CN101255431A - 编码代谢途径蛋白的谷氨酸棒杆菌基因 - Google Patents
编码代谢途径蛋白的谷氨酸棒杆菌基因 Download PDFInfo
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- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
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Abstract
描述了分离的编码新的谷氨酸棒杆菌MP蛋白的核酸分子,该分子被称为MP核酸分子。本发明也提供了反义核酸分子,含有MP核酸分子的重组表达载体,以及已导入表达载体的宿主细胞。本发明也进一步提供了分离的MP蛋白,MP突变蛋白,融合蛋白质,抗原肽,以及基于谷氨酸棒杆菌MP基因遗传工程提高由该生物体进行的所需化合物生产的方法。
Description
本申请是申请日为2000年12月22日,申请号为00819506.4的、发明名称和本发明相同的发明专利申请的分案申请。
相关申请
本申请是2000年6月23日提交的美国专利申请09/606,740的部分继续申请。本申请也是2000年6月23日提交的美国专利申请09/603,124的部分继续申请。本申请要求以下申请的优先权:1999年6月25日提交的美国临时专利申请60/141031,1999年6月2日提交的美国临时专利申请60/142101,1999年8月12日提交的美国临时专利申请60/148613,2000年3月9日提交的美国临时专利申请60/187970,以及1999年7月8日提交的德国专利申请19931420.9。上述申请的全部内容本文引为参考。
发明背景
细胞中天然存在的代谢过程中的特定产物和副产物,在包括食品、饲料、化妆品和制药产业在内的很多行业中有用。这些分子总称为“精细化学物质”,包括有机酸、蛋白源的和非蛋白源的氨基酸、核苷酸和核苷、脂质和脂肪酸、二醇、糖类、芳香族化合物、维生素和辅因子以及酶。可以通过大规模培养产生并分泌大量特定所需分子的细菌,最方便的制备这些产品。用于这一目的的一种特别有用的生物体就是谷氨酸棒杆菌(Corynebacterium glutamicum),一种革兰氏阳性非病原菌。通过菌株筛选,出现了许多产生大批所需化合物的突变株。然而,为改进特殊分子生产而进行的菌株筛选,是一个耗时并且困难的过程。
发明概述
本发明提供了新的细菌核酸分子,这些分子具有多种用途。这些用途包括鉴定可以产生精细化学物质(如赖氨酸和甲硫氨酸等氨基酸)的微生物、调节谷氨酸棒杆菌或者亲缘细菌中的精细化学物质的产生、谷氨酸棒杆菌或者亲缘细菌的分型和鉴定、作为绘制谷氨酸棒杆菌基因组图谱的参照点。这些新的核酸分子编码蛋白质,此处称为代谢途径(MP)蛋白质。
谷氨酸棒杆菌是一种革兰氏阳性需氧细菌,通常用于工大规模生产各种精细化学物质,也用于降解烃类(例如在石油泄漏中)和氧化萜品醇。因此,本发明的MP核酸分子,可用来鉴定能用于生产精细化学物质的微生物,例如通过发酵方法。调节本发明MP核酸分子的表达,或者修饰本发明MP核酸分子的序列,可以用于调节微生物中一种或者多种精细化学物质的生产(例如,提高棒杆菌或者短杆菌中一种或者多种精细化学物质的产生)。在一个优选实施方案中,本发明的MP基因与参与相同和不同代谢途径的一个或多个基因组合,以调节微生物生产一种或多种精细化学物质。
本发明MP核酸分子可用于鉴定一种微生物是否是谷氨酸棒杆菌或者其亲缘菌株,或者鉴定微生物混合群体中谷氨酸棒杆菌或者其亲缘菌株的存在。本发明提供了许多谷氨酸棒杆菌基因的核酸序列;在严格条件下,用探针探查从单一微生物或者混合微生物群体培养物中提取的基因组DNA,该探针覆盖了谷氨酸棒杆菌基因特有的一段区域,可以确定是否有该微生物存在。尽管谷氨酸棒杆菌本身是非病原性的,但是它与人体中的病原菌种有关,例如白喉棒状菌(Corynebacterium diphtheriae)(白喉致病原);探测这种微生物具有重大的临床实用性。
本发明MP核酸分子也可以用作绘制谷氨酸棒杆菌基因组图谱的参照点,或者绘制其亲缘菌株基因组图谱的参照点。相似的,这些分子,或者其变体或其部分,可以用作遗传工程棒杆菌或者短杆菌的遗传标记。
例如,本发明新核酸分子编码的MP蛋白可以进行某些精细化学物质代谢中的酶促步骤,所述精细化学物质包括氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子(nutraceutical)、核苷酸、核苷和海藻糖。考虑到可在谷氨酸棒杆菌中使用的克隆载体的实用性,例如在Sinskey etal.,美国专利号No.4,649,119中公开的,并且考虑到谷氨酸棒杆菌和亲缘短杆菌菌种(例如乳发酵短杆菌)的遗传操作技术(Yashihama et al,J.Bacteriol.162:591-597(1985);Katsumata et al.,J.Bacteriol.159:306-311(1984);以及Santamaria et al.,J.Gen.Microbiol.130:2237-2246(1984)),本发明的核酸分子可用于该生物体的遗传工程,使之成为一种或者多种精细化学物质更好的或者更有效的生产者。
精细化学物质的提高或有效生产可以是本发明基因操作的直接作用或这种基因操作的间接作用。具体而言,谷氨酸棒杆菌氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、核苷酸和海藻糖代谢途径的改变对这种生物生产一种或多种这些所需化合物具有直接的效应。例如,优化赖氨酸或甲硫氨酸生物合成途径蛋白活性或降低赖氨酸或甲硫氨酸分解途径蛋白活性可以导致由这种工程改造的生物生产赖氨酸或甲硫氨酸的产量或效率提高。这些代谢途径蛋白的改变也会对所需精细化学物质生产或效率有间接影响。例如,与生产所需分子必须的中间体竞争的反应可以被消除,或者生产所需化合物特定中间体必须的途径可以被优化。此外,氨基酸如赖氨酸或甲硫氨酸、维生素或核苷酸生物合成或降解的调控可以增加生产和分裂的能力,从而增加培养物中微生物的数量和/或生产能力,并增加所需精细化学物质的可能产量。
本发明的核酸和蛋白分子单独或与一种或多种相同或不同代谢途径的核酸和蛋白分子组合,可以用来直接提高谷氨酸棒杆菌中一种或多种所需精细化学物质(如甲硫氨酸或赖氨酸)的生产或生产效率。使用本领域已知的重组技术,一种或多种本发明的氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖的生物合成或降解酶可以被改变,从而调节其功能。例如,可以提高生物合成酶的效率,或破坏其别构控制区从而防止化合物生产的反馈抑制。类似地,降解酶可以通过置换、缺失或增加被缺失或修饰,从而其对所需化合物的降解活性降低,而不影响细胞的活力。在各种情况下,所需精细化学物质的总产量或产率均被提高。
本发明的蛋白质和核苷酸分子的改变也可能通过间接机制提高除氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖以外的其它精细化学物质的生产。任一种化合物的代谢必然与细胞内其它生物合成和降解途径关联,一种途径中的必需辅因子、中间体或底物可能由其它同类途径供给或受其限制。因此,通过调整一种或多种本发明蛋白的活性,另一种精细化学物质生物合成或降解途径活性的生产或效率可能会受到影响。例如,氨基酸可以作为所有蛋白质的结构单元,但其在细胞内的存在的水平可能会限制蛋白合成;因此,通过增加细胞内一种或多种氨基酸的生产效率或产率,诸如生物合成或降解蛋白的蛋白可以更容易合成。同样,代谢途径酶的改变使得特定副反应更有利或不利时,会导致一种或多种用作生产所需精细化学物质的中间体或底物的化合物过量生产或生产不足。
本发明提供了新的编码蛋白质的核酸分子,这种蛋白质在此处称作代谢途径(MP)蛋白质,它们能够完成对细胞正常功能重要的分子例如氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、核苷酸和核苷酸或海藻糖代谢过程中的酶促步骤。编码MP蛋白的核酸分子此处称为MP核酸分子。在优选实施方案中,MP蛋白单独或与相同或不同代谢途径的一种或多种蛋白组合,执行与以下一种或多种物质的代谢相关的酶促步骤:氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷和海藻糖。这些蛋白质的实例,包括那些在表1中列出的基因所编码的蛋白质。
因此,本发明的一个方面涉及,分离含有一段编码一种MP蛋白或者其生物活性部分的核酸序列的核酸分子(例如,cDNA,DNA,或者RNA),以及分离适合作为探测或扩增MP编码核酸(例如DNA或者RNA)的引物或者杂交探针的核酸片段。在特别优选的实施方案中,分离的核酸分子包含一段列在序列表中的序列号为奇数的核酸序列(例如,SEQ ID NO:1,SEQ ID NO:3,SEQ ID NO:5)或者一条这种核苷酸序列的编码区域或者其互补序列。在其他特别优选的实施方案中,分离的本发明核酸分子包含与序列表中的序列号为奇数的核苷酸序列(例如,SEQ ID NO:1,SEQ ID NO:3,SEQ ID NO:5)或者其部分有至少大约50%、51%、52%、53%、54%、55%、56%、57%、58%、59%或60%同源性,优选有至少大约61%、62%、63%、64%、65%、66%、67%、68%、69%或70%的同源性,更优选有至少大约71%,72%、73%、74%、75%、76%、77%、78%、79%或80%、81%、82%、83%、84%、85%、86%、87%、88%、89%或90%或91%、92%、93%、94%的同源性,甚至更优选的有至少大约95%,96%,97%,98%,99%、99.7%或者更高的同源性。在其他优选的实施方案中,已分离的核酸分子编码列在序列表中的偶数序列号氨基酸序列(例如,SEQ ID NO:2,SEQ ID NO:4,SEQ ID NO:6)。本发明优选的MP蛋白也优选具有至少一种此处描述的MP活性。
在另一个实施方案中,已分离的核酸分子编码一种蛋白质或者其部分,其中的蛋白质或者其部分包含一段氨基酸序列,该序列与本发明的氨基酸序列(例如,在序列表中偶数序列号的序列,如SEQ ID NO:2,SEQID NO:4,SEQ ID NO:6)有充分的同源性,例如,与本发明的氨基酸序列有充分的同源性而使得该蛋白质或者其部分具有MP活性。优选,核酸分子编码的蛋白质或者其部分,保持进行氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、核苷酸和核苷酸或海藻糖代谢途径中的酶促反应的能力。在一个实施方案中,核酸分子编码的蛋白质与本发明的氨基酸序列(例如,从序列表中的偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4,SEQ ID NO:6中选出的完整氨基酸序列)有至少大约50%、51%、52%、53%、54%、55%、56%、57%、58%、59%或60%同源性,优选有至少大约61%、62%、63%、64%、65%、66%、67%、68%、69%或70%的同源性,更优选有至少大约71%,72%、73%、74%、75%、76%、77%、78%、79%或80%、81%、82%、83%、84%、85%、86%、87%、88%、89%或90%或91%、92%、93%、94%的同源性,甚至更优选的有至少大约95%,96%,97%,98%,99%、99.7%或者更高的同源性。在另一个优选的实施方案中,蛋白质是全长的谷氨酸棒杆菌蛋白质,该蛋白质与本发明的全长氨基酸序列(由显示在相应序列表中的奇数序列号核酸序列(例如,SEQ ID NO:1,SEQ ID NO:3,SEQ ID NO:5)开放阅读框架编码的)基本同源。
在另一个优选的实施方案中,分离的核酸分子来自谷氨酸棒杆菌,并编码一种蛋白质(例如一种MP融合蛋白),该蛋白质包含一段生物活性区域,该区域与本发明的一种氨基酸序列(例如,序列表偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4,SEQ ID NO:6中的一个序列)有至少大约50%或者更高的同源性,并且该蛋白质能够催化氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、核苷酸和核苷酸或海藻糖代谢途径中的反应,或者拥有一种或者多种列在表1中的活性,并且该蛋白质还包含有一段编码异源多肽或者调节区域的异源核酸序列。
在另一个实施方案中,分离的核酸分子至少有15个核苷酸的长度,并且在严格条件下与含有本发明核苷酸序列(例如,在序列表中奇数序列号序列如SEQ ID NO:1,SEQ ID NO:3,SEQ ID NO:5)的核酸分子杂交。优选,分离的核酸分子与天然存在的核酸分子一致。更加优选,分离的核酸分子编码天然存在的谷氨酸棒杆菌MP蛋白,或者其生物活性部分。
本发明的另一个方面涉及载体,例如含有本发明核酸分子或者含有本发明核酸分子与相同或不同途径的一种或多种核酸分子组合的重组表达载体,和被引入这种载体的宿主细胞。在一个实施方案中,通过在合适的培养基中进行培养,这种宿主细胞被用于生产MP蛋白。然后可以从培养基或者宿主细胞中分离该MP蛋白。
另外,本发明的另一个方面涉及一种经过遗传改变的微生物,一种或多种MP基因已经被单独或与一种或多种相同或不同代谢途径的基因组合引入其中或者已经被改变。在一个实施方案中,通过单独引入作为转基因的编码一种或多种野生型或者突变型MP序列的本发明核酸分子或与一种或多种相同或不同代谢途径的核酸分子,改变了该微生物的基因组。在另一个实施方案中,改变了该微生物基因组中的一种或多种内源MP基因,例如,通过使用一种或多种已改变的MP基因进行同源重组而进行功能性破坏。在另一个实施方案中,该微生物中一种或多种内源的或者引入的MP基因(单独或与一种或多种相同或不同代谢途径的基因组合)通过一个或者多个点突变、缺失或者倒位而被改变,但是仍然能编码功能MP蛋白。在另一个实施方案中,改变一种或多种微生物MP基因(单独或与一种或多种相同或不同代谢途径的基因组合)的一个或者多个调节区域(例如,启动子、阻抑物或者诱导物),从而调节一种或多种MP基因的表达。在优选的实施方案中,微生物属于棒杆菌种或者短杆菌种,特别优选是谷氨酸棒杆菌。在优选的实施方案中,也使用微生物生产所需的化合物,例如氨基酸,特别优选是赖氨酸和甲硫氨酸。在特别优选的实施方案中,MP基因是metZ基因(SEQ ID NO:1)、metC基因(SEQ ID NO:3)或RXA00657基因(SEQ ID NO:5),单独或与一种或多种本发明的MP基因或与参与甲硫氨酸和/或赖氨酸代谢的一种或多种基因组合。
另一方面,本发明提供了一种鉴定受试者中白喉棒杆菌存在或者活性的方法。该方法包括对受试者中本发明的一种或者多种核酸或者氨基酸序列(例如,列在序列表中SEQ ID NO 1至122的序列)的检测,从而可以检测受试者中谷氨酸棒杆菌的存在或者活性。
