium callunae)
谷氨酸棒杆菌(Coynebacterium glutamicum)
百合花棒杆菌(Corynebacterium lilium)
栖糖蜜棒杆菌(Corynebacterium melassecola)
嗜热产氨棒杆菌(Corynebacterium thermoaminogenes)(Corynebacteriumefficiens)
力士棒杆菌(Corynebacterium herculis)
二歧短杆菌(Brevibacterium divaricatum)
黄色短杆菌(Brevibacterium flavum)
Brevibacterium immariophilum
乳发酵短杆菌(Brevibacterium lactofermentum)(谷氨酸棒杆菌)
玫瑰色短杆菌(Brevibacterium roseum)
解糖短杆菌(Brevibacterium saccharolyticum)
生硫短杆菌(Brevibacterium thiogenitalis)
产氨棒杆菌(Corynebacterium ammoniagenes)
白色短杆菌(Brevibacterium album)
蜡状短杆菌(Brevibacterium cerinum)
嗜氨微杆菌(Microbacterium ammoniaphilum)
棒状杆菌型细菌的具体实例如下:
嗜乙酰乙酸棒杆菌ATCC13870
醋谷棒杆菌ATCC15806
烷醇棒杆菌ATCC21511
美棒杆菌ATCC15991
谷氨酸棒杆菌ATCC13020,ATCC13032,ATCC13060,ATCC13869
百合花棒杆菌ATCC15990
栖糖蜜棒杆菌ATCC17965
Corynebacterium efficiens AJ12340(FERM BP-1539)
力士棒状杆菌ATCC13868
二歧短杆菌ATCC14020
黄色短杆菌ATCC13826,ATCC14067,AJ12418(FERM BP-2205)
Brevibacterium immariophilum ATCC14068
乳发酵短杆菌(谷氨酸棒杆菌)ATCC13869
玫瑰色短杆菌ATCC13825
解糖短杆菌ATCC14066
生硫短杆菌ATCC19240
产氨棒杆菌ATCC6871,ATCC6872
白色短杆菌ATCC15111
蜡状短杆菌ATCC15112
嗜氨微杆菌ATCC15354。
这些菌株可从美国典型培养物保藏中心(ATCC,地址:P.O.Box1549,Manassas,VA 20108,United States ofAmerica)获得。即,给予每个菌株唯一的登记号,该登记号列于ATCC的目录中。可以使用此登记号来定购菌株。根据布达佩斯条约的规定,将AJ12340菌株于1989年10月27日保藏在通商产业省工业技术院生命工学工业技术研究所(the National Institute of Bioscienceand Human-Technology,Agency of Industrial Science and Technology,Ministryof International Trade and Industry)(目前为,独立行政法人产业技术综合研究所特许微生物保藏中心(International Patent Organism Depositary,NationalInstitute of Advanced Industrial Science and Technology),地址为TsukubaCentral 6,1-1,Higashi 1-chome,Tsukuba-shi,Ibaraki-ken,305-5466,日本),并给予登记号FERM BP-1539。根据根据布达佩斯条约的规定,于1989年1月5日将AJ12418菌株保藏在通商产业省工业技术院生命工学工业技术研究所,并给予登记号FERM BP-2205。
在本发明中,″L-谷氨酸生产能力″指当在培养基中培养本发明的棒状杆菌型细菌时,导致在培养基中累积足够量的L-谷氨酸的能力。L-谷氨酸生产能力可以是培育本发明的棒状杆菌型细菌的亲本株的特性,或者是通过突变、基因重组等等所赋予或增强的特性。另外,可以通过向编码α-酮戊二酸脱氢酶复合物的E1o亚基的odhA基因中引入突变而赋予L-谷氨酸生产能力。
赋予L-谷氨酸生产能力的方法的例子包括增强编码L-谷氨酸生物合成酶的基因的表达。涉及L-谷氨酸生物合成的酶的例子包括谷氨酸脱氢酶、谷氨酰胺合成酶、谷氨酸合成酶、异柠檬酸脱氢酶、乌头酸水合酶、柠檬酸合酶、磷酸烯醇丙酮酸羧化酶、丙酮酸羧化酶、丙酮酸脱氢酶、丙酮酸激酶、磷酸烯醇丙酮酸合酶、烯醇化酶、磷酸甘油酸变位酶、磷酸甘油酸激酶、甘油醛-3-磷酸脱氢酶、磷酸丙糖异构酶、果糖二磷酸醛缩酶、磷酸果糖激酶、和磷酸葡糖异构酶。
这些基因表达的增强可以通过将包含这种基因的DNA片段整合到能够在棒状杆菌型细菌中自主复制的适当质粒中,并用所得质粒转化细菌细胞;通过同源重组、接合(conjugation)、移位(transposition)等等将这种基因整合到染色体中;或者将突变引入这种基因的启动子区来实现(WO/0018935)。
当上述基因通过质粒引入或者被整合在染色体上时,用于所述基因表达的启动子可以是任何启动子,只要它能够在棒状杆菌型细菌中起作用。这种启动子的例子包括lac启动子、trp启动子、trc启动子、PS2启动子、和pL启动子。也可以使用所述基因的天然启动子。
经修饰而使柠檬酸合成酶基因、磷酸烯醇丙酮酸羧化酶基因、和/或谷氨酸脱氢酶基因表达增强的微生物的例子包括JP2001-333769A(EP1078989A)、JP2000-106869A (EP955368A)、JP2000-189169A (EP952221A)、和JP2001-333769A (EP1078989A)中公开的那些微生物。
赋予L-谷氨酸生产能力的修饰包括减小或消除酶的活性,所述酶催化合成除L-谷氨酸之外的化合物并由L-谷氨酸生物合成途径分支的反应。这些酶的例子包括异柠檬酸裂解酶、乙酰磷酸转移酶、乙酸激酶、乙酰羟酸合酶、乙酰乳酸合酶、乙酰甲酸转移酶、乳酸脱氢酶、谷氨酸脱羧酶、和1-焦磷酸脱氢酶。
为了减小或消除上述酶的活性,可以将导致酶活性减小或丧失的突变或缺失引入到染色体上酶的基因中。这可以通过例如破坏染色体上编码酶的基因,或者通过修饰基因的表达控制序列如启动子和/或Shine Dargarno(SD)序列来实现。此外,可以通过引入导致氨基酸取代的错义突变、产生终止密码子的无义突变、或者在编码区中添加或缺失一个或两个核苷酸的移码突变、或者通过缺失基因的一部分来减小或消除这些酶的活性(Journal of biologicalChemistry 272:8611-8617(1997))。另外,可以通过构建其编码区被缺失的编码突变酶的基因,并通过同源重组用所得基因替换染色体基因来减小或消除这些酶的活性。
也可以通过筛选对有机酸类似物或呼吸作用抑制剂有抗性的菌株,或者通过筛选对细胞壁合成抑制剂敏感的菌株来赋予L-谷氨酸生产能力。这些方法的例子包括赋予对于苯并吡喃酮(benzopirone)或萘醌(Naphtoquinone)的抗性(JP56-1889A)、赋予对于HOQNO的抗性(JP56-140895A)、赋予对于α-酮丙二酸的抗性(JP57-2689A)、赋予对于胍的抗性(JP56-35981A)、和赋予对于青霉素的敏感性(JP04-88994A)。
这些细菌的具体例子包括下述菌株。
黄色短杆菌AJ11355(FERMP-5007;JP56-1889A)
谷氨酸棒杆菌AJ11355(FERMP-5020;JP56-1889A)
黄色短杆菌AJ1217(FERMP-4318;JP57-2689A)
谷氨酸棒杆菌AJ11218(FERMP-4319;JP57-2689A)
黄色短杆菌AJ11564(FERMP-5472;JP56-140895A)
黄色短杆菌AJ11439(FERMP-5136;JP56-35981A)
谷氨酸棒杆菌H7684(FERMBP-3004;JP04-88994A)
本发明的棒状杆菌型细菌可以通过向编码α-KGDH复合物E1o亚基的染色体odhA基因引入突变,并挑选几乎以与非突变菌株或野生型菌株相同的生长速率生长,其显示的α-KGDH活性小于非突变菌株或野生型菌株的活性的一半的那些菌株,从如前所述具有L-谷氨酸生产能力的棒状杆菌型细菌获得。或者,可以首先将这种突变引入到odhA基因,之后赋予L-氨基酸生产能力。
本发明中,α-KDGH活性指催化α-酮戊二酸(2-酮戊二酸)的氧化脱羧反应,产生丁二酰-CoA的活性。上述反应由α-KDGH复合物催化,所述α-KDGH复合物包括三种亚基,即α-酮戊二酸脱氢酶(E1o,ECl.2.4.2)、二氢硫辛酰胺-S-琥珀酰转移酶(E2o,EC:2.3.1.61)、和二氢硫辛酰胺脱氢酶(E3,EC:1.8.1.4)。也就是说,这三种亚基催化下述各个反应:
E1o亚基:
2-酮戊二酸+[二氢硫辛酰赖氨酸-残基琥珀酰转移酶]硫辛酰赖氨酸=[二氢硫辛酰赖氨酸-残基琥珀酰转移酶]S-琥珀酰二氢硫辛酰赖氨酸+CO2
E2o亚基:
CoA+酶N6-(S-琥珀酰二氢硫辛酰)赖氨酸=丁二酰-CoA+酶N6-(二氢硫辛酰)赖氨酸
E3亚基:
蛋白质N6-(二氢硫辛酰)赖氨酸+NAD+=蛋白质N6-(硫辛酰)赖氨酸+NADH+H+
″α-KDGH复合物″的活性指催化以上三种反应的总反应的活性。
在大肠杆菌(Escherichia coli)中,这三种亚基形成复合物。
在棒状杆菌型细菌的情况中,E1o亚基由odhA基因编码,E3亚基由lpd基因编码(GenBank登录号Y16642;SEQ ID NO:17)。E2o亚基是作为双功能蛋白与E1o亚基一起由odhA基因编码(Usuda et al.,Microbiology 1996 142,3347-3354),还是由GenBank数据库中登记为登录号NC_003450的NCg12126(SEQ ID NO:27)的单独基因所编码存在争议。因此,除了E1o亚基之外,odhA基因可编码E2o亚基。
本发明的棒状杆菌型细菌在染色体odhA基因中具有突变。odhA基因包括,例如,具有编码SEQ ID NO:10所示氨基酸序列的核苷酸序列的基因。odhA基因还包括例如具有编码SEQ ID NO:51所示氨基酸序列的核苷酸序列的基因。此外,尽管SEQ ID NO:9的核苷酸序列中551至553位的″gtg″是Val的密码子,但有可能翻译为Met,并可在该密码子启动翻译。因此,odhA基因还包括具有编码SEQ ID NO:10的氨基酸37至1257的氨基酸序列的核苷酸序列的基因。另外,odhA基因还可以是编码具有SEQ ID NO:10或51所示氨基酸序列,或SEQ ID NO:10的氨基酸37至1257的氨基酸序列的蛋白质的基因,其包括一个或几个氨基酸的取代、缺失、插入或添加,只要其显示α-KGDH复合物E1o亚基的活性。E1o亚基本身的活性可以用Massey etal.(Biochim.Biophys.Acta 38,447-460)的方法测定。此处所用″几个″优选为2至20个,更优选2至10个,特别优选2至5个。
更具体地,odhA基因优选具有SEQ ID NO:9的核苷酸443至4213的核苷酸序列,或SEQ ID NO:9的核苷酸551至4213的核苷酸序列。odhA基因可以具有SEQ ID NO:50所示的核苷酸序列。另外,由于odhA基因的核苷酸序列在棒状杆菌型细菌的菌种或菌株之间可以有所不同,因而odhA基因可以为在严紧条件下与具有SEQ ID NO:9核苷酸443至4213的核苷酸序列的多核苷酸、具有SEQ ID NO:50所示核苷酸序列、或SEQ ID NO:9的核苷酸551至4213的核苷酸序列的多核苷酸、或者由所述核苷酸序列制备的探针杂交的基因,只要所述基因编码的蛋白质显示α-KGDH复合物E1o亚基的活性。如此处所用的″严紧条件″为在此条件下形成所谓的特异性杂合体且不形成非特异性杂合体的条件。难以用任何的数值清楚地表示此条件。然而,严紧条件的例子包括在其条件下,DNA于Southern杂交通常洗涤的盐浓度下互相杂交的条件,即在1×SSC、0.1%SDS在60℃,优选0.1×SSC、0.1%SDS在60℃,更优选0.1×SS℃、0.1%SDS在68℃洗涤一次或优选2-3次。
本发明中,″突变被引入到染色体odhA基因中″包括其中染色体上的odhA基因被突变的odhA基因替换而产生基本相同的条件的情况,在该条件中突变被直接引入到染色体odhA基因中。
上述突变不仅可以在编码区引入,还可以在odhA基因的表达控制区引入。表达控制区包括,例如启动子、SD序列、和操纵子。odhA基因的表达控制区包括SEQ ID NO:9的核苷酸1至442的核苷酸序列。odhA基因的表达控制序列也可以通过基因分析软件如GENETYX或者通过GoIdstein et al.(Prokaryotic promoters in biotechnology.Biotechnol.Annu.Rev.,1995,1,105-128)中公开的方法鉴定。
″α-KGDH活性小于非突变或野生型菌株的α-KGDH活性的一半″优选指培养4小时的本发明棒状杆菌型细菌的每单位细胞(每单位蛋白质重量)的α-KGDH活性小于在相同条件下培养的棒状杆菌型细菌的非突变菌株或野生型菌株活性的一半。这种α-KGDH活性减小的棒状杆菌型细菌的例子包括其中每个细胞的α-KGDH分子数目减少的细菌,和其中每个α-KGDH分子的活性减小的细菌。用作对照的野生型棒状杆菌型细菌的例子包括乳发酵短杆菌ATCC13869菌株。本发明的棒状杆菌型细菌的胞内α-KGDH活性优选小于非突变或野生型菌株所述活性的40%,更优选小于其30%。虽然α-KGDH活性可以降低到不可检测的水平,但优选没有完全消除该活性。可以用Shiio et al.的方法(Isamu Shiio and Kyoko Ujigawa-Takeda,Agric.Biol.Chem.,44(8),1897-1904,1980)测定α-KGDH活性(α-KGDH复合物的活性)。
作为胞内α-KGDH活性减小的结果,棒状杆菌型细菌的L-谷氨酸生产能力提高。本发明中,″提高L-谷氨酸生产能力″优选指当在培养基中培养已在odhA基因中引入突变的棒状杆菌型细菌菌株时,由突变的棒状杆菌型细菌累积在培养基中的L-谷氨酸量大于由野生型或非突变菌株累积的量,或者突变的棒状杆菌型细菌生产L-谷氨酸的速率高于野生型或非突变菌株的生产速率。
短语“与野生型菌株或非突变菌株相比,细菌几乎以相同的生长速率生长”优选指本发明的细菌几乎以与野生型菌株如乳发酵短杆菌ATCC13869相同的生长速率生长。此处,″几乎以相同的生长速率″指野生型或非突变菌株生长速率的80%或更高,更优选90%、或更优选95%或更高。本发明的棒状杆菌型细菌在任何培养温度下都几乎以与野生型或非突变菌株相同的生长速率生长不是必需的。也就是说,在低温下显示足够的生长速率但在高温下显示降低的生长速率的棒状杆菌型细菌,如实施例中所示的GN-2-2菌株也包括于本发明的棒状杆菌型细菌中。
例如,可以通过在培养基中接种相同数目的各菌株的细胞,并在预定时间后比较活细胞数来比较本发明的菌株和野生型或非突变菌株的生长速率。当使用液体培养基时,可以根据例如预定时间后600nm波长的光密度(OD)或细菌细胞的重量计算活细胞数目。当使用固体培养基如平板培养基时,可以根据预定时间后出现的菌落数计算活细胞数目。
向染色体上的odhA基因引入突变的方法的例子包括用X-射线或紫外线照射棒状杆菌型细菌,以及用诱变剂如N-甲基-N’-硝基-N-亚硝基胍处理棒状杆菌型细菌。也可以通过制备突变的odhA基因,并通过同源重组技术用此突变的odhA基因替换染色体odhA基因将突变引入到染色体上的odhA基因中。这种突变的odhA基因可以通过易错PCR、DNA改组、或StEP-PCR(FirthAE,Patrick WM;Bioinformatics.2005 Jun 2;Statistics of protein libraryconstruction.),或重叠-延伸PCR(overlap-extension PCR)(Urban,A.,Neukirchen,S.and Jaeger,K.E.,A rapid and efficient method for site-directedmutagenesis using one-step overlap extension PCR.Nucleic Acids Res,25,2227-8.(1997))制备。
同源重组可以通过使用线性DNA的称为″Red-驱动的整合(Red-drivenintegration)″的方法(Datsenko,K.A.,PNAS,97(12),6640-6645,2000),或者通过使用具有温度敏感性复制起点的质粒的方法(USP6303383,和JP05-007491A)来进行。向染色体odhA基因引入突变也可以通过使用不能在棒状杆菌型细菌细胞中复制的质粒或能够通过接合转移到棒状杆菌型细菌的质粒来进行。
用于棒状杆菌型细菌的这种温度敏感性质粒的例子包括p48K和pSFKT2(JP2000-262288A)、pHSC4(法国专利Laid-open公布号2667875,1992和JP5-7491A)、pBS5T等等。这些质粒在棒状杆菌型细菌中可以在至少25℃的温度自主复制,但不能在37℃的温度自主复制。含有pHSC4的AJ12571菌株依据布达佩斯条约的规定于1991年8月26日保藏在通商产业省工业技术院生命工学工业技术研究所(National Institute of Bioscience and HumanTechnology,Agency of Industrial Science and Technology,Ministry ofInternational Trade and Industry)(目前为产业技术综合研究所特许微生物保藏中心International Patent Organism Depositary,National Institute of AdvancedIndustrial Science and Technology,其地址为Tsukuba Central 6,1-1,Higashi1-chome,Tsukuba-shi,Ibaraki-ken 305-5466,Japan),给予保藏号FERMBP-3524。
不能在棒状杆菌型细菌细胞中复制的质粒优选为可在埃希氏菌属的细菌细胞中复制的质粒,其例子包括pHSG299(Takara Bio)和pHSG399(TakaraBio)。能够通过接合转移到棒状杆菌型细菌的质粒的例子包括pK19mobsacB(J.Bacteriology 174:5462-65(1992))。
将突变的odhA基因插入到温度敏感性质粒或者不能在棒状杆菌型细菌中复制的质粒中,所获得的重组质粒用于转化棒状杆菌型细菌。转化可以通过常规方法进行。例如,可以使用用氯化钙处理感受体细胞以增加DNA通透性的方法,其已报导用于大肠杆菌(Mandel,M.and Higa,A.,J.MoI.Biol.,53,159(1970)),和使用从生长细胞制备的感受态细胞以引入DNA的方法,其已报导用于枯草芽孢杆菌(Bacillus subtilis)(Duncan,C.H.,Wilson,G.A.andYoung,RE.,Gene,1,153(1977))。除了这些方法外,还可以使用将重组DNA引入到原生质体-样或原生质球-样(protoplast-or spheroplast-like)受体细胞中的方法,其已报导可用于枯草芽孢杆菌、放线菌类(actinomytetes)和酵母(Chang,S.and Choen,S.N.,Molec.Gen.Genet.,168,111(1979);Bibb,M.J.,Ward,J.M.and Hopwood,O.A.,Nature,274,398(1978);Hinnen,.A.Hicks,J.B.and Fink,G.R.,Proc.Natl.Sci.USA,75,1929(1978))。此外,棒状杆菌型细菌的转化还可以通过电脉冲法进行(Sugimoto et al,JP2-207791A)。
在使用在棒状杆菌型细菌的细胞中无能的质粒的情况下,染色体上的野生型odhA基因由质粒上的突变odhA基因所替换。
在使用温度敏感性质粒的情况下,在热敏感性复制起点不起作用的温度(例如25℃)培养转化的菌株,以获得引入了所述质粒的菌株。然后,在高温下培养转化体以除去温度敏感性质粒,并涂布在含抗生素药物如卡那霉素的平板培养基上。虽然去除了所述质粒的菌株不能在含这种抗生素药物的平板上生长,但其中染色体odhA基因被突变的odhA基因替换的少数菌株能够生长并出现菌落。
在这种如上所述将含突变odhA基因的重组DNA整合到染色体DNA中的菌株中,重组DNA导致与原来存在于染色体上的odhA基因重组,并将染色体odhA基因与突变odhA基因的融合基因插入到染色体,以使重组DNA的其它部分(载体区段、温度敏感性复制起点和药物抗性标记)存在于融合基因之间。然后,为了在染色体DNA上只留下突变的odhA基因,将一个拷贝的odhA基因连同载体区段(包括温度敏感性复制起点和药物抗性标记)从染色体DNA去除。在这种情况下,将正常odhA基因留在染色体DNA上,并将突变的odhA基因从染色体DNA切除,或者相反,将突变的odhA基因留在染色体DNA上,将正常odhA基因从染色体DNA切除。然后,可以使用PCR、Southern杂交等选择在染色体上只留下突变odhA基因的菌株。
可以使用编码果聚糖蔗糖酶的sacB基因作为重组标志物来进行同源重组。编码果聚糖蔗糖酶的sacB基因用于有效选择菌株,在所述菌株中,染色体odhA基因被突变的odhA基因所替换,并从染色体消除(cure)载体部分(Schafer,A.et al.,Gene 145(1994)69-73)。也就是说,在棒状杆菌型细菌的情况中,当果聚糖蔗糖酶表达时,通过蔗糖同化产生的果聚糖对细菌来说变得致命,并由此细菌不能生长。因此,通过在含蔗糖平板上培养,可以选择其中在载体中的突变odhA基因和染色体odhA基因之间发生取代,并从中消除了载体的其它部分的菌株。
sacB基因的例子包括下述基因。
枯草芽孢杆菌(Bacillus subillus):sacB GenBank登录号X02730(SEQ IDNO:19)
解淀粉芽孢杆菌(Bacillus amyloliqufaciens):sacB GenBank登录号X52988
运动发酵单胞菌(Zymomonas mobilis):sacB GenBank登录号L33402
嗜热脂肪芽孢杆菌(Bacillus stearothermophilus):surB GenBank登录号U34874
Lactobacillus sanfranciscensis:frfA GenBank登录号AJ508391
Acetobacter xylinus:lsxA GenBank登录号AB034152
Gluconacetobacter diazotrophicus:lsdA GenBank登录号L41732
<2>本发明的突变的odhA基因
将要引入到染色体odhA基因中的突变没有具体限制,只要其为将α-KGDH活性降至低于非突变菌株或野生型菌株α-KGDH活性的一半,但不引起细菌生长延缓的突变。所述突变的具体例子包括下述突变。
(1)焦磷酸硫胺素结合区中的突变
该突变被引入到辅酶焦磷酸硫胺素的结合区。焦磷酸硫胺素结合区指相应于由odhA基因(SEQ ID NO:9)的核苷酸2498至2584编码区的区域。该区域的氨基酸序列如SEQ ID NO:10中氨基酸686至714所示。该区域中突变的例子包括导致一个或多个氨基酸残基缺失的突变,所述氨基酸残基选自下组:686位的Gly、687位的Leu、688位的Gly、713位的Asn、和714位的Asn。
也可以在odhA基因(SEQ ID NO:9)的核苷酸2534至2548的区域引入焦磷酸硫胺素结合区中的突变。该突变命名为GN型突变。GN型突变优选为导致一个或多个氨基酸残基缺失和/或取代的突变,所述氨基酸残基选自下组:SEQ ID NO:10中698位的Lys、699位的Leu、700位的Arg、701位的Gly、和702位的Tyr。这种突变的例子为缺失SEQ ID NO:10中701位的Gly的突变。具有该突变的突变odhA基因的例子包括具有SEQ ID NO:13的核苷酸443至4213的核苷酸序列的基因,和具有SEQ ID NOS:44、46或48的核苷酸443至4210的核苷酸序列的基因,和由这些基因编码的突变α-KGDH蛋白如SEQ ID NOS:14、45、47、和49所示。具有该突变的突变odhA基因的例子还包括具有SEQ ID NO:13的核苷酸551至4213的核苷酸序列的基因和具有SEQ ID NOS:44、46或48的核苷酸551至4210的核苷酸序列的基因,由这些基因编码的突变α-KGDH蛋白如SEQ ID NOS:14的氨基酸37至1255、和SEQ ID NOS:45、47、和49的氨基酸37至1256所示。在SEQ ID NO:13的核苷酸序列中引入在SEQ ID NO:9的2538位缺失″t″和在SEQ ID NO:9的2543至2547位缺失″ggcta″的突变。
另外,具有突变的突变odhA基因也是优选的,所述突变导致从SEQ IDNO:10中698位的Lys、699位的Leu、700位的Arg、和702位的Tyr组成的组中选择的一个或多个氨基酸残基被另一种氨基酸所替换。所述″另一种氨基酸″没有具体限制,只要其不同于原来的氨基酸,并选自天然氨基酸如Lys、Glu、Thr、Val、Leu、Ile、Ser、Asp、Asn、Gln、Arg、Cys、Met、Phe、Trp、Tyr、Gly、Ala、Pro、和His。然而,优选698位的Lys被除碱性氨基酸如Arg和His之外的氨基酸所替换,699位的Leu被除疏水性脂肪族氨基酸如Ile和Val之外的氨基酸所替换,700位的Arg被除碱性氨基酸如Lys和His之外的氨基酸所替换,702位的Tyr被除羟基氨基酸如Ser和Thr之外的氨基酸所替换。特别优选698位的Lys被疏水性脂肪族氨基酸如Ile、Leu、或Val所替换,699位的Leu被羟基氨基酸如Ser、Thr或Tyr所替换,700位的Arg被含硫氨基酸如Cys或Met所替换,702位的Tyr被疏水性脂肪族氨基酸如Ile、Leu、或Val所替换。
GN型突变可以同时导致701位的Gly的缺失和从SEQ ID NO:10中698位的Lys、699位的Leu、700位的Arg、和702位的Tyr组成的组中选择的一个或多个氨基酸残基的替换。这种突变odhA基因的例子包括具有SEQ IDNOS:44、46、和48的核苷酸443至4210的核苷酸序列的基因。由这些突变odhA基因编码的突变α-KGDH蛋白的氨基酸序列如SEQ ID NOS:45、47、和49所示。这种突变odhA基因的例子还包括具有SEQ ID NOS:44、46、和48的核苷酸551至4210的核苷酸序列的基因。由这些突变odhA基因编码的突变α-KGDH蛋白的氨基酸序列如SEQ ID NOS:45、47、和49的氨基酸37至1256所示。然而,在焦磷酸硫胺素结合区具有突变的突变odhA基因不限于这些例子。
(2)2-2型突变
这种类型的突变优选为引入到odhA基因(SEQ ID NO:9)的核苷酸1094至1111区域中的突变,其导致选自SEQ ID NO:10中选自218位的Asp、219位的Val、220位的Ile、221位的Asp、222位的Gly、223位的Lys、和224位的Pro的一个或多个氨基酸残基的缺失和/或取代。2-2型突变优选为在SEQID NO:10中221位缺失Asp的突变。具有此突变的odhA基因的例子包括具有SEQ ID NO:11的核苷酸443至4213的核苷酸序列的odhA基因,或者具有SEQ ID NO:11的核苷酸551至4213的核苷酸序列的odhA基因。由这些基因编码的突变α-KGDH的氨基酸序列分别如SEQ ID NO:12或SEQ ID NO:12的37至1256位氨基酸所示。SEQ ID NO:11的核苷酸序列具有在SEQ IDNO:9所示核苷酸序列中缺失1094至1098的″gacgt″并用″ggcc″替换1110至1111的″ag″的突变。
2-2型突变还优选为替换选自219位的Val、220位的Ile、221位的Asp、222位的Gly、和223位的Lys的一个或多个氨基酸的突变。虽然替换这些氨基酸的氨基酸没有具体限制,但优选用除脂肪族疏水性氨基酸如Ile和Leu之外的氨基酸替换219位的Val,用除脂肪族疏水性氨基酸如Leu之外的氨基酸替换220位的Ile,用除酸性氨基酸如Glu之外的氨基酸替换221位的Asp,以及用除简单氨基酸如Ala之外的氨基酸替换222位的Gly。更优选用碱性氨基酸如His、Arg和Lys替换219位的Val、220位的Ile、和221位的Asp,用氨基酸如Asp和Gln替换222位的Gly,以及用氨基酸如Gly和Ala替换223位的Lys。
然而,具有2-2型突变的突变odhA基因不限于这些例子。
(3)GN2-2型突变
该突变同时包括GN型突变和2-2型突变。具有此突变的odhA基因的例子包括具有SEQ ID NO:15的核苷酸443至4213的核苷酸序列的odhA基因,或者具有SEQ ID NO:15的核苷酸551至4213的核苷酸序列的odhA基因。由这些基因编码的突变α-KGDH的氨基酸序列分别如SEQ ID NO:16或SEQID NO:16的氨基酸37至1254所示。然而,具有GN2-2型突变的突变odhA基因不限于这些例子。
用于本发明的其他类型的突变odhA基因可以利用ygg突变株,如以下实施例5中所述的ATCC13869-L来筛选。也就是说,将随机突变引入到odhA基因中,并用于转化ygg突变菌株。从转化体选择几乎以与野生型菌株或非突变菌株相同的生长速率生长,并显示小于野生型菌株或非突变菌株的一半α-KGDH活性的菌株,之后测序。由此可以获得突变的odhA基因。
由上述突变odhA基因编码的α-KGDH复合物的突变E1o亚基是蛋白质,其通过与α-KGDH复合物的E2o和E3亚基形成复合物,具有小于野生型或非突变菌株α-KGDH活性一半的α-KGDH活性,并且在棒状杆菌型细菌中表达时不引起棒状杆菌型细菌严重的生长延缓。