另外,本发明的另一个方面涉及已分离出MP蛋白或者其部分,例如其生物活性部分。在一个优选的实施方案中,分离的MP蛋白或者其部分,单独或与一种或多种本发明的MP蛋白或与相同或不同代谢途径的一种或多种蛋白组合,可以催化氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢的一种或多种途径中的酶促反应。在另一个优选的实施方案中,已分离的MP蛋白或者其部分与本发明的一种氨基酸序列(例如,序列表偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4,SEQ ID NO:6中的一个序列)有足够高的同源性,使得该蛋白质或者其部分保持催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢的一种或多种途径中的酶促反应的能力。
本发明也提供了MP蛋白的分离制品。在优选的实施方案中,MP蛋白包含本发明的氨基酸序列(例如,序列表偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4,SEQ ID NO:6中的一个序列)。在另一个优选的实施方案中,本发明与分离的全长蛋白质有关,该蛋白质与本发明的完全氨基酸序列(序列表偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4,SEQ IDNO:6中的一个序列)(由显示在相应序列表中的序列号为奇数如SEQ IDNO:1,SEQ ID NO:3或SEQ ID NO:5的开放阅读框架编码)有相当高的同源性。此外,在另一个实施方案中,蛋白质与本发明的完全氨基酸序列(例如,序列表中偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4或SEQ ID NO:6)有至少大约50%、51%、52%、53%、54%、55%、56%、57%、58%、59%或60%同源性,优选有至少大约61%、62%、63%、64%、65%、66%、67%、68%、69%或70%的同源性,更优选有至少大约71%,72%、73%、74%、75%、76%、77%、78%、79%或80%、81%、82%、83%、84%、85%、86%、87%、88%、89%或90%或91%、92%、93%、94%的同源性,甚至更优选有至少大约95%,96%,97%,98%,99%、99.7%或者更高的同源性。在另一个实施方案中,分离的MP蛋白包含的氨基酸序列与本发明的一条氨基酸序列(例如,序列表中的偶数序列号序列如SEQ ID NO:2,SEQ ID NO:4,SEQ ID NO:6中的一个序列)有至少大约50%或者更高的同源性,并且单独或者与一种或多种本发明的MP蛋白或相同或不同代谢途径的任何蛋白组合,能够催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应,或者具有表1中列出的一种或者多种活性。
另外,分离的MP蛋白可以含有由核酸序列编码的氨基酸序列,该核酸序列与列在序列表中的偶数序列号的一个核苷酸序列杂交,例如在严格条件下杂交,或者与该核苷酸序列有至少大约50%、51%、52%、53%、54%、55%、56%、57%、58%、59%或60%同源性,优选有至少大约61%、62%、63%、64%、65%、66%、67%、68%、69%或70%的同源性,更优选有至少大约71%,72%、73%、74%、75%、76%、77%、78%、79%或80%、81%、82%、83%、84%、85%、86%、87%、88%、89%或90%或91%、92%、93%、94%的同源性,甚至更优选的有至少大约95%,96%,97%,98%,99%、99.7%或者更高的同源性。也优选MP蛋白的优选形式同样具有一种或者多种此处描述的生物活性。
MP多肽或者其生物活性部分,可以有效的连接到非MP多肽上而形成融合蛋白质。在优选的实施方案中,该融合蛋白质具有不同于单独MP蛋白本身的活性。在另外优选的实施方案中,该融合蛋白质被引入谷氨酸棒杆菌氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子代谢途径时,引起谷氨酸棒杆菌中所需精细化学物质产量、生产和/或生产效率的增加。在特别优选的实施方案中,把该融合蛋白整合进宿主细胞的氨基酸、微生物、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径,可以调节细胞中所需化合物的生产。
另一方面,本发明提供了筛选可调节MP蛋白活性的分子的方法。该分子通过与蛋白质分子本身或者底物相互作用,或者与MP蛋白的配偶体结合,或者通过调节本发明MP核酸分子的转录或者翻译来调节MP蛋白活性。
本发明的另一个方面涉及生产精细化学物质的方法。该方法涉及培养含有一种或多种载体的细胞,该载体指导本发明MP核酸分子单独表达,或与一种或多种本发明的MP核酸核酸分子或相同或不同代谢途径的任何核酸分子组合表达,从而产生精细化学物质。在一个优选的实施方案中,该方法还包含获得含有该载体细胞的步骤,在该步骤中,使用可以指导MP核酸分子表达的载体转染细胞。在另一个优选的实施方案中,该方法还包含从培养基中回收精细化学物质的步骤。在一个特别优选的实施方案中,细胞是棒杆菌种或者短杆菌种,或者选自列在表3中的那些菌株。在另一个优选实施方案中,MP基因是metZ基因(SEQ IDNO:1)、metC基因(SEQ ID NO:3)或RXA00657基因(SEQ ID NO:5),单独或与一种或多种本发明的MP基因或与参与甲硫氨酸和/或赖氨酸代谢的一种或多种基因组合。在另一个优选实施方案中,精细化学物质是氨基酸如L-赖氨酸和L-甲硫氨酸。
本发明的另一方面涉及调节微生物中一种分子产生的方法。这种方法包括使用调节MP蛋白活性或者MP核酸表达的药剂接触细胞,使得细胞的相关活性相对于缺少这种药剂时的活性发生了改变。在一个优选的实施方案中,调节谷氨酸棒杆菌细胞的一种或多种氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径,使得这种微生物所需精细化学物质的产量或者产生效率得到提高。调节MP蛋白活性的药剂,可以是刺激MP蛋白活性或者MP核酸表达的药剂。刺激MP蛋白活性或者MP核酸表达的药剂的实例,包括小分子、活性MP蛋白、以及编码已导入细胞的MP蛋白的核酸。抑制MP蛋白活性或者表达的药剂的实例包括小分子和反义MP核酸分子。
本发明的另一个方面,涉及调节细胞中所需化合物产量的方法,包括把野生型或者突变型MP基因导入细胞,单独或与一种获得多种本发明的MP核酸分子或相同或不同代谢途径的任何核酸分子组合,该基因或者保留在单独的质粒上,或者整合到宿主细胞基因组中。如果整合到宿主细胞基因组中,这种整合可以是任意的,或者是通过同源重组发生的,从而使得引入的拷贝取代天然基因,导致细胞中所需化合物的产生得到调节。在一个优选的实施方案中,该产量得到增加。在另一个优选的实施方案中,所说的精细化学物质是氨基酸。在一个特别优选的实施方案中,所说的氨基酸是L-赖氨酸和L-甲硫氨酸。在另一个优选实施方案中,所述基因是metZ基因(SEQ ID NO:1)、metC基因(SEQ IDNO:3)或RXA00657基因(SEQ ID NO:5),单独或与一种或多种本发明的MP核酸分子或与参与甲硫氨酸和/或赖氨酸代谢的一种或多种基因组合。
发明详述
本发明提供了MP核酸和蛋白质分子,它们参与谷氨酸棒杆菌中某些精细化学物质包括氨基酸如赖氨酸和甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷和海藻糖的代谢。本发明的分子可用于调节微生物如谷氨酸棒杆菌中精细化学物质的生产,这种调节,可以是直接的(例如在赖氨酸和甲硫氨酸生物合成蛋白的调节对该生物体的赖氨酸和甲硫氨酸生产或生产效率具有直接影响时),或者具有间接影响,但会导致所需化合物产量或生产效率的增加(例如当核苷酸生物合成蛋白活性的调节对细菌的有机酸或脂肪酸生产有影响,可能是由于生长改进或必需的辅因子、能量化合物或前体分子的供应增加)。MP分子可以单独使用,或与本发明的其他MP分子分子组合使用,或与相同或不同代谢途径(如赖氨酸或甲硫氨酸代谢)中的其他分子组合使用。在一个优选实施方案中,MP分子是metZ(SEQ ID NO:1)、metC(SEQ ID NO:3)或RXA00657(SEQ ID NO:5)核酸分子和由这些核酸分子编码的蛋白(分别是SEQID NO:2、SEQ ID NO:4和SEQ ID NO:6。本发明的各个方面进一步详细说明如下。
I.精细化学物质
“精细化学物质”这个词是本领域熟知的,包括生物体产生的在各种产业中使用的分子,例如但不仅仅局限于,制药、农业和化妆品产业。这种化合物包括有机酸,例如酒石酸、衣康酸和二氨基庚二酸,蛋白质源和非蛋白质源氨基酸,嘌呤碱基和嘧啶碱基,核苷,以及核苷酸(例如像是描述在Kuninaka,A.(1996)Nucleotides and related compounds,p.561-612,in Biotechnology vol.6,Rehm et al.,eds.VCH:Weinheim及其所含参考文献中的),脂质,饱和和不饱和脂肪酸(例如花生四烯酸),二醇(例如,丙烷二醇和丁烷二醇),芳香族化合物(例如,芳香胺、香草醛和靛),维生素和辅因子(参见Ullmann’s Encyclopedia of IndustrialChemistry,vol.A27,“Vitamins”,p.443-613(1996)VCH:Weinheim and referencestherein;and Ong,A.S.,Niki,E.&Packer,L.(1995)“Nutrition,Lipids,Health,andDisease”Proceedings of the UNESCO/Confederation of Scientific and TechnologicalAssociations in Malaysia,and the Society for Free Radical Research-Asia,held Sept.1-3,1994at Penang,Malaysia,AOCS Press,(1995)),酶,聚酮化合物(ployketides)(Cane et al.,(1998)Science 282:63-68),以及所有在Gutcho(1983)Chemicals by Fermentation,Noyes Data Corporation,ISBN:0818805086及其参考文献中描述的化学物质。某些这些精细化学物质的代谢和用途进一步详细说明如下。
A.氨基酸的代谢和用途
氨基酸包括所有蛋白质的基本结构单元,同样也是所有生物体正常细胞生物功能所必需的。“氨基酸”这个词是本领域熟知的。蛋白质源的氨基酸有20种,是蛋白质的结构单元,相互之间由肽键相连接,而非蛋白质源氨基酸(已知的有几百种)通常情况下不会出现在蛋白质中(参见Ulmann’s Encyclopedia of Industrial Chemistry,vol.A2,p.57-97VCH:Weinheim(1985))。虽然L-氨基酸通常是天然存在蛋白质中的唯一类型,但是氨基酸可以是D-或者L-光学构型。20种蛋白质源氨基酸中每一种的生物合成或者降解途径,都在原核细胞或者真核细胞中有各自的特点(例如参见Stryer,L.Biochemistry,3rd edition,pages 578-590(1988))。“必需”氨基酸(组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、苯丙氨酸、苏氨酸、色氨酸和缬氨酸)之所以这样命名,是因为这些氨基酸生物合成复杂通常是必需的营养条件,它们可以通过简单的生物合成途径转化为其余的11种“非必需”氨基酸(丙氨酸、精氨酸、天冬酰胺、天冬氨酸、半胱氨酸、谷氨酸、谷氨酰胺、甘氨酸、脯氨酸、丝氨酸、酪氨酸)。虽然高等生物确实具有合成一些这种氨基酸的能力,但是为了正常的蛋白质合成必须从饮食中补充必需氨基酸。
它们除了在蛋白质合成中的功能,这些氨基酸就其自身来说是有趣的化学物质,并且它们中的很多在食品、饲料、化学、化妆品、农业和制药产业中具有各种应用。赖氨酸在营养方面不仅对于人类是一种重要的氨基酸,而且对于像是家禽和猪这样单胃动物也是重要的。谷氨酸是最常用的风味添加剂(谷氨酸单钠,MSG),并且广泛应用于整个食品产业,如同天冬氨酸、甘氨酸、半胱氨酸一样。甘氨酸、L-甲硫氨酸和色氨酸全部用于制药产业。谷氨酰胺、缬氨酸、亮氨酸、异亮氨酸、组氨酸、精氨酸、脯氨酸、丝氨酸和丙氨酸都应用在制药产业和化妆品产业中。苏氨酸、色氨酸和D/L-甲硫氨酸是常用的饲料添加剂(Leuchtenberger,W.(1996)Amino aids-technical production and use,p.466-502in Rehm et al.(eds.)Biocemistry vol.6,chapter 14a,VCH:Weinheim)。另外,这些氨基酸作为合成氨基酸和蛋白质合成的前体也是很有用的,例如N-乙酰半胱氨酸,S-羧甲基-L-半胱氨酸,(S)-5-羟色氨酸,以及其他在Ulmann’s Encyclopedia of Industrial Chemistry,vol.A2,p.57-97VCH:Weinheim,1985中描述的分子。
在能够产生天然氨基酸的生物体中,例如细菌,这些天然氨基酸的生物合成已经了解得很充分(细菌氨基酸的生物合成及其调节,参见Umbarger,H.E.(1978)Ann.Rev.Biochem.47:533-606)。天冬氨酸由α-酮戊二酸还原型氨化合成,后者是柠檬酸循环的中间体。谷氨酰胺、脯氨酸和精氨酸都是由谷氨酸依次产生的。丝氨酸的生物合成是一个三步的过程,开始于3-磷酸甘油酸(糖酵解的中间体),经过氧化作用、转氨作用、水解作用各步骤之后,终止于该氨基酸。半胱氨酸和甘氨酸都由丝氨酸产生;前者由高半胱氨酸与丝氨酸缩合而成,后者是把侧链β-碳原子转移到四氢叶酸得到的,该反应是由丝氨酸羟甲基转移酶催化的。苯丙氨酸和酪氨酸,由4-磷酸赤藓糖和磷酸烯醇丙酮酸在一条9步的生物合成途径中合成,它们是糖酵解途径和戊糖磷酸途径的前体,这两条途径只是在合成预苯酸之后不同。色氨酸也可以由这两种初始分子产生,但是其合成是一个11步的途径。酪氨酸也可以由苯丙氨酸合成,其反应是由苯丙氨酸羟化酶催化的。丙氨酸、缬氨酸和亮氨酸都是糖酵解终产物丙酮酸的生物合成产物。天冬氨酸由草酰乙酸合成,后者是柠檬酸循环的中间体。天冬酰胺、甲硫氨酸、苏氨酸和赖氨酸都由天冬氨酸转化而成。异亮氨酸由苏氨酸形成。
甲硫氨酸生物合成途径已经在不同生物体中进行了研究。第一步即高丝氨酸的酰化在所有生物体中是相同的,即便转移酰基的来源不同。大肠杆菌和相关物种使用琥珀酰辅酶A(Michaeli,S.And Ron,E.Z.(19810Mol.Gen.Genet.182,349-354),而酿酒酵母(Langin,T.,et al.(1986)Gene 49,283-293)、黄色短杆菌(Miyajima,R.And Shiio,I.(1973)J.Biochem.73,1061-1068;Ozaki,H and Shiio,I.(1982)J.Biochem.91,1163-1171)、谷氨酸棒杆菌(Park,S.D.et al.(1998)Mol.Cells 8,286-294)和迈氏钩端螺旋体(Belfaiza,J.et al.(1998)180,250-255;Bourhy,P.,et al.(1997)J.Bacteriol.179,4396-4398)使用乙酰辅酶A作为酰基供体。由酰基高丝氨酸形成高半胱氨酸有两种不同途径。大肠杆菌使用转硫途径,通过胱硫醚γ合酶(metB的产物)和胱硫醚β裂合酶(metC的产物)催化。酿酒酵母(Cherest,H.andSurdin-Kerjan,Y.(1992)Genetics 130,51-58),黄色短杆菌(Ozaki,H.andShiio,I.(1982)J.Biochem.91,1163-1171),铜绿假单胞菌(Foglino,M.,et al.(1995)Microbiology 141,431-439),和迈氏钩端螺旋体(Belfaiza,J.,et al.(1998)J.Bacteriol.180,250-255)利用直接硫化氢解途径,由酰基高丝氨酸硫化氢解酶催化。不像密切相关的黄色短杆菌仅利用直接硫化氢解途径,在谷氨酸棒杆菌的抽提物中检测到了转硫途径的酶活性,在该生物中该途径被认为是甲硫氨酸生物合成的路径(Hwang,B-J.,et al(1999)Mol.Cells 9,300-308;Kase,H.and Nakayama,K.(1974)Agr.Biol.Chem.38,2021-2030;Park,S.-D.,et al.1998)Mol.Cells 8,286-294)。
尽管已经分离到参与谷氨酸棒杆菌甲硫氨酸生物合成的某些酶,关于谷氨酸棒杆菌中甲硫氨酸生物合成的信息仍非常有限。从该生物中只分离出了metA和metB基因。为了理解谷氨酸棒杆菌中甲硫氨酸的生物合成途径,我们分离和鉴定了谷氨酸棒杆菌的metC基因(SEQ ID NO:3)和metZ(也称为metY)基因(SEQ ID NO:1)(见表1)。
超出细胞蛋白质合成所需的氨基酸是不能储存的,而是被降解后为细胞主要代谢途径提供中间体(评论参见Stryer,L.Biochemistry 3rd ed.Ch.21“Amino Acid Degradation and the Urea Cycle”p.495-516(1988))。尽管细胞能转化多余的氨基酸为有用的代谢中间体,但是产生氨基酸要消耗很多的能量、前体分子和合成所需的酶。因此用反馈抑制来调节氨基酸的生物合成是不令人吃惊的,特殊氨基酸的存在可以减慢或者完全停止其自身的产生(对于氨基酸生物合成途径反馈机制的评论,参见Stryer,L.Biochemistry 3rd ed.Ch.24“Biosynthesis of Amino Acid and Heme”p.575-600(1988))。因此,任何特定氨基酸的产量都被细胞内存在的氨基酸数量所限制。
B.维生素、辅因子和营养因子的代谢和用途
维生素、辅因子和营养因子包括另一组分子,虽然其他生物,例如细菌,可以合成这些分子,但是高等动物失去了合成它们的能力而只能摄取。这些分子或者其本身是生物活性物质,或者是生物活性物质的前体,该生物活性物质可以是电子载体或者各种代谢途径的中间体。除了其营养价值,这些化合物作为色素、抗氧化剂和催化剂或者其他加工助剂也有重大的工业价值。(对于这些化合物结构、活性和工业应用的评述,参见例如,Ullmann’s Encyclopedia of Industrial Chemistry,“Vitamins”vol.A27,p.443-613VCH:Weinheim 1996.)“维生素”这个词是本领域熟知的,包含了生物体正常功能所需但是又不能自身合成的营养素。维生素可以包括辅因子和营养因子化合物。术语“辅因子”包含了进行正常酶活性所需的非蛋白质化合物。这些化合物可以是无机的或者有机的;本发明的辅因子分子优选是有机的。“营养因子”这个词包含了对植物和动物,特别是人体有益的饮食增补剂。这些分子的实例是维生素、抗氧化剂和某些脂质(例如多饱和脂肪酸)。
在能够产生这些分子的生物体,例如细菌中这些分子的生物合成,大部分已经被鉴定(Ullman’s Encyclopedia of Industrial Chemistry,“Vitamins”vol.A27,p.443-613,VCH:Weinheim,1996;Michal,G.(1999)Biochemical Pathways:An Atlas of Biochemistry and Molecular Biology,John Wiley&Sons;Ong,A.S.,Niki,E.&Packer,L.(1995)“Nutrition,Lipids,Health,and Disease”Proceedings of the UNESCO/Confederation ofScientific and Technological Associations in Malaysia,and the Society forFree Radical Research-Asia,held Sept.1-3,1994at Penang,Malaysia,AOCS Press:Champaign,IL X,374S)
硫胺素(维生素B1)是由嘧啶和噻唑经化学连接产生的。核黄素(维生素B2)由5’-三磷酸鸟嘌呤核苷和5’-磷酸核糖合成。核黄素依次用于合成黄素单核苷酸(FMN)和黄素腺嘌呤二核苷酸(FAD)。合称为“维生素B6”的一组化合物(例如,吡哆醇、吡哆胺,5’-磷酸吡哆醛,以及商品化的盐酸吡哆醛)都是共同结构单元5-羟基-6-甲基吡啶的衍生物。泛酸盐(泛酸,(R)-(+)-N-(2,4-二羟基-3,3-二甲基-1-氧代丁基)-β-丙氨酸)可由化学合成或者发酵得到。泛酸盐生物合成的最后一步包括ATP驱动的β-丙氨酸和泛解酸的缩合。负责转化成泛解酸和β-丙氨酸酶,以及缩合成泛酸盐的酶都是已知的。泛酸盐的代谢活性形式是辅酶A,其生物合成过程是5个酶促步骤。泛酸盐、5’-磷酸吡哆醛、半胱氨酸和ATP是辅酶A的前体。这些酶不仅催化泛酸盐的形成,也催化(R)-泛解酸、(R)-pantolacton,(R)-泛醇(维生素原B5)泛酰巯基乙胺(及其衍生物)的产生。
在微生物中由前体分子庚二酰辅酶A到生物素的生物合成研究得很详细,并且所涉及的几个基因已被鉴定。很多相应的蛋白质也被发现参与了铁簇(Fe-cluster)的合成,并且是nifS家族蛋白质成员。硫辛酸来自辛酸,在能量代谢中用作辅酶,可以成为丙酮酸脱氢酶复合物和α-酮戊二酸脱氢酶复合物的一部分。叶酸盐是一组叶酸的衍生物,依次来自L-谷氨酸、对氨基苯甲酸和6-甲基蝶呤。起始于代谢中间体5’-三磷酸鸟嘌呤(GTP)、L-谷氨酸和对氨基苯甲酸的叶酸及其衍生物的生物合成,在某些微生物中有详细的研究。
类咕啉(例如钴胺素,以及特别是维生素B12)和卟啉都属于以四吡咯环体系为特征的化学物质。维生素B12的生物合成是这样的复杂,以至于还没有彻底了解其特征,但是许多涉及的酶和底物现在已知。烟酸(烟酸盐)和烟碱是吡啶底衍生物,也被称作“尼亚新”。尼亚新是重要辅酶NAD(烟酰胺腺嘌呤二核苷酸)和NADP(烟酰胺腺嘌呤二核苷酸磷酸)及其还原形式的前体。
尽管有些这样的化合物也可以用大规模微生物培养生产,例如核黄素、维生素B6、泛酸和生物素,但是大规模生产这些化合物很大程度还依赖于非细胞化学体系。只有维生素B12,由于其合成的复杂性,只能用发酵生产。体外方法需要相当多的物质和时间投入,经常花费很大。
C.嘌呤、嘧啶、核苷和核苷酸的代谢和用途
嘌呤和嘧啶代谢基因及其相应的蛋白质,是肿瘤疾病治疗和病毒感染治疗重要的目标物。术语“嘌呤”和“嘧啶”,包含了作为核酸、辅酶和核苷酸组成的含氮碱基。术语“核苷酸”包含核酸分子基本结构单元,核酸分子由含氮碱基、戊糖(对于RNA,该戊糖是核糖;对于DNA,该戊糖是脱氧核糖)和磷酸组成。术语“核苷”包含了作为核苷酸前体的分子,但是缺少核苷酸所具有的磷酸部分。通过抑制这些分子的生物合成,或者抑制为合成核酸分子而进行的移动,可能会抑制RNA和DNA的合成;通过定向肿瘤细胞的方式来抑制该活性,肿瘤细胞分裂和复制的能量可能会得到抑制。另外,有的核苷酸不用于形成核酸,而是用作能量储存(例如AMP)或者辅酶(例如FAD和NAD)。
有些出版物描述了通过影响嘌呤和/或嘧啶的代谢,这些化学物质作为这些医学指征的使用(例如,Christopherson,R.I.and Lyons,S.D.(1990)“Potent inhibitors of de novo pyrimidine and purine biosynthesis aschemotherapeutic agents.”Med.Res.Reviews 10:505-548)。涉及嘌呤和嘧啶代谢酶类的研究,集中在可以使用的新药开发上面,例如,作为免疫抑制剂或者抗增生剂(Smith,J.L.,(1995)“Enzyme in nucleotidesynthesis.”Curr.Opin.Struct.Biol.5:752-757;(1995)Biochem Soc.Transact.23:877-902)。然而,嘌呤和嘧啶碱基,核苷和核苷酸还具有另外的作用:作为许多精细化学物质生物合成的中间体(例如,硫胺素、S-腺苷甲硫氨酸、叶酸、或者核黄素),作为细胞能量载体(例如ATP或者GTP),而作为化学物质本身,通常用作风味增强剂(例如IMP或者GMP)或者几种医学应用(参见,例如,Kuninaka,A.(1996)Nucleotidesand Related Compounds in Biotechnology vol.6,Rehm et al.,eds.VCH:Weinheim,,p.561-612)。同样,涉及嘌呤、嘧啶、核苷或者核苷酸代谢的酶,日渐成为开发出的用作保护农作物的化学物质的作用目标,这些化学物质包括杀真菌剂、除草剂和杀虫剂。
细菌中这些化合物的代谢具有特征(评论参见,例如Zalkin,H.andDixon,J.E.(1992)“de novo purine nucleotide biosynthesis”,in:Progress in NucleicAcid Research and Molecular Biology,vol.42,Academic Press:,p.259-287;andMichal,G.(1999)“Nucleotides and Nucleosides”,Chapter 8in:BiochemicalPathways:An Atlas of Biochemistry and Molecular Biology,Wiley:New York)。嘌呤代谢一直是重点研究课题,而且它是细胞正常功能所必需的。高等动物中受损的嘌呤代谢能够造成严重的疾病,例如痛风。嘌呤核苷酸由5’-磷酸核糖合成,通过一系列步骤,经过中间体5’-磷酸次黄嘌呤核苷(IMP),最终产生5’-单磷酸鸟嘌呤(GMP)和5’-单磷酸腺嘌呤(AMP),并由它们形成用作核苷酸的三磷酸形式。这些化合物也用作能量储存,其降解为细胞中各种不同的生化过程提供能量。嘧啶的生物合成,是通过由5’-磷酸核糖形成5’-磷酸尿嘧啶核苷(UMP)。UMP接下来转变成5’-三磷酸胞嘧啶(CTP)。所有这些核苷酸的脱氧形式都是经过一步还原反应产生的,由核苷酸的二磷酸核糖形式到核苷酸的二磷酸脱氧核糖形式。一经磷酸化,这些分子就可以参与DNA的合成了。
D.海藻糖的代谢和用途
海藻糖包括两个葡萄糖分子,通过α,α-1,1连接。通常在食品产业中用作增甜剂、干燥食品或者冷冻食品添加剂,以及饮料当中。而且,它也应用在制药、化妆品和生物技术产业(参见,例如Nishimoto et al.,(1998)U.S.Patent No.5,759,610;Singer,M.A.and Lindquist,S.(1998)Trends Biotech.16:460-467;Paiva,C.L.A.and Panek,A.D.(1996)Biotech.Ann.Rev.2:293-314;and Shiosaka,M.(1997)J.Japan 172:97-102)。很多微生物中的酶可以产生海藻糖,并将其天然释放到周围培养基中,可以使用技术上熟知的方法从中进行收集。
II.本发明的元件和方法
本发明至少部分是建立在发现新分子的基础上的,此处将其称作MP核酸和蛋白质分子(见表1),它们在一种或多种细胞代谢途径中起作用或发挥功能。在一个实施方案中,MP分子催化一种或多种氨基酸如赖氨酸或甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应。在优选实施方案中,本发明的一种或多种谷氨酸棒杆菌氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的MP分子活性,单独或与相同或不同代谢途径(如甲硫氨酸或赖氨酸代谢)的分子组合,对用该微生物生产所需精细化学物质有影响。在一个特别优选的实施方案中,本发明MP分子的活性被调节,使得本发明的MP蛋白参与的代谢途径效率或产量被调节,这会直接或间接影响谷氨酸棒杆菌中一种或者多种精细化学物质的生产和生产效率。在优选实施方案中,精细化学物质是氨基酸如赖氨酸或甲硫氨酸。在另一个优选实施方案中,MP分子是metZ、metY和/或RXA00657(见表1)。
术语“MP蛋白”或者“MP多肽”包含了在一种或多种氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中起作用如催化其中的酶促反应的蛋白质。MP蛋白的实例包括那些由列在序列表中序列号为奇数的MP基因编码的蛋白质。术语“MP基因”或者“MP核酸序列”包含了编码MP蛋白的核酸序列,后者包含编码区域以及相应的非翻译的5’和3’序列区域。MP基因的实例包括那些列在表1中的基因。术语“生产”或者“生产力”是本领域熟知的,包含了在给定时间和给定发酵体积内,发酵产物(例如,所需精细化学物质)的浓度(例如,每小时每升千克产物)。术语“生产效率”包含了,要达到特定的生产水平所需的时间(例如,需要多长时间才能使细胞达到特定的精细化学物质)。术语“收益”“产物/碳收益”是本领域熟知的,包含了把碳源转化成产物(例如精细化学物质)的效率。例如,经常写作千克产物每千克碳源。通过提高化合物的收益或者生产,可增加回收分子的数量,或者增加在给定时间内给定数量的培养物中该化合物有用回收分子的数量。术语“生物合成”或者“生物合成途径”是本领域熟知的,包含了在细胞中,从中间化合物经过可能是多步并且是高度调控的过程,合成化合物,特别是有机化合物。术语“降解”或者一条“降解途径”是本领域熟知的,包含了在细胞中,经过可能是多步并且是高度调控的过程,把化合物,优选是有机化合物,分解为降解产物(一般而言,是更小或者复杂性更小的分子)。术语“代谢”是本领域熟知的,包含了生物体中所发生的生化反应的全部。因而,特殊化合物的代谢(例如,像是甘氨酸这样的氨基酸代谢)包括细胞中与该化合物相关的全部生物合成、修饰和降解途径。
本发明的MP分子可以与一种或多种本发明的MP分子或一种或多种相同或不同代谢途径的分子组合,以增加所需精细化学物质的产量。在优选实施方案中,精细化学物质是氨基酸如赖氨酸和甲硫氨酸。或者,此外,不需要的负产物通过MP分子或其他代谢分子(如参与赖氨酸或甲硫氨酸代谢的分子)的组合或破坏而得以减少。与相同或不同代谢途径的其他分子组合的MP分子可以改变器核苷酸序列和相应氨基酸序列,以改变其生理条件下的活性,从而增加所需精细化学物质的产量和/或产率。在再一个实施方案中,原始形式或如上所述改变形式的MP分子可以与相同或不同代谢途径的其他分子组合,这些分子的核苷酸序列发生了改变,导致所需精细化学物质如甲硫氨酸或赖氨酸等氨基酸的产量和/或产率增加。