认为即便是一个或几个氨基酸被其它氨基酸所替换,突变E1o亚基的这些特性也将保持,所述一个或几个氨基酸是除上述GN型或2-2型突变体中被替换的特定氨基酸之外的氨基酸,例如不影响酶活性的氨基酸。因此,在以上提到的突变E1o亚基蛋白的氨基酸序列(例如,SEQ ID NOS:12、14、16、45、47和49,SEQ ID NOS:12、45、47和49的氨基酸37至1256,SEQ IDNO:14的氨基酸37至1255,或者SEQ ID NO:16的氨基酸37至1254)中,可以替换除GN型或2-2型突变体中被取代的特定氨基酸之外的其它一个或几个氨基酸,只要所编码的蛋白质通过与α-KGDH E2o亚基和E3亚基蛋白一起形成复合物,显示小于野生型α-KGDH复合物一半的α-KGDH活性。此处″几个″优选指2至20个,更优选2至10个,并特别优选2至5个。这种氨基酸取代可以是通过源于细菌个体差异和菌种差异的天然存在的突变所引起的取代,其中odhA基因由所述细菌衍生。例如,具有编码氨基酸序列SEQID NO:51的核苷酸序列的突变odhA基因也可用于本发明,其中对应于上述GN-型突变或2-2型突变的氨基酸被缺失和/或取代。氨基酸序列SEQ ID NO:51中对应于GN-型突变或2-2型突变的这些氨基酸可以很容易地通过比对SEQ ID NOS:10和51的氨基酸序列来鉴定。上述取代优选为保守性取代。在芳香族氨基酸的情况中,保守性取代指phe、trp、和tyr之间的相互取代。在疏水性氨基酸的情况中,保守性取代指leu、ile、和val之间的相互取代。在极性氨基酸的情况中,保守性取代指gln和asn之间的相互取代。在碱性氨基酸的情况中,保守性取代指arg、lys、和his之间的相互取代。在酸性氨基酸的情况中,保守性取代为asp和glu之间的相互取代。在含羟基氨基酸的情况中,保守性取代指ser和thr之间的取代。保守性取代还包括:ser或thr取代ala,gln、his或lys取代arg;glu、gln、lys、his或asp取代asn;asn、glu或gln取代asp;ser或ala取代cys;asn、glu、lys、his、asp或arg取代gln;asn、gln、lys或asp取代glu;val取代gly;asn、lys、gln、arg或tyr取代his;leu、met、val或phe取代ile;ile、met、val或phe取代leu;asn、glu、gln、his或arg取代lys;ile、leu、val或phe取代met;trp、tyr、met、ile或leu取代phe;thr或ala取代ser;ser或ala取代thr;phe或tyr取代trp;his、phe、或trp取代tyr;以及met、ile、或leu取代val。
当E1o亚基和E2o亚基由单个odhA基因编码时,α-KGDH复合物可以包括SEQ ID NO:10(或SEQ ID NO:10的氨基酸37至1257)的蛋白质和SEQID NO:18的蛋白质。当E1o亚基由odhA基因编码且E2o亚基由SEQ ID NO:27的基因编码时,α-KGDH复合物可以包括SEQ ID NO:10的蛋白质、SEQID NO:18的蛋白质、和SEQ ID NO:28的蛋白质。
本发明的突变odhA基因是编码上述突变α-KGDH E1o亚基的基因。本发明的突变odhA基因可以是在严紧条件下,与各自具有SEQ ID NO:11、13、或15的核苷酸443至4213的核苷酸序列的多核苷酸,与各自具有SEQ ID NO:44、46或48的核苷酸443至4210的核苷酸序列的多核苷酸,与SEQ ID NO:11、13、或15的核苷酸551至4213的核苷酸序列,或与SEQ ID NO:44、46、或48的核苷酸443至4210的核苷酸序列,或与从所述序列制备的探针杂交的基因,只要该基因编码通过与E2o亚基和E3亚基蛋白形成复合物,显示小于野生型α-KGDH复合物一半的α-KGDH活性的蛋白质。严紧条件的例子包括在其条件下,DNAs于Southem杂交通常洗涤的盐浓度下互相杂交的条件,即在1×SSC、0.1%SDS在60℃,优选0.1×SSC、0.1%SDS在60℃,更优选0.1×SSC、0.1%SDS在68℃的条件下洗涤一次或优选2-3次。
<3>生产L-谷氨酸的方法
可以通过在培养基中培养本发明的棒状杆菌型细菌,导致L-谷氨酸在培养基中和/或在细菌细胞中累积,并从培养基和/或细菌细胞收集L-谷氨酸来生产L-谷氨酸。在本发明的生产方法中,优选通过例如在25至40℃培养本发明的棒状杆菌型细菌8至120小时来生产L-谷氨酸。如果在低温下显示足够的生长速率但在高温下显示减小的生长速率的菌株,如实施例中所示的GN-2-2菌株用于L-谷氨酸生产,则优选在低温下如25至30℃培养这种菌株8至30小时,以使所述菌株能够生长;然后在高温下如34至40℃将所得细菌细胞保温16至48小时,使所述菌株能够生产L-谷氨酸。
所述培养基可以是包含碳源、氮源、无机盐、和任选有机微量营养物如氨基酸和维生素的常规培养基。合成培养基或天然培养基均可使用。可以使用任何种类的碳源和氮源,只要它们能够被要培养的菌株所利用。
糖类如葡萄糖、丙三醇、果糖、蔗糖、麦芽糖、甘露糖、半乳糖、淀粉水解物、和糖蜜可以用作碳源。此外,有机酸如醋酸和柠檬酸、以及醇如乙醇也可以单独或组合作为碳源使用。氨、铵盐如硫酸铵、碳酸铵、氯化铵、磷酸铵、和乙酸铵、硝酸盐等可以用作氮源。氨基酸、维生素、脂肪酸、核酸、包含蛋白胨的物质、酪蛋白氨基酸、酵母提取物、和大豆蛋白分解产物可以以少量使用作为有机营养物。当使用其生长需要氨基酸等的营养缺陷型突变株时,优选添加这种所需的营养物。磷酸盐、镁盐、钙盐、铁盐、锰盐等可以作为无机盐使用。
优选通过控制发酵温度并将培养基的pH调节至3到9来进行需氧培养。培养期间pH降低时,通过添加碱如碳酸钙或氨气来中和培养基。培养约10至约120小时使培养基中累积大量的L-谷氨酸。
此外,可以使用液体培养基进行培养,所述液体培养基被调节至生产的L-谷氨酸结晶并析出的条件。L-谷氨酸结晶的条件包括pH5.0至4.0,优选pH4.5至4.0,更优选pH4.3至4.0,特别优选pH4.0(EP1233069、EP1233070)。
培养完成后,可以通过常规方法从培养基收集L-谷氨酸。例如可以通过从培养基去除细菌细胞并浓缩L-谷氨酸或者通过使用离子交换层析来收集L-谷氨酸。当在L-谷氨酸结晶并析出的条件下进行培养时,可以例如通过离心或过滤来收集结晶的L-谷氨酸。在这种情况下,也可以在溶解的L-谷氨酸结晶后收集溶解在培养基中的L-谷氨酸。
实施例
以下本发明将通过参考以下非限制性实施例更具体地解释本发明。
实施例1
<1>携带sacB基因的载体的构建
(A)pBS3的构建
使用枯草芽孢杆菌的染色体DNA作为模板,SEQ ID NOS:21和22的寡核苷酸作为引物通过PCR,获得了sacB基因(SEQ ID NO:19)。按照以下程序用LAtaq(可由TaKaRa获得)进行PCR:94℃预变性5分钟的一个循环;94℃变性30秒、49℃退火30秒、72℃延伸2分钟的25个循环。用常规方法纯化所得PCR产物,然后用BglII和BamHI消化并平端化。将所述片段插入到已用AvaII消化且平端化的pHSG299中。所得DNA用于转化大肠杆菌JM109(可由Takara Bio获得)的感受态细胞。然后,将转化的细菌细胞涂布到含25μg/ml卡那霉素(以下缩写为″Km″)的LB琼脂平板上,保温一夜。此后,选择出现的菌落作为转化体。从所得转化体分离质粒,并将具有目的PCR产物插入物的质粒命名为pBS3。图1显示构建pBS3的方法。
(B)pBS4S的构建
使用不引起氨基酸取代的交换(cross-over)PCR来破坏pBS3上卡那霉素抗性基因中的SmaI识别位点。首先使用pBS3作为模板,SEQ ID NOS:23和24的合成DNAs作为引物进行PCR,以获得卡那霉素抗性基因的N末端片段。另一方面,为了获得卡那霉素抗性基因的C末端片段,使用pBS3作为模板,SEQ ID NOS:25和26的合成DNAs作为引物进行PCR。按照以下程序用Pyrobest DNA聚合酶(可由TaKaRa Bio获得)进行PCR:98℃预变性5分钟;及98℃变性10秒、57℃退火30秒、72℃延伸1分钟的25个循环。SEQ ID NOS:24和25相互之间部分互补,而且不包含SmaI识别位点。然后,为了获得不含SmaI识别位点的突变卡那霉素抗性基因的全长片段,将上述N末端和C末端基因产物以基本上等摩尔的量互相混合。使用所述基因产物作为模板,SEQ ID NOS:23和26的合成DNAs作为引物进行PCR,以获得引入了突变的Km抗性基因。按照以下程序用Pyrobest DNA聚合酶(可由TaKaRaBio获得)进行PCR:98℃预变性5分钟;及98℃变性10秒、57℃退火30秒、72℃延伸1.5分钟的25个循环。
用常规方法纯化PCR产物,接下来用BanII消化,然后插入到pBS3的上述BanII识别位点中。所得质粒用于转化大肠杆菌JM109(可由Takara Bio获得)的感受态细胞。即,将转化的细菌细胞涂布到包含25μg/ml卡那霉素的LB琼脂培养基上,并保温一夜。此后,选择出现的菌落作为转化体。从所得转化体分离质粒,并将具有目的PCR产物插入物的质粒命名为pBS4S。图2显示构建pBS4S的方法。
<2>将odhA突变(GN-型)引入到谷氨酸棒杆菌ATCC13869菌株中
编码棒状杆菌型细菌α-酮戊二酸脱氢酶的odhA的序列已有报导(Microbiology 142,3347-3354(1996),GenBank登录号D84102)。
本发明的发明人成功培育的生产L-谷氨酸的细菌菌株GN的核苷酸序列分析显示,该菌株在odhA基因(SEQ ID NO:9)的核苷酸序列中具有核苷酸2538和核苷酸2543至2547的缺失,如表1所示。表1还显示了由突变odhA基因编码的突变E1o亚基的氨基酸序列。
表1
菌株 | odhA基因的核苷酸序列 |
ATCC13869 | GCT AAG CTG CGTGGC TAC GAC GTC GGA GGC ACC ATC |
OAGN | GCT AAG C-G CGT--- --C GAC GTC GGA GGC ACC ATC |
菌株 | E1o亚基的氨基酸序列 |
ATCC13869 | Ala Lys Leu Arg Gly Tyr Asp Val Gly Gly Thr Ile |
OAGN | Ala Lys Arg Val--- ---Asp Val Gly Gly Thr Ile |
然后,将该突变odhA基因引入到谷氨酸棒杆菌2256菌株(ATCC13869)中并评估。首先制备用于将这些突变引入到谷氨酸棒杆菌2256菌株(ATCC13869)中的质粒。使用Bacterial Genomic DNA Purif.试剂盒(由MSTechnosystems Co.,Ltd.生产)从上述GN菌株提取染色体DNA,使用所获得的染色体DNA作为模板,SEQ ID NOS:1和2的寡核苷酸作为引物进行PCR。按照以下程序使用Pyrobest聚合酶(可由TaKaRa Bio获得)进行PCR以扩增约2.75kb的片段:98℃变性10秒、50℃退火30秒、72℃延伸3分钟的30个循环。所述引物在5’端包含BamHI识别序列,并设计用于扩增GenBank登录号D84102序列的核苷酸1521至4270的区域。扩增的片段用BamHI消化,并连接到用BamHI消化的pBS4S载体(Ligation kit Ver.2,使用可从Takara Bio获得的产品),由此获得质粒pBSOAGN(图3显示构建方法)。
通过电脉冲法(JP02-207791A)将pBSOAGN导入到谷氨酸棒杆菌ATCC13869菌株中,将所获得的细菌细胞涂布到含25μg/ml卡那霉素的CM-Dex琼脂培养基(5g/l葡萄糖、10g/l多蛋白胨、10g/l酵母提取物、1g/lKH2PO4、0.4g/l MgSO4·7H2O、0.01g/l FeSO4·7H2O、0.01g/l MnSO4·4-5H2O、3g/l尿素、1.2g/l大豆蛋白水解物、和20g/l琼脂,pH调节到7.5)。25℃培养后,通过使用PCR确认出现的菌落为一次重组菌株(once recombinantstrain),在此菌株中,pBSOAGN通过同源重组整合在染色体上。使用候选菌株的染色体DNA作为模板,具有特异于pBS4S的序列(SEQ ID NO:3)的核苷酸序列的寡核苷酸和具有特异于染色体上的序列(SEQ ID NO:4)的核苷酸序列的寡核苷酸作为引物进行PCR。也就是说,由于pBS4S的序列在非重组菌株的染色体上不存在,因而如果所述候选菌株不是重组菌株,则没有片段通过PCR被扩增。将所获得的一次重组菌株在25℃在含25μg/ml卡那霉素的CM-Dex液体培养基中培养一天,适当稀释所得培养物并涂布到S10平板上,所述S10平板具有上述CM-Dex培养基的组成,其中只是用10g/l的蔗糖替换5g/l的葡萄糖。选择了几个生长在S10平板上并显示卡那霉素敏感性的菌株,然后通过Sanger的方法(J.MoI.Biol.,143,161(1980))确认odhA序列的核苷酸序列。使用BigDye终止物测序试剂盒(由Applied Biysystems生产)通过遗传Analyzer ABI310(由Applied Biosystems生产)分析核苷酸序列。将如此获得的携带GN-型突变odhA基因的菌株命名为ATCC13869 OAGN。
<3>将2-2型突变odhA基因导入谷氨酸棒杆菌ATCC13869菌株中
本发明的发明人成功培育的生产L-谷氨酸的″2-2″菌株的核苷酸序列分析显示,该菌株在odhA基因(SEQ ID NO:9)中包含核苷酸1094至1098的缺失和以″ggcc″替换处于1110至1111的″ag″,如表2所示。表2还显示了由突变odhA基因编码的突变E1o亚基的氨基酸序列。
表2
菌株 | odhA基因的核苷酸序列 |
ATCC13869 | AAC TCC TAC GAC GTC ATC GAC GGC AAG CCA ACC CTG |
OA2-2 | AAC TCC TAC---CAT CGA CGG CAG GCC CCA ACC CTG |
菌株 | E1o亚基的氨基酸序列 |
ATCC13869 | Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro Thr Leu |
OA2-2 | Asn Ser Tyr---His Arg Arg Gln Ala Pro Thr Leu |
然后,将该突变的odhA基因导入到谷氨酸棒杆菌2256菌株(ATCC13869)中并评估。首先制备用于将这些突变引入到谷氨酸棒杆菌2256中的质粒。用Bacterial Genomic DNA Purif.试剂盒(由MS Technosystems Co.,Ltd.生产)从2-2菌株分离染色体DNA,使用此染色体DNA作为模板,SEQ ID NOS:5和6的寡核苷酸作为引物进行PCR。按照以下程序使用TaKaRa Ex Taq(可由TaKaRa Bio获得)进行PCR以扩增约2.0kb的片段:94℃变性30秒、55℃退火10秒、72℃延伸2分钟的25个循环。所述引物在5’端包含BamHI识别序列,该引物设计用于扩增GenBank登录号D84102核苷酸序列的核苷酸51至2150区域。扩增的片段用BamHI消化,并使用TakaraBio的Ligation kit Ver.2连接到经BamHI消化的pBS4S载体,由此获得质粒pBSOA2-2(图4显示构建图)。实施与实施例1<2>中所述相同的步骤以获得携带2-2型突变odhA基因的菌株(ATCC13869 OA2-2菌株)。
<4>将2-2型突变引入到ATCC13869 OAGN菌株的GN型odhA基因中
按照与实施例1<2>中所述相同的方法,将实施例1<3>中构建的质粒pBSOA2-2导入实施例1<2>中制备的ATCC13869 OAGN菌株中,以获得odhA基因中同时携带GN型和2-2型突变的ATCC13869菌株。以下由2-2型突变和GN型突变组成的双突变也称为GN2-2突变。
虽然在实施例1<2>至<4>中,所述突变株使用通过PCR扩增的突变odhA基因制备,所述PCR利用来自GN菌株或2-2菌株的染色体DNA作为模板,但它们也可以通过使用突变的odhA基因制备,所述突变的odhA基因通过定点诱变技术如使用Mutan-Super Express Km试剂盒(Takara Bio)的技术获得。例如,可以如以下所述获得具有类似于实施例1<2>中制备的pBSOAGN的结构的质粒。即,使用SEQ ID NOS:2和5的合成DNAs作为引物,野生型odhA基因的染色体DNA作为模板,通过PCR制备odhA基因片段,并将所得PCR片段克隆到Mutan-Super Express Km试剂盒附带的质粒pKF19k的BamHI识别位点。接下来,使用所得质粒模板和具有磷酸化的5’-末端的SEQ ID NO:7的合成DNA引物和Mutan-Super Express Km试剂盒附带的选择引物进行另一PCR。用所得PCR产物转化sup0大肠杆菌,例如MV1184菌株(可由Takara Bio获得),结果构建了包含突变odhA基因片段的质粒。最后,用BamHI消化所述片段并插入到pBS4S载体中,由此可以构建与pBSOAGN类似的质粒。
类似地,为了获得包含2-2型突变的odhA基因,可以用具有磷酸化的5’-末端的SEQ ID NO:8的合成DNA代替上述方法中具有磷酸化的5’-末端的SEQ ID NO:7的合成DNA来构建类似于pBSOA2-2的质粒。
此外,为了获得包含GN2-2双突变的odhA基因,从上述类似于pBSOAGN的质粒切除或PCR扩增包含GN-型突变的片段,并用于替换上述类似于pBSOA2-2的质粒上的相应片段。
实施例 2
菌株ATCC13869 OAGN、OA2-2、和OAGN2-2降解谷氨酸的能力的对比
虽然携带野生型odhA基因的菌株能够同化培养基中的L-谷氨酸,但推测由于odhA突变而削弱或消除了α-KGDH活性的菌株也具有减小的降解谷氨酸的能力。然后,使用实施例1<2>至<4>中制备的各odhA突变株ATCC13869 OAGN、OA2-2、和OAGN2-2测定降解谷氨酸的能力。各菌株均在CM-Dex平板上在25℃培养一天,然后接种到液体培养基中,所述液体培养基由20g/l谷氨酸钠、2.64g/l(NH4)2SO4、0.5g/l KH2PO4、0.5g/l K2HPO4、0.25g/l MgSO4·7H2O、0.01g/l FeSO4·7H2O、0.01g/l MnSO4·4-5H2O、0.01g/lCaCl2、0.02mg/l CuSO4、40g/l MOPS、0.03g/l原儿茶酸(protocatechinic acid)、200μg/l维生素B1、和300μg/l生物素(用NaOH调节至pH6.7)组成,之后在25℃和34℃培养50小时。分别测定起点、和培养25小时和50小时后谷氨酸的量,并比较25℃和34℃培养的菌株之间降解的(degraded)谷氨酸的量。表3显示结果。特别地,GN2-2突变株显示降解的谷氨酸的量明显减小,尤其是当培养温度为34℃时。这些结果表明引入GN2-2突变能够有效地减小α-KGDH活性,具有此突变的菌株优选用于L-谷氨酸生产。
表3:降解的谷氨酸的量
菌株 | 25小时 | 50小时 |
25℃ 34℃ | 25℃ 34℃ |
ATCC13869 | 15 15 | 15.4 15.4 |
OAGN | 3.8 5.8 | 9.4 10.0 |
OA2-2 | 11.7 15.1 | 15.4 15.4 |
OAGN2-2 | 0.7 0.5 | 4.9 1.0 |
(单位:g/l)
实施例3
ATCC13869 OAGN、OA2-2、和OAGN2-2菌株α-KGDH活性的对比
菌株ATCC13869 OAGN、OA2-2、和OAGN2-2的α-KGDH活性如下所述使用实施例4中自培养起始4小时后收集的培养液测定。所述活性按照Agric.Biol.Che m.,44(8),p1897(1980)中所述方法测定。具体地,在用0.2%氯化钠洗涤细菌细胞后,将其悬浮在含30%甘油的100mM TES-NaOH(pH7.5)缓冲溶液中。用Bioruptor(Olympus)超声处理细菌细胞,然后离心以去除未破裂的细菌细胞,之后使用Sephadex-G25(Amersham Pharmacia)用相同缓冲液凝胶过滤。将如此获得的制备物用作粗酶溶液。将所述粗制酶溶液添加到含100mM TES-NaOH (pH7.7)、5mM MgCl2、0.2mM CoA、0.3mM辅羧酶(cocarboxylase)、1mMα-酮戊二酸、3mM L-半胱氨酸、和1mM乙酰吡啶-腺嘌呤-二核苷酸的反应体系,用Hitachi分光光度计U-2001在31.5℃在365nm测定吸收。在粗制酶溶液中蛋白浓度的测量中,使用蛋白质测试(Bio-Rad)。牛血清白蛋白用作标准蛋白质。
表4显示测定α-KGDH活性的结果。与ATCC13869相比,引入GN型突变、2-2型突变、和GN2-2型突变导致α-KGDH活性减小。特别地,在引入GN2-2型突变的菌株的情况中,观测到相当大的活性减小。这表明GN型突变、2-2型突变、和GN2-2型突变是减小α-KGDH活性的突变。
表4:α-KGDH活性
菌株 | 培养温度25℃ | 培养温度34℃ |
ATCC13869OAGNOA2-2OAGN2-2 | 0.0360.0010.0070.001> | 0.0210.0020.0090.001> |
(单位:ΔAbs/min/mg蛋白质)
实施例4
菌株ATCC13869 OAGN、OA2-2、和OAGN2-2的L-谷氨酸生产能力的对比
在发酵釜培养中检验菌株ATCC13869 OAGN、OA2-2、和OAGN2-2的L-谷氨酸生产能力。首先,将上述四种菌株在25℃在CM-Dex琼脂培养基上培养一天,并将所得的细菌细胞接种在300ml灭菌的种子培养基(seed medium)中,所述种子培养基含60g/l葡萄糖、1.54g/l H3PO4、1.45g/l KOH、0.9g/lMgSO4·7H2O、0.01g/l FeSO4·7H2O、670μg/l维生素B1、3,200μg/l生物素、0.28g/l DL-Met、1.54g/l大豆蛋白水解物、和0.1ml/l消泡剂AZ-20R,一直培养到糖被完全耗尽。在培养期间,以1/l VVM通气来搅拌培养基,以使溶解氧的浓度保持在不小于5%。用氨气将培养期间的pH控制到pH7.2。然后,将30ml所得种子培养物接种到270ml含80g/l葡萄糖、3.46g/l KH2PO4、1.0g/l MgSO4·7H2O、0.01g/l FeSO4·7H2O、0.01g/l MnSO4·4-5H2O、230μg/l维生素B1、525μg/l生物素、0.35g/l大豆蛋白水解物、和0.2ml/l消泡剂AZ-20R的主培养基中,在25℃或34℃培养。培养期间如上所述进行通气。用氨气将培养期间的pH控制在pH7.3。
表5显示自培养起始约7.5小时后累积的L-谷氨酸的量。图5、6和7分别显示L-谷氨酸累积、细菌细胞量、和残余糖分的时间进程。ATCC13869菌株不累积L-谷氨酸,而OAGN和OAGN2-2突变菌株累积L-谷氨酸。特别地,当这些菌株在34℃培养时,所累积的L-谷氨酸的量显著增加。每种引入突变的菌株都显示几乎与野生型菌株相同的生长速率。
由这些结果证实GN型突变、2-2型突变、和GN2-2型突变对增加L-谷氨酸生产是有效的。发现在这些突变当中,GN2-2型突变对L-谷氨酸生产有最显著的影响。
表5:引入odhA突变的菌株所生产的L-谷氨酸(GIu)的量
菌株 | 25℃Glu(g/L) | 34℃Glu(g/L) |
ATCC13869OAGNOA2-2OAGN2-2 | 0.240.180.590.38 | 0.131.884.705.13 |
实施例5
用yggB突变菌株筛选odhA突变菌株
<L30型yggB突变菌株的构建>
首先,由ATCC13869菌株构建在yggB基因中具有突变的突变菌株。L30型突变为在yggB基因(SEQ ID NO:29)中用″T″替换1768位的″C″的突变。具有L30型突变的突变yggB基因如SEQ ID NO:31所示,由该基因编码的氨基酸序列如SEQ ID NO:32所示。所述突变yggB基因以与实施例1中相同的方法构建。即,使用SEQ ID NOS:33和34的引物和ATCC13869菌株的染色体DNA模板通过PCR制备编码ygg基因N-末端部分的片段。以类似的方式使用SEQ ID NOS:35和36的引物和ATCC13869菌株的染色体DNA模板通过PCR制备编码ygg基因C-末端部分的片段。随后,利用SEQ ID NOS:37和35的引物以及等量的N-末端片段和C-末端片段的混合物模板通过PCR获得包含L30型突变的yggB基因片段。用SacI消化所得PCR产物并连接到SacI消化的pBS4S,由此获得用于引入所述突变的质粒(pBS4 yggB-L)。将所得pBS4 yggB-L整合到ATCC13869菌株的染色体中,然后按照与实施例1中类似的方法将其从所述菌株消除。测定所得卡那霉素抗性菌株yggB基因的核苷酸序列,并选择其中yggB基因被L30型替换的菌株。将具有SEQ ID NO:31的yggB基因的菌株命名为ATCC13869-L菌株。该菌株可用于odhA突变基因的筛选。
<ygg、odhA双突变菌株的构建>
然后,将表6所示的各种突变引入到ATCC13869-L菌株的染色体odhA基因中。表6中显示了各菌株相应于SEQ ID NO:9的核苷酸2528至2562区域的核苷酸序列。表7中显示了各菌株相应于SEQ ID NO:10的氨基酸696至707区域的氨基酸序列。
引入具有核苷酸序列SEQ ID NO:42的odhA基因的L30sucA8菌株可以如下获得。使用SEQ ID NOS:2和5的引物通过PCR制备突变的odhA基因片段。所得片段用BamHI消化并连接到Mutan-Super Express Km(Takara Bio)附带的质粒pKF19m的BamHI位点。然后,使用具有磷酸化的5’-末端的引物SEQ ID NO:38和Mutan-Super Express Km的选择引物进行PCR,所得PCR产物用于转化sup0-大肠杆菌菌株如MV1184菌株,以得到包含突变odhA片段的质粒。将该片段插入到pBS4S质粒中,按照与实施例1中类似的方法用所得质粒转化ATCC13869-L菌株,由此获得所述质粒整合到其染色体中的菌株。然后,从这些菌株中选择对蔗糖有抗性且对卡那霉素敏感的菌株。测定所选择菌株的odhA基因的核苷酸序列,并选择其中由于odhA基因中的移码突变而使α-KGDH功能缺陷的菌株作为ATCC13869-L30sucA8 (odhA8)菌株。其它的odhA突变菌株可以使用yggB突变菌株通过类似的方法获得。
引入了具有核苷酸序列SEQ ID NO:44的突变odhA基因的sucA801菌株可以通过与上述类似的方法获得,所述方法中,用具有磷酸化的5’-末端的引物SEQ ID NO:39代替引物SEQ ID NO:38。
引入了具有核苷酸序列SEQ ID NO:46的突变odhA基因的sucA805菌株可以通过与上述类似的方法获得,所述方法中,用具有磷酸化的5’-末端的引物SEQ ID NO:40代替引物SEQ ID NO:38。
引入了具有核苷酸序列SEQ ID NO:48的突变odhA基因的sucA77菌株可以通过与上述类似的方法获得,所述方法中,用具有磷酸化的5’-末端的引物SEQ ID NO:41代替引物SEQ ID NO:38。
L30sucA8菌株没有胞内α-KGDH,因为sucA8突变是引起α-KGDH蛋白不成熟截断的移码突变。另一方面,sucA801菌株、sucA805菌株和sucA77菌株具有减小的但有一些α-KGDH活性,因为这些突变不是移码突变,而且不引起α-KGDH蛋白的不成熟截断。
表6:odhA突变基因的部分核苷酸序列
菌株 | odhA基因的核苷酸序列 |
ATCC13869-L | CTG GCT AAG CTG CGT GGC TAC GAC GTC GGA GGC ACC |
L30sucA8 | CTG GCT AAG CTG CGT C GAC GTC GGA GGC ACC |
L30sucA801 | CTG GCT AAG CTG CGT CTC GAC GTC GGA GGC ACC |
L30sucA805 | CTG GCT AAA AGC TGC GTC GAC GTC GGA GGC ACC |
L30sucA77 | CTG GCT ATA AGC TGC GTC GAC GTC GGA GGC ACC |
表7:odhA突变体的氨基酸序列
菌株 | E1o亚基的氨基酸序列 |
野生的 | Leu Ala Lys Leu Arg Gly Tyr Asp Val Gly Gly Thr |
L30sucA8(ΔsucA) | Leu Ala Lys Leu Arg Arg Arg Arg Arg His |
L30sucA801 | Leu Ala Lys Leu Arg---Leu Asp Val Gly Gly Thr |
L30sucA805 | Leu Ala Lys Ser Cys---Val Asp Val Gly Gly Thr |
L30sucA77 | Leu Ala Ile Ser Cys---Val Asp Val Gly Gly Thr |
<使用携带各种突变odhA基因的菌株生产L-谷氨酸>
通过在坂口(Sakaguchi)烧瓶中培养所得odhA突变菌株来评估这些菌株的L-谷氨酸生产力。将表6列出的各种菌株在CM-Dex琼脂培养基上在31.5℃培养过夜,然后将1/6的培养物转移到20ml含60g/l葡萄糖、22.5g/l(NH4)2SO4、1g/l KH2PO4、0.4g/l MgSO4·7H2O、0.01g/l FeSO4·7H2O、0.01g/lMnSO4·4-5H2O、200μg/l维生素B1、0.48g/l大豆蛋白水解物、和300μg/l生物素(用KOH调节至pH 8.0)的培养基中,添加CaCO3并在31.5℃、115rpm搅动培养。培养19小时后累积的L-谷氨酸的量如表8所示。sucA801、sucA805、和sucA77菌株显示高于ATCC13869-L菌株和sucA8菌株的L-谷氨酸生产力,其中ATCC13869-L菌株携带野生型odhA基因,而SucA8菌株携带含有移码突变的odhA基因。这些结果显示,通过将突变引入到odhA基因的焦磷酸硫胺素结合区附近区域中以调节α-KGDH活性,从而有效地生产了L-谷氨酸。
表8:odhA突变菌株的L-谷氨酸生产
菌株 | L-谷氨酸(g/L) |
ATCC13869-L | 4.9 |
L30sucA8 | 19.8 |
L30sucA801 | 22.1 |
L30sucA805 | 23.8 |
L30sucA77 | 21.6 |
序列表
<110>味之素株式会社(Ajinomoto Co.,Inc.)