在另一个实施方案中,本发明的MP分子单独或与相同或不同代谢添加中的一种或多种分子组合,能够调节微生物中,例如谷氨酸棒杆菌中,所需化合物例如精细化学物质的产生。使用重组遗传技术可以操作本发明的一种或者多种氨基酸如赖氨酸或甲硫氨酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖生物合成或降解酶,从而调节其活性。例如,可以提高生物合成酶的效率,或破坏其别构控制区从而防止化合物生产的反馈抑制。类似地,降解酶可以通过置换、缺失或增加被缺失或修饰,从而其对所需化合物的降解活性降低,而不影响细胞的活力。在各种情况下,所需精细化学物质的总产量或产率均被提高。
本发明的蛋白质和核苷酸分子的改变也可能提高除氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖以外的其它精细化学物质的生产。任一种化合物的代谢必然与细胞内其它生物合成和降解途径关联,一种途径中的必需辅因子、中间体或底物可能由其它同类途径供给或受其限制。因此,通过调整一种或多种本发明蛋白的活性,另一种精细化学物质生物合成或降解途径活性的生产或效率可能会受到影响。例如,氨基酸可以作为所有蛋白质的结构单元,但其在细胞内的存在的水平可能会限制蛋白合成;因此,通过增加细胞内一种或多种氨基酸的生产效率或产率,诸如生物合成或降解蛋白的蛋白可以更容易合成。同样,代谢途径酶的改变使得特定副反应更有利或不利时,会导致一种或多种用作生产所需精细化学物质的中间体或底物的化合物过量生产或生产不足。
本发明的分离核酸序列,包含在谷氨酸棒杆菌菌株的基因组中,该菌株可由美国典型培养物保藏中心获得,保藏号ATCC 13032。分离的谷氨酸棒杆菌MP DNA核酸序列,以及预测的谷氨酸棒杆菌MP蛋白氨基酸序列,在序列表中分别以奇数序列号和偶数序列号列出。进行了计算机分析,并将这些核酸序列分类和/或鉴定为编码代谢途径蛋白质的序列,如参与甲硫氨酸或赖氨酸代谢途径的蛋白。
本发明也与这样的蛋白质有关,该蛋白质的氨基酸序列与本发明的氨基酸序列有充分的同源性(例如,序列表中偶数序列号的序列)。如此处所用的那样,具有与挑选出的氨基酸序列有充分同源性的氨基酸序列的蛋白质,与挑选出的氨基酸序列,例如挑选出的氨基酸全序列,有至少大约50%同源性。具有与挑选出的氨基酸序列有很大同源性的氨基酸序列的蛋白质,也可以与挑选出的氨基酸序列有至少大约50%、51%、52%、53%、54%、55%、56%、57%、58%、59%或60%同源性,优选有至少大约61%、62%、63%、64%、65%、66%、67%、68%、69%或70%的同源性,更优选有至少大约71%,72%、73%、74%、75%、76%、77%、78%、79%或80%、81%、82%、83%、84%、85%、86%、87%、88%、89%或90%或91%、92%、93%、94%的同源性,甚至更优选的有至少大约95%,96%,97%,98%,99%、99.7%或者更高的同源性。
本发明的MP蛋白或者其生物活性部分或其片段,单独或与一种或多种相同或不同代谢途径的蛋白组合,能够催化一种或多种氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应,或者具有表1中列出的一种或者多种活性(如甲硫氨酸或赖氨酸生物合成代谢)。
以下各部分更加详细地描述了本发明的各个方面:
A.分离的核酸分子
本发明的一个方面涉及分离的编码MP多肽或者其生物活性部分的核酸分子,以及足够用作杂交探针或者引物的核酸分子片段,这些片段用于鉴定或者扩增编码MP的核酸(例如MP DNA)。如此处所用的那样,术语“核酸分子”的意思是包含DNA分子(例如,cDNA或者基因组DNA)和RNA分子(例如mRNA),以及由核苷酸类似物产生的DNA或者RNA类似物。该术语也包括位于基因编码区域3’和5’末端的非翻译序列:编码区域5’末端上游序列的至少100个核苷酸,和基因编码区域3’末端下游序列的至少20个核苷酸。核酸分子可以是单链的或者双链的,但是优选是双链DNA。“分离的”核酸分子,是指那些与存在于核酸天然来源中的其他核酸分子相互分离的核酸分子。优选,“分离的”核酸不含有天然位于生物体基因组DNA中核酸两侧的序列(例如,位于核酸5’和3’末端的序列),核酸就是从该生物体中获得的。例如,在各种实施方案中,分离的MP核酸分子可以含有少于大约5kb,4kb,3kb,2kb,1kb,0.5kb或者0.1kb的核苷酸序列,该序列天然位于细胞基因组DNA核酸分子的两侧,核酸就是从这些细胞(例如,谷氨酸棒杆菌细胞)中获得的。另外,“分离的”核酸分子,例如DNA分子,当用重组技术生产时可以基本上不含有其他细胞物质或者培养基,当化学合成时可以不含化学前体或者其他化学物质。
本发明核酸分子,例如序列表中奇数序列号的核苷酸序列,或者其部分,可以通过标准分子生物学技术和此处提供的序列信息分离得到。例如,谷氨酸棒杆菌MP DNA可以从谷氨酸棒杆菌文库中,使用序列表中奇数序列号序列中一个序列的全部或者其部分作为杂交探针,以及标准杂交技术(例如,像是描述在Sambrook,J.,Fritsh,E.F.,and Maniatis,T.Molecular Cloning:A Laboratory Manual.2nd,ed.Cold Spring HarborLaboratory,Cold Spring Harbor Laboratory Press,Cold Spring Harbor,NY,1989中的)分离得到。另外,包含一条本发明核酸序列(例如,序列表中奇数序列号的核苷酸序列)全部或者一部分的核酸分子,可以通过聚合酶链式反应,使用基于该序列设计的寡聚核苷酸引物,分离得到(例如,包含一条本发明核酸序列(例如,序列表中奇数序列号的核苷酸序列)全部或者一部分的核酸分子,可以通过聚合酶链式反应,使用基于该相同序列设计的寡聚核苷酸引物,分离得到)。例如,mRNA可以从正常内皮细胞分离得到(例如,使用Chirgwin et al.(1979)Biochemistry 18:5294-5299中的硫氰酸胍提取方法),DNA可以通过逆转录酶(例如,Gibco/BRL,Bethesda,MD提供的Moloney MLV逆转录酶;或者SeikagakuAmerica,Inc.,St.Peterburg,FL提供的AMV逆转录酶)制备。为聚合酶链式反应合成的寡聚核苷酸引物,可以基于序列表中列出的一条核苷酸序列设计。本发明的核酸,可以使用cDNA或者作为另一种选择的基因组DNA作模板,合适的寡聚核苷酸引物,根据标准PCR扩增技术来扩增。这样扩增出的核酸,可以克隆到合适的载体中,并用DNA序列分析辨别其特征。另外,与MP核苷酸序列相对应的寡聚核苷酸,可以用标准合成技术准备,例如使用自动DNA合成仪。
在一个优选的实施方案中,分离的本发明核酸分子包含序列表中列出的一条核苷酸序列。本发明的核酸序列,正如在序列表中列出的那些,与本发明的谷氨酸棒杆菌MP DNA是一致的。这些DNA包含编码MP蛋白的序列(即“编码区域”,显示在每条序列表中奇数序列号序列中),以及5’非编码序列和3’非编码序列,也显示在每条序列表中奇数序列号序列中。作为另一种选择,核酸分子可以只包含序列表中核酸序列的编码区域。
为了该申请的目的,可以理解序列表中列出的一些MP核酸和氨基酸序列,都有一个用于识别的RXA,RXN,RXS或者RXC编号,“RXA”,“RXN”,“RXS”,或者“RXC”后面有5个数字(即,RXA,RXN,RXS,或者RXC)。每条核酸序列最多包含三部分:5’上游区域,编码区域,下游区域。三个区域的每个部分,都用相同的RXA,RXN,RXS,或者RXC名称确定以消除混淆。于是叙述“序列表中的一条奇数编码的序列”,是指序列表中的任何核酸序列,这些序列也可以用它们不同的RXA,RXN,RXS,或者RXC名称相互区分。每条这种序列的编码区域都被翻译成相应的氨基酸序列,这些序列也列在序列表中,为紧随相应核酸序列之后偶数序列号。例如,RXA00115的编码区域列在SEQ ID NO:69,而它编码的氨基酸序列列在SEQ ID NO:70。本发明的核酸分子序列,与其编码的氨基酸分子,用相同的RXA,RXN,RXS,或者RXC名称表示,使得它们容易相互联系。例如,称为RXA00115,RXN00403和RXS03158的氨基酸序列,分别是RXA00115,RXN00403和RXS03158核酸分子核苷酸序列编码区域的翻译。本发明RXA,RXN,RXS和RXC核苷酸和氨基酸序列之间的对应,以及它们被指定的序列号列在表1中。
本发明的几个基因是“F-标明的基因”。F-标明的基因包括那些列在表1中并在RXA,RXN,RXS,或者RXC标明前有“F”的基因。例如,SEQ ID NO:77,像在表1中表示的那样,被指定为“F RXA00254”,就是一个F-标明的基因。
表1中列出的还有metZ(或metY)和metC基因(分别为SEQ ID NO:1和SEQ ID NO:3)。metZ和metC基因编码的相应氨基酸序列分别称为SEQ ID NO:2和SEQ ID NO:5。
在一个实施方案中,本发明的核酸分子不包含汇编在表2中的那些谷氨酸棒杆菌分子。
在另一个优选的实施方案中,分离的本发明的核酸分子,包含那些是本发明核苷酸序列(例如,序列表中奇数序列号序列)或者其部分的互补分子的核酸分子。与本发明一条核苷酸序列充分互补的核酸分子,是指该分子与序列表中列出的一条核苷酸序列(例如,奇数序列号序列)充分互补,因此它可以与本发明的一条核苷酸序列杂交,从而形成稳定的双螺旋。
同样在另一个优选的实施方案中,分离的本发明的核酸分子,包含这样的核苷酸序列,该序列与本发明的核苷酸序列(例如,序列表中奇数序列号序列)或者其部分,有至少大约50%,51%,52%,53%,54%,55%,56%,57%,58%,59%或者60%的同源性,优选的有至少大约61%,62%,63%,64%,65%,66%,67%,68%,69%或者70%的同源性,更优选的有至少大约71%,72%,73%,74%,75%,76%,77%,78%,79%或者80%,81%,82%,83%,84%,85%,86%,87%,88%,88%,89%或者90%,或者91%,92%,93%,94%,以及甚至更优选的有至少大约95%,96%,97%,98%,99%,99.7%或者更高的同源性。以上引用范围(例如,70-90%一致性或者80-95%一致性)中间的范围和一致性值,也包含在本发明中。例如,包含了这样的一致性值范围,这些范围是上面引用的上限和/或下限值的组合。在另一种优选的实施方案中,本发明分离的核酸分子包括这样的核苷酸序列,该序列可以与本发明的一条核苷酸序列或者其部分进行杂交,例如,在严格条件下杂交。
另外,本发明核酸分子可能只包含序列表中奇数序列号序列编码区域的一部分,例如,可以用作探针或者引物的片段,或者编码MP蛋白生物活性部分的片段。由谷氨酸棒杆菌MP基因克隆出的核苷酸序列,容许产生探针和引物,这些探针和引物的设计是用于鉴定和/或克隆其他细胞类型或者其他生物体中的MP同系物,以及其他棒杆菌或者亲缘物种中的MP同系物。探针/引物典型的包括相当纯化的寡聚核苷酸。寡聚核苷酸典型的包括这样一段核苷酸序列的区域,该区域在严格杂交条件下,与本发明核苷酸序列(例如,序列表中奇数序列号序列)的有义链,这些序列的反义序列,或者其天然存在的突变体的至少大约12个,优选的大约25个,更优选的大约40,50,或者75个连续核苷酸杂交。基于本发明核苷酸序列的引物,可以用于克隆MP同系物的PCR反应。基于MP核苷酸序列的探针,可以用于探测相同的或者同源蛋白的转录或者基因组序列。在一个优选的实施方案中,探针更是包括另外的附着标记基团,例如标记基团可以是放射性同位素、荧光化合物、酶或者酶的辅因子。这种探针可以用作诊断检测试剂盒的一部分,该试剂盒用于鉴定错误表达MP蛋白的细胞,像是通过测定样本细胞中MP编码核酸的水平,例如,检测MP mRNA的水平,或者测定基因组MP基因是否发生了突变或者缺失。
在一个实施方案中,本发明核酸分子编码一种蛋白质或者其部分,该蛋白质或者其部分的氨基酸序列与本发明的氨基酸序列(例如,序列表中偶数序列号序列)有充分的同源性,从而使得该蛋白质或者其部分可以催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应。如此处所用的那样,术语“充分的同源性”是指蛋白质或者其部分的氨基酸序列,含有最小数目的与本发明氨基酸序列一致的或者等价的(例如,具有与序列表偶数序列号序列中的氨基酸残基相似侧链的氨基酸残基)氨基酸残基,从而使得该蛋白质或者其部分,能够催化谷氨酸棒杆菌中氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应。这种代谢途径蛋白质成员,像这里描述的那样,其功能是催化一种或多种氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖的生物合成或降解。这里也描述了这种活性的实例。因而,“MP蛋白的功能”对于一种或多种这种代谢途径有作用,和/或直接或者间接影响一种或者多种精细化学物质的产量、生产和/或生产效率。MP蛋白活性的实例在表1中列出。
在另一个实施方案中,蛋白质与本发明的全部氨基酸序列(例如,序列表中偶数序列号序列)有至少大约50%,51%,52%,53%,54%,55%,56%,57%,58%,59%或者60%的同源性,优选的有至少大约61%,62%,63%,64%,65%,66%,67%,68%,69%或者70%的同源性,更优选的有至少大约71%,72%,73%,74%,75%,76%,77%,78%,79%或者80%,81%,82%,83%,84%,85%,86%,87%,88%,88%,89%或者90%,或者91%,92%,93%,94%,以及甚至更优选的有至少大约95%,96%,97%,98%,99%,99.7%或者更高的同源性。
本发明MP核酸分子编码蛋白质的部分,优选是MP蛋白的生物活性部分。如此处所用的那样,术语“MP蛋白的生物活性部分”的意思是包含MP蛋白这样的部分,例如结构域/基元,该部分能够催化谷氨酸棒杆菌中一种或多种氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应或具有表1中所列活性。可以进行一种酶活性分析,以确定MP蛋白或者其生物活性部分是否可以催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应。这种分析方法对于本领域技术人员来说是熟知的,在范例的实例8中有详细的描述。
编码MP蛋白生物活性部分的额外的核酸片段,可以通过以下方法制备,分离本发明氨基酸序列(例如,序列表中偶数序列号序列)的一部分,表达MP蛋白或者多肽的编码部分(例如,通过体外重组表达),并且估算MP蛋白或者多肽编码部分的活性。