<120>C453-C5043
<130>生产L-谷氨酸的微生物和生产L-谷氨酸的方法
<150>JP2004-264458
<151>2004-09-10
<160>51
<170>PatentIn version 3.1
<210>1
<211>40
<212>DNA
<213>人工的
<220>
<223>引物
<400>1
gccgggatcc tccggtgaat tcctgcgtac catgtctcgc 40
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gccgggatcc ctgtgtgatt cacactgcat aaggccctct 40
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gcttccggct cgtatgttgt gtgg 24
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<212>DNA
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gatcgtgacc gcacagattc 20
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gccgggatcc ccatcgccgc catccctgat ggtttcaatc 40
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gccgggatcc ggccctggcc tgcggcggtg tcgatggcgg 40
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gctaagcgcg tcgacgtcgg aggcaccatc cac 33
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<220>
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<400>8
ctcctaccat cgacggcagg ccccaaccct gatcgtgcct ga 42
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<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4213)
<223>
<400>9
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Asn Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag cttccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac gac 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr Asp
205 210 215
gtc atc gac ggc aag cca acc ctg atc gtg cct gag cac atc aac ctg 1144
Val Ile Asp Gly Lys Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu
220 225 230
ggc ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc 1192
Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val
235 240 245 250
gta gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc 1240
Val Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu
255 260 265
gca gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc 1288
Ala Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr
270 275 280
atg gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc 1336
Met Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly
285 290 295
atc ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc 1384
Ile Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr
300 305 310
atc atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct 1432
Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala
315 320 325 330
tcc gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc 1480
Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile
335 340 345
acc tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa 1528
Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu
350 355 360
ttc ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat 1576
Phe Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp
365 370 375
gag atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca 1624
Glu Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala
380 385 390
cag gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag 1672
Gln Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln
395 400 405 410
ctc att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac 1720
Leu Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn
415 420 425
cca ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac 1768
Pro Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp
430 435 440
ctc gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc 1816
Leu Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr
445 450 455
ttc agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag 1864
Phe Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu
460 465 470
gta ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa 1912
Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu
475 480 485 490
tac acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc 1960
Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg
495 500 505
ctc gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc 2008
Leu Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile
510 515 520
ctg cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc 2056
Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr
525 530 535
aag tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc 2104
Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu
540 545 550
atc cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc 2152
Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu
555 560 565 570
gac gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg 2200
Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu
575 580 585
ttc aac atc gtg ggc aag cca ctg gcatcc atc ttc aac gag ttt gaa 2248
Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu
590 595 600
ggc caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac 2296
Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr
605 610 615
cac ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag 2344
His Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu
620 625 630
atc aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac 2392
Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn
635 640 645 650
cca gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag 2440
Pro Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys
655 660 665
ggc gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct 2488
Gly Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala
670 675 680
gca ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aag 2536
Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys
685 690 695
ctg cgt ggc tac gac gtc gga ggc acc atc cac atc gtg gtg aac aac 2584
Leu Arg Gly Tyr Asp Val Gly Gly Thr Ile His Ile Val Val Asn Asn
700 705 710
cag atc ggc ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac 2632
Gln Ile Gly Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr
715 720 725 730
gca acc gac tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat 2680
Ala Thr Asp Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn
735 740 745
ggt gat gac cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag 2728
Gly Asp Asp Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu
750 755 760
tac cgt cgt cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac 2776
Tyr Arg Arg Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr
765 770 775
cgc ctc cgc ggc cac aac gaa gct gat gat cct tcc atg acc cag cca 2824
Arg Leu Arg Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro
780 785 790
aag atg tat gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac 2872
Lys Met Tyr Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr
795 800 805 810
acc gaa gac ctg ctc gga cgt gga gac ctc tcc aac gaa gat gca gaa 2920
Thr Glu Asp Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu
815 820 825
gca gtc gtc cgc gac ttc cac gac cag atg gaa tct g tg ttc aac gaa 2968
Ala Val Val Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu
830 835 840
gtc aag gaa ggc ggc aag aag cag gct gag gca cag acc ggc atc acc 3016
Val Lys Glu Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr
845 850 855
ggc tcc cag aag ctt cca cac ggc ctt gag acc aac atc tcc cgt gaa 3064
Gly Ser Gln Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu
860 865 870
gag ctc ctg gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc 3112
Glu Leu Leu Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe
875 880 885 890
aac tac cac cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct 3160
Asn Tyr His Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser
895 900 905
gtc acc gaa ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc 3208
Val Thr Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe
910 915 920
ggt tcc ctg gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat 3256
Gly Ser Leu Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp
925 930 935
tcc cgc cgc ggt acc ttc acc cag cgc cac gca gtt gcc atc gac cca 3304
Ser Arg Arg Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro
940 945 950
gcg acc gct gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag 3352
Ala Thr Ala Glu Glu Phe Asn Pro Leu His Glu Leu Ala Gln Ser Lys
955 960 965 970
ggc aac aac ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac 3400
Gly Asn Asn Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr
975 980 985
gca ggc atg ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc 3448
Ala Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Ash Glu Asp Set
990 995 1000
gtc gtt gca tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct 3493
Val Val Ala Trp Glu Ala Gln Phe Gly Asp Phe Ala Ash Gly Ala
1005 1010 1015
cag acc atc atc gat gag tac gtc tcc tca ggc gaa gct aag tgg 3538
Gln Thr Ile Ile Asp Glu Tyr Val Ser Ser Gly Glu Ala Lys Trp
1020 1025 1030
ggc cag acc tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa 3583
Gly Gln Thr Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu
1035 1040 1045
ggc cag ggc cca gac cac tct tcc gca cgt atc gag cgc ttc ctg 3628
Gly Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu
1050 1055 1060
cag ctg tgc gct gag ggt tcc atg act gtt gct cag cca tcc acc 3673
Gln Leu Cys Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr
1065 1070 1075
cca gca aac cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac 3718
Pro Ala Ash His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp
1080 1085 1090
ctg aag cgt cca ctg gtt atc ttc acc ccg aag tcc atg ctg cgt 3763
Leu Lys Arg Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg
1095 1100 1105
aac aag gct gct gcc tcc gca cca gaa gac ttc act gag gtc acc 3208
Asn Lys Ala Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr
1110 1115 1120
aag ttc caa tcc gtg atc gac gat cca aac gtt gca gat gca gcc 3853
Lys Phe Gln Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala
1125 1130 1135
aag gtg aag aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa 3898
Lys Val Lys Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu
1140 1145 1150
ttg gca aag cgc aag gag aag gac gga cgc gac gac atc gcg atc 3943
Leu Ala Lys Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile
1155 1160 1165
gtt cgt atc gaa atg ctc cac cca att ccg ttc aac cgc atc tcc 3988
Val Arg Ile Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser
1170 1175 1180
gag gct ctt gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt 4033
Glu Ala Leu Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val
1185 1190 1195
cag gat gag cca gca aac cag ggc cca tgg ccg ttc tac cag gag 4078
Gln Asp Glu Pro Ala Ash Gln Gly Pro Trp Pro Phe Tyr Gln Glu
1200 1205 1210
cac ctc cca gag ctg atc ccg aac atg cca aag atg cgc cgc gtt 4123
His Leu Pro Glu Leu Ile Pro Ash Met Pro Lys Met Arg Arg Val
1215 1220 1225
tcc cgc cgc gct cag tcc tcc acc gca act ggt gtt gct aag gtg 4168
Ser Arg Arg Ala Gln Ser Ser Thr Ala Thr Gly Val Ala Lys Val
1230 1235 1240
cac cag ctg gag gag aag cag ctt atc gac gag gct ttc gag gct 4213
His Gln Leu Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala
1245 1250 1255
taagtcttta tagtcctgca ctagcctaga gggccttatg cagtgtgaat cacacagcat 4273
aaggcccttt ttgctgccgt ggttgcctaa ggtggaaggc atgaaacgaa tctgtgcggt 4333
cacgatctct tcagtacttt tgctaagtgg ctgctcctcc acttccacca cgcagctcga 4393
g 4394
<210>10
<211>1257
<212>PRT
<213>谷氨酸棒杆菌
<400>10
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu Hi sIle Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr Hi sIle Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 510
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Leu Arg Gly Tyr Asp Val
690 695 700
Gly Gly Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr
705 710 715 720
Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys
725 730 735
Ala Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala
740 745 750
Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly
755 760 765
Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn
770 775 780
Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile
785 790 795 800
Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly
805 810 815
Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe
820 825 830
His Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys
835 840 845
Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro
850 855 860
His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly
865 870 875 880
Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val
885 890 895
Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile
900 905 910
Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser
915 920 925
Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe
930 935 940
Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe
945 950 955 960
Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe
965 970 975
Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu
980 985 990
Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala
995 1000 1005
Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu
1010 1015 1020
Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu
1025 1030 1035
Ile Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His
1040 1045 1050
Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly
1055 1060 1065
Ser Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His
1070 1075 1080
Leu Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val
1085 1090 1095
Ile Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser
1100 1105 1110
Ala Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile
1115 1120 1125
Asp Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met
1130 1135 1140
Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu
1145 1150 1155
Lys Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu
1160 1165 1170
His Pro Ile Pro Phe Ash Arg Ile Ser Glu Ala Leu Ala Gly Tyr
1175 1180 1185
Pro Ash Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn
1190 1195 1200
Gln Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile
1205 1210 1215
Pro Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser
1220 1225 1230
Ser Thr Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys
1235 1240 1245
Gln Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>11
<211>4391
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4213)
<223>
<400>11
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Ash Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Set Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac cat 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr His
205 210 215
cga cgg cag gcc cca acc ctg atc gtg cct gag cac atc aac ctg ggc 1144
Arg Arg Gln Ala Pro Thr Leu Ile Val Pro Glu Hi s Ile Asn Leu Gly
220 225 230
ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc gta 1192
Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val
235 240 245 250
gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc gca 1240
Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala
255 260 265
gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc atg 1288
Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met
270 275 280
gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc atc 1336
Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile
285 290 295
ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc atc 1384
Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile
300 305 3l0
atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct tcc 1432
Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser
315 320 325 330
gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc acc 1480
Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr
335 340 345
tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa ttc 1528
Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe
350 355 360
ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat gag 1576
Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu
365 370 375
atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca cag 1624
Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln
380 385 390
gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag ctc 1672
Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu
395 400 405 410
att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac cca 1720
Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro
415 420 425
ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac ctc 1768
Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp Leu
430 435 440
gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc ttc 1816
Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe
445 450 455
agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag gta 1864
Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val
460 465 470
ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa tac 1912
Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr
475 480 485 490
acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc ctc 1960
Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu
495 500 505
gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc ctg 2008
Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu
510 515 520
cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc aag 2056
Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys
525 530 535
tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc atc 2104
Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile
540 545 550
cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc gac 2152
Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp
555 560 565 570
gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg ttc 2200
Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu Phe
575 580 585
aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa ggc 2248
Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly
590 595 600
caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac cac 2296
Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His
605 610 615
ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag atc 2344
Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile
620 625 630
aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac cca 2392
Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro
635 640 645 650
gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag ggc 2440
Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly
655 660 665
gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct gca 2488
Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala Ala
670 675 680
ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aag ctg 2536
Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Leu
685 690 695
cgt ggc tac gac gtc gga ggc acc atc cac atc gtg gtg aac aac cag 2584
Arg Gly Tyr Asp Val Gly Gly Thr Ile His Ile Val Val Asn Asn Gln
700 705 710
atc ggc ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac gca 2632
Ile Gly Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala
715 720 725 730
acc gac tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat ggt 2680
Thr Asp Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn Gly
735 740 745
gat gac cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag tac 2728
Asp Asp Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr
750 755 760
cgt cgt cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac cgc 2776
Arg Arg Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg
765 770 775
ctc cgc ggc cac aac gaa gct gat gat cct tcc atg acc cag cca aag 2824
Leu Arg Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys
780 785 790
atg tat gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac acc 2872
Met Tyr Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr
795 800 805 810
gaa gac ctg ctc gga cgt gga gac ctc tcc aac gaa gatgca gaa gca 2920
Glu Asp Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala
815 820 825
gtc gtc cgc gac ttc cac gac cag atg gaa tct gtg ttc aac gaa gtc 2968
Val Val Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu Val
830 835 840
aag gaa ggc ggc aag aag cag gct gag gca cag acc ggc atc acc ggc 3016
Lys Glu Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly
845 850 855
tcc cag aag ctt cca cac ggc ctt gag acc aac atc tcc cgt gaa gag 3064
Ser Gln Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu
860 865 870
ctc ctg gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc aac 3112
Leu Leu Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn
875 880 885 890
tac cac cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct gtc 3160
Tyr His Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val
895 900 905
acc gaa ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt 3208
Thr Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly
910 915 920
tcc ctg gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc 3256
Ser Leu Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser
925 930 935
cgc cgc ggt acc ttc acc cag cgc cac gca gtt gcc atc gac cca gcg 3304
Arg Arg Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala
940 945 950
acc gct gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag ggc 3352
Thr Ala Glu Glu Phe Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly
955 960 965 970
aac aac ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac gca 3400
Asn Asn Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala
975 980 985
ggc atg ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc gtc 3448
Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val
990 995 1000
gtt gca tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct cag 3493
Val Ala Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln
1005 1010 1015
acc atc atc gat gag tac gtc tcc tca ggc gaa gct aag tgg ggc 3538
Thr Ile Ile Asp Glu Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly
1020 1025 1030
cag acc tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa ggc 3583
Gln Thr Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu Gly
1035 l040 1045
cag ggc cca gac cac tct tcc gca cgt arc gag cgc ttc ctg cag 3628
Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln
1050 1055 1060
ctg tgc gct gag ggt tcc atg act gtt gct cag cca tcc acc cca 3673
Leu Cys Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr Pro
1065 1070 1075
gca aac cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac ctg 3718
Ala Asn His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp Leu
1080 1085 1090
aag cgt cca ctg gtt arc ttc acc ccg aag tcc atg ctg cgt aac 3763
Lys Arg Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg Asn
1095 1100 1105
aag gct gct gcc tcc gca cca gaa gac ttc act gag gtc acc aag 3808
Lys Ala Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr Lys
1110 1115 1120
ttc caa tcc gtg atc gac gat cca aac gtt gca gat gca gcc aag 3853
Phe Gln Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala Lys
1125 1130 1135
gtg aag aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa ttg 3898
Val Lys Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu
1140 1145 1150
gca aag cgc aag gag aag gac gga cgc gac gac atc gcg atc gtt 3943
Ala Lys Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile Val
1155 1160 1165
cgt atc gaa atg ctc cac cca att ccg ttc aac cgc atc tcc gag 3988
Arg Ile Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser Glu
1170 1175 1180
gct ctt gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt cag 4033
Ala Leu Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val Gln
1185 1190 1195
gat gag cca gca aac cag ggc cca tgg ccg ttc tac cag gag cac 4078
Asp Glu Pro Ala Asn Gln Gly Pro Trp Pro Phe Tyr Gln Glu His
1200 1205 1210
ctc cca gag ctg atc ccg aac atg cca aag atg cgc cgc gtt tcc 4123
Leu Pro Glu Leu Ile Pro Asn Met Pro Lys Met Arg Arg Val Ser
1215 1220 1225
cgc cgc gct cag tcc tcc acc gca act ggt gtt gct aag gtg cac 4168
Arg Arg Ala Gln Ser Ser Thr Ala Thr Gly Val Ala Lys Val His
1230 1235 1240
cag ctg gag gag aag cag ctt atc gac gag gct ttc gag gct taa 4213
Gln Leu Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala
1245 1250 1255
gtctttatag tcctgcacta gcctagaggg ccttatgcag tgtgaatcac acagcataag 4273
gccctttttg ctgccgtggt tgcctaaggt ggaaggcatg aaacgaatct gtgcggtcac 4333
gatctcttca gtacttttgc taagtggctg ctcctccact tccaccacgc agctcgag 4391
<210>12
<211>1256
<212>PRT
<213>谷氨酸棒杆菌
<400>12
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Ash Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr His Arg Arg Gln Ala Pro Thr
210 215 220
Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu Pro
225 230 235 240
Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu Thr
245 250 255
Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile Val
260 265 270
Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly Val
275 280 285
Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser Val
290 295 300
Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser Met
305 310 315 320
Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala Glu
325 330 335
Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His Arg
340 345 350
Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser Arg
355 360 365
Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met Asn
370 375 380
Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr Gly
385 390 395 400
Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg Ser
405 410 415
Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln Pro
420 425 430
Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His Ser
435 440 445
Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe Gly
450 455 460
Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg Ala
465 470 475 480
Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp Arg
485 490 495
Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro Lys
500 505 510
Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala Ala
515 520 525
Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys Arg
530 535 540
Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser Ala
545 550 555 560
Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly Met
565 570 575
Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys Pro
580 585 590
Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly Gln
595 600 605
Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly Gln
610 615 620
His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr Ala
625 630 635 640
Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile Val
645 650 655
Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr Val
660 665 670
Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly Ile
675 680 685
Val Pro Glu Thr Ile Asn Leu Ala Lys Leu Arg Gly Tyr Asp Val Gly
690 695 700
Gly Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr
705 710 715 720
Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys Ala
725 730 735
Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val
740 745 750
Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys
755 760 765
Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn Glu
770 775 780
Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr
785 790 795 800
Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly Arg
805 810 815
Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe His
820 825 830
Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys Lys
835 840 845
Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro His
850 855 860
Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly Gln
865 870 875 880
Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val Ala
885 890 895
Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile Asp
900 905 910
Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser Gly
915 920 925
Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr
930 935 940
Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe Asn
945 950 955 960
Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe Leu
965 970 975
Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr
980 985 990
Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala Gln
995 1000 1005
Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr
1010 1015 1020
Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu Ile
1025 1030 1035
Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His Ser
1040 1045 1050
Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly Ser
1055 1060 1065
Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His Leu