因为遗传密码子的简并性,以及由此可以编码得到和本发明核苷酸序列编码蛋白质相同的MP蛋白,所以本发明进一步包含不同于本发明核苷酸序列(例如,序列表中奇数序列号序列)(和其部分)的核酸分子。在另一个实施方案中,分离的本发明的核酸分子具有这样的核苷酸序列,该序列编码具有序列表中列出的氨基酸序列(例如,偶数序列号)的蛋白质。同样在另一个实施方案中,本发明核酸分子编码全长的谷氨酸棒杆菌蛋白质,该蛋白质与本发明的氨基酸序列(由序列表中奇数序列号开放阅读框架编码)有充分的同源性。
在一个实施方案中,本发明的序列并不意味着包括以前技术上已知的序列,例如那些列在表2中的在本发明以前就可获得的Genbank序列,这对于本领域技术人员来说是可以理解的。在一个实施方案中,本发明包含这样的核苷酸序列和氨基酸序列,该序列与本发明的核苷酸序列和氨基酸序列有一定百分比的一致性,该百分比大于技术上已知的序列(例如,表2列出的Genbank序列(或者该序列编码的蛋白质))与本发明的核苷酸序列和氨基酸序列一致性的百分比。例如,本发明包含与标明为RXA00657(SEQ ID NO:5)的核苷酸序列有大于和/或至少45%一致性的核苷酸序列。本领域技术人员,通过检查表4中列出的对于每个特定序列给出的3个最高符合的GPA-计算百分比一致性,以及经过从百分之一百中减去最高的GPA-计算百分比一致性,可以计算任何本发明特定序列百分比一致性的低端域值。本领域技术人员也可以意识到,其百分比一致性大于如此计算出的低端域值(例如,至少约50%,51%,52%,53%,54%,55%,56%,57%,58%,59%或者60%,优选的至少大约61%,62%,63%,64%,65%,66%,67%,68%,69%或者70%,更优选的至少大约71%,72%,73%,74%,75%,76%,77%,78%,79%或者80%,81%,82%,83%,84%,85%,86%,87%,88%,88%,89%或者90%,或者91%,92%,93%,94%,以及甚至更优选的至少大约95%,96%,97%,98%,99%或者更高的一致性)的核酸和氨基酸序列,也是包含在本发明中的。
本领域技术人员可以意识到,除了在序列表中以奇数序列号列出的谷氨酸棒杆菌MP核苷酸序列之外,导致MP蛋白氨基酸序列改变的DNA多态性可以在一定群体(例如谷氨酸棒杆菌群体)中存在。这种MP基因的遗传多态性,可以由于自然条件的变异而在一个群体的不同个体中存在。如此处所用的那样,术语“基因”和“重组基因”是指含有编码MP蛋白的开放阅读框架的核酸分子,优选的MP蛋白是谷氨酸棒杆菌MP蛋白。这种自然条件的变异典型的可以造成MP基因核苷酸序列1-5%的变化。任何以及全部由于自然条件的变异造成的,并且不改变MP蛋白功能活性的,这种核苷酸的变化,以及引起的MP氨基酸的多态性,都属于本发明范围之内。
相应天然变体的核酸分子,和本发明谷氨酸棒杆菌MP DNA的非谷氨酸棒杆菌同源物,可以基于此处公开的它们与谷氨酸棒杆菌MP核酸分子的同源性,使用谷氨酸棒杆菌DNA或者其部分作为杂交探针,在严格杂交条件下根据标准杂交技术分离得到。因此,在另一个实施方案中,分离的本发明核酸分子的长度至少有15个核苷酸,在严格条件下与含有序列表奇数序列号核苷酸序列的核酸分子杂交。在其他实施方案中,核酸分子的长度至少有30,50,100,250或者更多个核苷酸。如此处所用的那样,术语“在严格条件下杂交”的意思是描述这样的杂交和清洗的条件,在该条件下彼此之间有至少60%同源性的核苷酸序列相互之间保持典型的杂交。优选,这种条件是序列之间有至少大约65%,更优选的有至少大约70%,以及甚至更优选的有至少大约75或者更高的同源性,相互之间保持典型的杂交。这种严格条件对于本领域技术人员是已知的,可以在Ausubel et al.,Current Protocols in Molecular Biology,JohnWiley&Sons,N.Y.(1989),6.3.1-6.3.6中找到。一种优选的但不是限制的严格杂交条件是,在6X氯化钠/柠檬酸钠(SSC)中大约45℃进行杂交,然后用0.2X SSC,0.1%SDS在50-65℃清洗一次或者多次。优选,分离的本发明的核酸分子,在严格杂交条件下与本发明的核苷酸序列杂交,相当于得到天然存在的核酸分子。如此处所用的那样,“天然存在的”核酸分子是指具有自然中存在的核苷酸序列(例如,编码天然蛋白质)的RNA或者DNA分子。在一个实施方案中,核酸编码天然谷氨酸棒杆菌MP蛋白。
本领域技术人员可以进一步意识到,除了群体中存在的天然存在的MP序列变体以外,可以通过突变把改变引入本发明核苷酸序列中,从而导致被编码MP蛋白的氨基酸序列的改变,而不改变MP蛋白的功能。例如,可以在本发明核苷酸序列中,进行可以导致“非必需”氨基酸残基的氨基酸取代的核苷酸取代。“非必需”氨基酸残基是指这样的残基,该残基可以在MP蛋白的野生型序列(例如,序列表中偶数序列号序列)中发生改变,而不改变MP蛋白的活性,而“必需”氨基酸残基是MP蛋白活性所必需的。然而,其他氨基酸残基(例如,那些在MP活性结构域中非保守的或者只是半保守的氨基酸残基)可能对于活性不是必需的,因此也可以在不改变MP活性的情况下被改变。
因此,本发明的另一个方面涉及编码这样的MP蛋白的核酸分子,该MP蛋白含有对MP活性非必需的氨基酸残基的变化。这些蛋白质的氨基酸序列不同于序列表中偶数序列号序列,但仍然保持至少一种此处描述的MP活性。在一个实施方案中,分离的核酸分子包含一段编码蛋白质的核苷酸序列,其中该蛋白质的氨基酸序列与本发明的氨基酸序列有至少大约50%的同源性,并且能够催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应,或者具有表1中列出的一种或者多种活性。优选,核酸分子编码的蛋白质与本发明的氨基酸序列,有至少大约50%,51%,52%,53%,54%,55%,56%,57%,58%,59%或者60%的同源性,优选的有至少大约61%,62%,63%,64%,65%,66%,67%,68%,69%或者70%的同源性,更优选的有至少大约71%,72%,73%,74%,75%,76%,77%,78%,79%或者80%,81%,82%,83%,84%,85%,86%,87%,88%,88%,89%或者90%,或者91%,92%,93%,94%,以及甚至更优选的有至少大约95%,96%,97%,98%,99%,99.7%或者更高的同源性。。
为了确定两种氨基酸序列(例如,本发明的一种氨基酸序列与其突变体形式)或者两种核酸序列的同源性百分比,出于最适宜比较的目的,对序列进行序列对比(例如,为了与其他蛋白质或者核酸进行最适宜的序列对比,可以在一种蛋白质或者核酸的序列中引入间隙)。然后比较相应氨基酸位置的氨基酸残基或者核酸位置的核苷酸。当一条序列(例如,本发明的一条氨基酸序列)中的一个位置被与其他序列(例如,氨基酸序列的突变体形式)相应位置相同的氨基酸残基或者核苷酸占据时,该分子在这个位置是同源的(即,如此处所用的氨基酸或者核酸“同源性”与氨基酸或者核酸的“一致性”是相同的)。两条序列之间的百分比同源性,是一个相同位置数目被序列均分的函数(即,%一致性=相同位置的#/全部位置的#x 100)。
分离的与本发明蛋白质序列(例如,序列表中偶数序列号序列)同源的编码MP蛋白质的核酸分子,可以通过向本发明核苷酸序列中引入一个或者多个核苷酸取代、插入、缺失而产生,从而在编码蛋白质中引入一个或者多个氨基酸取代、插入、缺失。可以使用标准技术,例如定点诱变和PCR介导的诱变,在本发明核苷酸序列中引入突变。优选,保守的氨基酸取代是在一个或者多个预期的非必需氨基酸残基进行的。“保守的氨基酸取代”是指氨基酸残基被具有相似侧链的氨基酸残基所取代。具有相似侧链的氨基酸残基家族,在技术上有规定。这些家族包括,具有碱性侧链的氨基酸(例如,赖氨酸、精氨酸、组氨酸),具有酸性侧链的氨基酸(例如,天冬氨酸、谷氨酸),具有无电荷极性侧链的氨基酸(例如,甘氨酸、天冬氨酸、谷氨酰胺、丝氨酸、苏氨酸、酪氨酸、半胱氨酸),具有非极性侧链的氨基酸(例如,丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸、色氨酸),具有β-支链侧链的氨基酸(例如,苏氨酸、缬氨酸、异亮氨酸),以及具有芳香组侧链的氨基酸(例如,酪氨酸、苯丙氨酸、色氨酸、组氨酸)。因此,预期的MP蛋白中的非必需氨基酸残基,优选的被同一侧链家族中的其他氨基酸取代。另外,在另一个实施方案中,可以在MP编码序列全长或者部分,随机的引入突变,例如通过饱和诱变,根据此处描述的鉴定具有MP活性突变体的MP活性,筛选出得到的突变体。在一条序列表中奇数序列号核苷酸序列诱变之后,被编码蛋白质可以重组表达,蛋白质活性也可以,例如使用此处描述的分析(参见范例的实例8),得到确定。
除了以上描述的编码MP蛋白质的核酸分子以外,本发明的另一方面还与分离的反义核酸分子有关。“反义”核酸包括与编码蛋白质的“有义”核酸互补的核苷酸序列,例如与双链DNA分子编码链互补,或者与mRNA序列互补。因此,反义核酸可以通过氢键与有义核酸连接。反义核酸可以与全部MP编码链互补,也可以仅与其部分互补。在一个实施方案中,反义核酸分子,与编码MP蛋白的核苷酸序列编码链的“编码区域”反义。术语“编码区域”是指包含翻译成氨基酸残基的密码子的核苷酸序列区域(例如,SEQ ID NO.1(metZ)的全部编码区域包括363至1673核苷酸)。在另一个实施方案中,反义核酸分子,与编码MP的核苷酸序列编码链的反义。术语“非编码区域”是指编码区域两侧不翻译成氨基酸的5’和3’序列(即5’和3’非翻译区域)。
考虑到此处公布的编码MP的编码链序列(例如,序列表中列出的奇数序列号序列),本发明反义核酸可以根据Watson和Crick的碱基配对规则进行设计。反义核酸分子可以与MP mRNA的全部编码区域互补,但是更优选是这样的寡聚核苷酸,该寡聚核苷酸仅与MP mRNA的编码区域或者非编码区域的部分是反义的。例如,反义寡聚核苷酸可以与MP mRNA翻译起始位置附近的区域互补。例如,反义寡聚核苷酸的长度可以是5,10,15,20,25,30,35,40,45或者50个核苷酸。本发明的反义核酸分子,可以使用技术上已知的程序,通过化学合成或者酶促连接反应构建。可以使用天然存在的核苷酸或者各种经过修饰的核苷酸,化学合成反义核酸(例如反义寡聚核苷酸),那些经过修饰的核苷酸,是为了增加分子的生物稳定性,或者为了增加反义核酸与有义核酸之间形成双螺旋的物理稳定性而设计的,例如可以使用硫代磷酸衍生物和吖啶取代的核苷酸。可用于产生反义核酸的经修饰的核苷酸的实例包括,5-氟尿嘧啶,5-溴尿嘧啶,5-氯尿嘧啶,5-碘尿嘧啶,次黄嘌呤,黄嘌呤,4-乙酰胞嘧啶,5-(羧基羟基甲基)尿嘧啶,5-羧甲基氨基甲基-2-硫尿嘧啶,5-羧甲基氨基甲基尿嘧啶,二氢尿嘧啶,beta-D-半乳糖基肌苷,N6-异戊基腺嘌呤,1-甲基鸟嘌呤,1-甲基肌苷,2,2-二甲基鸟嘌呤,2-甲基腺嘌呤,2-甲基鸟嘌呤,3-甲基胞嘧啶,5-甲基胞嘧啶,N6-腺嘌呤,7-甲基鸟嘌呤,5-甲基氨基甲基尿嘧啶,5-甲氧基氨基甲基尿嘧啶-2-硫代尿嘧啶,beta-D-甘露糖基queosine,5’-甲氧基羧基甲基尿嘧啶,5-甲氧基尿嘧啶,2-甲基硫代-N6-异戊基腺嘌呤,尿嘧啶-5-含氧乙酸(v),wybutoxosine,假尿嘧啶,queosine,2-硫代胞嘧啶,5-甲基-2-硫代尿嘧啶,2-硫代尿嘧啶,4-硫代尿嘧啶,5-甲基尿嘧啶,尿嘧啶-5-含氧乙酸甲酯,尿嘧啶-5-含氧乙酸(v),5-甲基-2-硫代尿嘧啶,3-(3-氨基-3-N-2-羧基丙基)尿嘧啶,(acp3)w,以及2,6-二氨基嘌呤。另外,反义核酸可以使用表达载体生物合成,其中核酸被反义方向亚克隆到表达载体中(即,由插入核酸转录的RNA,相对于插入的目的核酸是反义方向的,以下部分有进一步叙述)。
本发明的反义核酸分子,被典型的施用于细胞或者在原位产生,从而它们可以与编码MP蛋白的细胞mRNA和/或基因组DNA杂交或者结合,进而抑制蛋白质的表达,例如,抑制转录和/或翻译。杂交可以通过常规核苷酸互补性而形成稳定的双螺旋,或者,例如,当反义核酸分子结合DNA双螺旋时,它与双螺旋的大沟发生特殊相互作用。反义分子可以被修饰,从而使得该分子可以与受体或者与特定细胞表面表达的抗原特异性结合,例如,反义核酸分子与多肽或者抗体的结合,该抗体与细胞表面受体或者抗原结合。反义核酸分子也可以使用此处描述的载体递送至细胞。为了得到细胞内足够浓度的反义分子,这样的载体是优选,即在该载体中,反义核酸分子被置于原核、病毒或者真核启动子的控制之下。
而在另一个实施方案中,本发明的反义核酸分子是一种α-异头物核酸分子。α-异头物核酸分子与互补的RNA形成特异的双链杂交体,杂交体中两股链走向彼此平行,这与通常的β-单元相反(Gaultier et al.(1987)Nucleic Acids.Res.15:6625-6641)。反义核酸分子也可以包含2’-o-甲基核糖核苷酸(Inoue et al.(1987)NucleicAcids.Res.15:6131-3148)或者化学RNA-DNA类似物(Inoue et al.(1987)FEBS Lett.215:327-330)。
而在另一个实施方案中,本发明的反义核酸分子是核酶。核酶是催化型RNA分子,具有核糖核酸酶活性,能够切割单链核酸,例如mRNA,它具有与单链核酸互补的区域。因此,核酶(例如,锤头核酶(描述于Haselhoffand Gerlach(1988)Nature 334:585-591))可以用于催化切割MP mRNA转录物,从而抑制MP mRNA的翻译。对于MP编码核酸分子有特异性的核酶,可以基于此处公布的MP DNA核苷酸序列(即SEQ IDNO:1(metZ))来设计。例如,可以构建四膜虫属L-19IVS RNA的衍生物,其活性位点的核苷酸序列与被切割的MP-编码mRNA的核苷酸序列是互补的。参见,例如,Cech et al.U.S.Patent No.4,987,071和Cech et al.U.S.PatentNo.5,116,742。另外,MP mRNA可以用于RNA分子库中筛选具有特异核酶活性的催化型RNA。参见,例如,Bartel,D.and Szostak,J.W.(1993)Science 261:1411-1418。
另外,通过把与MP核苷酸序列调节区域(例如,MP启动子和/或增强子)互补的核苷酸序列作为目标,形成三螺旋结构,可以抑制MP基因的表达,该三螺旋结构可以阻止MP基因在目的细胞中的转录。一般参见,Helene,C.(1991)Anticancer Drug Des.6(6):569-84;Helene,C.etal.(1992)Ann.N.Y.Acad.Sci.660:27-36;and Maher,L.J.(1992)Bioassays 14(12):807-15。