1070 1075 1080
Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val Ile
1085 1090 1095
Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser Ala
1100 1105 1110
Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile Asp
1115 1120 1125
Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met Leu
1130 1135 1140
Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys
1145 1150 1155
Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu His
1160 1165 1170
Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr Pro
1175 1180 1185
Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn Gln
1190 1195 1200
Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile Pro
1205 1210 1215
Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser Ser
1220 1225 1230
Thr Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys Gln
1235 1240 1245
Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>13
<211>4388
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4213)
<223>
<400>13
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg Hi s Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Asn Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac gac 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr Asp
205 210 215
gtc atc gac ggc aag cca acc ctg atc gtg cct gag cac atc aac ctg 1144
Val Ile Asp Gly Lys Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu
220 225 230
ggc ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc 1192
Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val
235 240 245 250
gta gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc 1240
Val Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu
255 260 265
gca gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc 1288
Ala Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr
270 275 280
atg gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc 1336
Met Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly
285 290 295
atc ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc 1384
Ile Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr
300 305 310
atc atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct 1432
Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala
315 320 325 330
tcc gaa gac cgc ctt gca gag ctc ggc gtt gga aag cttgtc acc atc 1480
Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile
335 340 345
cc tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa 1528
Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu
350 355 360
ttc ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat 1576
Phe Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp
365 370 375
gag atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca 1624
Glu Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala
380 385 390
cag gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag 1672
Gln Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln
395 400 405 410
ctc att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac 1720
Leu Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn
415 420 425
cca ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac 1768
Pro Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp
430 435 440
ctc gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc 1816
Leu Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr
445 450 455
ttc agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag 1864
Phe Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu
460 465 470
gta ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa 1912
Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu
475 480 485 490
tac acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc 1960
Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg
495 500 505
ctc gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc 2008
Leu Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile
510 515 520
ctg cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc 2056
Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr
525 530 535
aag tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc 2104
Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu
540 545 550
atc cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc 2152
Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu
555 560 565 570
gac gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg 2200
Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu
575 580 585
ttc aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa 2248
Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu
590 595 600
ggc caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac 2296
Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr
605 610 615
cac ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag 2344
His Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu
620 625 630
atc aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac 2392
Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn
635 640 645 650
cca gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag 2440
Pro Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys
655 660 665
ggc gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct 2488
Gly Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala
670 675 680
gca ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aag 2536
Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys
685 690 695
cgc gtc gac gtc gga ggc acc atc cac atc gtg gtg aac aac cag atc 2584
Arg Val Asp Val Gly Gly Thr Ile His Ile Val Val Asn Asn Gln Ile
700 705 710
ggc ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac gca acc 2632
Gly Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr
715 720 725 730
gac tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat ggt gat 2680
Asp Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn Gly Asp
735 740 745
gac cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag tac cgt 2728
Asp Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg
750 755 760
cgt cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac cgc ctc 2776
Arg Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu
765 770 775
cgc ggc cac aac gaa gct gat gat cct tcc atg acc cag eca aag atg 2824
Arg Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met
780 785 790
tat gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac acc gaa 2872
Tyr Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu
795 800 805 810
gac ctg ctc gga cgt gga gac ctc tcc aac gaa gat gca gaa gca gtc 2920
Asp Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val
815 820 825
gtc cgc gac ttc cac gac cag atg gaa tct gtg ttc aac gaa gtc aag 2968
Val Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys
830 835 840
gaa ggc ggc aag aag cag gct gag gca cag acc ggc atc acc ggc tcc 3016
Glu Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser
845 850 855
cag aag ctt cca cac ggc ctt gag acc aac atc tcc cgt gaa gag ctc 3064
Gln Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu
860 865 870
ctg gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc aac tac 3112
Leu Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr
875 880 885 890
cac cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct gtc acc 3160
His Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr
895 900 905
gaa ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt tcc 3208
Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser
910 915 920
ctg gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc cgc 3256
Leu Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg
925 930 935
cgc ggt acc ttc acc cag cgc cac gca gtt gcc atc gac cca gcg acc 3304
Arg Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr
940 945 950
gct gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag ggc aac 3352
Ala Glu GluPhe Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn
955 960 965 970
aac ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac gca ggc 3400
Asn Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly
975 980 985
atg ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc gtc gtt 3448
Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val
990 995 1000
gca tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct cag acc 3493
Ala Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr
1005 1010 1015
atc atc gat gag tac gtc tcc tca ggc gaa gct aag tgg ggc cag 3538
Ile Ile Asp Glu Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly Gln
1020 1025 1030
acc tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa ggc cag 3583
Thr Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu Gly Gln
1035 1040 1045
ggc cca gac cac tct tcc gca cgt atc gag cgc ttc ctg cag ctg 3628
Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu
1050 1055 1060
tgc gct gag ggt tcc atg act gtt gct cag cca tcc acc cca gca 3673
Cys Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr Pro Ala
1065 1070 1075
aac cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac ctg aag 3718
Asn His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp Leu Lys
1080 1085 1090
cgt cca ctg gtt atc ttc acc ccg aag tcc atg ctg cgt aac aag 3763
Arg Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg Asn Lys
1095 1100 1105
gct gct gcc tcc gca cca gaa gac ttc act gag gtc acc aag ttc 3808
Ala Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr Lys Phe
1110 1115 1120
caa tcc gtg atc gac gat cca aac gtt gca gat gca gcc aag gtg 3853
Gln Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala Lys Val
1125 1130 1135
aag aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa ttg gca 3898
Lys Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala
1140 1145 1150
aag cgc aag gag aag gac gga cgc gac gac atc gcg arc gtt cgt 3943
Lys Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile Val Arg
1155 1160 1165
atc gaa atg ctc cac cca att ccg ttc aac cgc atc tcc gag gct 3988
Ile Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala
1170 1175 1180
ctt gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt cag gat 4033
Leu Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val Gln Asp
1185 1190 1195
gag cca gca aac cag ggc cca tgg ccg ttc tac cag gag cac ctc 4078
Glu Pro Ala Asn Gln Gly Pro Trp Pro Phe Tyr Gln Glu His Leu
1200 1205 1210
cca gag ctg atc ccg aac atg cca aag atg cgc cgc gtt tcc cgc 4123
Pro Glu Leu Ile Pro Asn Met Pro Lys Met Arg Arg Val Ser Arg
1215 1220 1225
cgc gct cag tcc tcc acc gca act ggt gtt gct aag gtg cac cag 4168
Arg Ala Gln Ser Ser Thr Ala Thr Gly Val Ala Lys Val His Gln
1230 1235 1240
ctg gag gag aag cag ctt atc gac gag gct ttc gag gct taa gtc 4213
Leu Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala Val
1245 1250 1255
tttatagtcc tgcactagcc tagagggcct tatgcagtgt gaatcacaca gcataaggcc 4273
ctttttgctg ccgtggttgc ctaaggtgga aggcatgaaa cgaatctgtg cggtcacgat 4333
ctcttcagta cttttgctaa gtggctgctc ctccacttcc accacgcagc tcgag 4388
<210>14
<211>1255
<212>PRT
<213>谷氨酸棒杆菌
<400>14
Met Leu Gln Leu Gly Leu Arg His Ash Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu AlaIle Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr SerThr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 510
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Arg Val Asp Val Gly Gly
690 695 700
Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr Pro
705 710 715 720
Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys Ala Phe
725 730 735
Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val Val
740 745 750
Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys Asp
755 760 765
Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn Glu Ala
770 775 780
Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr Gly
785 790 795 800
Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly Arg Gly
805 810 815
Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe His Asp
820 825 830
Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys Lys Gln
835 840 845
Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro His Gly
850 855 860
Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly Gln Ala
865 870 875 880
Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val Ala Pro
885 890 895
Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile Asp Trp
900 905 910
Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser Gly Arg
915 920 925
Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr Gln
930 935 940
Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe Asn Pro
945 950 955 960
Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe Leu Val
965 970 975
Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr Gly
980 985 990
Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala Gln Phe
995 1000 1005
Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr Val
1010 1015 1020
Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu Ile Leu
1025 1030 1035
Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His Ser Ser
1040 1045 1050
Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly Ser Met
1055 1060 1065
Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His Leu Leu
1070 1075 1080
Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val Ile Phe
1085 1090 1095
Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser Ala Pro
1100 1105 1110
Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile Asp Asp
1115 1120 1125
Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met Leu Val
1130 1135 1140
Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys Asp
1145 1150 1155
Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu His Pro
1160 1165 1170
Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr Pro Asn
1175 1180 1185
Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn Gln Gly
1190 1195 1200
Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile Pro Asn
1205 1210 1215
Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser Ser Thr
1220 1225 1230
Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys Gln Leu
1235 1240 1245
Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>15
<211>4385
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4213)
<223>
<400>15
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Ash Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac cat 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr His
205 210 215
cga cgg cag gcc cca acc ctg atc gtg cct gag cac atc aac ctg ggc 1144
Arg Arg Gln Ala Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly
220 225 230
ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc gta 1192
Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val
235 240 245 250
gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc gca 1240
Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala
255 260 265
gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc atg 1288
Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met
270 275 280
gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc atc 1336
Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile
285 290 295
ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc atc 1384
Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile
300 305 310
atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct tcc 1432
Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser
315 320 325 330
gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc acc 1480
Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr
335 340 345
tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa ttc 1528
Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe
350 355 360
ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gatgag 1576
Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu
365 370 375
atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca cag 1624
Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln
380 385 390
gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag ctc 1672
Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu
395 400 405 410
att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac cca 1720
Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro
415 420 425
ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac ctc 1768
Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp Leu
430 435 440
gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc ttc 1816
Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe
445 450 455
agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag gta 1864
Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val
460 465 470
ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa tac 1912
Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr
475 480 485 490
acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc ctc 1960
Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu
495 500 505
gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc ctg 2008
Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu
510 515 520
cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc aag 2056
Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys
525 530 535
tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc atc 2104
Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile
540 545 550
cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc gac 2152
Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp
555 560 565 570
gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg ttc 2200
Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu Phe
575 580 585
aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa ggc 2248
Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly
590 595 600
caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac cac 2296
Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His
605 610 615
ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag atc 2344
Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile
620 625 630
aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac cca 2392
Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro
635 640 645 650
gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag ggc 2440
Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly
655 660 665
gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct gca 2488
Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala Ala
670 675 680
ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aag cgc 2536
Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Arg
685 690 695
gtc gac gtc gga ggc acc atc cac atc gtg gtg aac aac cag atc ggc 2584
Val Asp Val Gly Gly Thr Ile His Ile Val Val AsnAsn Gln Ile Gly
700 705 710
ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac gca acc gac 2632
Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp
715 720 725 730
tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat ggt gat gac 2680
Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp
735 740 745
cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag tac cgt cgt 2728
Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg
750 755 760
cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac cgc ctc cgc 2776
Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg
765 770 775
ggc cac aac gaa gct gat gat cct tcc atg acc cag cca aag atg tat 2824
Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr
780 785 790
gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac acc gaa gac 2872
Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp
795 800 805 810
ctg ctc gga cgt gga gac ctc tcc aac gaa gat gca gaa gca gtc gtc 2920
Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val
815 820 825
cgc gac ttc cac gac cag atg gaa tct gtg ttc aac gaa gtc aag gaa 2968
Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu
830 835 840
ggc ggc aag aag cag gct gag gca cag acc ggc atc acc ggc tcc cag 3016
Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln
845 850 855
aag ctt cca cac ggc ctt gag acc aac atctcc cgt gaa gag ctc ctg 3064
Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu
860 865 870
gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc aac tac cac 3112
Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His
875 880 885 890
cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct gtc acc gaa 3160
Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu
895 900 905
ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt tcc ctg 3208
Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu
910 915 920
gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc cgc cgc 3256
Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg
925 930 935
ggt acc ttc acc cag cgc cac gca gtt gcc atc gac cca gcg acc gct 3304
Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala
940 945 950
gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag ggc aac aac 3352
Glu Glu Phe Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn
955 960 965 970
ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac gca ggc atg 3400
Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met
975 980 985
ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc gtc gtt gca 3448
Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala
990 995 1000
tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct cag acc atc 3493
Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile
1005 1010 1015
atc gat gag tac gtc tcc tca ggc gaa gct aag tgg ggc cag acc 3538
Ile Asp Glu Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr
1020 1025 1030
tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa ggc cag ggc 3583
Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly
1035 1040 1045
cca gac cac tct tcc gca cgt atc gag cgc ttc ctg cag ctg tgc 3628
Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys
1050 1055 1060
gct gag ggt tcc atg act gtt gct cag cca tcc acc cca gca aac 3673
Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn
1065 1070 1075
cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac ctg aag cgt 3718
His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg
1080 1085 1090
cca ctg gtt atc ttc acc ccg aag tcc atg ctg cgt aac aag gct 3763
Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala
1095 1100 1105
gct gcc tcc gca cca gaa gac ttc act gag gtc acc aag ttc caa 3808
Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln
1110 1115 1120
tcc gtg atc gac gat cca aac gtt gca gat gca gcc aag gtg aag 3853
Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys
1125 1130 1135
aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa ttg gca aag 3898
Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys
1140 1145 1150
cgc aag gag aag gac gga cgc gac gac atc gcg atc gtt cgt atc 3943
Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile
1155 1160 1165
gaa atg ctc cac cca att ccg ttc aac cgc atc tcc gag gct ctt 3988
Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu
1170 1175 1180
gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt cag gat gag 4033
Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu
1185 1190 1195
cca gca aac cag ggc cca tgg ccg ttc tac cag gag cac ctc cca 4078
Pro Ala Asn Gln Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro
1200 1205 1210
gag ctg atc ccg aac atg cca aag atg cgc cgc gtt tcc cgc cgc 4123
Glu Leu Ile Pro Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg
1215 1220 1225
gct cag tcc tcc acc gca act ggt gtt gct aag gtg cac cag ctg 4168
Ala Gln Set Ser Thr Ala Thr Gly Val Ala Lys Val His Gln Leu
1230 1235 1240
gag gag aag cag ctt atc gac gag gct ttc gag gct taa gtc ttt 4213
Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala Val Phe
1245 1250 1255
atagtcctgc actagcctag agggccttat gcagtgtgaa tcacacagca taaggccctt 4273
tttgctgccg tggttgccta aggtggaagg catgaaacga atctgtgcgg tcacgatctc 4333
ttcagtactt ttgctaagtg gctgctcctc cacttccacc acgcagctcg ag 4385
<210>16
<211>1254
<212>PRT
<213>谷氨酸棒杆菌
<400>16
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys lle Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Ash Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr His Arg Arg Gln Ala Pro Thr
210 215 220
Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu Pro
225 230 235 240
Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu Thr
245 250 255
Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile Val
260 265 270
Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly Val
275 280 285
Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser Val
290 295 300
Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser Met
305 310 315 320
Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala Glu
325 330 335
Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His Arg
340 345 350
Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser Arg
355 360 365
Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met Asn
370 375 380
Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr Gly
385 390 395 400
Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg Ser
405 410 415
Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln Pro
420 425 430
Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His Ser
435 440 445
Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe Gly
450 455 460
Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg Ala
465 470 475 480
Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp Arg
485 490 495
Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro Lys
500 505 510
Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala Ala
515 520 525
Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys Arg
530 535 540
Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser Ala
545 550 555 560
Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly Met
565 570 575
Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys Pro
580 585 590
Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly Gln
595 600 605
Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly Gln
610 615 620
His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr Ala
625 630 635 640
Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile Val
645 650 655
Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr Val
660 665 670
Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly Ile
675 680 685
Val Pro Glu Thr Ile Asn Leu Ala Lys Arg Val Asp Val Gly Gly Thr
690 695 700
Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr Pro Asp
705 710 715 720
Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys Ala Phe Gly
725 730 735
Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val Val Trp
740 745 750
Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys Asp Val
755 760 765
Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn Glu Ala Asp
770 775 780
Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr Gly Arg
785 790 795 800
Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly Arg Gly Asp
805 810 815
Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe His Asp Gln
820 825 830
Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys Lys Gln Ala
835 840 845
Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro His Gly Leu
850 855 860
Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly Gln Ala Phe
865 870 875 880
Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val Ala Pro Val
885 890 895
Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile Asp Trp Ala
900 905 910
Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser Gly Arg Leu
915 920 925
Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr Gln Arg
930 935 940
His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe Asn Pro Leu
945 950 955 960
His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe Leu Val Tyr
965 970 975
Asn Ser Ala Leu Thr GluTyr Ala Gly Met Gly Phe Glu Tyr Gly Tyr
980 985 990
Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala Gln Phe Gly
995 1000 1005
Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr Val Ser
1010 1015 1020
Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu Ile Leu Leu
1025 1030 1035
Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His Ser Ser Ala
1040 1045 1050
Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly Ser MetThr
1055 1060 1065
Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His Leu Leu Arg
1070 1075 1080
Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val Ile Phe Thr
1085 1090 1095
Pro Lys Ser Met Leu ArgAsn Lys Ala Ala Ala Ser Ala Pro Glu