本发明另一方面涉及参与甲硫氨酸和/或赖氨酸代谢的基因组合和该基因组合在本发明方法中的用途。优选的组合是metZ和metC、metB(编码胱硫醚合酶)、metA(编码高丝氨酸O乙酰转移酶)、metE(编码甲硫氨酸合酶)、metH(编码甲硫氨酸合酶)、hom(编码高丝氨酸脱氢酶)、asd(编码天冬氨酸半醛脱氢酶)、lysC/ask(编码天冬氨酸激酶)和rxa00657(此处称为SEQ ID NO:5)、dapA(编码二氢吡啶二羧酸合成酶)、dpaB(编码二氢吡啶二羧酸还原酶的基因)、dapC(编码2,3,4,5-四氢吡啶-2-羧化物N-琥珀酰转移酶的基因),dapD/argD(编码乙酰鸟氨酸转氨酶的基因),dapE(编码琥珀酰二氨基庚二酸脱琥珀酰酶的基因),dapF(编码二氨基庚二酸差向异构酶的基因),lysA(编码二氨基庚二酸脱羧酶的基因),ddh(编码二氨基庚二酸脱氢酶的基因),lysE(赖氨酸外泌蛋白的基因),lysG(编码外泌调节物的基因),hsk(编码高丝氨酸激酶的基因)以及参与添补反应的基因如ppc(编码磷酸烯醇丙酮酸羧激酶的基因),ppcK(编码磷酸烯醇丙酮酸羧激酶的基因),pycA(编码丙酮酸羧化酶的基因),accD,accA,accB,accC(编码乙酰辅酶A羧化酶的基因),以及戊糖磷酸途径的基因,编码葡萄糖-6-磷酸脱氢酶的gpdh基因,opcA,pgdh(编码6-磷酸葡萄糖酸脱氢酶的基因),ta(编码转醛醇酶的基因),tk(编码转酮醇酶的基因),pgl(编码6-磷酸葡萄糖酸内酯酶的基因),rlpe(编码核酮糖磷酸3-差向异构酶的基因)、rpe(编码核酮糖磷酸5-差向异构酶的基因)的组合或上述戊糖磷酸途径基因或其他本发明MP基因的组合。
所述基因可以改变其核苷酸序列和相应氨基酸序列,产生在生理条件下活性发生改变的衍生物,从而增加所需精细化学物质如甲硫氨酸和赖氨酸等氨基酸的产量和/或产率。编码天冬氨酸激酶的ask基因的核苷酸序列的改变或衍生物是熟知的。这些改变导致赖氨酸和苏氨酸的反馈抑制消除,从而消除了对赖氨酸过量产生的抑制。在优选的实施方案中,metZ基因或metZ基因的改变形式与ask、hom、metA和metH或这些基因的衍生物组合用于棒杆菌中。在另一个优选实施方案中,metZ基因或metZ基因的改变形式的组合与ask、hom、metA和metE或这些基因的衍生物组合,或者metZ与ask、hom、metA和metE或这些基因的衍生物组合用于棒杆菌中,并且硫源如硫酸盐、硫代硫酸盐、亚硫酸盐和更加还原的硫源如硫化氢和硫醚和衍生物用于培养基中。硫源如甲基硫醇、甲烷磺酸、硫代羟乙酸、硫代氰酸盐、硫脲、含硫氨基酸如半胱氨酸和其他含硫化合物也可使用。本发明另一方面涉及上述基因组合在棒杆菌中的用途,所述菌株在导入所述基因之前或之后,通过辐射或本领域技术人员熟知的诱变剂进行诱变,并对高浓度目标精细化学物质如赖氨酸或甲硫氨酸或所需精细化学物质的类似物如甲硫氨酸类似物乙硫氨酸、甲基甲硫氨酸等进行选择。在另一个实施方案中,上述基因组合可以在具有特定基因破坏的棒杆菌菌株中表达。优选的基因破坏是是编码促使碳向不需要的代谢物流动的蛋白质。在甲硫氨酸是所需精细化学物质时,形成赖氨酸就是不利的。在这种情况下,上述基因的组合应当在lysA基因(编码二氨基庚二酸脱羧酶)或ddh基因(编码催化四氢吡啶二羧酸向内消旋二氨基庚二酸转变的内消旋二氨基庚二酸脱氢酶)被破坏的棒杆菌菌株中进行。在优选的实施方案中,上述基因的有利组合被改变使得其基因产物不受终产物或形成所需精细化学物质的生物合成途径的代谢物反馈抑制。在所需精细化学物质是甲硫氨酸时,该基因组合可以在已经用诱变剂或辐射处理的菌株中表达,并对上述抗性选择。此外,该菌株应当生长在含有一种或多种上述硫源的培养基中。
在另一个优选实施方案中,从谷氨酸棒杆菌基因组中鉴定到一个推定的转录调控蛋白。该基因被称为RXA00657。RXA00657的核苷酸序列为SEQ ID NO:5。RXA00657的氨基酸序列为SEQ ID NO:6。当RXA00657以及实施例中所述的上游和下游调控区被克隆到能够在谷氨酸棒杆菌中复制的载体中并转化从而在产赖氨酸菌株如ATCC13286中表达时,该菌株相对于用缺少上述RXA00657核苷酸片段的相同质粒转化的菌株产生更多的赖氨酸。除了观察到上述菌株中赖氨酸效价增加外,由产生的赖氨酸的摩尔数与消耗的蔗糖摩尔数比较确定的选择性也增加了(见实施例14)。RXA00657的过量表达与其他直接参与赖氨酸特异性途径的基因如lysC,dapA,dapB,dapC,dapD,dapF,ddh,LysE,LysG和lysR的过量表达组合产生的赖氨酸较仅有RXA00657时增加了。
B.重组表达载体和宿主细胞
本发明的另一方面涉及载体,优选是含有编码MP蛋白(或者其部分)的核酸或有至少一个编码MP蛋白的基因的基因组合的表达载体。如此处所用的那样,术语“载体”是指能够连接其他核酸,并对其进行运输的核酸分子。一种类型的载体是“质粒”,质粒是指环形双链DNA环,其中连接有额外的DNA片段。另一种类型的载体是病毒载体,其中额外的DNA片段可以连接到病毒基因组中。某些载体可以在它们被引入的宿主细胞中进行自主复制(例如,具有细菌复制起点的细菌载体,以及附加型哺乳动物载体)。其他的载体(例如,非附加型哺乳动物载体)一经引入宿主细胞就会整合到宿主细胞的基因组中,从而与宿主基因组一同复制。另外,某些载体能够指导与之相连接的基因的表达。这些载体此处称作“表达载体”。总之,重组DNA技术使用的表达载体经常是质粒形式。在本说明中,“质粒”和“载体”可以交换使用,因为质粒是最常使用的载体形式。然而,本发明有意包括这些表达载体的其他形式,例如病毒载体(例如,复制缺陷型逆转录病毒,腺病毒和腺伴随病毒),它们具有相同的功能。
本发明重组表达载体包括本发明的核酸,该核酸在宿主细胞中以适合核酸表达的形式存在,这就意味着重组表达载体含有一条或者多条调节序列,这些序列是基于用作表达的宿主细胞选出的,它们被可行的连接到要表达的核酸序列上。在重组表达载体中,“可行的连接”的意思是指,感兴趣核苷酸序列与调节序列以允许核苷酸序列表达的方式进行连接(例如,在体外转录/翻译系统中,或者在载体被引入的宿主细胞中)。术语“调节序列”的意思是包括启动子、增强子和其他表达控制元素(例如,聚腺苷酸化信号)。这种调节序列在,例如,Goeddel;GeneExpression Technology:Methods in Enzymology 185,Academic Press,SanDiego,CA(1990)中有描述。调节序列包括,那些在很多类型宿主细胞中,指导核苷酸序列组成型表达的序列,以及那些在某些宿主细胞中,指导核苷酸序列表达的序列。优选的调节序列是,例如,像是cos-,tac-,trp-,tet-,trp-tet-,lpp-,lac-,lpp-lac-,lacIq-,T7-,T5-,T3-,gal-,trc-,ara-,SP6-,arny-,SPO2-,λ-PR-或者λPL这样的启动子,这些启动子优选的使用在细菌中。另外的调节序列是,例如,酵母和真菌的启动子,例如ADCl,MFα,AC,P-60,CYCl,GAPDH,TEF,rp28,ADH,植物的启动子,例如,CaMV/35S,SSU,OCS,lib4,usp,STLS1,B33,nos或者ubiquitin-或phaseolin-启动子。也可以使用人造启动子。这些对于本领域技术人员是可以意识到的,即表达载体的设计依赖于这些因素:用于转化的宿主细胞的选择,所需蛋白质的表达水平等。本发明的表达载体可以引入宿主细胞,从而产生此处描述的核酸所编码的蛋白质或者多肽,包括融合蛋白质或者多肽(例如,MP蛋白、MP蛋白的突变形式、融合蛋白质等)。
可以设计本发明的重组表达载体,用于在原核或者真核细胞中表达MP蛋白。例如,MP基因可以在以下细胞中表达,像是谷氨酸棒杆菌这样的细菌细胞,昆虫细胞(使用杆状病毒表达载体),酵母和其他真菌细胞(参见Romanos,M.A.et al.(1992)“Foreign gene expression in yeast:areview”,Yeast 8:423-488;van den Hondel,C.A.M.J.J.et al.(1991)“Heterologous gene expression in filamentous fungi”in:More GeneManipulations in Fungi,J.W.Bennet&L.L.Lasure,eds.,p.396-428:Academic Press:San Diego;and van den Hondel,C.A.M.J.J.&Punt,P.J.(1991)“Gene transfer systems and vector development for filamentous fungi,in:Applied Molecular Genetics ofFungi,Peberdy,J.F.et al.,eds.,p.1-28,Cambridge University Press:Cambridge)藻类或者多细胞植物细胞(参见Schmidt,R.and Willmitzer,L.(1998)High efficiency Agrobacteriumtumefaciens-mediated transformation of Arabidopsis thaliana leaf andcotyledon explants”Plant Cell Rep.:583-586),或者哺乳动物细胞。适当的宿主细胞在Goeddel,Gene Expression Technology:Methods inEnzymology 185,Academic Press,San Diego,CA(1990)中有进一步论述。另外,重组表达载体可以在体外转录和翻译,例如使用T7启动子调节序列和T7聚合酶。
原核细胞中的蛋白质表达,经常是由含有组成型或者诱导型启动子的载体进行的,这些启动子指导融合蛋白质或者非融合蛋白质的表达。融合载体在编码蛋白质上添加一定数目的氨基酸,通常是在重组蛋白质的氨基末端,但也可以在C末端,或与蛋白中的合适区域融合。这种融合载体具有3个典型用途:1)增加重组蛋白质的表达;2)增加重组蛋白质的溶解性;和3)用作亲和纯化的配基,帮助融合蛋白纯化。在融合表达载体中,蛋白质切割位点经常是被引入到融合部分与重组蛋白质的结合处,使得在纯化出融合蛋白质之后,能够把重组蛋白质与融合部分分离开。这种酶,以及它们的同源识别序列,包括Xa因子、凝血酶和肠激酶。
典型的融合表达载体包括pGEX(Pharmacia Biotech Inc;Smith,D.B.and Johnson,K.S.(1988)Gene 67:31-40),pMAL(New England Biolabs,Beverly,MA)和pRIT5(Pharmacia,Piscataway,NJ),它们分别与目标重组蛋白融合了谷光甘肽S-转移酶(GST),麦芽糖E结合蛋白,或者蛋白质A。在一个实施方案中,MP蛋白编码序列被克隆到pGEX表达载体中,产生一个编码融合蛋白的载体,该载体从N-末端到C-末端包括,GST-凝血酶切割位点-X蛋白质。融合蛋白可以使用谷光甘肽-琼脂糖树脂,通过亲和层析纯化。与GST分离开的重组MP蛋白,可以通过用凝血酶切割融合蛋白质得到。
合适的大肠杆菌诱导型非融合表达载体的实例包括,pTrc(Amann etal.,(1988)Gene 69:301-315),pLG338,pACYC184,pBR322,pUC18,pUC19,pKC30,pRep4,pSH1,pSH2,pPLc236,pMBL24,pLG200,pUR290,pIN-III 113-B1,λgt11,pBdC1,和pET 11d(Studier et al.,Gene ExpressionTechnology:Methods in Enzymology 185,Academic Press,San Diego,California(1990)60-89;and Pouwels et al.,eds.(1985)Cloning Vectors.Elsevier:New York IBSN 0444904018)。pTrc载体的目标基因表达,依赖于杂交trp-lac融合启动子的宿主RNA聚合酶的转录。pET 11d载体的目标基因表达,依赖于共表达的病毒RNA聚合酶(T7gn1)介导的T7gn10-1ac融合启动子的转录。该病毒聚合酶由宿主菌株BL21(DE3)或者HMS 174(DE3)中驻留的λ噬菌体提供,该噬菌体含有在lacUV 5启动子转录控制下的T7gn1基因。对于其他种类细菌的转化,可以选择合适的载体。例如,已知质粒pIJ101,pIJ364,pIJ702和pIJ361转化链霉菌是有效的,而质粒pUB110,pC194,或者pBD214适合转化杆状菌种。有助于把遗传信息转入棒状杆菌的几种质粒包括pHM1519,pBL1,pSA77或者pAJ667(Pouwels et al.,eds.(1985)Cloning Vectors,Elsevier:NewYork IBSN 0444904018)。
一种最大限度增大重组蛋白表达的方案是,在宿主细胞中表达这样的蛋白质,该蛋白质具有不会减弱的蛋白酶剪切重组蛋白质的能力(Gottesman,S.,Gene Expression Technology:Methods in Enzymology 185,Academic Press,San Diego,California(1990)119-128)。另一种方案是改变插入表达载体中核酸的核酸序列,使得每个氨基酸的密码子都是所选用于表达的细菌优先使用的,例如谷氨酸棒杆菌(Wada et al.(1992)Nucleic Acids Res.20:2111-2118)。本发明核酸序列的这种改变,是可以通过标准DNA合成技术进行的。
在另一个实施方案中,MP蛋白表达载体是酵母表达载体。酵母S.cerivisae用于表达的载体的实例包括,pYepSec1(Baldari,et al.,(1987)Embo J.6:229-234),2μ,pAG-1,Yep6,Yep13,pEMBK Ye23,pMFa(Kurjan and Herskowitz,(1982)Cell 30:933-943),pJRY88(Schultz et al.,(1987)Gene 54:113-123),和pYES2(Invitrogen Corporation,San Diego,CA)。用于构建适合在其他真菌中,例如丝状真菌中,使用的载体的载体和方法,包括那些详述于下列文献中的:van den Hondel,C.A.M.J.J.&Punt,P.J.(1991)“Gene transfer systems and vector development forfilamentous fungi,in:Applied Molecular Genetics of Fungi,J.F.Peberdy,etal.,eds.,p.1-28,Cambridge University Press:Cambridge,and Pouwels et al.,eds.(1985)Cloning Vectors,Elsevier:New York IBSN 0444904018)。
另外,本发明MP蛋白可以使用杆状病毒表达载体在昆虫细胞中表达。在培养的昆虫细胞(例如Sf9细胞)中,用于表达蛋白质的杆状病毒载体包括,pAC系列(Smith et al.(1983)Mol.Cell Biol.3:2156-2165)和pVL系列(Lucklow and Summer(1989)Virology 170:31-39)。