1100 1105 1110
Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile Asp Asp Pro
1115 1120 1125
Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met Leu Val Ser
1130 1135 1140
Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys Asp Gly
1145 1150 1155
Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu Hi s Pro Ile
1160 1165 1170
Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr Pro Asn Ala
1175 1180 1185
Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn Gln Gly Pro
1190 1195 1200
Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile Pro Asn Met
1205 1210 1215
Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser Ser ThrAla
1220 1225 1230
Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys Gln Leu Ile
1235 1240 1245
Asp Glu Ala Phe Glu Ala
1250
<210>17
<211>1800
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(211)..(1620)
<223>
<400>17
gatcctccgc attcagtaat ctttcccaca gcatcggctt gttctaaaac agcacggtgc 60
ttagaccgta cataaaacac ctgcatttct gtgcccattt gggcccggat gtgggtgttt 120
ttcattttct tccactctaa aattaagtat ggaaaaccaa ccgcacccgg atgcacgaca 180
atgacccact aaacacgtat ccttgaatgc gtg act gaa cat tat gac gta gta 234
Met Thr Glu His Tyr Asp Val Val
1 5
gta ctc gga gcc ggc ccc ggt ggc tat gtc tcc gcc atc cgt gca gcg 282
Val Leu Gly Ala Gly Pro Gly Gly Tyr Val Ser Ala Ile Arg Ala Ala
10 15 20
cag ctt ggc aag aag gtt gct gta att gag aag cag tac tgg ggt ggt 330
Gln Leu Gly Lys Lys Val Ala Val Ile Glu Lys Gln Tyr Trp Gly Gly
25 30 35 40
gtt tgc cta aac gtg ggc tgc att cct tcc aag tct ctg atc aaa aac 378
Val Cys Leu Asn Val Gly Cys Ile Pro Ser Lys Ser Leu Ile Lys Asn
45 50 55
gct gaa gtt gcc cat acc ttt acc cat gag aag aag acc ttc ggc atc 426
Ala Glu Val Ala His Thr Phe Thr His Glu Lys Lys Thr Phe Gly Ile
60 65 70
aat ggc gaa gtg acc ttc aac tat gag gat gct cac aag cgt tcc cgt 474
Asn Gly Glu Val Thr Phe Asn Tyr Glu Asp Ala His Lys Arg Ser Arg
75 80 85
ggc gtt tcc gac aag atc gtt gga ggc gtt cat tac ttg atg aag aag 522
Gly Val Ser Asp Lys Ile Val Gly Gly Val His Tyr Leu Met Lys Lys
90 95 100
aac aag atc atc gaa att cat ggt ctt gga aac ttc aag gat gct aag 570
Asn Lys Ile Ile Glu Ile His Gly Leu Gly Asn Phe Lys Asp Ala Lys
105 110 115 120
act ctt gag gtc acc gac ggt aag gat gct ggc aag acc atc acc ttt 618
Thr Leu Glu Val Thr Asp Gly Lys Asp Ala Gly Lys Thr Ile Thr Phe
125 130 135
gat gac tgc atc atc gca acc ggt tcg gta gtc aac acc ctc cgt ggc 666
Asp Asp Cys Ile lle Ala Thr Gly Ser Val Val Asn Thr Leu Arg Gly
140 145 150
gtt gac ttc tca gag aac gtt gtg tct ttt gaa gag cag att ctt aac 714
Val Asp Phe Ser Glu Asn Val Val Ser Phe Glu Glu Gln Ile Leu Asn
155 160 165
cct gtt gcg cca aag aag atg gtc att gtt ggt gca ggc gca att gga 762
Pro Val Ala Pro Lys Lys Met Val Ile Val Gly Ala Gly Ala Ile Gly
170 175 180
atg gaa ttc gcc tac gtt ctt ggt aac tac ggt gta gat gta acc gtc 810
Met Glu Phe Ala Tyr Val Leu Gly Asn Tyr Gly Val Asp Val Thr Val
185 190 195 200
atc gag ttc atg gat cgt gtg ctt cca aatgaa gat gct gaa gtc tcc 858
Ile Glu Phe Met Asp Arg Val Leu Pro Asn Glu Asp Ala Glu Val Ser
205 210 215
aag gtt att gca aag gcc tac aag aag atg ggc gtt aag ctt ctt cct 906
Lys Val Ile Ala Lys Ala Tyr Lys Lys Met Gly Val Lys Leu Leu Pro
220 225 230
ggc cat gca acc act gct gtt cgg gac aac ggt gac ttt gtc gag gtt 954
Gly His Ala Thr Thr Ala Val Arg Asp Asn Gly Asp Phe Val Glu Val
235 240 245
gat tac cag aag aag ggc tct gac aag aca gag act ctt act gtt gat 1002
Asp Tyr Gln Lys Lys Gly Ser Asp Lys Thr Glu Thr Leu Thr Val Asp
250 255 260
cga gtc atg gtt tcc gtt ggt ttc cgt cca cgc gtt gag gga ttt ggt 1050
Arg Val Met Val Ser Val Gly Phe Arg Pro Arg Val Glu Gly Phe Gly
265 270 275 280
ctt gaa aac act ggc gtt aag ctc acc gag cgt ggc gca atc gag atc 1098
Leu Glu Asn Thr Gly Val Lys Leu Thr Glu Arg Gly Ala Ile Glu Ile
285 290 295
gat gat tac atg cgt acc aac gtc gat ggc att tac gcc atc ggt gac 1146
Asp Asp Tyr Met Arg Thr Asn Val Asp Gly Ile Tyr Ala Ile Gly Asp
300 305 310
gtg acc gcc aag ctt cag ctt gct cac gtc gca gaa gca cag ggc att 1194
Val Thr Ala Lys Leu Gln Leu Ala His Val Ala Glu Ala Gln Gly Ile
315 320 325
gtt gcc gca gag act att gct ggt gca gaa act cag act ctt ggt gat 1242
Val Ala Ala Glu Thr Ile Ala Gly Ala Glu Thr Gln Thr Leu Gly Asp
330 335 340
tac atg atg atg cca cgt gca acc ttc tgc aac cca cag gtt tct tcc 1290
Tyr Met Met Met Pro Arg Ala Thr Phe Cys Asn Pro Gln Val Ser Ser
345 350 355 360
ttt ggt tac acc gaa gag cag gcc aag gag aag tgg cca gat cgt gag 1338
Phe Gly Tyr Thr Glu Glu Gln Ala Lys Glu Lys Trp Pro Asp Arg Glu
365 370 375
atc aag gtt gct tcc ctc cca ttc tct gca aac ggt aaa gca gtt ggc 1386
Ile Lys Val Ala Ser Leu Pro Phe Ser Ala Asn Gly Lys Ala Val Gly
380 385 390
ctg gca gaa act gat ggt ttc gca aag atc gtt gct gat gca gaa ttc 1434
Leu Ala Glu Thr Asp Gly Phe Ala Lys Ile Val Ala Asp Ala Glu Phe
395 400 405
ggt gag ctg ctc ggt gca cac ctg gtt gga gca aat gca tca gag ctc 1482
Gly Glu Leu Leu Gly Ala His Leu Val Gly Ala Asn Ala Ser Glu Leu
410 415 420
atc aat gaa ttg gtg ctt gct cag aac tgg gat ctc acc act gaa gag 1530
Ile Asn Glu Leu Val Leu Ala Gln Asn Trp Asp Leu Thr Thr Glu Glu
425 430 435 440
atc tct cgt agc gtc cat att cac cca acg cta tct gag gca gtt aag 1578
Ile Ser Arg Ser Val His Ile His Pro Thr Leu Ser Glu Ala Val Lys
445 450 455
gaa gct gca cac ggt atc tct gga cac atg atc aac ttc tag 1620
Glu Ala Ala His Gly Ile Ser Gly His Met Ile Asn Phe
460 465
aatccacctc gttggccctg tttctgtatg gaaacagggc caaaaccgat tttcaatcca 1680
aaccgagtgc tggttagtgc tcatactcac acaaactttt cgattatcaa agagaattat 1740
ttctaaaatt cggtatcgtc taagaaatga gtttgccaat agctcagcat caaaatgctg 1800
<210>18
<211>469
<212>PRT
<213>谷氨酸棒杆菌
<400>18
Met Thr Glu His Tyr Asp Val Val Val Leu Gly Ala Gly Pro Gly Gly
1 5 10 15
Tyr Val Ser Ala Ile Arg Ala Ala Gln Leu Gly Lys Lys Val Ala Val
20 25 30
Ile Glu Lys Gln Tyr Trp Gly Gly Val Cys Leu Asn Val Gly Cys Ile
35 40 45
Pro Ser Lys Ser Leu Ile Lys Asn Ala Glu Val Ala His Thr Phe Thr
50 55 60
His Glu Lys Lys Thr Phe Gly Ile Asn Gly Glu Val Thr Phe Asn Tyr
65 70 75 80
Glu Asp Ala His Lys Arg Ser Arg Gly Val Ser Asp Lys Ile Val Gly
85 90 95
Gly Val His Tyr Leu Met Lys Lys Asn Lys Ile Ile Glu Ile His Gly
100 105 110
Leu Gly Asn Phe Lys Asp Ala Lys Thr Leu Glu Val Thr Asp Gly Lys
115 120 125
Asp Ala Gly Lys Thr Ile Thr Phe Asp Asp Cys Ile Ile Ala Thr Gly
130 135 140
Ser Val Val Asn Thr Leu Arg Gly Val Asp Phe Ser Glu Asn Val Val
145 150 155 160
Ser Phe Glu Glu Gln Ile Leu Asn Pro Val Ala Pro Lys Lys Met Val
165 170 175
Ile Val Gly Ala Gly Ala Ile Gly Met Glu Phe Ala Tyr Val Leu Gly
180 185 190
Asn Tyr Gly Val Asp Val Thr Val Ile Glu Phe Met Asp Arg Val Leu
195 200 205
Pro Asn Glu Asp Ala Glu Val Ser Lys Val Ile Ala Lys Ala Tyr Lys
210 215 220
Lys Met Gly Val Lys Leu Leu Pro Gly His Ala Thr Thr Ala Val Arg
225 230 235 240
Asp Asn Gly Asp Phe Val Glu Val Asp Tyr Gln Lys Lys Gly Ser Asp
245 250 255
Lys Thr Glu Thr Leu Thr Val Asp Arg Val Met Val Ser Val Gly Phe
260 265 270
Arg Pro Arg Val Glu Gly Phe Gly Leu Glu Ash Thr Gly Val Lys Leu
275 280 285
Thr Glu Arg Gly Ala Ile Glu Ile Asp Asp Tyr Met Arg Thr Asn Val
290 295 300
Asp Gly Ile Tyr Ala Ile Gly Asp Val Thr Ala Lys Leu Gln Leu Ala
305 310 315 320
His Val Ala Glu Ala Gln Gly Ile Val Ala Ala Glu Thr Ile Ala Gly
325 330 335
Ala Glu Thr Gln Thr Leu Gly Asp Tyr Met Met Met Pro Arg Ala Thr
340 345 350
Phe Cys Asn Pro Gln Val Ser Ser Phe Gly Tyr Thr Glu Glu Gln Ala
355 360 365
Lys Glu Lys Trp Pro Asp Arg Glu Ile Lys Val Ala Ser Leu Pro Phe
370 375 380
Ser Ala Asn Gly Lys Ala Val Gly Leu Ala Glu Thr Asp Gly Phe Ala
385 390 395 400
Lys Ile Val Ala Asp Ala Glu Phe Gly Glu Leu Leu Gly Ala His Leu
405 410 415
Val Gly Ala Asn Ala Ser Glu Leu Ile Asn Glu Leu Val Leu Ala Gln
420 425 430
Asn Tro Asp Leu Thr Thr Glu Glu Ile Ser Arg Ser Val His Ile His
435 440 445
Pro Thr Leu Ser Glu Ala Val Lys Glu Ala Ala His Gly Ile Ser Gly
450 455 460
His Met Ile Asn Phe
465
<210>19
<211>2014
<212>DNA
<213>枯草芽孢杆菌
<220>
<221>CDS
<222>(464)..(1885)
<223>
<400>19
gatccttttt aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat 60
tatgatattt tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat 120
aaaaaataca gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct 180
gcaaatcctt ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa 240
acgagagtct aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc 300
aggcaagacc taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat 360
tttaggtctt tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag 420
aagcagaccg ctaacacagt acataaaaaa ggagacatga acg atg aac atc aaa 475
Met Asn Ile Lys
1
aag ttt gca aaa caa gca aca gta tta acc ttt act acc gca ctg ctg 523
Lys Phe Ala Lys Gln Ala Thr Val Leu Thr Phe Thr Thr Ala Leu Leu
5 10 15 20
gca gga ggc gca act caa gcg ttt gcg aaa gaa acg aac caa aag cca 571
Ala Gly Gly Ala Thr Gln Ala Phe Ala Lys Glu Thr Asn Gln Lys Pro
25 30 35
tat aag gaa aca tac ggc att tcc cat att aca cgc cat gat atg ctg 619
Tyr Lys Glu Thr Tyr Gly Ile Ser His Ile Thr Arg His Asp Met Leu
40 45 50
caa atc cct gaa cag caa aaa aat gaa aaa tat caa gtt cct gaa ttc 667
Gln Ile Pro Glu Gln Gln Lys Asn Glu Lys Tyr Gln Val Pro Glu Phe
55 60 65
gat tcg tcc aca att aaa aat atc tct tct gca aaa ggc ctg gac gtt 715
Asp Ser Ser Thr Ile Lys Asn Ile Ser Ser Ala Lys Gly Leu Asp Val
70 75 80
tgg gac agc tgg cca tta caa aac gct gac ggc act gtc gca aac tat 763
Trp Asp Ser Trp Pro Leu Gln Asn Ala Asp Gly Thr Val Ala Asn Tyr
85 90 95 100
cac ggc tac cac atc gtc ttt gca tta gcc gga gat cct aaa aat gcg 811
His Gly Tyr His Ile Val Phe Ala Leu Ala Gly Asp Pro Lys Asn Ala
105 110 115
gat gac aca tcg att tac atg ttc tat caa aaa gtc ggc gaa act tct 859
Asp Asp Thr Ser Ile Tyr Met Phe Tyr Gln Lys Val Gly Glu Thr Ser
120 125 130
att gac agc tgg aaa aac gct ggc cgc gtc ttt aaa gac agc gac aaa 907
Ile Asp Ser Trp Lys Asn Ala Gly Arg Val Phe Lys Asp Ser Asp Lys
135 140 145
ttc gat gca aat gat tct atc cta aaa gac caa aca caa gaa tgg tca 955
Phe Asp Ala Asn Asp Ser Ile Leu Lys Asp Gln Thr Gln Glu Trp Ser
150 155 160
ggt tca gcc aca ttt aca tct gac gga aaa atc cgt tta ttc tac act 1003
Gly Ser Ala Thr Phe Thr Ser Asp Gly Lys Ile Arg Leu Phe Tyr Thr
165 170 175 180
gat ttc tcc ggt aaa cat tac ggc aaa caa aca ctg aca act gca caa 1051
Asp Phe Ser Gly Lys His Tyr Gly Lys Gln Thr Leu Thr Thr Ala Gln
185 190 195
gtt aac gta tca gca tca gac agc tct ttg aac atc aac ggt gta gag 1099
Val Asn Val Ser Ala Ser Asp Ser Ser Leu Asn Ile Asn Gly Val Glu
200 205 210
gat tat aaa tca atc ttt gac ggt gac gga aaa acg tat caa aat gta 1147
Asp Tyr Lys Ser Ile Phe Asp Gly Asp Gly Lys Thr Tyr Gln Asn Val
215 220 225
cag cag ttc atc gat gaa ggc aac tac agc tca ggc gac aac cat acg 1195
Gln Gln Phe Ile Asp Glu Gly Asn Tyr Ser Ser Gly Asp Asn His Thr
230 235 240
ctg aga gat cct cac tac gta gaa gat aaa ggc cac aaa tac tta gta 1243
Leu Arg Asp Pro His Tyr Val Glu Asp Lys Gly His Lys Tyr Leu Val
245 250 255 260
ttt gaa gca aac act gga act gaa gat ggc tac caa ggc gaa gaa tct 1291
Phe Glu Ala Asn Thr Gly Thr Glu Asp Gly Tyr Gln Gly Glu Glu Ser
265 270 275
tta ttt aac aaa gca tac tat ggc aaa agc aca tca ttc ttc cgt caa 1339
Leu Phe Asn Lys Ala Tyr Tyr Gly Lys Ser Thr Ser Phe Phe Arg Gln
280 285 290
gaa agt caa aaa ctt ctg caa agc gat aaa aaa cgc acg gct gag tta 1387
Glu Ser Gln Lys Leu Leu Gln Ser Asp Lys Lys Arg Thr Ala Glu Leu
295 300 305
gca aac ggc gct ctc ggt atg att gag cta aac gat gat tac aca ctg 1435
Ala Asn Gly Ala Leu Gly Met Ile Glu Leu Asn Asp Asp Tyr Thr Leu
310 315 320
aaa aaa gtg atg aaa ccg ctg att gca tct aac aca gta aca gat gaa 1483
Lys Lys Val Met Lys Pro Leu Ile Ala Ser Asn Thr Val Thr Asp Glu
325 330 335 340
att gaa cgc gcg aac gtc ttt aaa atg aac ggc aaa tgg tac ctg ttc 1531
Ile Glu Arg Ala Asn Val Phe Lys Met Asn Gly Lys Trp Tyr Leu Phe
345 350 355
act gac tcc cgc gga tca aaa atg acg att gac ggc att acg tct aac 1579
Thr Asp Ser Arg Gly Ser Lys Met Thr Ile Asp Gly Ile Thr Ser Asn
360 365 370
gat att tac atg ctt ggt tat gtt tct aat tct tta act ggc cca tac 1627
Asp Ile Tyr Met Leu Gly Tyr Val Ser Asn Ser Leu Thr Gly Pro Tyr
375 380 385
aag ccg ctg aac aaa act ggc ctt gtg tta aaa atg gat ctt gat cct 1675
Lys Pro Leu Asn Lys Thr Gly Leu Val Leu Lys Met Asp Leu Asp Pro
390 395 400
aac gat gta acc ttt act tac tca cac ttc gct gta cct caa gcg aaa 1723
Asn Asp Val Thr Phe Thr Tyr Ser His Phe Ala Val Pro Gln Ala Lys
405 410 415 420
gga aac aat gtc gtg att aca agc tat atg aca aac aga gga ttc tac 1771
Gly Asn Asn Val Val Ile Thr Ser Tyr Met Thr Asn Arg Gly Phe Tyr
425 430 435
gca gac aaa caa tca acg ttt gcg cca agc ttc ctg ctg aac atc aaa 1819
Ala Asp Lys Gln Ser Thr Phe Ala Pro Ser Phe Leu Leu Asn Ile Lys
440 445 450
ggc aag aaa aca tct gtt gtc aaa gac agc atc ctt gaa caa gga caa 1867
Gly Lys Lys Thr Ser Val Val Lys Asp Ser Ile Leu Glu Gln Gly Gln
455 460 465
tta aca gtt aac aaa taa aaacgcaaaa gaaaatgccg atatcctatt 1915
Leu Thr Val Asn Lys
470
ggcattttct tttatttctt atcaacataa aggtgaatcc catatgaact atataaaagc 1975
aggcaaatgg ctaaccgtat tcctaacctt tt gaagatc 2014
<210>20
<211>473
<212>PRT
<213>枯草芽孢杆菌
<400>20
Met Asn Ile Lys Lys Phe Ala Lys Gln Ala Thr Val Leu Thr Phe Thr
1 5 10 15
Thr Ala Leu Leu Ala Gly Gly Ala Thr Gln Ala Phe Ala Lys Glu Thr
20 25 30
Asn Gln Lys Pro Tyr Lys Glu Thr Tyr Gly Ile Ser His Ile Thr Arg
35 40 45
His Asp Met Leu Gln Ile Pro Glu Gln Gln Lys Asn Glu Lys Tyr Gln
50 55 60
Val Pro Glu Phe Asp Ser Ser Thr Ile Lys Asn Ile Ser Ser Ala Lys
65 70 75 80
Gly Leu Asp Val Trp Asp Ser Trp Pro Leu Gln Asn Ala Asp Gly Thr
85 90 95
Val Ala Asn Tyr His Gly Tyr His Ile Val Phe Ala Leu Ala Gly Asp
100 105 110
Pro Lys Asn Ala Asp Asp Thr Ser Ile Tyr Met Phe Tyr Gln Lys Val
115 120 125
Gly Glu Thr Ser Ile Asp Ser Trp Lys Asn Ala Gly Arg Val Phe Lys
130 135 140
Asp Ser Asp Lys Phe Asp Ala Asn Asp Ser Ile Leu Lys Asp Gln Thr
145 150 155 160
Gln Glu Trp Ser Gly Ser Ala Thr Phe Thr Ser Asp Gly Lys Ile Arg
165 170 175
Leu Phe Tyr Thr Asp Phe Ser Gly Lys His Tyr Gly Lys Gln Thr Leu
180 185 190
Thr Thr Ala Gln Val Asn Val Ser Ala Ser Asp Ser Ser Leu Asn Ile
195 200 205
Asn Gly Val Glu Asp Tyr Lys Ser Ile Phe Asp Gly Asp Gly Lys Thr
210 215 220
Tyr Gln Asn Val Gln Gln Phe Ile Asp Glu Gly Asn Tyr Ser Ser Gly
225 230 235 240
Asp Asn His Thr Leu Arg Asp Pro His Tyr Val Glu Asp Lys Gly His
245 250 255
Lys Tyr Leu Val Phe Glu Ala Asn Thr Gly Thr Glu Asp Gly Tyr Gln
260 265 270
Gly Glu Glu Ser Leu Phe Asn Lys Ala Tyr Tyr Gly Lys Ser Thr Ser
275 280 285
Phe Phe Arg Gln Glu Ser Gln Lys Leu Leu Gln Ser Asp Lys Lys Arg
290 295 300
Thr Ala Glu Leu Ala Asn Gly Ala Leu Gly Met Ile Glu Leu Asn Asp
305 310 315 320
Asp Tyr Thr Leu Lys Lys Val Met Lys Pro Leu Ile Ala Ser Asn Thr
325 330 335
Val Thr Asp Glu Ile Glu Arg Ala Asn Val Phe Lys Met Asn Gly Lys
340 345 350
Trp Tyr Leu Phe Thr Asp Ser Arg Gly Ser Lys Met Thr Ile Asp Gly
355 360 365
Ile Thr Ser Asn Asp Ile Tyr Met Leu Gly Tyr Val Ser Asn Ser Leu
370 375 380
Thr Gly Pro Tyr Lys Pro Leu Asn Lys Thr Gly Leu Val Leu Lys Met
385 390 395 400
Asp Leu Asp Pro Asn Asp Val Thr Phe Thr Tyr Ser His Phe Ala Val
405 410 415
Pro Gln Ala Lys Gly Asn Asn Val Val Ile Thr Ser Tyr Met Thr Asn
420 425 430
Arg Gly Phe Tyr Ala Asp Lys Gln Ser Thr Phe Ala Pro Ser Phe Leu
435 440 445
Leu Asn Ile Lys Gly Lys Lys Thr Ser Val Val Lys Asp Ser Ile Leu
450 455 460
Glu Gln Gly Gln Leu Thr Val Asn Lys
465 470
<210>21
<211>24
<212>DNA
<213>人工的
<220>
<223>引物
<400>21
cgggat cctttttaacccat caca 24
<210>22
<211>29
<212>DNA
<213>人工的
<220>
<223>引物
<400>22
gaagatcttc aaaaggttag gaatacggt 29
<210>23
<211>23
<212>DNA
<213>人工的
<220>
<223>引物
<400>23
ccttttgaag atcgaccagt tgg 23
<210>24
<211>44
<212>DNA
<213>人工的
<220>
<223>引物
<400>24
tacctggaat gctgttttcc cagggatcgc agtggtgagt aacc 44
<210>25
<211>28
<212>DNA
<213>人工的
<220>
<223>引物
<400>25
cctgggaaaa cagcat tcca ggtattag 28
<210>26
<211>23
<212>DNA
<213>人工的
<220>
<223>引物
<400>26
tgcaggtcga ctctagagga tcc 23
<210>27
<211>2028
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(2028)
<223>
<400>27
atg gcg ttc tcc gta gag atg ccc gag ctg ggc gaa tca gta acc gaa 48
Met Ala Phe Ser Val Glu Met Pro Glu Leu Gly Glu Ser Val Thr Glu
1 5 10 15
ggc acg atc acc cag tgg ttg aag tct gtt ggt gac act gtt gag gta 96
Gly Thr Ile Thr Gln Trp Leu Lys Ser Val Gly Asp Thr Val Glu Val
20 25 30
gat gag ccg ttg ctc gag gtc tca act gac aag gtc gac acc gag att 144
Asp Glu Pro Leu Leu Glu Val Ser Thr Asp Lys Val Asp Thr Glu Ile
35 40 45
ccc tct cct gtc gcc ggt gtc atc cta gag att aag gct gaa gag gat 192
Pro Ser Pro Val Ala Gly Val Ile Leu Glu Ile Lys Ala Glu Glu Asp
50 55 60
gac acc gtc gac gtc ggc ggt gtc att gca ata atc ggc gat gct gat 240
Asp Thr Val Asp Val Gly Gly Val Ile Ala Ile Ile Gly Asp Ala Asp
65 70 75 80
gag act cct gcc aac gaa gct cct gcc gac gag gca cca gct cct gcc 288
Glu Thr Pro Ala Asn Glu Ala Pro Ala Asp Glu Ala Pro Ala Pro Ala
85 90 95
gaa gag gaa gaa cca gtt aag gaa gag cca aag aag gag gca gct cct 336
Glu Glu Glu Glu Pro Val Lys Glu Glu Pro Lys Lys Glu Ala Ala Pro
100 105 110
gaa gct cca gca gca act ggc gcc gca acc gat gtg gaa atg cca gaa 384
Glu Ala Pro Ala Ala Thr Gly Ala Ala Thr Asp Val Glu Met Pro Glu
115 120 125
ctc ggc gaa tcc gtc acc gaa ggc acc att acc cag tgg ctc aag gct 432
Leu Gly Glu Ser Val Thr Glu Gly Thr Ile Thr Gln Trp Leu Lys Ala
130 135 140
gtc ggc gac acc gtcgaa gta gac gaa cca ctt ctt gag gtc tcc acc 480
Val Gly Asp Thr Val Glu Val Asp Glu Pro Leu Leu Glu Val Ser Thr
145 150 155 160
gac aag gtc gac acc gaa atc cca tcc cca gta gca ggc acc atc gtg 528
Asp Lys Val Asp Thr Glu Ile Pro Ser Pro Val Ala Gly Thr Ile Val
165 170 175
gag atc ctt gca gac gaa gac gac acc gtc gac gtc ggc gca gtc atc 576
Glu Ile Leu Ala Asp Glu Asp Asp Thr Val Asp Val Gly Ala Val Ile
180 185 190
gcc cgc atc ggt gac gca aac gca gct gca gca cct gcc gaa gag gaa 624
Ala Arg Ile Gly Asp Ala Asn Ala Ala Ala Ala Pro Ala Glu Glu Glu
195 200 205
gca gct cct gcc gaa gag gaa gaa cca gtt aag gaa gag cca aag aag 672
Ala Ala Pro Ala Glu Glu Glu Glu Pro Val Lys Glu Glu Pro Lys Lys
210 215 220
gag gca gct cct gaa gct cca gca gca act ggc gcc gca acc gat gtg 720
Glu Ala Ala Pro Glu Ala Pro Ala Ala Thr Gly Ala Ala Thr Asp Val
225 230 235 240
gaa atg cca gaa ctc ggc gaa tcc gtc acc gaa ggc acc att acc cag 768
Glu Met Pro Glu Leu Gly Glu Ser Val Thr Glu Gly Thr Ile Thr Gln
245 250 255
tgg ctc aag gct gtc ggc gac acc gtc gaa gta gac gaa cca ctt ctt 816
Trp Leu Lys Ala Val Gly Asp Thr Val Glu Val Asp Glu Pro Leu Leu
260 265 270
gag gtc tcc acc gac aag gtc gac acc gaa atc cca tcc cca gta gca 864
Glu Val Ser Thr Asp Lys Val Asp Thr Glu Ile Pro Ser Pro Val Ala
275 280 285
ggc acc atc gtg gag atc ctt gca gac gaa gac gac acc gtc gac gtc 912
Gly Thr Ile Val Glu Ile Leu Ala Asp Glu Asp Asp Thr Val Asp Val
290 295 300
ggc gca gtc atc gcc cgc atc ggt gac gca aac gca gct gca gca cct 960
Gly Ala Val Ile Ala Arg Ile Gly Asp Ala Asn Ala Ala Ala Ala Pro
305 310 315 320
gcc gaa gag gaa gca gct cct gcc gaa gag gag gaa cca gtt aag gaa 1008
Ala Glu Glu Glu Ala Ala Pro Ala Glu Glu Glu Glu Pro Val Lys Glu
325 330 335
gag cca aag aag gaa gag ccc aag aag gaa gag ccc aag aag gaa gca 1056
Glu Pro Lys Lys Glu Glu Pro Lys Lys Glu Glu Pro Lys Lys Glu Ala
340 345 350
gct act aca cct gct gcg gca tcc gca act gtg tcc gct tct ggc gac 1104
Ala Thr Thr Pro Ala Ala Ala Ser Ala Thr Val Ser Ala Ser Gly Asp
355 360 365
aac gtt cca tac gtc acc cca ctg gtg cgc aag ctt gct gaa aag cac 1152
Asn Val Pro Tyr Val Thr Pro Leu Val Arg Lys Leu Ala Glu Lys His
370 375 380
ggc gtt gac ttg aac acc gtg acc ggt acc ggt atc ggt ggc cgt atc 1200
Gly Val Asp Leu Asn Thr Val Thr Gly Thr Gly Ile Gly Gly Arg Ile
385 390 395 400
cgc aag cag gat gtt ttg gct gct gcg aac ggc gag gct gca cct gct 1248
Arg Lys Gln Asp Val Leu Ala Ala Ala Asn Gly Glu Ala Ala Pro Ala
405 410 415
gag gct gct gct cct gtt tcc gct tgg tcc act aag tct gtt gac cct 1296
Glu Ala Ala Ala Pro Val Ser Ala Trp Ser Thr Lys Ser Val Asp Pro
420 425 430
gag aag gct aag ctc cgt ggt acc act cag aag gtc aac cgc atc cgt 1344
Glu Lys Ala Lys Leu Arg Gly Thr Thr Gln Lys Val Asn Arg Ile Arg
435 440 445
gag atc acc gcg atg aag acc gtc gag gct ctg cag att tct gct cag 1392
Glu Ile Thr Ala Met Lys Thr Val Glu Ala Leu Gln Ile Ser Ala Gln
450 455 460
ctc acc cag ctg cac gag gtc gat atg act cgc gtt gct gag ctg cgt 1440
Leu Thr Gln Leu His Glu Val Asp Met Thr Arg Val Ala Glu Leu Arg
465 470 475 480
aag aag aac aag ccc gcg ttc atc gag aag cac ggt gtg aac ctc act 1488
Lys Lys Asn Lys Pro Ala Phe Ile Glu Lys His Gly Val Asn Leu Thr
485 490 495
tac ctg cca ttc ttc gtg aag gca gtt gtc gag gct ttg gtt tcc cat 1536
Tyr Leu Pro Phe Phe Val Lys Ala Val Val Glu Ala Leu Val Ser His
500 505 510
cca aac gtc aac gcg tct ttc aac gcg aag acc aag gag atg acc tac 1584
Pro Asn Val Asn Ala Ser Phe Asn Ala Lys Thr Lys Glu Met Thr Tyr
515 520 525
cac tcc tcc gtt aac ctc tcc atc gct gtt gat acc cca gct ggt ctg 1632
His Ser Ser Val Asn Leu Ser Ile Ala Val Asp Thr Pro Ala Gly Leu
530 535 540
ttg acc cca gtc att cac gat gct cag gat ctc tcc atc cca gag atc 1680
Leu Thr Pro Val Ile His Asp Ala Gln Asp Leu Ser Ile Pro Glu Ile
545 550 555 560
gca aag gca att gtt gac ctg gct gat cgt tca cgc aac aac aag ctg 1728
Ala Lys Ala Ile Val Asp Leu Ala Asp Arg Ser Arg Asn Asn Lys Leu
565 570 575
aag cca aac gat ctg tcc ggt ggc acc ttc acc atc acc aac att ggt 1776
Lys Pro Asn Asp Leu Ser Gly Gly Thr Phe Thr Ile Thr Asn Ile Gly
580 585 590
tct gaa ggc gca ctg tct gat acc cca atc ctg gtt cca cca cag gct 1824
Ser Glu Gly Ala Leu Ser Asp Thr Pro Ile Leu Val Pro Pro Gln Ala
595 600 605
ggc atc ttg ggc acc ggc gcg atc gtg aag cgt cca gtt gtc atc acc 1872
Gly Ile Leu Gly Thr Gly Ala Ile Val Lys Arg Pro Val Val Ile Thr
610 615 620
gag gat gga att gat tcc atc gcg atc cgt cag atg gtc ttc cta cca 1920
Glu Asp Gly Ile Asp Ser Ile Ala Ile Arg Gln Met Val Phe Leu Pro
625 630 635 640
ctg acc tac gac cac cag gtt gta gat ggc gca gat gct ggt cgc ttc 1968
Leu Thr Tyr Asp His Gln Val Val Asp Gly Ala Asp Ala Gly Arg Phe
645 650 655
ctg acc acc atc aag gac cgc ctt gag acc gct aac ttc gaa ggc gat 2016
Leu Thr Thr Ile Lys Asp Arg Leu Glu Thr Ala Asn Phe Glu Gly Asp
660 665 670
ctg cag ctc taa 2028
Leu Gln Leu
675
<210>28
<211>675
<212>PRT
<213>谷氨酸棒杆菌
<400>28
Met Ala Phe Ser Val Glu Met Pro Glu Leu Gly Glu Ser Val Thr Glu
1 5 10 15
Gly Thr Ile Thr Gln Trp Leu Lys Ser Val Gly Asp Thr Val Glu Val
20 25 30
Asp Glu Pro Leu Leu Glu Val Ser Thr Asp Lys Val Asp Thr Glu Ile
35 40 45
Pro Ser Pro Val Ala Gly Val Ile Leu Glu Ile Lys Ala Glu Glu Asp
50 55 60
Asp Thr Val Asp Val Gly Gly Val Ile Ala Ile Ile Gly Asp Ala Asp
65 70 75 80
Glu Thr Pro Ala Asn Glu Ala Pro Ala Asp Glu Ala Pro Ala Pro Ala
85 90 95
Glu Glu Glu Glu Pro Val Lys Glu Glu Pro Lys Lys Glu Ala Ala Pro
100 105 110
Glu Ala Pro Ala Ala Thr Gly Ala Ala ThrAsp Val Glu Met Pro Glu
115 120 125
Leu Gly Glu Ser Val Thr Glu Gly Thr Ile Thr Gln Trp Leu Lys Ala
130 135 140
Val Gly Asp Thr Val Glu Val Asp Glu Pro Leu Leu Glu Val Ser Thr
145 150 155 160
Asp Lys Val Asp Thr Glu Ile Pro Ser Pro Val Ala Gly Thr Ile Val
165 170 175
Glu Ile Leu Ala Asp Glu Asp Asp Thr Val Asp Val Gly Ala Val Ile
180 185 190
Ala Arg Ile Gly Asp Ala Asn Ala Ala Ala Ala Pro Ala Glu Glu Glu
195 200 205
Ala Ala Pro Ala Glu Glu Glu Glu Pro Val Lys Glu Glu Pro Lys Lys
210 215 220
Glu Ala Ala Pro Glu Ala Pro Ala Ala Thr Gly Ala Ala Thr Asp Val
225 230 235 240
Glu Met Pro Glu Leu Gly Glu Ser Val Thr Glu Gly Thr Ile Thr Gln
245 250 255
Trp Leu Lys Ala Val Gly Asp Thr Val Glu Val Asp Glu Pro Leu Leu
260 265 270
Glu Val Ser Thr Asp Lys Val Asp Thr Glu Ile Pro Ser Pro Val Ala
275 280 285
Gly Thr Ile Val Glu Ile Leu Ala Asp Glu Asp Asp Thr Val Asp Val
290 295 300
Gly Ala Val Ile Ala Arg Ile Gly Asp Ala Asn Ala Ala Ala Ala Pro
305 310 315 320
Ala Glu Glu Glu Ala Ala Pro Ala Glu Glu Glu Glu Pro Val Lys Glu
325 330 335
Glu Pro Lys Lys Glu Glu Pro Lys Lys Glu Glu Pro Lys Lys Glu Ala
340 345 350
Ala Thr Thr Pro Ala Ala Ala Ser Ala Thr Val Ser Ala Ser Gly Asp
355 360 365
Asn Val Pro Tyr Val Thr Pro Leu Val Arg Lys Leu Ala Glu Lys His
370 375 380
Gly Val Asp Leu Asn Thr Val Thr Gly Thr Gly Ile Gly Gly Arg Ile
385 390 395 400
Arg Lys Gln Asp Val Leu Ala Ala Ala Asn Gly Glu Ala Ala Pro Ala
405 410 415
Glu Ala Ala Ala Pro Val Ser Ala Trp Ser Thr Lys Ser Val Asp Pro
420 425 430
Glu Lys Ala Lys Leu Arg Gly Thr Thr Gln Lys Val Asn Arg Ile Arg
435 440 445
Glu Ile Thr Ala Met Lys Thr Val Glu Ala Leu Gln Ile Ser Ala Gln
450 455 460
Leu Thr Gln Leu His Glu Val Asp Met Thr Arg Val Ala Glu Leu Arg
465 470 475 480
Lys Lys Asn Lys Pro Ala Phe Ile Glu Lys His Gly Val Asn Leu Thr
485 490 495
Tyr Leu Pro Phe Phe Val Lys Ala Val Val Glu Ala Leu Val Ser His
500 505 510
Pro Asn Val Asn Ala Ser Phe Asn Ala Lys Thr Lys Glu Met Thr Tyr
515 520 525
His Ser Ser Val Asn Leu Ser Ile Ala Val Asp Thr Pro Ala Gly Leu
530 535 540
Leu Thr Pro Val Ile His Asp Ala Gln Asp Leu Ser Ile Pro Glu Ile
545 550 555 560
Ala Lys Ala Ile Val Asp Leu Ala Asp Arg Ser Arg Asn Asn Lys Leu
565 570 575
Lys Pro Asn Asp Leu Ser Gly Gly Thr Phe Thr Ile Thr Asn Ile Gly
580 585 590
Ser Glu Gly Ala Leu Ser Asp Thr Pro Ile Leu Val Pro Pro Gln Ala
595 600 605
Gly Ile Leu Gly Thr Gly Ala Ile Val Lys Arg Pro Val Val Ile Thr
610 615 620
Glu Asp Gly Ile Asp Ser Ile Ala Ile Arg Gln Met Val Phe Leu Pro
625 630 635 640
Leu Thr Tyr Asp His Gln Val Val Asp Gly Ala Asp Ala Gly Arg Phe
645 650 655
Leu Thr Thr Ile Lys Asp Arg Leu Glu Thr Ala Asn Phe Glu Gly Asp
660 665 670
Leu Gln Leu
675
<210>29
<211>3481
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1437)..(3035)
<223>yggB
<400>29
gatcttgccg atttcggcag gatcaatgtc ggcgtgaatg atcttggcat caggtgcgaa 60
agtgtcaacg tcaccggtga cgcggtcatc aaagcgggag ccgatagcaa tcagcaggtc 120
gctgcgctgc agtgcaccaa cagcggacac agtgccatgc atgcctggca tacccatgtg 180
cagctcgtgg gactctggga aggttcccag cgccatcaat gtggtgacaa ctggaatgcc 240
ggtgtgctca gcgaacgcac gaagctcttc gtgggcatca gccttgataa cgccgccgcc 300
aacgtaaagg acaggcttct tagactcacc gatcagtttg acagcctgct caatctgtcg 360
agcatgcggt gttgaaactg ggcggtagcc tggcaggtcg atctttggtg gccagacgaa 420
atccaattca gcgttctgaa catccttggg gatatccact agaacaggac cagggcgacc 480
agtaatcgcg aggtggaatg cctcagccaa tgcctgtgga atgtcgttgg ggttggtgac 540
catgaagttg tgcttggtca ctggcatggt gatgccgcgg atatcggctt cctggaaagc 600
atcggtaccc agcaggctac ttccgacctg gccggtgatg gcaaccatgg gaacggagtc 660
caagtttgca tcagcgattg gggtaaccaa gttggttgcg cctgggccag aggttgcaat 720
gcagacgcca acgcgtccag taacctgcgc gtagccggtt gctgcgtggc ctgcgccctg 780
ctcgtggcgc actaggacgt ggcgcacctt tgtggaggaa tagagcgggt catacaccgg 840
tagcaccgca ccaccaggaa taccgaacac gatgtcggcg ttaagctcct cgagcgatcg 900
aacaattgcc tgtgcacctg tcatccgctc aggggcggcg gatcgaccac ggcttgcaac 960
cgtggcggga gtgggctgtt gagaagctgc cacattcacg actttctggc tcctttacta 1020
aataaggatt ttcacaggac ccgtccaagc caagccgatt tcaactcagc ctaaagacaa 1080
agccctcatt taaaattgtt ccgacgcgga tgcgtgtgca cgcagtgcga cagatgtctg 1140
ttgcaaagtt ggctacttgg gtcataacca acaagaaagc cctcgttcca acactgtggt 1200
gagtgttgtc gagggcgctt gacgagacga cttggaaggc cgttacggca ggcgccgcgc 1260
ggttactact acaagtcgaa taatggtcat ggtgtgtcat gctacacaca tcgagtttcc 1320
aattccacaa cgcacgaaaa ttcccacccc caaaactccc ccacttcggt taaggaatca 1380
ggattctcac aaagttcagg caggctcccg ctacttttca gcgctaatct tggctc atg 1439
Met
1
att tta ggc gta ccc att caa tat ttg ctc tat tca ttg tgg aat tgg 1487
Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn Trp
5 10 15
att gtc gat acc ggt ttt gat gta gca att atc ctg gtc ttg gcg ttt 1535
Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala Phe
20 25 30
ttg att cca cgt atc ggc cga ctg gcc atg cgt att atc aag cag cga 1583
Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Gln Arg
35 40 45
gtg gag tct gca gcc gat gcg gac acc act aag aac cag ctc gcg ttc 1631
Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala Phe
50 55 60 65
gct ggc gtt ggc gtt tat atc gcg caa att gtg gcg ttt ttc atg ctt 1679
Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met Leu
70 75 80
gcc gtc tcc gcg atg cag gct ttt ggt ttc tct ctc gcg ggc gct gcg 1727
Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala Ala
85 90 95
att ccg gca acc att gcg tca gct gcc att ggt ctt ggt gcg cag tcg 1775
Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln Ser
100 105 110
att gtt gcg gac ttc ttg gcc gga ttt ttc atc ctg acg gaa aag caa 1823
Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys Gln
115 120 125
ttc ggc gtg ggt gac tgg gtg cgc ttt gag ggc aac ggc atc gtt gtt 1871
Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val Val
130 135 140 145
gaa ggc acc gtc att gag atc acc atg cgc gcg acc aaa att cgc acg 1919
Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg Thr
150 155 160
att gca caa gag acc gtg atc atc ccg aac tcc acg gcg aaa gtg tgc 1967
Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val Cys
165 170 175
atc aac aat tct aat aac tgg tcg cgt gcg gtt gtc gtt att ccg atc 2015
Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro Ile
180 185 190
ccc atg ttg ggt tct gaa aac atc aca gat gtc atc gcg cgc tct gaa 2063
Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser Glu
195 200 205
gct gcg act cgt cgc gca ctt ggc cag gag aaa atc gca ccg gaa atc 2111
Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu lle
210 215 220 225
ctc ggt gaa ctc gat gtg cac cca gcc acg gaa gtc aca ccg cca acg 2159
Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro Thr
230 235 240
gtg gtc ggc atg ccg tgg atg gtc acc atg cgt ttc ctc gtg caa gtc 2207
Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln Val
245 250 255
acc gcc ggc aat caa tgg ctg gtc gaa cgc gcc atc cgc aca gaa atc 2255
Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu Ile
260 265 270
atc aac gaa ttc tgg gaa gaa tac ggc agc gca acc act aca tcg gga 2303
Ile Asn Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser Gly
275 280 285
acc ctc att gat tcc tta cac gtt gag cat gaa gag cca aag acc tcg 2351
Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr Ser
290 295 300 305
ctt atc gac gcc tcc ccc cag gct ctt aag gaa ccg aag ccg gag gct 2399
Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu Ala
310 315 320
gcg gcg acg gtt gca tcg cta gct gca tcg tct aac gac gat gca gac 2447
Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala Asp
325 330 335
aat gca gac gcc tcg gcg atc aat gca ggc aat cca gag aag gaa ctt 2495
Asn Ala Asp Ala Ser Ala Ile Ash Ala Gly Asn Pro Glu Lys Glu Leu
340 345 350
gat tcc gat gtg ctg gaa caa gaa ctc tcc agc gaa gaa ccg gaa gaa 2543
Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu Glu
355 360 365
aca gca aaa cca gat cac tct ctc cga ggc ttc ttc cgc act gat tac 2591
Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp Tyr
370 375 380 385
tac cca aat cgg tgg cag aag atc ctg tcg ttt ggc gga cgt gtc cgc 2639
Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val Arg
390 395 400
atg agc act tcc ctg ttg ttg ggt gcg ctg ctc ttg ctg tca cta ttt 2687
Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu Phe
405 410 415
aag gtc atg act gtg gaa cca agt gag aat tgg caa aac tcc agt gga 2735
Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser Gly
420 425 430
tgg ctg tca cca agc act gcc acc tca act gcg gtg acc acc tcc gaa 2783
Trp Leu Ser Pro 8er Thr Ala Thr Ser Thr Ala Val Thr Thr Ser Glu
435 440 445
act tcc gcg cca gca agc acg cct tcg atg aca gtg ccc act acg gtg 2831
Thr Ser Ala Pro Ala Ser Thr Pro Ser Met Thr Val Pro Thr Thr Val
450 455 460 465
gag gag acc cca acg atg gaa tct agc gtc gaa acg cag cag gaa acc 2879
Glu Glu Thr Pro Thr Met Glu Ser Ser Val Glu Thr Gln Gln Glu Thr
470 475 480
tca acc cct gca acc gca acg ccc cag cga gcc gac acc atc gaa ccg 2927
Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr lle Glu Pro
485 490 495
acc gag gaa gcc acg tcg cag gag gaa acg act gca tcg cag acg cag 2975
Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr Gln
500 505 510
tct cca gca gtg gaa gca cca acc gcg gtc caa gaa aca gtt gcg ccg 3023
Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala Pro
515 520 525
acg tcc acc cct taggacgctg attacagacg tgtcccattt ctttactact 3075
Thr Ser Thr Pro
530
attggaaatt atgagttcag acgcagaaaa ggcatccgtg gagctttccg aaaaatttca 3135
cccagaacgc acccatattt tgggcgccgt tgtttttggc ctgatctcat tattagtcat 3195
cggcgcagcc cctcagtacc tgttttggct gctcgcgctc cctgtcatct tcggttactg 3255
ggttctaaaa tcatccacga tcgttgatga acagggcatc accgcaaact acgccttcaa 3315
gggcaaaaag gttgtggcct gggaagacct cgcaggaatc ggattcaagg gtgcccgcac 3375
tttcgctcgc accacctccg atgcagaagt caccctcccc ggcgtcacct tcaactccct 3435
tccccgcctt gaagctgctt cccacggccg catccccgat gcgatc 3481
<210>30
<211>533
<212>PRT
<213>谷氨酸棒杆菌
<400>30
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Gln
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Asn Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Ala Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Ala Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Ser Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210>31
<211>3481
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1437)..(3035)
<223>
<400>31
gatcttgccg atttcggcag gatcaatgtc ggcgtgaatg atcttggcat caggtgcgaa 60
agtgtcaacg tcaccggtga cgcggtcatc aaagcgggag ccgatagcaa tcagcaggtc 120
gctgcgctgc agtgcaccaa cagcggacac agtgccatgc atgcctggca tacccatgtg 180
cagctcgtgg gactctggga aggttcccag cgccatcaat gtggtgacaa ctggaatgcc 240
ggtgtgctca gcgaacgcac gaagctcttc gtgggcatca gccttgataa cgccgccgcc 300
aacgtaaagg acaggcttct tagactcacc gatcagtttg acagcctgct caatctgtcg 360
agcatgcggt gttgaaactg ggcggtagcc tggcaggtcg atctttggtg gccagacgaa 420
atccaattca gcgttctgaa catccttggg gatatccact agaacaggac cagggcgacc 480
agtaatcgcg aggtggaatg cctcagccaa tgcctgtgga atgtcgttgg ggttggtgac 540
catgaagttg tgcttggtca ctggcatggt gatgccgcgg atatcggctt cctggaaagc 600
atcggtaccc agcaggctac ttccgacctg gccggtgatg gcaaccatgg gaacggagtc 660
caagtttgca tcagcgattg gggtaaccaa gttggttgcg cctgggccag aggttgcaat 720
gcagacgcca acgcgtccag taacctgcgc gtagccggtt gctgcgtggc ctgcgccctg 780
ctcgtggcgc actaggacgt ggcgcacctt tgtggaggaa tagagcgggt catacaccgg 840
tagcaccgca ccaccaggaa taccgaacac gatgtcggcg ttaagctcct cgagcgatcg 900
aacaattgcc tgtgcacctg tcatccgctc aggggcggcg gatcgaccac ggcttgcaac 960
cgtggcggga gtgggctgtt gagaagctgc cacattcacg actttctggc tcctttacta 1020
aataaggatt ttcacaggac ccgtccaagc caagccgatt tcaactcagc ctaaagacaa 1080
agccctcatt taaaattgtt ccgacgcgga tgcgtgtgca cgcagtgcga cagatgtctg 1140
ttgcaaagtt ggctacttgg gtcataacca acaagaaagc cctcgttcca acactgtggt 1200
gagtgttgtc gagggcgctt gacgagacga cttggaaggc cgttacggca ggcgccgcgc 1260
ggttactact acaagtcgaa taatggtcat ggtgtgtcat gctacacaca tcgagtttcc 1320
aattccacaa cgcacgaaaa ttcccacccc caaaactccc ccacttcggt taaggaatca 1380
ggattctcac aaagttcagg caggctcccg ctacttttca gcgctaatct tggctc atg 1439
Met
1
att tta ggc gta ccc att caa tat ttg ctc tat tca ttg tgg aat tgg 1487
Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Ash Trp
5 10 15
att gtc gat acc ggt ttt gat gta gca att atc ctg gtc ttg gcg ttt 1535
Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala Phe
20 25 30
ttg att cca cgt atc ggc cga ctg gcc atg cgt att atc aag cag cga 1583
Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Gln Arg
35 40 45
gtg gag tct gca gcc gat gcg gac acc act aag aac cag ctc gcg ttc 163l
Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala Phe
50 55 60 65
gct ggc gtt ggc gtt tat atc gcg caa att gtg gcg ttt ttc atg ctt 1679
Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met Leu
70 75 80
gcc gtc tcc gcg atg cag gct ttt ggt ttc tot ctc gcg ggc gct gcg 1727
Ala Val Ser Ala Met Gln Ala Phe Gly Phe Sar Leu Ala Gly Ala Ala
85 90 95
att ccg gca acc att gcg tca gct gcc att ggt ctt ggt gtg cag tcg 1775
Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Val Gln Ser
100 105 110
att gtt gcg gac ttc ttg gcc gga ttt ttc atc ctg acg gaa aag caa 1823
Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys Gln
115 120 125
ttc ggc gtg ggt gac tgg gtg cgc ttt gag ggc aac ggc atc gtt gtt 1871
Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val Val
130 135 140 145
gaa ggc acc gtc att gag atc acc atg cgc gcg acc aaa att cgc acg 1919
Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg Thr
150 155 160
att gca caa gag acc gtg atc atc ccg aac tcc acg gcg aaa gtg tgc 1967
Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val Cys
165 170 175
atc aac aat tct aat aac tgg tcg cgt gcg gtt gtc gtt att ccg atc 2015
Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro Ile
180 185 190
ccc atg ttg ggt tct gaa aac atc aca gat gtc atc gcg cgc tct gaa 2063
Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser Glu
195 200 205
gct gcg act cgt cgc gca ctt ggc cag gag aaa atc gca ccg gaa atc 2111
Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu Ile
210 215 220 225
ctc ggt gaa ctc gat gtg cac cca gcc acg gaa gtc aca ccg cca acg 2159
Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro Thr
230 235 240
gtg gtc ggc atg ccg tgg atg gtc acc atg cgt ttc ctc gtg caa gtc 2207
Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln Val
245 250 255
acc gcc ggc aat caa tgg ctg gtc gaa cgc gcc atc cgc aca gaa atc 2255
Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu Ile
260 265 270
atc aac gaa ttc tgg gaa gaa tac ggc agc gca acc act aca tcg gga 2303
Ile Asn Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser Gly
275 280 285
acc ctc att gat tcc tta cac gtt gag cat gaa gag cca aag acc tcg 2351
Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr Ser
290 295 300 305
ctt atc gac gcc tcc ccc cag gct ctt aag gaa ccg aag ccg gag gct 2399
Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu Ala
310 315 320
gcg gcg acg gtt gca tcg cta gct gca tcg tct aac gac gat gca gac 2447
Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala Asp
325 330 335
aat gca gac gcc tcg gcg atc aat gca ggc aat cca gag aag gaa ctt 2495
Asn Ala Asp Ala Ser Ala Ile Asn Ala Gly Asn Pro Glu Lys Glu Leu
340 345 350
gat tcc gat gtg ctg gaa caa gaa ctc tcc agc gaa gaa ccg gaa gaa 2543
Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu Glu
355 360 365
aca gca aaa cca gat cac tct ctc cga ggc ttc ttc cgc act gat tac 2591
Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp Tyr
370 375 380 385
tac cca aat cgg tgg cag aag atc ctg tcg ttt ggc gga cgt gtc cgc 2639
Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val Arg
390 395 400
atg agc act tcc ctg ttg ttg ggt gcg ctg ctc ttg ctg tca cta ttt 2687
Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu Phe
405 410 415
aag gtc atg act gtg gaa cca agt gag aat tgg caa aac tcc agt gga 2735
Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser Gly
420 425 430
tgg ctg tca cca agc act gcc acc tca act gcg gtg acc acc tcc gaa 2783
Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser Glu
435 440 445
act tcc gcg cca gca agc acg cct tcg atg aca gtg ccc act acg gtg 2831
Thr Ser Ala Pro Ala Ser Thr Pro Ser Met Thr Val Pro Thr Thr Val
450 455 460 465
gag gag acc cca acg atg gaa tct agc gtc gaa acg cag cag gaa acc 2879
Glu Glu Thr Pro Thr Met Glu Ser Ser Val Glu Thr Gln Gln Glu Thr
470 475 480
tca acc cct gca acc gca acg ccc cag cga gcc gac acc atc gaa ccg 2927
Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu Pro
485 490 495
acc gag gaa gcc acg tcg cag gag gaa acg act gca tcg cag acg cag 2975
Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr Gln
500 505 510
tct cca gca gtg gaa gca cca acc gcg gtc caa gaa aca gtt gcg ccg 3023
Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala Pro
515 520 525
acg tcc acc cct taggacgctg attacagacg tgtcccattt ctttactact 3075
Thr Ser Thr Pro
530
attggaaatt atgagttcag acgcagaaaa ggcatccgtg gagctttccg aaaaatttca 3135
cccagaacgc acccatattt tgggcgccgt tgtttttggc ctgatctcat tattagtcat 3195
cggcgcagcc cctcagtacc tgttttggct gctcgcgctc cctgtcatct tcggttactg 3255
ggttctaaaa tcatccacga tcgttgatga acagggcatc accgcaaact acgccttcaa 3315
gggcaaaaag gttgtggcct gggaagacct cgcaggaatc ggattcaagg gtgcccgcac 3375
tttcgctcgc accacctccg atgcagaagt caccctcccc ggcgtcacct tcaactccct 3435
tccccgcctt gaagctgctt cccacggccg catccccgat gcgatc 3481
<210>32
<211>533
<212>PRT
<213>谷氨酸棒杆菌
<400>32
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Gln
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Val Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Asn Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Ala Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Ala Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Ser Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Glu Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210>33
<211>20
<212>DNA
<213>人工的DNA
<220>
<223>yggB N-末端引物
<400>33
ggctgttgag aagctgccac 20
<210>34
<211>29
<212>DNA
<213>人工的DNA
<220>
<223>yggB N-末端引物2
<400>34
ccgcaacaat cgact gcaca coaagacca 29
<210>35
<211>28
<212>DNA
<213>人工的DNA
<220>
<223>yggB C-末端引物1
<400>35
gggagctcca cggcatgccg accaccgt 28
<210>36
<211>29
<212>DNA
<213>人工的DNA
<220>
<223>yggB C-末端引物2
<400>36
tggtcttggt gtgcagtcga ttgttgcgg 29
<210> 37
<211> 28
<212>DNA
<213>人工的DNA
<220>
<223>yggB 2nd引物
<400>37
gggagctcga ctttctggct cctttact 28
<210>38
<211>28
<212>DNA
<213>人工的DNA
<220>
<223>sucA8引物
<400>38
ctgcgtcgac gtcggaggca ccatccac 28
<210>39
<211> 30
<212>DNA
<213>人工的DNA
<220>
<223>sucA801引物
<400>39
ctgcgtctcg acgtcggagg caccatccac 30
<210>40
<211>36
<212>DNA
<213>人工的DNA
<220>
<223>sucA805引物
<400>40
gctaaaagct gcgtcgacgt cggaggcacc atccac 36
<210>41
<211>36
<212>DNA
<213>人工的DNA
<220>
<223>sucA77
<400>41
gctataagct gcgtcgacgt cggaggcacc atccac 36
<210>42
<211>4389
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(2542)
<223>
<400>42
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcat cgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc tt ggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtct gct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln0 1 5 10cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520Pro Thr Thr Asn Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser75 80 85 90gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys ProAla Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala ArgLeu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His IleIle Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac gac 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr Asp
205 210 215
gtc atc gac ggc aag cca acc ctg atc gtg cct gag cac atc aac ctg 1144
Val Ile Asp Gly Lys Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu
220 225 230
ggc ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc 1192
Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val
235 240 245 250
gta gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc 1240
Val Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu
255 260 265
gca gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc 1288
Ala Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr
270 275 280
atggat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc 1336
Met Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly
285 290 295
atc ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc 1384
Ile Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr
300 305 310
atc atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct 1432
Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala
315 320 325 330
tcc gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc 1480
Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile
335 340 345
acc tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa 1528
Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu
350 355 360
ttc ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat 1576
Phe Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp
365 370 375
gag atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca 1624
Glu Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala
380 385 390
cag gac gtt cca aac acc ggt gtt gat aag aacacc cgc gtc at g cag 1672
Gln Asp Val Pro Asn Thr Gly Val Asp LysAsn Thr Arg Val Met Gln
395 400 405 4l0
ctc att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac 1720
Leu Il e Glu Ala Tyr Arg Ser Arg Gly Hi s Leu Ile Ala Asp Thr Asn
4l5 420 425
cca ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac 1768
Pro Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp
430 435 440
ctc gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc 1816
Leu Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr
445 450 455
ttc agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag 1864
Phe Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu
460 465 470
gta ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa 1912
Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu
475 480 485 490
tac acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc 1960
Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg
495 500 505
ctc gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc 2008
Leu Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile
510 515 520
ctg cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc 2056
Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr
525 530 535
aag tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc 2104
Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu
540 545 550
atc cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc 2152
Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu
555 560 565 570
gac gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg 2200
Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu
575 580 585
ttc aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa 2248
Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu
590 595 600
ggc caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac 2296
Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr
605 610 615
cac ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag 2344
His Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu
620 625 630
atc aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac 2392
Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn
635 640 645 650
cca gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag 2440
Pro Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys
655 660 665
ggc gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct 2488
Gly Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala
670 675 680
gca ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aag 2536
Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys
685 690 695
ctg cgt cgacgtcgga ggcaccatcc acatcgtggt gaacaaccag atcggcttca 2592
Leu Arg
700
ccaccacccc agactccagc cgctccatgc actacgcaac cgactacgcc aaggcattcg 2652
gctgcccagt cttccacgtc aatggtgatg acccagaggc agttgtctgg gttggccagc 2712
tggcaaccga gtaccgtcgt cgcttcggca aggacgtctt catcgacctc gtttgctacc 2772
gcctccgcgg ccacaacgaa gctgatgatc cttccatgac ccagccaaag atgtatgagc 2832
tcatcaccgg ccgcgagacc gttcgtgctc agtacaccga agacctgctc ggacgtggag 2892
acctctccaa cgaagatgca gaagcagtcg tccgcgactt ccacgaccag atggaatctg 2952
tgttcaacga agtcaaggaa ggcggcaaga agcaggctga ggcacagacc ggcatcaccg 3012
gctcccagaa gcttccacac ggccttgaga ccaacatctc ccgtgaagag ctcctggaac 3072
tgggacaggc tttcgccaac accccagaag gcttcaacta ccacccacgt gtggctccag 3132
ttgctaagaa gcgcgtctcc tctgtcaccg aaggtggcat cgactgggca tggggcgagc 3192
tcctcgcctt cggttccctg gctaactccg gccgcttggt tcgccttgca ggtgaagatt 3252
cccgccgcgg taccttcacc cagcgccacg cagttgccat cgacccagcg accgctgaag 3312
agttcaaccc actccacgag cttgcacagt ccaagggcaa caacggtaag ttcctggtct 3372
acaactccgc actgaccgag tacgcaggca tgggcttcga gtacggctac tccgtaggaa 3432
acgaagactc cgtcgttgca tgggaagcac agttcggcga cttcgccaac ggcgctcaga 3492
ccatcatcga tgagtacgtc tcctcaggcg aagctaagtg gggccagacc tccaagctga 3552
tccttctgct gcctcacggc tacgaaggcc agggcccaga ccactcttcc gcacgtatcg 3612
agcgcttcct gcagctgtgc gctgagggtt ccatgactgt tgctcagcca tccaccccag 3672
caaaccactt ccacctgctg cgtcgtcacg ctctgtccga cctgaagcgt ccactggtta 3732
tcttcacccc gaagtccatg ctgcgtaaca aggctgctgc ctccgcacca gaagacttca 3792
ctgaggtcac caagttccaa tccgtgatcg acgatccaaa cgttgcagat gcagccaagg 3852
tgaagaaggt catgctggtc tccggcaagc tgtactacga attggcaaag cgcaaggaga 3912
aggacggacg cgacgacatc gcgatcgttc gtatcgaaat gctccaccca attccgttca 3972
accgcatctc cgaggctctt gccggctacc ctaacgctga ggaagtcctc ttcgttcagg 4032
atgagccagc aaaccagggc ccatggccgt tctaccagga gcacctccca gagctgatcc 4092
cgaacatgcc aaagatgcgc cgcgtttccc gccgcgctca gtcctccacc gcaactggtg 4152
ttgctaaggt gcaccagctg gaggagaagc agcttatcga cgaggctttc gaggcttaag 4212
tctttatagt cctgcactag cctagagggc cttatgcagt gtgaatcaca cagcataagg 4272
ccctttttgc tgccgtggtt gcctaaggtg gaaggcatga aacgaatctg tgcggtcacg 4332
atctcttcag tacttttgct aagtggctgc tcctccactt ccaccacgca gctcgag 4389
<210>43
<211>700
<212>PRT
<213>谷氨酸棒杆菌
<400>43
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Ara Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 510
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 6 65 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Leu Arg
690 695 700
<210>44
<211>4391
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4210)
<223>sucA801
<400>44
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Asn Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac gac 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr Asp
205 210 215
gtc atc gac ggc aag cca acc ctg atc gtg cct gag cac atc aac ctg 1144
Val Ile Asp Gly Lys Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu
220 225 230
ggc ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc 1192
Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val
235 240 245 250
gta gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc 1240
Val Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu
255 260 265
gca gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc 1288
Ala Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr
270 275 280
atg gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc 1336
Met Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly
285 290 295
atc ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc 1384
Ile Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr
300 305 310
atc atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct 1432
Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala
315 320 325 330
tcc gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc 1480
Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile
335 340 345
acc tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa 1528
Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu
350 355 360
ttc ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat 1576
Phe Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp
365 370 375
gag atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca 1624
Glu Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala
380 385 390
cag gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag 1672
Gln Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln
395 400 405 410
ctc att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac 1720
Leu Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn
415 420 425
cca ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac 1768
Pro Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp
430 435 440
ctc gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc 1816
Leu Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr
445 450 455
ttc agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag 1864
Phe Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu
460 465 470
gta ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa 1912
Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu
475 480 485 490
tac acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc 1960
Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg
495 500 505
ctc gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc 2008
Leu Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile
510 515 520
ctg cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc 2056
Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr
525 530 535
aag tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc 2104
Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu
540 545 550
atc cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc 2152
Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu
555 560 565 570
gac gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg 2200
Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu
575 580 585
ttc aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa 2248
Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu
590 595 600
ggc caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac 2296
Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr
605 610 615
cac ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag 2344
His Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu
620 625 630
atc aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac 2392
Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn
635 640 645 650
cca gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag 2440
Pro Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys
655 660 665
ggc gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct 2488
Gly Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala
670 675 680
gca ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aag 2536
Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys
685 690 695
ctg cgt ctc gac gtc gga ggc acc atc cac atc gtg gtg aac aac cag 2584
Leu Arg Leu Asp Val Gly Gly Thr Ile His Ile Val Val Asn Asn Gln
700 705 710
atc ggc ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac gca 2632
Ile G1y Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala
715 720 725 730
acc gac tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat ggt 2680
Thr Asp Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn Gly
735 740 745
gat gac cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag tac 2728
Asp Asp Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr
750 755 760
cgt cgt cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac cgc 2776
Arg Arg Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg
765 770 775
ctc cgc ggc cac aac gaa gct gat gat cct tcc atg acc cag cca aag 2824
Leu Arg Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys
780 785 790
atg tat gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac acc 2872
Met Tyr Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr
795 800 805 810
gaa gac ctg ctc gga cgt gga gac ctc tcc aac gaa gat gca gaa gca 2920
Glu Asp Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala
815 820 825
gtc gtc cgc gac ttc cac gac cag atg gaa tct gtg ttc aac gaa gtc 2968
Val Val Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu Val
830 835 840
aag gaa ggc ggc aag aag cag gct gag gca cag acc ggc atc acc ggc 3016
Lys Glu Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly
845 850 855
tcc cag aag ctt cca cac ggc ctt gag acc aac atc tcc cgt gaa gag 3064
Ser Gln Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu
860 865 870
ctc ctg gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc aac 3112
Leu Leu Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn
875 880 885 890
tac cac cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct gtc 3160
Tyr His Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val
895 900 905
acc gaa ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt 3208
Thr Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly
910 915 920
tcc ctg gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc 3256
Ser Leu Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser
925 930 935
cgc cgc ggt acc ttc acc cag cgc cac gca gtt gcc atc gac cca gcg 3304
Arg Arg Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala
940 945 950
acc gct gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag ggc 3352
Thr Ala Glu Glu Phe Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly
955 960 965 970
aac aac ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac gca 3400
Asn Asn Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala
975 980 985
ggc atg ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc gtc 3448
Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val
990 995 1000
gtt gca tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct cag 3493
Val Ala Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln
1005 1010 1015