在另一个实施方案中,本发明MP蛋白可以在单细胞植物细胞(例如藻类)中表达,或者在高等植物(例如种子植物,像是作物植物)的植物细胞中表达。植物表达载体的实例包括那些详述于下列文献中的:Becker,D.,Kemper,E.,Schell,J.and Masterson,R.(1992)″New plantbinary vectors with selectable markers located proximal to the left borde″Plant Mol.Biol.20:1195-1197;和Bevan,M.W.(1984)″BinaryAgrobacterium vectors for plant transformation”,Nucl.Acid.Res.12:8711-8721,包括pLGV23,pGHlac+,pBIN19,pAK2004和pDH51(Pouwels et al.,eds.(1985)Cloning Vectors.Elsevier:New York IBSN 0444904018)。
也是在另一个实施方案中,本发明核酸使用哺乳动物表达载体在哺乳动物细胞中表达。哺乳动物表达载体的实例包括pCDM8(Seed,B.(1987)Nature 329:840)和pMT2PC(Kaufman et al.(1987)EMBO J.6:187-195)。表达载体的控制功能,当时用在哺乳动物中时,经常是由病毒调节元素提供的。例如,通常使用的启动子来自多形瘤、腺病毒2、巨细胞病毒和猿猴病毒40。其他对于原核细胞和真核细胞都合适的表达体系,参见Sambrook,J.,Fritsh,E.F.,and Maniatis,T.Molecular Cloning:ALaboratory Manual.2nd,ed.Cold Spring Harbor Laboratory,Cold SpringHarbor Laboratory Press,Cold Spring Harbor,NY,1989的16章和17章。
在另一个实施方案中,重组的哺乳动物表达载体,能够指导特定细胞类型中优选核酸的表达(例如,组织特异性调节元素被用于表达核酸)。组织特异性调节元素在技术上是已知的。合适的组织特异性启动子的实例包括但不局限于,白蛋白启动子(肝脏特异;Pinkert et al.(1987)Gene Dev.1:268-277),淋巴特异启动子(Calame and Eaton(1988)Adv.Immunol.43:235-275),T-细胞受体的特殊启动子(Winoto and Baltimore(1989)EMBO J.8:729-933)和免疫球蛋白的特殊启动子(Banerji et al.(1983)Cell 33:729-740;Queen and Baltimore(1983)Cell 33:741-748),神经元特异的启动子(例如神经丝启动子;Byrne and Ruddle(1989)PANS86:5473-5477),胰腺特异的启动子(Edlund et al.(1985)Science 230:912-916),以及乳腺特异的启动子(例如乳汁乳清启动子;U.S.PatentNo.4,873,316和European Application Publication No.264,166)。也包括发育调节的启动子,例如鼠类hox启动子(Kessel and Gruss(1990)Science249:374-379)和α-胎蛋白启动子(Campes and Tilghman(1989)Genes Dev.3:537-546)。
本发明此外还提供了含有本发明DNA分子的重组表达载体,该DNA分子以反义方向克隆在表达载体中。也就是说,DNA分子可以可操作性的按以下方式连接到调节序列上,即允许与MP mRNA反义的RNA分子表达(通过DNA分子的转录)的方式。可以选择那些在各种细胞类型中指导反义RNA分子连续表达的调节序列,,例如病毒启动子和/或增强子,或者可以选择指导连续的、组织特异的或者细胞类型特异的反义RNA表达的调节序列,作为调节序列。反义表达载体可以以重组质粒、噬菌粒或者减毒病毒的形式存在,在其中反义核酸在高效调节区域的控制下产生,其活性可以通过引入载体的细胞类型来确定。关于使用反义基因调节基因表达,可以参见Weintraub,H.et al.,Antisense RNA as amolecular tool for genetics analysis,Review-Trends in Genetics,Vol.1(1)1986。
本发明的另一方面,涉及被引入本发明重组表达载体的宿主细胞的。术语“宿主细胞”和“重组宿主细胞”在此处可以交替使用。该术语应该理解为,不仅指被挑选的特定细胞,而且也指这些细胞的后代或者可能的后代。因为突变或者环境影响会使得某些修饰发生在成功的传代中,这些后代细胞实际上不可能与母细胞完全相同,但是也包含在此处使用的术语范围之内。
宿主细胞可以是任何原核或者真核细胞。例如,MP蛋白可以在像是谷氨酸棒杆菌这样的细菌细胞中、昆虫细胞中、酵母细胞中或者哺乳动物细胞(例如中国大鼠卵巢细胞(CHO)或者COS细胞)中表达。其他合适的宿主细胞,对于本领域技术人员来说是熟知的。可以用作本发明核酸和蛋白质分子宿主细胞的谷氨酸棒杆菌亲缘微生物,在表3中列出。
载体DNA可以通过常规转化或者转染技术,引入原核或者真核细胞。如此处所用的那样,术语“转化”和“转染”的意思是指各种本领域熟知的,把外源核酸(例如,线性DNA或者RNA(例如,线性载体或者没有载体的单独基因结构))或者以载体形式存在的核酸(例如,质粒、噬菌体、噬菌粒、噬菌粒、转座子或者其他DNA)转入宿主细胞的技术,包括磷酸钙或者氯化钙共沉淀,DEAE-右旋糖苷介导的转染,脂质转染,或者电传孔。转化或者转染宿主细胞的合适方法,可以在Sambrook,et al.(Molecular Cloning:ALaboratory Manual.2nd,ed.,ColdSpring Harbor Laboratory,Cold Spring Harbor Laboratory Press,ColdSpring Harbor,NY,1989),以及其他实验室手册上找到。
已知,为了稳定的转染哺乳动物细胞,依靠使用的表达载体和转染技术,只有一小部分可以把外源DNA整合到其自身基因组中。为了鉴定和筛选这些整合体,编码筛选标记(例如,对抗生素的抗性)的基因通常与感兴趣的基因被一同引入宿主细胞。优选的筛选标记包括那些能赋予药物抗性的标记,例如G418、潮霉素和氨甲蝶呤。编码筛选标记的核酸,可以与MP蛋白在同一个载体上被引入宿主细胞,或者在单独的载体上引入宿主细胞。经被引入核酸稳定转染的细胞,可以使用药物筛选鉴定(例如,与筛选标记基因合并的细胞可以存活,而其他细胞则死掉)。
为了创造同源重组微生物,制备含有至少部分MP基因的载体,该基因具有缺失、添加或者取代,从而改变,例如功能性破坏,MP基因。优选的该是谷氨酸棒杆菌MP基因,但是它也可以是来自亲缘细菌的同源物,甚至是来自哺乳动物、酵母或者昆虫。在一个优选的实施方案中,设计载体,使得根据同源重组,内源MP基因被功能性破坏(即,不在编码功能蛋白质;也称作“敲除”载体)。另外,可以设计载体,使得根据同源重组,内源MP基因被发生突变或者改变,但是仍编码功能蛋白质(例如,改变上游调节区域,从而改变内源MP基因的表达)。在同源重组载体中,被改变的MP基因部分,在其5’和3’末端侧面连接有多余的MP核酸,使得同源重组可以发生在载体携带的外源MP基因和微生物的内源MP基因之间。多余的侧面连接的MP核酸具有足够的长度,可以与内源基因成功的发生同源重组。典型的,载体中含有几千个碱基的侧链DNA(5’和3’末端)(参见,例如,Thomas,K.R.,and Capecchi,M.R.(1987)Cell 51:503for a description of homologous recombinationvectors)。引入微生物(例如电传孔)和细胞的载体,选择那些其中引入的MP基因与内源MP基因,使用技术上已知的技术可以同源重组的。
在另一个实施方案中,可以产生含有所选择系统的重组微生物,该系统允许调节引入基因的表达。例如,包含的MP基因在载体中处于lac操纵子的控制之下,使得MP基因只能在IPTG存在时表达。这种调节系统在技术上是熟知的。
在另一个实施方案中,宿主细胞中的内源MP基因被破坏(例如,通过同源重组或者其他技术上已知的遗传方法),使得其蛋白质产物的表达不能发生。在另一个实施方案中,宿主细胞中的内源的或者引入的MP基因,经一个或者多个点突变、缺失或者倒置而改变,但是仍旧编码功能MP蛋白。而在另一个实施方案中,微生物MP基因的一个或者多个调节区域(例如,启动子、阻抑物或者诱导子)被改变(例如,通过缺失、剪切、倒置或者点突变),使得MP基因的表达得到调节。本领域技术人员可以意识到,含有不止一个所述MP基因和蛋白质修饰的宿主细胞,使用本发明的方法可以很容易的产生,这些细胞也包含在本发明中。
本发明宿主细胞,例如培养的原核或者真核宿主细胞,可以用于产生(例如表达)MP蛋白。因此,本发明进一步提供了,使用本发明宿主细胞产生MP蛋白的方法。在一个实施方案中,该方法包括在合适的培养基中培养本发明的宿主细胞(其中引入了编码MP蛋白的重组表达载体,或者其基因组中引入了编码野生型或者改变的MP蛋白的基因),直到产生MP蛋白。在另一个实施方案中,该方法进一步包括从培养基或者宿主细胞中分离MP蛋白。
C.分离的MP蛋白
本发明的另一方面涉及分离的MP蛋白及其生物活性部分的。“分离的”或者“纯化的”蛋白,或者其生物活性部分,当使用重组DNA技术生产时基本上没有细胞物质,当化学合成时基本上没有化学前体或者其他化学物质。术语“基本上不含细胞物质”包括这样的MP蛋白制备,其中蛋白质被从天然或者重组产生该蛋白质的细胞的细胞组分中分离出。在一个实施方案中,术语“基本上不含细胞物质”包括制备含有至少大约30%(干重)非MP蛋白(此处也称作“污染蛋白质”)的MP蛋白,更优选的含有少于大约20%的非MP蛋白,甚至更优选的含有少于大约10%的非MP蛋白,最优选的含有少于大约5%的非MP蛋白。当MP蛋白或者其生物活性部分经重组产生时,优选是基本上不含培养基,即培养基少于制备蛋白质体积的大约20%,优选的少于10%,最优选的少于大约5%。术语“基本上不含化学前体或者其他化学物质”包括这样的MP蛋白制备,其中蛋白质被从参与蛋白质合成的化学前体或者其他化学物质中分离出。在一个实施方案中,术语“基本上不含化学前体或者其他化学物质”包括制备含有至少大约30%(干重)化学前体或者非MP化学物质的MP蛋白,更优选的含有少于大约20%的化学前体或者非MP化学物质,甚至更优选的含有少于大约10%的化学前体或者非MP化学物质,最优选的含有少于大约5%的化学前体或者非MP化学物质。在一个优选的实施方案中,分离的蛋白质或者其生物活性部分,不含有来自获得MP蛋白的同一生物体的污染蛋白质。这种蛋白质典型的是由重组表达产生,例如像谷氨酸棒杆菌这样微生物中的谷氨酸棒杆菌MP蛋白的重组表达。
分离的本发明的MP蛋白或者其生物活性部分,能够催化氨基酸、维生素、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应或者具有一种或者多种列在表1中的活性。在一个优选的实施方案中,蛋白质或者其部分含有这样的氨基酸序列,该序列与本发明的氨基酸序列(例如,序列表偶数序列号序列中的一个序列)有充分的同源性,使得该蛋白质或者其生物活性部分,能够催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应。蛋白质的部分,优选是指此处描述的生物活性部分。在另一个优选的实施方案中,本发明的MP蛋白具有在序列表中以偶数序列号列出的氨基酸序列。在另一个优选的实施方案中,MP蛋白具有由核苷酸序列编码的氨基酸序列,该核苷酸序列与本发明的核苷酸序列(例如,序列表奇数序列号序列中的一个序列)杂交,例如在严格条件下杂交。在另一个优选的实施方案中,MP蛋白具有由这样的核苷酸序列编码的氨基酸序列,该核苷酸序列与本发明的一条核酸序列或者其部分,有至少大约50%,51%,52%,53%,54%,55%,56%,57%,58%,59%或者60%的同源性,优选的有至少大约61%,62%,63%,64%,65%,66%,67%,68%,69%或者70%的同源性,更优选的有至少大约71%,72%,73%,74%,75%,76%,77%,78%,79%或者80%,81%,82%,83%,84%,85%,86%,87%,88%,88%,89%或者90%,或者91%,92%,93%,94%,以及甚至更优选的有至少大约95%,96%,97%,98%,99%或者更高的同源性。介于上面引述的值之间的范围或者一致性值(例如,70-90%的一致性或者80-95%的一致性),也有意的包含在本发明中。例如,有意的包含了这样的一致性值范围,这些范围是上面引用的上限和/或下限值的组合。本发明优选的MP蛋白也优选的具有至少一种此处描述的MP活性。例如,一种本发明优选的MP蛋白包含这样的核苷酸序列编码的氨基酸序列,该核苷酸序列与本发明的核苷酸序列杂交,例如在严格条件下杂交,并且该序列能够催化氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖代谢途径中的酶促反应,或者具有一种或者多种列在表1中的活性。
在其他实施方案中,MP蛋白与本发明的氨基酸序列(例如,序列表偶数序列号序列中的一个序列)有充分的同源性,并且具有本发明氨基酸序列蛋白质的功能活性,正如以上I部分详细描述的那样,其氨基酸序列由于天然改变或者突变而有所不同。因此,在另一个实施方案中,MP蛋白是这样的蛋白质,它具有的氨基酸序列与本发明的完全氨基酸序列,有至少大约50%,51%,52%,53%,54%,55%,56%,57%,58%,59%或者60%的同源性,优选的有至少大约61%,62%,63%,64%,65%,66%,67%,68%,69%或者70%的同源性,更优选的有至少大约71%,72%,73%,74%,75%,76%,77%,78%,79%或者80%,81%,82%,83%,84%,85%,86%,87%,88%,88%,89%或者90%,或者91%,92%,93%,94%,以及甚至更优选的有至少大约95%,96%,97%,98%,99%或者更高的同源性,并且具有至少一种此处描述的MP活性。介于上面引述的值之间的范围或者一致性值(例如,70-90%的一致性或者80-95%的一致性),也有意的包含在本发明中。例如,有意的包含了这样的一致性值范围,这些范围是上面引用的上限和/或下限值的组合。在另一个实施方案中,本发明与这样的全长谷氨酸棒杆菌蛋白质有关,该蛋白质与本发明的氨基酸序列有充分的同源性。
MP蛋白的生物活性部分包含这样的多肽,该多肽含有来自MP蛋白氨基酸序列的氨基酸序列,例如,序列表偶数序列号的氨基酸序列或者与MP蛋白同源蛋白质的氨基酸序列,该部分含有比全长MP蛋白或者全长MP蛋白同源蛋白质更少的氨基酸,并且表现出至少一种MP蛋白活性。典型的生物活性部分(肽,例如,氨基酸长度为像是5,10,15,20,30,35,36,37,38,39,40,50,100或者更多的肽)包括一个具有至少一种MP蛋白活性的结构域或者基元。另外,其他生物活性部分,其中蛋白质的其他部分已被删除,可以通过重组技术制备,并鉴定其此处描述的一种或者多种活性。优选的MP蛋白的生物活性部分,含有一个或者多个挑选的具有生物活性的结构域/基元或者其部分。