acc atc atc gat gag tac gtc tcc tca ggc gaa gct aag tgg ggc 3538
Thr Ile Ile Asp Glu Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly
1020 1025 1030
cag acc tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa ggc 3583
Gln Thr Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu Gly
1035 1040 1045
cag ggc cca gac cac tct tcc gca cgt atc gag cgc ttc ctg cag 3628
Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln
1050 1055 1060
ctg tgc gct gag ggt tcc atg act gtt gct cag cca tcc acc cca 3673
Leu Cys Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr Pro
1065 1070 1075
gca aac cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac ctg 3718
Ala Asn His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp Leu
1080 1085 1090
aag cgt cca ctg gtt atc ttc acc ccg aag tcc atg ctg cgt aac 3763
Lys Arg Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg Asn
1095 1100 1105
aag gct gct gcc tcc gca cca gaa gac ttc act gag gtc acc aag 3808
Lys Ala Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr Lys
1110 1115 1120
ttc caa tcc gtg atc gac gat cca aac gtt gca gat gca gcc aag 3853
Phe Gln Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala Lys
1125 1130 1135
gtg aag aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa ttg 3898
Val Lys Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu
1140 1145 1150
gca aag cgc aag gag aag gac gga cgc gac gac atc gcg atc gtt 3943
Ala Lys Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile Val
1155 1160 1165
cgt atc gaa atg ctc cac cca att ccg ttc aac cgc atc tcc gag 3988
Arg Ile Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser Glu
1170 1175 1180
gct ctt gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt cag 4033
Ala Leu Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val Gln
1185 1190 1195
gat gag cca gca aac cag ggc cca tgg ccg ttc tac cag gag cac 4078
Asp Glu Pro Ala Asn Gln Gly Pro Trp Pro Phe Tyr Gln Glu His
1200 1205 1210
ctc cca gag ctg atc ccg aac atg cca aag atg cgc cgc gtt tcc 4123
Leu Pro Glu Leu Ile Pro Asn Met Pro Lys Met Arg Arg Val Ser
1215 1220 1225
cgc cgc gct cag tcc tcc acc gca act ggt gtt gct aag gtg cac 4168
Arg Arg Ala Gln Ser Ser Thr Ala Thr Gly Val Ala Lys Val His
1230 1235 1240
cag ctg gag gag aag cag ctt atc gac gag gct ttc gag gct 4210
Gln Leu Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala
1245 1250 1255
taagtcttta tagtcctgca ctagcctaga gggccttatg cagtgtgaat cacacagcat 4270
aaggcccttt ttgctgccgt ggttgcctaa ggtggaaggc atgaaacgaa tctgtgcggt 4330
cacgatctct tcagtacttt tgctaagtgg ctgctcctcc acttccacca cgcagctcga 4390
g 4391
<210>45
<211>1256
<212>PRT
<213>谷氨酸棒杆菌
<400>45
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val AIa Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 5l0
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Leu Arg Leu Asp Val Gly
690 695 700
Gly Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr
705 710 715 720
Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys Ala
725 730 735
Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val
740 745 750
Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys
755 760 765
Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn Glu
770 775 780
Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr
785 790 795 800
Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly Arg
805 810 815
Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Va1 Arg Asp Phe His
820 825 830
Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys Lys
835 840 845
Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro His
850 855 860
Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly Gln
865 870 875 880
Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val Ala
885 890 895
Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile Asp
900 905 910
Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser Gly
915 920 925
Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr
930 935 940
Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe Asn
945 950 955 960
Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe Leu
965 970 975
Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr
980 985 990
Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala Gln
995 1000 1005
Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr
1010 1015 1020
Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu Ile
1025 1030 1035
Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His Ser
1040 1045 1050
Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly Ser
1055 1060 1065
Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His Leu
1070 1075 1080
Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val Ile
1085 1090 1095
Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser Ala
1100 1105 1110
Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile Asp
1115 1120 1125
Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met Leu
1130 1135 1140
Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys
1145 1150 1155
Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu His
1160 1165 1170
Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr Pro
1175 1180 1185
Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Aan Gln
1190 1195 1200
Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile Pro
1205 1210 1215
Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser Ser
1220 1225 1230
Thr Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys Gln
1235 1240 1245
Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>46
<211>4391
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4210)
<223>sucA805
<400>46
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Asn Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac gac 1096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr Asp
205 210 215
gtc atc gac ggc aag cca acc ctg atc gtg cct gag cac atc aac ctg 1144
Val Ile Asp Gly Lys Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu
220 225 230
ggc ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc 1192
Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val
235 240 245 250
gta gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc 1240
Val Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu
255 260 265
gca gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc 1288
Ala Ala Tyr Glu Asp Ile Val ThT Arg Ser Arg Lys Gly Lys Leu Thr
270 275 280
atg gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc 1336
Met Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly
285 290 295
atc ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc 1384
Ile Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr
300 305 310
atc atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct 1432
Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala
315 320 325 330
tcc gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc 1480
Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile
335 340 345
acc tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa 1528
Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu
350 355 360
ttc ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat 1576
Phe Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp
365 370 375
gag atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca 1624
Glu Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala
380 385 390
cag gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag 1672
Gln Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln
395 400 405 410
ctc att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac 1720
Leu Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn
415 420 425
cca ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac 1768
Pro Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp
430 435 440
ctc gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc 1816
Leu Asp Ile Glu Thr Hi s Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr
445 450 455
ttc agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag 1864
Phe Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu
460 465 470
gta ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa 1912
Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu
475 480 485 490
tac acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc 1960
Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg
495 500 505
ctc gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc 2008
Leu Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile
510 515 520
ctg cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc 2056
Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr
525 530 535
aag tac gtc ggc cag aag cgc ttc tcc ctc gaa ggt gca gaa gct ctc 2104
Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu
540 545 550
atc cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc 2152
Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu
555 560 565 570
gac gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg 2200
Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu
575 580 585
ttc aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa 2248
Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu
590 595 600
ggc caa atg gag cag ggc cag atc ggt ggc tcc ggt gac gtg aag tac 2296
Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr
605 610 615
cac ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag 2344
His Leu Gly Ser Glu Gly Gln Hi s Leu Gln Met Phe Gly Asp Gly Glu
620 625 630
atc aag gtctcc ct g act gct aac ccg tcc cac ctg gaa gct gtt aac 2392
Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn
635 640 645 650
cca gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag 2440
Pro Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys
655 660 665
ggc gta gac ggc aag act gtt gtg cca ctg ctg ctc cac ggt gac gct 2488
Gly Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala
670 675 680
gca ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct aaa 2536
Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Lys
685 690 695
agc tgc gtc gac gtc gga ggc acc atc cac atc gtg gtg aac aac cag 2584
Ser Cys Val Asp Val Gly Gly Thr Ile His Ile Val Val Asn Asn Gln
700 705 710
atc ggc ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac gca 2632
Ile Gly Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala
715 720 725 730
acc gac tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat ggt 2680
Thr Asp Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn Gly
735 740 745
gat gac cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag tac 2728
Asp Asp Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr
750 755 760
cgt cgt cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac cgc 2776
Arg Arg Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg
765 770 775
ctc cgc ggc cac aac gaa gct gat gat cct tcc atg acc cag cca aag 2824
Leu Arg Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys
780 785 790
atg tat gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac acc 2872
Met Tyr Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr
795 800 805 810
gaa gac ctg ctc gga cgt gga gac ctc tcc aac gaa gat gca gaa gca 2920
Glu Asp Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala
815 820 825
gtc gtc cgc gac ttc cac gac cag atg gaa tct gtg ttc aac gaa gtc 2968
Val Val Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu Val
830 835 840
aag gaa ggc ggc aag aag cag gct gag gca cag acc ggc atc acc ggc 3016
Lys Glu Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly
845 850 855
tcc cag aag ctt cca cac ggc ctt gag acc aac atc tcc cgt gaa gag 3064
Ser Gln Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu
860 865 870
ctc ctg gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc aac 3112
Leu Leu Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn
875 880 885 890
tac cac cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct gtc 3160
Tyr His Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val
895 900 905
acc gaa ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt 3208
Thr Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly
910 915 920
tcc ctg gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc 3256
Ser Leu Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser
925 930 935
cgc cgc ggt acc ttc acc cag cgc cacgca gtt gcc atc gac cca gcg 3304
Arg Arg Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala
940 945 950
acc gct gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag ggc 3352
Thr Ala Glu Glu Phe Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly
955 960 965 970
aac aac ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac gca 3400
Asn Asn Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala
975 980 985
ggc atg ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc gtc 3448
Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val
990 995 1000
gtt gca tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct cag 3493
Val Ala Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln
1005 1010 1015
acc atc atc gat gag tac gtc tcc tca ggc gaa gct aag tgg ggc 3538
Thr Ile Ile Asp Glu Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly
1020 1025 1030
cag acc tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa ggc 3583
Gln Thr Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu Gly
1035 1040 1045
cag ggc cca gac cac tct tcc gca cgt atc gag cgc ttc ctg cag 3628
Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln
1050 1055 1060
ctg tgc gct gag ggt tcc atg act gtt gct cag cca tcc acc cca 3673
Leu Cys Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr Pro
1065 1070 1075
gca aac cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac ctg 3718
Ala Asn His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp Leu
1080 1085 1090
aag cgt cca ctg gtt atc ttc acc ccg aag tcc atg ctg cgt aac 3763
Lys Arg Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg Asn
1095 1100 1105
aag gct gct gcc tcc gca cca gaa gac ttc act gag gtc acc aag 3808
Lys Ala Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr Lys
1110 1115 1120
ttc caa tcc gtg atc gac gat cca aac gtt gca gat gca gcc aag 3853
Phe Gln Set Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala Lys
1125 1130 1135
gtg aag aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa ttg 3898
Val Lys Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu
1140 1145 1150
gca aag cgc aag gag aag gac gga cgc gac gac atc gcg atc gtt 3943
Ala Lys Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile Val
1155 1160 1165
cgt atc gaa atg ctc cac cca att ccg ttc aac cgc atc tcc gag 3988
Arg Ile Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser Glu
1170 1175 1180
gct ctt gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt cag 4033
Ala Leu Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val Gln
1185 1190 1195
gat gag cca gca aac cag ggc cca tgg ccg ttc tac cag gag cac 4078
Asp Glu Pro Ala Asn Gln Gly Pro Trp Pro Phe Tyr Gln Glu His
1200 1205 1210
ctc cca gag ctg atc ccg aac atg cca aag atg cgc cgc gtt tcc 4123
Leu Pro Glu Leu Ile Pro Asn Met Pro Lys Met Arg Arg Val Ser
1215 1220 1225
cgc cgc gct cag tcc tcc acc gca act ggt gtt gct aag gtg cac 4168
Arg Arg Ala Gln Ser Ser Thr Ala Thr Gly Val Ala Lys Val His
1230 1235 1240
cag ctg gag gag aag cag ctt atc gac gag gct ttc gag gct 4210
Gln Leu Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala
1245 1250 1255
taagtcttta tagtcctgca ctagcctaga gggccttatg cagtgtgaat cacacagcat 4270
aaggcccttt ttgctgccgt ggttgcctaa ggtggaaggc atgaaacgaa tctgtgcggt 4330
cacgatctct tcagtacttt tgctaagtgg ctgctcctcc acttccacca cgcagctcga 4390
4391
g
<210>47
<211>1256
<212>PRT
<213>谷氨酸棒杆菌
<400>47
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 5l0
Lys Pro Thr Gln Ala Gl uGln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Ser Cys Val Asp Val Gly
690 695 700
Gly Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr
705 710 715 720
Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys Ala
725 730 735
Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val
740 745 750
Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys
755 760 765
Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn Glu
770 775 780
Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr
785 790 795 800
Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly Arg
805 810 815
Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe His
820 825 830
Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys Lys
835 840 845
Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro His
850 855 860
Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly Gln
865 870 875 880
Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val Ala
885 890 895
Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile Asp
900 905 910
Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser Gly
915 920 925
Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr
930 935 940
Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe Asn
945 950 955 960
Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe Leu
965 970 975
Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr
980 985 990
Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala Gln
995 1000 1005
Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr
1010 1015 1020
Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu Ile
1025 1030 1035
Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His Ser
1040 1045 1050
Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly Ser
1055 1060 1065
Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His Leu
1070 1075 1080
Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val Ile
1085 1090 1095
Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser Ala
1100 1105 1110
Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile Asp
1115 1120 1125
Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met Leu
1130 1135 1140
Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys
1145 1150 1155
Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu His
1160 1165 1170
Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr Pro
1175 1180 1185
Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn Gln
1190 1195 1200
Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile Pro
1205 1210 1215
Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser Ser
1220 1225 1230
Thr Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys Gln
1235 1240 1245
Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>48
<211>4391
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(443)..(4210)
<223>
<400>48
gtcgacaagc aaaatcgaag cggcagcacg ccgcgtcgga gccttaaacg ccatcgccgc 60
catccctgat ggtttcaatc atcaagtcgg tgaacgcggg cgcaacctgt catccggaca 120
gcgccaactg atcgcgctgg cgcgcgccga actcatcgag ccttccatca tgcttctcga 180
cgaagccacc tccaccctcg accccgccac cgaagccgtt atcctcaacg cctccgatcg 240
agtcactaag ggacgcacca gcatcatcgt cgcgcaccgc ttggcaaccg ctaaaagggc 300
cgaccgtatt cttgttgttg aacaaggacg tatcattgag gacggatctc acgacgcgtt 360
gttgtctgct aacggcacct acgcccgcat gtggcattta atggcctgac acgttatttt 420
taggagaact gtcaacaaat ta atg cta caa ctg ggg ctt agg cat aat cag 472
Met Leu Gln Leu Gly Leu Arg His Asn Gln
1 5 10
cca acg acc aac gtt aca gtg gat aaa ata aag ctc aat aaa ccc tca 520
Pro Thr Thr Asn Val Thr Val Asp Lys Ile Lys Leu Asn Lys Pro Ser
15 20 25
aga agc aag gaa aag agg cga gta cct gcc gtg agc agc gct agt act 568
Arg Ser Lys Glu Lys Arg Arg Val Pro Ala Val Ser Ser Ala Ser Thr
30 35 40
ttc ggc cag aat gcg tgg ctg gta gac gag atg ttc cag cag ttc cag 616
Phe Gly Gln Asn Ala Trp Leu Val Asp Glu Met Phe Gln Gln Phe Gln
45 50 55
aag gac ccc aag tcc gtg gac aag gaa tgg aga gaa ctc ttt gag gcg 664
Lys Asp Pro Lys Ser Val Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala
60 65 70
cag ggg gga cca aat gct acc ccc gct aca aca gaa gca cag cct tca 712
Gln Gly Gly Pro Asn Ala Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser
75 80 85 90
gcg ccc aag gag tct gcg aaa cca gca cca aag gct gcc cct gca gcc 760
Ala Pro Lys Glu Ser Ala Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala
95 100 105
aag gca gca ccg cgc gta gaa acc aag ccg gcc gcc aag acc gcc cct 808
Lys Ala Ala Pro Arg Val Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro
110 115 120
aag gcc aag gag tcc tca gtg cca cag caa cct aag ctt ccg gag cca 856
Lys Ala Lys Glu Ser Ser Val Pro Gln Gln Pro Lys Leu Pro Glu Pro
125 130 135
gga caa acc cca atc agg ggt att ttc aag tcc atc gcg aag aac atg 904
Gly Gln Thr Pro Ile Arg Gly Ile Phe Lys Ser Ile Ala Lys Asn Met
140 145 150
gat atc tcc ctg gaa atc cca acc gca acc tcg gtt cgc gat atg cca 952
Asp Ile Ser Leu Glu Ile Pro Thr Ala Thr Ser Val Arg Asp Met Pro
155 160 165 170
gct cgc ctc atg ttc gaa aac cgc gcg atg gtc aac gat cag ctc aag 1000
Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn Asp Gln Leu Lys
175 180 185
cgc acc cgc ggt ggc aag atc tcc ttc acc cac atc att ggc tac gcc 1048
Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile Ile Gly Tyr Ala
190 195 200
atg gtg aag gca gtc atg gct cac ccg gac atg aac aac tcc tac gac l096
Met Val Lys Ala Val Met Ala His Pro Asp Met Asn Asn Ser Tyr Asp
205 210 215
gtc atc gac ggc aag cca acc ctg atc gtg cct gag cac atc aac ctg 1144
Val Ile Asp Gly Lys Pro Thr Leu Ile Val Pro Glu His Ile Asn Leu
220 225 230
ggc ctt gcc atc gac ctt cct cag aag gac ggc tcc cgc gca ctt gtc 1192
Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser Arg Ala Leu Val
235 240 245 250
gta gca gcc atc aag gaa acc gag aag atg aac ttc tcc gag ttc ctc 1240
Val Ala Ala Ile Lys Glu Thr Glu Lys Met Asn Phe Ser Glu Phe Leu
255 260 265
gca gca tac gaa gac atc gtg aca cgc tcc cgc aag ggc aag ctc acc 1288
Ala Ala Tyr Glu Asp Ile Val Thr Arg Ser Arg Lys Gly Lys Leu Thr
270 275 280
atg gat gac tac cag ggc gtt acc gtt tcc ttg acc aac cca ggt ggc 1336
Met Asp Asp Tyr Gln Gly Val Thr Val Ser Leu Thr Asn Pro Gly Gly
285 290 295
atc ggt acc cgc cac tct gtc cca cgt ctg acc aag ggc cag ggc acc 1384
Ile Gly Thr Arg His Ser Val Pro Arg Leu Thr Lys Gly Gln Gly Thr
300 305 3l0
atc atc ggt gtc ggt tcc atg gat tac cca gca gag ttc cag ggc gct 1432
Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala
315 320 325 330
tcc gaa gac cgc ctt gca gag ctc ggc gtt gga aag ctt gtc acc atc 1480
Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys Leu Val Thr Ile
335 340 345
acc tcc acc tac gat cac cgc gtg atc cag ggt gct gtg tcc ggt gaa 1528
Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala Val Ser Gly Glu
350 355 360
ttc ctg cgt acc atg tct cgc ctg ctc acc gat gat tcc ttc tgg gat 1576
Phe Leu Arg Thr Met Ser Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp
365 370 375
gag atc ttc gac gca atg aac gtt cct tac acc cca atg cgt tgg gca l624
Glu Ile Phe Asp Ala Met Asn Val Pro Tyr Thr Pro Met Arg Trp Ala
380 385 390
cag gac gtt cca aac acc ggt gtt gat aag aac acc cgc gtc atg cag 1672
Gln Asp Val Pro Asn Thr Gly Val Asp Lys Asn Thr Arg Val Met Gln
395 400 405 410
ctc att gag gca tac cgc tcc cgt gga cac ctc atc gct gac acc aac l720
Leu Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile Ala Asp Thr Asn
415 420 425
cca ctt tca tgg gtt cag cct ggc atg cca gtt cca gac cac cgc gac 1768
Pro Leu Ser Trp Val Gln Pro Gly Met Pro Val Pro Asp His Arg Asp
430 435 440
ctc gac atc gag acc cac agc ctg acc atc tgg gat ctg gac cgt acc 1816
Leu Asp Ile Glu Thr His Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr
445 450 455
ttc agc gtc ggt ggc ttc ggc ggc aag gag acc atg acc ctg cgc gag 1864
Phe Ser Val Gly Gly Phe Gly Gly Lys Glu Thr Met Thr Leu Arg Glu
460 465 470
gta ctg tcc cgc ctg cgc gct gcc tac acc ttg aag gtc ggc tcc gaa 1912
Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu
475 480 485 490
tac acc cac atc ctg gac cgc gac gag cgc acc tgg ctg cag gac cgc 1960
Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg
495 500 505
ctc gaa gcc gga atg cca aag cca acc cag gca gag cag aag tac atc 2008
Leu Glu Ala Gly Met Pro Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile
510 515 520
ctg cag aag ctg aac gcc gca gag gct ttc gag aac ttc ctg cag acc 2056
Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr
525 530 535
aag tac gtc ggc cag aag cgc ttctcc ctc gaa ggt gca gaa gct ctc 2104
Lys Tyr Val Gly GlnLys Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu
540 545 550
atc cca ctg atg gac tcc gcc atc gac acc gcc gca ggc cag ggc ctc 2152
Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu
555 560 565 570
gac gaa gtt gtc atc ggt atg cca cac cgt ggt cgc ctc aac gtg ctg 2200
Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg Leu Asn Val Leu
575 580 585
ttc aac atc gtg ggc aag cca ctg gca tcc atc ttc aac gag ttt gaa 2248
Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu
590 595 600
ggc caa atg gag cag ggc cagatc ggt ggc tcc ggt gac gtg aag tac 2296
Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr
605 610 615
cac ctc ggt tcc gaa ggc cag cac ctg cag atg ttc ggc gac ggc gag 2344
His Leu Gly Ser Glu Gly Gln His Leu Gln Met Phe Gly Asp Gly Glu
620 625 630
atc aag gtc tcc ctg act gct aac ccg tcc cac ctg gaa gct gtt aac 2392
Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu Glu Ala Val Asn
635 640 645 650
cca gtg atg gaa ggt atc gtc cgc gca aag cag gac tac ctg gac aag 2440
Pro Val Met Glu Gly Ile Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys
655 660 665
ggc gta gac ggc aag act gtt gtg cca ctg ctg ctc eac ggt gac gct 2488
Gly Val Asp Gly Lys Thr Val Val Pro Leu Leu Leu His Gly Asp Ala
670 675 680
gca ttc gca ggc ctg ggc atc gtg cca gaa acc atc aac ctg gct ata 2536
Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile Asn Leu Ala Ile
685 690 695
agc tgc gtc gac gtc gga ggc acc atc cac atc gtg gtg aac aac cag 2584
Ser Cys Val Asp Val Gly Gly Thr Ile His Ile Val Val Asn Asn Gln
700 705 710
atc ggc ttc acc acc acc cca gac tcc agc cgc tcc atg cac tac gca 2632
Ile Gly Phe Thr Thr Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala
715 720 725 730
acc gac tac gcc aag gca ttc ggc tgc cca gtc ttc cac gtc aat ggt 2680
Thr Asp Tyr Ala Lys Ala Phe Gly Cys Pro Val Phe His Val Asn Gly
735 740 745
gat gac cca gag gca gtt gtc tgg gtt ggc cag ctg gca acc gag tac 2728
Asp Asp Pro Glu Ala Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr
750 755 760
cgt cgt cgc ttc ggc aag gac gtc ttc atc gac ctc gtt tgc tac cgc 2776
Arg Arg Arg Phe Gly Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg
765 770 775
ctc cgc ggc cac aac gaa gct gat gat cct tcc atg acc cag cca aag 2824
Leu Arg Gly His Asn Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys
780 785 790
atg tat gag ctc atc acc ggc cgc gag acc gtt cgt gct cag tac acc 2872
Met Tyr Glu Leu Ile Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr
795 800 805 810
gaa gac ctg ctc gga cgt gga gac ctc tcc aac gaa gat gca gaa gca 2920
Glu Asp Leu Leu Gly Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala
815 820 825
gtc gtc cgc gac ttc cac gac cag atg gaa tct gtg ttc aac gaa gtc 2968
Val Val Arg Asp Phe His Asp Gln Met Glu Ser Val Phe Asn Glu Val
830 835 840
aag gaa ggc ggc aag aag cag gct gag gca cag acc ggc atc acc ggc 3016
Lys Glu Gly Gly Lys Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly
845 850 855
tcc cag aag ctt cca cac ggc ctt gag acc aac atc tcc cgt gaa gag 3064
Ser Gln Lys Leu Pro His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu
860 865 870
ctc ctg gaa ctg gga cag gct ttc gcc aac acc cca gaa ggc ttc aac 3112
Leu Leu Glu Leu Gly Gln Ala Phe Ala Asn Thr Pro Glu Gly phe Asn
875 880 885 890
tac cac cca cgt gtg gct cca gtt gct aag aag cgc gtc tcc tct gtc 3160
Tyr His Pro Arg Val Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val
895 900 905
acc gaa ggt ggc atc gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt 3208
Thr Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly
910 915 920
tcc ctg gct aac tcc ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc 3256
Ser Leu Ala Asn Ser Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser
925 930 935
cgc cgc ggt acc ttc acc cag cgc cac gca gtt gcc atc gac cca gcg 3304
Arg Arg Gly Thr Phe Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala
940 945 950
acc gct gaa gag ttc aac cca ctc cac gag ctt gca cag tcc aag ggc 3352
Thr Ala Glu Glu Phe Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly
955 960 965 970
aac aac ggt aag ttc ctg gtc tac aac tcc gca ctg acc gag tac gca 3400
Asn Asn Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala
975 980 985
ggc atg ggc ttc gag tac ggc tac tcc gta gga aac gaa gac tcc gtc 3448
Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Val
990 995 1000
gtt gca tgg gaa gca cag ttc ggc gac ttc gcc aac ggc gct cag 3493
Val Ala Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln
1005 1010 1015
acc atc atc gat gag tac gtc tcc tca ggc gaa gct aag tgg ggc 3538
Thr Ile Ile Asp Glu Tyr Val Ser Ser Gly GLu Ala Lys Trp Gly
1020 1025 1030
cag acc tcc aag ctg atc ctt ctg ctg cct cac ggc tac gaa ggc 3583
Gln Thr Ser Lys Leu Ile Leu Leu Leu Pro His Gly Tyr Glu Gly
1035 1040 1045
cag ggc cca gac cac tct tcc gca cgt atc gag cgc ttc ctg cag 3628
Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln
1050 1055 1060
ctg tgc gct gag ggt tcc atg act gtt gct cag cca tcc acc cca 3673
Leu Cys Ala Glu Gly Ser Met Thr Val Ala Gln Pro Ser Thr Pro
1065 1070 1075
gca aac cac ttc cac ctg ctg cgt cgt cac gct ctg tcc gac ctg 3718
Ala Asn His Phe His Leu Leu Arg Arg His Ala Leu Ser Asp Leu
1080 1085 1090
aag cgt cca ctg gtt atc ttc acc ccg aag tcc atg ctg cgt aac 3763
Lys Arg Pro Leu Val Ile Phe Thr Pro Lys Ser Met Leu Arg Asn
1095 1100 1105
aag gct gct gcc tcc gca cca gaa gac ttc act gag gtc acc aag 3808
Lys Ala Ala Ala Ser Ala Pro Glu Asp Phe Thr Glu Val Thr Lys
1110 1115 1120
ttc caa tcc gtg atc gac gat cca aac gtt gca gat gca gcc aag 3853
Phe Gln Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ala Lys
1125 1130 1135
gtg aag aag gtc atg ctg gtc tcc ggc aag ctg tac tac gaa ttg 3898
Val Lys Lys Val Met Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu
1140 1145 1150
gca aag cgc aag gag aag gac gga cgc gac gac atc gcg atc gtt 3943
Ala Lys Arg Lys Glu Lys Asp Gly Arg Asp Asp Ile Ala Ile Val
1155 1160 1165
cgt atc gaa atg ctc cac cca att ccg ttc aac cgc atc tcc gag 3988
Arg Ile Glu Met Leu His Pro Ile Pro Phe Asn Arg Ile Ser Glu
1170 1175 1180
gct ctt gcc ggc tac cct aac gct gag gaa gtc ctc ttc gtt cag 4033
Ala Leu Ala Gly Tyr Pro Asn Ala Glu Glu Val Leu Phe Val Gln
1185 1190 1195
gat gag cca gca aac cag ggc cca tgg ccg ttc tac cag gag cac 4078
Asp Glu Pro Ala Asn Gln Gly Pro Trp Pro Phe Tyr Gln Glu His
1200 1205 1210
ctc cca gag ctg atc ccg aac atg cca aag atg cgc cgc gtt tcc 4123
Leu Pro Glu Leu Ile Pro Asn Met Pro Lys Met Arg Arg Val Ser
1215 1220 1225
cgc cgc gct cag tcc tcc acc gca act ggt gtt gct aag gtg cac 4168
Arg Arg Ala Gln Ser Ser Thr Ala Thr Gly Val Ala Lys Val His
1230 1235 1240
cag ctg gag gag aag cag ctt atc gac gag gct ttc gag gct 4210
Gln Leu Glu Glu Lys Gln Leu Ile Asp Glu Ala Phe Glu Ala
1245 1250 1255
taagtcttta tagtcctgca ctagcctaga gggccttatg cagtgtgaat cacacagcat 4270
aaggcccttt ttgctgccgt ggttgcctaa ggtggaaggc atgaaacgaa tctgtgcggt 4330
cacgatctct tcagtacttt tgctaagtgg ctgctcctcc acttccacca cgcagctcga 4390
g 4391
<210>49
<211>1256
<212>PRT
<213>谷氨酸棒杆菌
<400>49
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Ile Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Ala
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Ala Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
155 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Thr Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 3l5 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Ser Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Ser Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 510
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Ile Ser Cys Val Asp Val Gly
690 695 700
Gly Thr Ile HisIle Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr
705 7l0 715 720
Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys Ala
725 730 735
Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val
740 745 750
Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys
755 760 765
Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn Glu
770 775 780
Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr
785 790 795 800
Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly Arg
805 810 815
Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe His
820 825 830
Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys Lys
835 840 845
Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro His
850 855 860
Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly Gln
865 870 875 880
Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val Ala
885 890 895
Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile Asp
900 905 910
Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser Gly
915 920 925
Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr
930 935 940
Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe Asn
945 950 955 960
Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe Leu
965 970 975
Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr
980 985 990
Gly Tyr Ser Val Gly Asn Glu Asp Ser Val Val Ala Trp Glu Ala Gln
995 1000 1005
Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr
1010 1015 1020
Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu Ile
1025 1030 1035
Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His Ser
1040 1045 1050
Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly Ser
1055 1060 1065
Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His Leu
1070 1075 1080
Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val Ile
1085 1090 1095
Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser Ala
1100 1105 1110
Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile Asp
1115 1120 1125
Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met Leu
1130 1135 1140
Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys
1145 1150 1155
Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu His
1160 1165 1170
Pro Ile Pro Phe Asn Arg Ile Ser G1u Ala Leu Ala Gly Tyr Pro
1175 1180 1185
Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn Gln
1190 1195 1200
Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile Pro
1205 1210 1215
Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser Ser
1220 1225 1230
Thr Ala Thr Gly Mal Ala Lys Val His Gln Leu Glu Glu Lys Gln
1235 1240 1245
Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>50
<211>3774
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1).(3774)
<223>
<400>50
atg cta caa ctg ggg ctt agg cat aat cag cca acg acc aac gtt aca 48
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
gtg gat aaa aca aag ctc aat aaa ccc tca aga agc aag gaa aag agg 96
Val Asp Lys Thr Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
cga gta cct gcc gtg agc agc gct agt act ttc ggc cag aat gcg tgg 144
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
ctg gta gac gag atg ttc cag cag ttc cag aag gac ccc aag tcc gtg 192
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
gac aag gaa tgg aga gaa ctc ttt gag gcg cag ggg gga cca aat act 240
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Thr
65 70 75 80
acc ccc gct aca aca gaa gca cag cct tca gcg ccc aag gag tct gcg 288
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
aaa cca gca cca aag gct gcc cct gca gcc aag gca gca ccg cgc gta 336
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
gaa acc aag ccg gcc gac aag acc gcc cct aag gcc aag gag tcc tca 384
Glu Thr Lys Pro Ala Asp Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
gtg cca cag caa cct aag ctt ccg gag cca gga cas acc cca atc agg 432
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
ggt att ttc aag tcc atc gcg aag aac atg gat atc tcc ctg gaa atc 480
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
cca acc gca acc tcg gtt cgc gat atg cca gct cgc ctc atg ttc gaa 528
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
aac cgc gcg atg gtc aac gat cag ctc aag cgc acc cgc ggt ggc aag 576
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
atc tcc ttc acc cac atc att ggc tac gcc atg gtg aag gca gtc atg 624
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
gct cac ccg gac atg aac aac tcc tac gac gtc atc gac ggc aag cca 672
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
acc ctg atc gtg cct gag cac atc aac ctg ggc ctt gct atc gac ctt 720
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
cct cag aag gac ggc tcc cgc gca ctt gtc gta gca gcc atc aag gaa 768
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
acc gag aag atg aac ttc tcc gag ttc ctc gca gcc tac gaa gac atc 8l6
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
gtg gca cgc tcc cgc aag ggc aag ctc acc atg gat gac tac cag ggc 864
Val Ala Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
gtt acc gtt tcc ttg acc aac cca ggt ggc atc ggt acc cgc cac tct 912
Val Thr Val Ser Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser
290 295 300
gtt cca cgt cta acc aag ggc cag ggc acc atc atc ggt gtc ggt tcc 960
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
atg gat tac cca gca gag ttc cag ggc gct tca gaa gac cgc ctt gca 1008
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
gag ctc ggc gtt ggc aaa ctt gtc acc atc acc tcc acc tac gat cac 1056
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
cgc gtg atc cag ggt gct gtg tcc ggt gaa ttc ctg cgc acc atg tct 1104
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
cgc ctg ctc acc gat gat tcc ttc tgg gat gag atc ttc gac gca atg 1152
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
aac gtt cct tac acc cca atg cgt tgg gca cag gac gtt cca aac acc 1200
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
ggt gtt gat aag aac acc cgc gtc atg cag ctc att gag gca tac cgc 1248
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 410 415
tcc cgt gga cac ctc atc gct gac acc aac cca ctt tca tgg gtt cag 1296
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
cct ggc atg cca gtt cca gac cac cgc gac ctc gac atc gag acc cac 1344
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
aac ctg acc atc tgg gat ctg gac cgt acc ttc aac gtc ggt ggc ttc 1392
Asn Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Asn Val Gly Gly Phe
450 455 460
ggc ggc aag gag acc atg acc ctg cgc gag gta ctg tcc cgc ctc cgc 1440
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
gct gcg tac acc ctc aag gtc ggc tcc gaa tac acc cac atc ctg gac 1488
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
cgc gac gag cgc acc tgg ctg cag gac cgc ctc gag gcc gga atg cca 1536
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 510
aag cca acc cag gca gag cag aag tac atc ctg cag aag ctg aac gcc 1584
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
gcg gag gct ttc gag aac ttc ctg cag acc aag tac gtc ggc cag aag 1632
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
cgc ttc tcc ctc gaa ggt gca gaa gca ctt atc cca ctg atg gac tcc 1680
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
gcc atc gac acc gcc gca ggc caa ggc ctc gac gaa gtt gtc atc ggt 1728
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
atg cca cac cgt ggt cgc ctc aac gtg ctg ttc aac atc gtg ggc aag 1776
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
cca ctg gca tcc atc ttc aac gag ttt gaa ggc caa atg gag cag ggc 1824
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
cag atc ggt ggc tcc ggt gac gtg aag tac cac ctc ggt tcc gaa ggc 1872
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
cag cac ctg cag atg ttc ggc gac ggc gag atc aag gtc tcc ctg act 1920
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
gct aac ccg tcc cac ctg gaa gct gtt aac cca gtg atg gaa ggt atc 1968
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
gtc cgc gca aag cag gac tac ctg gac aag ggc gta gac ggc aag act 2016
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
gtt gtg cca ctg ctg ctc cac ggt gac gct gca ttc gca ggc ctg ggc 2064
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
atc gtg cca gaa acc atc aac ctg gct aag ctg cgt ggc tac gac gtc 2112
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Leu Arg Gly Tyr Asp Val
690 695 700
ggc ggc acc atc cac atc gtg gtg aac aac cag atc ggc ttc acc acc 2160
Gly Gly Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr
705 710 715 720
acc cca gac tcc agc cgc tcc atg cac tac gca acc gac tac gcc aag 2208
Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys
725 730 735
gca ttc ggc tgc cca gtc ttc cac gtc aac ggc gac gac cca gag gca 2256
Ala Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala
740 745 750
gtt gtc tgg gtt ggc cag ctg gcc acc gag tac cgt cgt cgc ttc ggc 2304
Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly
755 760 765
aag gac gtc ttc atc gac ctc gtc tgc tac cgc ctc cgc ggc cac aac 2352
Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn
770 775 780
gaa gct gat gat cct tcc atg acc cag cca aag atg tat gag ctc atc 2400
Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile
785 790 795 800
acc ggc cgc gag acc gtt cgt gct cag tac acc gaa gac ctg ctc gga 2448
Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly
805 810 815
cgt gga gac ctc tcc aac gaa gat gca gaa gca gtc gtc cgc gac ttc 2496
Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe
820 825 830
cac gac cag atg gaa tct gtg ttc aac gaa gtc aag gaa ggc ggc aag 2544
His Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys
835 840 845
aag cag gct gag gca cag acc ggc atc acc ggc tcc cag aag ctt cca 2592
Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro
850 855 860
cac ggc ctt gag acc aac atc tcc cgt gaa gag ctc ctg gaa ctg gga 2640
His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly
865 870 875 880
cag gct ttc gcc aac acc cca gaa ggc ttc aac tac cac cca cgt gtg 2688
Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val
885 890 895
gct ccc gtt gct aag aag cgc gtc tcc tct gtc acc gaa ggt ggc atc 2736
Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile
900 905 910
gac tgg gca tgg ggc gag ctc ctc gcc ttc ggt tcc ctg gct aac tcc 2784
Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser
915 920 925
ggc cgc ttg gtt cgc ctt gca ggt gaa gat tcc cgc cgc ggt acc ttc 2832
Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe
930 935 940
acc cag cgc cac gca gtt gcc atc gac cca gcg acc gct gaa gag ttc 2880
Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe
945 950 955 960
aac cca ctc cac gag ctt gca cag tcc aag ggc aac aac ggt aag ttc 2928
Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe
965 970 975
ctg gtc tac aac t cc gca ctg acc gag tac gca ggc atg ggc ttc gag 2976
Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu
980 985 990
tac ggc tac tcc gta gga aac gaa gac tcc atc gtt gca tgg gaa gca 3024
Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Ile Val Ala Trp Glu Ala
995 1000 1005
cag ttc ggc gac ttc gcc aac ggc gct cag acc atc atc gat gag 3069
Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Il e Asp Glu
1010 1015 1020
tac gtc tcc tca ggc gaa gct aag tgg ggc cag acc tcc aag ctg 3114
Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu
1025 1030 1035
atc ctt ctg ctg cct cac ggc tac gaa ggc cag ggc cca gac cac 3159
Ile Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His
1040 1045 1050
tct tcc gca cgt atc gag cgc ttc ctg cag ctg tgc gct gag ggt 3204
Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly
1055 1060 1065
tcc atg act gtt gct cag cca tcc acc cca gca aac cac ttc cac 3249
Ser Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His
1070 1075 1080
cta ctg cgt cgt cac gct ctg tcc gac ctg aag cgt cca ctg gtt 3294
Leu Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg ProLeu Val
1085 1090 1095
atc ttc acc ccg aag tcc atg ctg cgt aac aag gct gct gcc tcc 3339
Ile Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser
1100 1105 1110
gca cca gaa gac ttc act gag gtc acc aag ttc cag tcc gtg atc 3384
Ala Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile
1115 1120 1125
aac gat cca aac gtt gca gat gca gcc aag gtg aag aag gtc atg 3429
Asn Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met
1130 1135 1140
ctg gtc tcc ggc aag ctg tac tac gaa ttg gca aag cgc aag gag 3474
Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu
1145 1150 1155
aag gac gga cgc gac gac atc gcg atc gtt cgt atc gaa atg ctc 3519
Lys Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu
1160 1165 1170
cac cca att ccg ttc aac cgc atc tcc gag gct ctt gcc ggc tac 3564
His Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr
1175 1180 1185
cct aac gct gag gaa gtc ctc ttc gtt cag gat gag cca gca aac 3609
Pro Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn
1190 1195 1200
cag ggc cca tgg ccg ttc tac cag gag cac ctc cca gag ctg atc 3654
Gln Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile
1205 1210 1215
ccg aac atg cca aag atg cgc cgc gtt tcc cgc cgc gct cag tcc 3699
Pro Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser
1220 1225 1230
tcc acc gca act ggt gtt gcc aag gtg cac cag ctg gag gag aag 3744
Ser Thr Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys
1235 1240 1245
cag ctt atc gac gag gct ttc gag gct taa 3774
Gln Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
<210>51
<211>1257
<212>PRT
<213>谷氨酸棒杆菌
<400>51
Met Leu Gln Leu Gly Leu Arg His Asn Gln Pro Thr Thr Asn Val Thr
1 5 10 15
Val Asp Lys Thr Lys Leu Asn Lys Pro Ser Arg Ser Lys Glu Lys Arg
20 25 30
Arg Val Pro Ala Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp
35 40 45
Leu Val Asp Glu Met Phe Gln Gln Phe Gln Lys Asp Pro Lys Ser Val
50 55 60
Asp Lys Glu Trp Arg Glu Leu Phe Glu Ala Gln Gly Gly Pro Asn Thr
65 70 75 80
Thr Pro Ala Thr Thr Glu Ala Gln Pro Ser Ala Pro Lys Glu Ser Ala
85 90 95
Lys Pro Ala Pro Lys Ala Ala Pro Ala Ala Lys Ala Ala Pro Arg Val
100 105 110
Glu Thr Lys Pro Ala Asp Lys Thr Ala Pro Lys Ala Lys Glu Ser Ser
115 120 125
Val Pro Gln Gln Pro Lys Leu Pro Glu Pro Gly Gln Thr Pro Ile Arg
130 135 140
Gly Ile Phe Lys Ser Ile Ala Lys Asn Met Asp Ile Ser Leu Glu Ile
145 150 155 160
Pro Thr Ala Thr Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu
165 170 175
Asn Arg Ala Met Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys
180 185 190
Ile Ser Phe Thr His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met
195 200 205
Ala His Pro Asp Met Asn Asn Ser Tyr Asp Val Ile Asp Gly Lys Pro
210 215 220
Thr Leu Ile Val Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu
225 230 235 240
Pro Gln Lys Asp Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu
245 250 255
Thr Glu Lys Met Asn Phe Ser Glu Phe Leu Ala Ala Tyr Glu Asp Ile
260 265 270
Val Ala Arg Ser Arg Lys Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly
275 280 285
Val Thr Val Ser Leu Thr Asn Pro Gly Gly IIe Gly Thr Arg His Ser
290 295 300
Val Pro Arg Leu Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser
305 310 315 320
Met Asp Tyr Pro Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala
325 330 335
Glu Leu Gly Val Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His
340 345 350
Arg Val Ile Gln Gly Ala Val Ser Gly Glu Phe Leu Arg Thr Met Ser
355 360 365
Arg Leu Leu Thr Asp Asp Ser Phe Trp Asp Glu Ile Phe Asp Ala Met
370 375 380
Asn Val Pro Tyr Thr Pro Met Arg Trp Ala Gln Asp Val Pro Asn Thr
385 390 395 400
Gly Val Asp Lys Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg
405 4l0 415
Ser Arg Gly His Leu Ile Ala Asp Thr Asn Pro Leu Ser Trp Val Gln
420 425 430
Pro Gly Met Pro Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His
435 440 445
Asn Leu Thr Ile Trp Asp Leu Asp Arg Thr Phe Asn Val Gly Gly Phe
450 455 460
Gly Gly Lys Glu Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg
465 470 475 480
Ala Ala Tyr Thr Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp
485 490 495
Arg Asp Glu Arg Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro
500 505 5l0
Lys Pro Thr Gln Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala
515 520 525
Ala Glu Ala Phe Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys
530 535 540
Arg Phe Ser Leu Glu Gly Ala Glu Ala Leu Ile Pro Leu Met Asp Ser
545 550 555 560
Ala Ile Asp Thr Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly
565 570 575
Met Pro His Arg Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys
580 585 590
Pro Leu Ala Ser Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly
595 600 605
Gln Ile Gly Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly
610 615 620
Gln His Leu Gln Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr
625 630 635 640
Ala Asn Pro Ser His Leu Glu Ala Val Asn Pro Val Met Glu Gly Ile
645 650 655
Val Arg Ala Lys Gln Asp Tyr Leu Asp Lys Gly Val Asp Gly Lys Thr
660 665 670
Val Val Pro Leu Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly
675 680 685
Ile Val Pro Glu Thr Ile Asn Leu Ala Lys Leu Arg Gly Tyr Asp Val
690 695 700
Gly Gly Thr Ile His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr
705 710 7l5 720
Thr Pro Asp Ser Ser Arg Ser Met His Tyr Ala Thr Asp Tyr Ala Lys
725 730 735
Ala Phe Gly Cys Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala
740 745 750
Val Val Trp Val Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly
755 760 765
Lys Asp Val Phe Ile Asp Leu Val Cys Tyr Arg Leu Arg Gly His Asn
770 775 780
Glu Ala Asp Asp Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile
785 790 795 80O
Thr Gly Arg Glu Thr Val Arg Ala Gln Tyr Thr Glu Asp Leu Leu Gly
805 810 815
Arg Gly Asp Leu Ser Asn Glu Asp Ala Glu Ala Val Val Arg Asp Phe
820 825 830
His Asp Gln Met Glu Ser Val Phe Asn Glu Val Lys Glu Gly Gly Lys
835 840 845
Lys Gln Ala Glu Ala Gln Thr Gly Ile Thr Gly Ser Gln Lys Leu Pro
850 855 860
His Gly Leu Glu Thr Asn Ile Ser Arg Glu Glu Leu Leu Glu Leu Gly
865 870 875 880
Gln Ala Phe Ala Asn Thr Pro Glu Gly Phe Asn Tyr His Pro Arg Val
885 890 895
Ala Pro Val Ala Lys Lys Arg Val Ser Ser Val Thr Glu Gly Gly Ile
900 905 910
Asp Trp Ala Trp Gly Glu Leu Leu Ala Phe Gly Ser Leu Ala Asn Ser
915 920 925
Gly Arg Leu Val Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe
930 935 940
Thr Gln Arg His Ala Val Ala Ile Asp Pro Ala Thr Ala Glu Glu Phe
945 950 955 960
Asn Pro Leu His Glu Leu Ala Gln Ser Lys Gly Asn Asn Gly Lys Phe
965 970 975
Leu Val Tyr Asn Ser Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu
980 985 990
Tyr Gly Tyr Ser Val Gly Asn Glu Asp Ser Ile Val Ala Trp Glu Ala
995 1000 1005
Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu
1010 1015 1020
Tyr Val Ser Ser Gly Glu Ala Lys Trp Gly Gln Thr Ser Lys Leu
1025 1030 1035
Ile Leu Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro Asp His
1040 1045 1050
Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Cys Ala Glu Gly
1055 1060 1065
Ser Met Thr Val Ala Gln Pro Ser Thr Pro Ala Asn His Phe His
1070 1075 1080
Leu Leu Arg Arg His Ala Leu Ser Asp Leu Lys Arg Pro Leu Val
1085 1090 1095
Ile Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala Ala Ser
1100 1105 1110
Ala Pro Glu Asp Phe Thr Glu Val Thr Lys Phe Gln Ser Val Ile
1115 1120 1125
Asn Asp Pro Asn Val Ala Asp Ala Ala Lys Val Lys Lys Val Met
1130 1135 1140
Leu Val Ser Gly Lys Leu Tyr Tyr Glu Leu Ala Lys Arg Lys Glu
1145 1150 1155
Lys Asp Gly Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu
1160 1165 1170
His Pro Ile Pro Phe Asn Arg Ile Ser Glu Ala Leu Ala Gly Tyr
1175 1180 1185
Pro Asn Ala Glu Glu Val Leu Phe Val Gln Asp Glu Pro Ala Asn
1190 1195 1200
Gln Gly Pro Trp Pro Phe Tyr Gln Glu His Leu Pro Glu Leu Ile
1205 12l0 1215
Pro Asn Met Pro Lys Met Arg Arg Val Ser Arg Arg Ala Gln Ser
1220 1225 1230
Ser Thr Ala Thr Gly Val Ala Lys Val His Gln Leu Glu Glu Lys
1235 1240 1245
Gln Leu Ile Asp Glu Ala Phe Glu Ala
1250 1255
Claims (13)
1.生产L-谷氨酸的棒状杆菌型细菌,其包含
a)胞内α-酮戊二酸脱氢酶活性,所述活性小于非突变或野生型菌株的一半,和
b)在染色体odhA基因的编码区或表达控制区中的突变,所述染色体odhA基因编码α-酮戊二酸脱氢酶复合物的Elo亚基,
其中所述细菌几乎以与非突变或野生型菌株相同的生长速率生长。
2.根据权利要求1的棒状杆菌型细菌,其中所述odhA基因编码选自下组的蛋白质:
(A)包含SEQ ID NO:10的氨基酸序列的蛋白质,
(B)包含SEQ ID NO:10的氨基酸序列的蛋白质,其中在所述蛋白质中取代、缺失、插入、或添加了一个或几个氨基酸,并且其中所述蛋白质具有α-酮戊二酸脱氢酶复合物的Elo亚基的活性,
(C)包含SEQ ID NO:10的氨基酸37至1257的蛋白质,和
(D)包含SEQ ID NO:10的氨基酸37至1257的蛋白质,其中在所述蛋白质中取代、缺失、插入、或添加了一个或几个氨基酸,并且其中所述蛋白质具有α-酮戊二酸脱氢酶复合物的Elo亚基的活性。
3.根据权利要求2的棒状杆菌型细菌,其中所述几个氨基酸是2至20个氨基酸。
4.根据权利要求1的棒状杆菌型细菌,其中所述odhA基因选自下组:
(a)包含SEQ ID NO:9的核苷酸443至4213的基因,
(b)能够在严紧条件下与以下物质杂交的基因:包含SEQ ID NO:9的核苷酸443至4213的多核苷酸或由包含SEQ ID NO:9的核苷酸443至4213的多核苷酸制备的探针,并且其中所述基因编码蛋白质,该蛋白质具有α-酮戊二酸脱氢酶复合物的Elo亚基的活性,
(c)包含SEQ ID NO:9的核苷酸551至4213的基因,和
(d)能够在严紧条件下与以下物质杂交的基因:包含SEQ ID NO:9的核苷酸551至4213的多核苷酸或由包含SEQ ID NO:9的核苷酸551至4213的多核苷酸制备的探针,并且其中所述基因编码蛋白质,该蛋白质具有α-酮戊二酸脱氢酶复合物的Elo亚基的活性。
5.根据权利要求1至4中任一项的棒状杆菌型细菌,其中所述突变被引入到编码焦磷酸硫胺素结合区的区域。
6.根据权利要求5的棒状杆菌型细菌,其中所述突变被引入到包含SEQID NO:9的核苷酸2534至2548的区域。
7.根据权利要求5的棒状杆菌型细菌,其中所述突变包括在包含SEQ IDNO:10的氨基酸698至702的区域中缺失一个或多个氨基酸。
8.根据权利要求5的棒状杆菌型细菌,其中所述突变包括氨基酸的替换,所述氨基酸从下组选择:SEQ ID NO:10所示氨基酸序列中698位的Lys、699位的Leu、700位的Arg、702位的Tyr、及其组合。
9.根据权利要求1至4中任一项的棒状杆菌型细菌,其中所述突变被引入到包含SEQ ID NO:9的核苷酸1094至1111的区域。
10.根据权利要求9的棒状杆菌型细菌,其中所述突变包括在包含SEQID NO:10的氨基酸218至224的区域中缺失一个或多个氨基酸。
11.产生L-谷氨酸的方法,包括:
a)在培养基中培养根据权利要求1至10中任一项的棒状杆菌型细菌,和
b)从培养基和/或所述细菌收集L-谷氨酸。
12.编码突变α-酮戊二酸脱氢酶的基因,选自下组:
(a)包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸443至4213的基因,或者包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸443至4210的基因,
(b)能够在严紧条件下与以下物质杂交的基因:包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸443至4213的多核苷酸、包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸443至4210的多核苷酸、由从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸443至4213制备的探针、或者由从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸443至4210制备的探针,并且其中所述基因编码具有α-酮戊二酸脱氢酶活性的蛋白质,通过与E2o和E3亚基形成复合物,该蛋白质的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半,
(c)包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸551至4213的基因,或者包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸551至4210的基因,
(d)能够在严紧条件下与以下物质杂交的基因:包含从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸551至4213的多核苷酸、包含从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸551至4210的多核苷酸、由从SEQ ID NO:11、13和15组成的组中选择的多核苷酸的核苷酸551至4213制备的探针、或者由从SEQ ID NO:44、46和48组成的组中选择的多核苷酸的核苷酸551至4210制备的探针,并且其中所述基因编码具有α-酮戊二酸脱氢酶活性的蛋白质,通过与E2o和E3亚基形成复合物,该蛋白质的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半。
13.突变α-酮戊二酸脱氢酶,选自下组:
(a)从SEQ ID NO:12、14、16、45、47和49组成的组中选择的蛋白质,
(b)从SEQ ID NO:12、14、16、45、47和49组成的组中选择的蛋白质,其中在所述蛋白质中取代、缺失或添加了一个或几个氨基酸,并且其中通过与E2o和E3亚基形成复合物,所述蛋白质显示的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半,
(c)包含从SEQ ID NO:12、45、47和49组成的组中选择的氨基酸序列的氨基酸37至1256的蛋白质,包含氨基酸序列SEQ ID NO:14的氨基酸37至1255的蛋白质,或者包含氨基酸序列SEQ ID NO:16的氨基酸37至1254的蛋白质,和
(d)包含从SEQ ID NO:12、45、47和49组成的组中选择的氨基酸序列的氨基酸37至1256的蛋白质,包含氨基酸序列SEQ ID NO:14的氨基酸37至1255的蛋白质,或者包含氨基酸序列SEQ ID NO:16的氨基酸37至1254的蛋白质,其中在所述蛋白质中取代、缺失、或添加了一个或几个氨基酸,并且其中通过与E2o和E3亚基形成复合物,所述蛋白质显示的α-酮戊二酸脱氢酶活性小于野生型或非突变菌株的α-酮戊二酸脱氢酶活性的一半。
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CN108250278A (zh) * | 2016-12-29 | 2018-07-06 | 廊坊梅花生物技术开发有限公司 | 生产l-谷氨酸的菌株和生产l-谷氨酸的方法 |
CN113637699A (zh) * | 2020-04-27 | 2021-11-12 | 中国科学院分子植物科学卓越创新中心 | 一种提高氨基酸产生菌生产能力的方法 |
TWI834985B (zh) * | 2020-09-09 | 2024-03-11 | 南韓商Cj第一製糖股份有限公司 | 用於生產左旋麩胺酸的重組微生物及使用其生產左旋麩胺酸的方法 |
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KR100948246B1 (ko) * | 2004-12-28 | 2010-03-18 | 아지노모토 가부시키가이샤 | L-글루탐산 생산 미생물 및 l-글루탐산의 제조방법 |
JP2010041920A (ja) | 2006-12-19 | 2010-02-25 | Ajinomoto Co Inc | L−アミノ酸の製造法 |
JP2010130899A (ja) | 2007-03-14 | 2010-06-17 | Ajinomoto Co Inc | L−グルタミン酸系アミノ酸生産微生物及びアミノ酸の製造法 |
WO2009103547A1 (en) | 2008-02-21 | 2009-08-27 | Basf Se | Process for the production of gamma-aminobutyric acid |
KR20140091742A (ko) | 2011-11-11 | 2014-07-22 | 아지노모토 가부시키가이샤 | 발효법에 의한 목적 물질의 제조법 |
JP2016165225A (ja) | 2013-07-09 | 2016-09-15 | 味の素株式会社 | 有用物質の製造方法 |
JP2016192903A (ja) | 2013-09-17 | 2016-11-17 | 味の素株式会社 | 海藻由来バイオマスからのl−アミノ酸の製造方法 |
KR101547651B1 (ko) * | 2014-06-03 | 2015-08-26 | 씨제이제일제당 주식회사 | O-숙시닐호모세린 또는 숙신산의 생산능을 갖는 미생물 및 이를 이용한 숙신산 또는 o-숙시닐호모세린의 생산 방법 |
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KR102665227B1 (ko) * | 2021-06-30 | 2024-05-10 | 씨제이제일제당 주식회사 | 고농도 l-글루탐산을 생산하기 위한 균주 및 이를 이용한 l-글루탐산 생산방법 |
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CN108250278A (zh) * | 2016-12-29 | 2018-07-06 | 廊坊梅花生物技术开发有限公司 | 生产l-谷氨酸的菌株和生产l-谷氨酸的方法 |
CN108250278B (zh) * | 2016-12-29 | 2021-06-22 | 廊坊梅花生物技术开发有限公司 | 生产l-谷氨酸的菌株和生产l-谷氨酸的方法 |
CN113637699A (zh) * | 2020-04-27 | 2021-11-12 | 中国科学院分子植物科学卓越创新中心 | 一种提高氨基酸产生菌生产能力的方法 |
CN113637699B (zh) * | 2020-04-27 | 2023-05-30 | 中国科学院分子植物科学卓越创新中心 | 一种提高氨基酸产生菌生产能力的方法 |
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