MP蛋白优选的通过重组DNA技术生产。例如,把编码蛋白质的核酸分子克隆到表达载体中(如上所述),将表达载体引入宿主细胞(如上所述)并在宿主细胞中表达MP蛋白。然后按照合适的纯化方案,使用标准蛋白质纯化技术,从细胞中分离MP蛋白。除了重组表达,可以使用标准肽合成技术化学合成MP蛋白、多肽或者肽。另外,天然MP蛋白可以从细胞(例如内皮细胞)中分离,例如使用抗-MP抗体,该抗体可以使用本发明的MP蛋白或者其部分通过标准技术产生。
本发明也提供了MP嵌合蛋白或者融合蛋白。如此处所用的,MP“嵌合蛋白”或者“融合蛋白”含有可操作性连接到非MP多肽上的MP多肽。“MP多肽”是指含有MP相关氨基酸序列的多肽,而“非MP蛋白”是指含有这样的蛋白质相关氨基酸序列的多肽,该蛋白质与MP蛋白没有基本的同源性,例如,来自相同或者不同生物体的与MP蛋白不同的蛋白质。在融合蛋白质中,术语“可操作性连接”的意思是指,MP蛋白与非MP蛋白相互之间是符合读框的融合。非MP多肽可以融合到MP多肽的N-末端或者C-末端。例如,在一个实施方案中,融合蛋白质是DST-MP融合蛋白,其中MP序列融合到GST序列的C-末端。该融合蛋白质有助于重组MP蛋白的纯化。在另一个实施方案中,融合蛋白质是在其N-末端有异源信号序列的MP蛋白。在某些宿主细胞(例如哺乳动物宿主细胞)中,通过使用异源信号序列可以增加MP蛋白的表达和/或分泌。
优选,本发明的嵌合蛋白或者融合蛋白通过标准重组DNA技术产生。例如,依照常规技术编码不同多肽序列的DNA片段被符合读框的连接在一起,例如,使用平头末端或者交错末端的末端连接,使用限制性酶进行消化以提供合适的末端,使用粘性末端补平作为合适的末端,使用碱性磷酸酶处理以避免不合需要的连接,以及使用酶促连接。在另一个实施方案中,可以使用常规技术包括自动DNA合成仪合成融合基因。另外,可以使用锚引物进行基因片段的PCR扩增,锚引物可以增加两条连续基因片段之间的互补的突出端,连续基因可以随后进行退火和再扩增而产生嵌合基因序列(参见,例如,Current Protocols in MolecularBiology,eds.Ausubel et al.John Wiley&Sons:1992)。另外,很多已经编码融合部分(例如GST多肽)的表达载体是商业提供的。MP-编码核酸可以被克隆到这种表达载体中,使得融合部分符合读框的连接到MP蛋白上。
MP蛋白的同源物可以通过突变产生,例如MP蛋白的不连续点突变或者剪切。如此处所用的,术语“同源物”是指MP蛋白的变体形式,它们可以用作MP蛋白活性的激动剂或者拮抗物。MP蛋白的激动剂可以基本上具有MP蛋白相同的或者部分的生物活性。MP蛋白的拮抗物可以抑制MP蛋白天然存在形式的一种或者多种活性,例如,通过与包含MP蛋白的MP系统的下游或者上游成员竞争性结合。因此,本发明的谷氨酸棒杆菌MP蛋白及其同源物,可以调节一条或者多条糖类转运途径的活性,或者调节MP蛋白在该微生物中发挥作用的细胞内信号传导途径的活性。
在另外的实施方案中,MP蛋白的同源物可以通过筛选MP蛋白突变体的组合文库,例如剪切突变体,来鉴定MP蛋白激动剂或者拮抗剂活性。在一个实施方案中,MP变体的多样性文库通过在核酸水平上组合性突变而产生,并由多样性基因文库编码。MP变体的多样性文库可以通过,例如,把合成的寡聚核苷酸混合物酶促连接到基因序列中,使得潜在MP序列的简并集合作为单个多肽,或者其中含有MP序列集合的更大的融合蛋白质(例如为了噬菌体展示)的集合,是可以表达的。有各种方法可以用于从简并寡聚核苷酸序列,产生潜在MP同源物文库。可以用自动DNA合成仪进行简并基因序列的化学合成,然后合成基因被连接到合适的表达载体中。基因简并集合的使用,允许混合的提供编码所需潜在MP序列集合的全部序列。合成简并寡聚核苷酸的方法在技术上是已知的(参见,例如,Narang,S.A.(1983)Tetrahedron 39:3;Itakura et al.(1984)Annu.Rev.Biochem.53:323;Itakura et al.(1984)Science 198:1056;lke et al.(1983)NucleicAcidRes.11:477)。
另外,编码MP蛋白片段的文库,可用于产生MP片段的多样性群体,该群体用于筛选并挑选MP蛋白的同源物。在一个实施方案中,编码序列片段的文库可以这样产生,即在大约每分子只产生一个切口的条件下,用核酸酶处理MP编码序列的双链PCR片段,变性双链DNA,复性DNA以形成双链DNA,该双链DNA可以包含从不同有切口的产物形成的有义/反义对,在重新形成的双螺旋中通过S1核酸酶处理除去单链部分,以及把最后得到的片段文库连接到表达载体中。通过该方法,可以得到编码N-末端、C-末端和不同大小MP蛋白中间片段的表达文库。
筛选由点突变或者剪切得到的组合文库中的基因产物的许多技术,以及筛选cDNA文库中具有所挑选特性基因产物的技术,在技术上都是已知的。这些技术都适用于由MP同源物组合突变得到的基因文库的快速筛选。筛选大型基因库的应用最广泛的技术,能够用于高产量分析,包括把基因组文库克隆到可复制表达载体中,用得到的载体文库转化载体文库,以及在一定条件下表达组合基因,在该条件下所需活性的检测有助于编码被检测产物基因的载体的分离。回归系综突变(REM),一种增加文库中功能突变体频率的新技术,可以与筛选分析一起用于鉴定MP同源物(Arkin and Yourvan(1992)PANS 89:7811-7815;Delgrave et al.(1993)Protein Engineering 6(3):327-331)。
在另一个实施方案中,使用技术上已知的方法,基于细胞的分析可以用于分析多样性MP文库。
D.本发明的应用和方法
此处描述的核酸分子、蛋白质、蛋白质同源物、融合蛋白质、引物、载体和宿主细胞,可以应用于下述一种或者多种方法中:鉴定谷氨酸棒杆菌和亲缘微生物;绘制谷氨酸棒杆菌亲缘生物体的基因组图谱;鉴定和定位谷氨酸棒杆菌的感兴趣序列;进化研究;确定MP蛋白的功能必需区域;MP蛋白活性调节;MP途径活性调节;所需化合物,例如精细化学物质的细胞生产的调节。
本发明MP核酸分子具有各种用途。首先,它们可以用于鉴定一种生物体是否是谷氨酸棒杆菌或者其近亲生物体。它们也可以用于鉴定混合微生物群体中谷氨酸棒杆菌或者其亲缘生物体的存在。本发明提供了许多谷氨酸棒杆菌基因的核酸序列;在严格条件下,使用跨越对谷氨酸棒杆菌特异基因的探针,探测从单一或者混合微生物培养物中提取的基因组DNA,可以确定该生物体是否存在。尽管谷氨酸棒杆菌本身是非致病性的,但是它与致病种类相关,例如白喉棒杆菌。白喉棒杆菌是白喉的致病源,白喉是一种发展迅速、急性、发烧的感染,它涉及局部病状和系统病状。得这种疾病时,上呼吸道发生局部病变,并且包括上皮细胞坏死性损伤;细菌分泌毒素,毒素从病变处散布到身体易受感染的末梢组织。这些组织包括心脏、肌肉、外周神经、肾上腺、肾脏、肝脏和脾脏,在其中由于蛋白质合成被抑制而造成的变质性改变,会导致该疾病的系统病状。白喉在世界许多地区保持高发病率,这些地区包括非洲、亚洲、东欧和前苏联的独立国家。从1990年起,在后两个地区白喉的持续流行,导致了至少5,000人死亡。
在一个实施方案中,本发明与鉴定受试者中白喉棒杆菌存在或者活性的方法有关。该方法包括鉴定受试者中本发明的一条或者多条核酸或者氨基酸序列(例如,分别列在序列表中的奇数或者偶数序列号序列),从而检测受试者中白喉棒杆菌的存在或者活性。谷氨酸棒杆菌和白喉棒杆菌是有亲缘关系的细菌,谷氨酸棒杆菌中的许多核酸和蛋白质分子是白喉棒杆菌中核酸和蛋白质分子的同源物,因此也可以用于检测受试者中的白喉棒杆菌。
本发明的核酸和蛋白质分子也可以用作基因组特定区域的标记。这不仅在绘制基因组图谱时有用,而且可以用于谷氨酸棒杆菌蛋白质功能研究。例如,为了鉴定特定谷氨酸棒杆菌DNA结合蛋白与之结合的基因组区域,可以消化谷氨酸棒杆菌基因组,将片段与DNA结合蛋白孵育。与蛋白质结合的片段可以进一步用本发明的核酸分子探测,优选使用易检测标记;这些核酸分子与基因组片段的结合,可以定位片段在谷氨酸棒杆菌基因组图谱上的位置,而且,当使用不同的酶进行多次操作时,有助于快速确定蛋白质与之结合的核酸序列。另外,本发明核酸分子可以与亲缘种类有充分的同源性,使得这些核酸分子可以作为构建亲缘细菌基因组图谱的标记,例如乳发酵短杆菌。
本发明的MP核酸分子又可以用于进化和蛋白质结构研究。本发明分子参与的糖类摄取系统,被各种各样的细菌所使用;通过比较本发明核酸分子序列和那些在其他生物体中编码相似酶的核酸分子序列,可以估算生物体的进化相关性。类似的,这种比较允许估算保守序列区域和非保守序列区域,这可以有助于确定蛋白质中对于酶功能必需的区域。这种类型的确定对于蛋白质工程研究是有价值的,并且可以指示那些蛋白质可以忍受突变而不失去功能。
本发明MP核酸分子的操作可以导致具有与野生型MP蛋白不同功能的MP蛋白的产生。可以提高这些蛋白质的效率或者活性,可以使之以比通常更多的数目出现在细胞中,或者降低其效率或者活性。
本发明也提供了筛选可调节MP蛋白活性的分子的方法,这些分子或者通过与蛋白质本身或者底物相互作用,或者与MP蛋白的配偶体结合,或者通过调节本发明MP核酸分子的转录或者翻译来调节MP蛋白活性。在该方法中,表达一种或者多种MP蛋白的微生物,与一种或者多种试验化合物接触,并且评估每种测试化合物对于MP蛋白活性或者表达水平的作用。
当需要从谷氨酸棒杆菌的大规模发酵培养物中分离的所需精细化学物质是氨基酸、维生素、辅因子、营养因子、核苷酸、核苷或海藻糖时,通过重组遗传机制调节一种或多种本发明蛋白的活性效率或活性可以直接影响这些精细化学物质中的一种。例如,对所需氨基酸生物合成途径中的酶而言,该酶活性或效率的提高(包括存在多个拷贝的基因)应当导致所需氨基酸的生产或生产效率增加。对于其合成与所需氨基酸的生物合成竞争的氨基酸生物合成途径中的酶,该酶活性或效率的降低(包括基因缺失)会导致所需氨基酸生产或生产效率的增加,是由于对中间体化合物和/或能量的竞争减少。对于所需氨基酸降解途径的酶,该酶活性或效率的降低会导致所需产物的产量或生产效率更高,这是由于其降解减少了。最后,对所需氨基酸生物合成相关酶进行诱变使得该酶不再被反馈抑制,这会导致所需氨基酸的产量或生产效率提高。对本发明的维生素、辅因子、营养因子、核苷酸、核苷和海藻糖代谢相关的生物合成和降解酶来说,同样如此。
类似地,当所需精细化学物质不是上述化合物之一时,本发明的一种蛋白活性的调节仍有可能影响谷氨酸棒杆菌大规模培养生产该化合物的效率和/或产量。任何生物体的代谢途径都是密切关联的,一种途径使用的中间体经常由不同的途径供给。酶表达和功能可以根据不同代谢过程化合物的细胞水平来调节,基础生产必需的分子如氨基酸和核苷酸的细胞水平对大规模培养中的微生物活力具有重大影响。因此,调节一种氨基酸生物合成酶使得其对反馈抑制不再有反应或其效率或转变提高,会导致一种或多种氨基酸细胞水平增加。结果,该增加的氨基酸供给不但增加对蛋白质合成必需分子的供应,也会增加用作多种其它生物合成途径中的中间体和前体的分子的供应。如果细胞内特定氨基酸有限,增加其生产也会增加细胞进行多种其它代谢反应的能力,并使细胞更有效地生产各种蛋白,可能会增加大规模培养中的细胞总生产速率或存活能力。活力增加提高了发酵培养物中能够产生所需精细化学物质的细胞的数量,从而增加该化合物的产量。通过调节本发明降解酶活性,使得该酶不再催化对所需化合物生物合成重要,或使大规模培养物中的细胞生长和增殖更有效的细胞化合物的降解或催化效率降低,也会存在类似情况。应当强调的是,优化本发明某些分子的降解活性或降低生物合成活性也会对谷氨酸棒杆菌生产某些精细化学物质有正面作用。例如,通过降低与所需化合物生物合成途径竞争一种或多种中间体的途径中生物合成酶的活性效率,更多的中间体可以用于所需物质的转化。类似情形下需要提高一种或多种本发明蛋白的降解能力或效率。
前面提到的导致所需化合物产量增加的MP蛋白诱变方案的列表,并不意味着仅局限于此;这些诱变方案的变化对于本领域普通技术人员来说是很明白的。经过这些机制,本发明的核酸和蛋白质分子可以用于产生表达突变MP核酸和蛋白质分子的谷氨酸棒杆菌或者其亲缘菌株,从而增加所需化合物的产量、生产和/或生产效率。该所需化合物可以是谷氨酸棒杆菌的任何天然产物,这包括生物合成途径的最终产物和天然存在代谢途径的中间体,以及不是在谷氨酸棒杆菌代谢中天然存在但是由本发明谷氨酸棒杆菌菌株产生的分子。在谷氨酸棒杆菌中生产的优选化合物是L-赖氨酸和L-甲硫氨酸。
在一个实施方案中,由谷氨酸棒杆菌中分离出编码甲硫氨酸生物合成途径的第三个酶胱硫醚β裂合酶的metC基因。该基因的翻译产物与来自其他生物的metC基因没有明显的同源性。导入含有metC基因的质粒到谷氨酸棒杆菌中导致胱硫醚β裂合酶活性增加5倍。现被称为MetC的该蛋白产物编码35574道尔顿的蛋白产物,由325个氨基酸组成,与以前报导的aec基因(Rossol,I and Puhler,A.(1992)J.Bacteriology 174,2968-2977)只有两个氨基酸不同。象aecD基因一样,以多拷贝存在时,metC基因赋予对赖氨酸的毒性类似物S-(β-氨基乙基)-半胱氨酸的抗性。但是,遗传和生化证据揭示,metC基因的天然活性是介导谷氨酸棒杆菌中的甲硫氨酸生物合成。构建了metC突变菌株,该菌株显示甲硫氨酸原养型。突变菌株完全失去了对S-(β-氨基乙基)-半胱氨酸的抗性。这些结果显示,除了另一个生物合成途径转硫作用外,直接硫化氢解途径在谷氨酸棒杆菌中作为平行的甲硫氨酸生物合成途径起作用。
在另一个实施方案中,附加的硫化氢解途径显示是由O乙酰高丝氨酸硫化氢解酶催化。分离到了相应的metZ(或metY)基因和酶(分别为SEQ ID NO:1和SEQ ID NO:2)证实了该途径的存在。在真核生物中,真菌和酵母已经报导同时具有转硫作用和直接硫化氢解途径。迄今为止,还未发现具有两种途径的原核生物。与大肠杆菌仅具有一个赖氨酸生物合成途径不同,谷氨酸棒杆菌具有该氨基酸的两个平行的生物合成途径。在这方面,甲硫氨酸生物合成途径类似于赖氨酸。
metZ基因位于metA的上游,后者编码催化甲硫氨酸生物合成第一步的酶(Park,S.-D.,et al(1998)Mol.Cells 8,286-294)。对metA的上游核下游进行了测序以鉴定其他met基因。似乎metZ和metA形成一个操纵子。编码MetA和MetZ的基因的表达导致相应多肽的过量产生。
令人吃惊的是,metZ克隆可以补充大肠杆菌metB突变株营养缺陷型。这显示metZ蛋白产物催化的步骤可以跨过metB蛋白产物催化的步骤。MetZ也被破坏,突变菌株显示甲硫氨酸原养型。也构建了谷氨酸棒杆菌metB和metZ双重突变体。该双重突变体是甲硫氨酸营养缺陷型。因此,metZ编码的蛋白催化O-乙酰-高丝氨酸到高半胱氨酸的反应,这是甲硫氨酸生物合成的硫化氢解途径的一个步骤。谷氨酸棒杆菌同时具有甲硫氨酸生物合成的转硫作用和硫化氢解途径
导入metZ至谷氨酸棒杆菌表达47000道尔顿的蛋白。同时导入metZ和metA至谷氨酸棒杆菌在凝胶电泳中显示有metA和metZ蛋白。如果棒杆菌菌株是赖氨酸的过量生产者,导入含有metA和metZ的质粒导致较低的赖氨酸效价,但检测到高半胱氨酸和甲硫氨酸积累。
在另一个实施方案中,metA和metZ与hom基因一起被导入到谷氨酸棒杆菌中,hom基因编码高丝氨酸脱氢酶,催化天冬氨酸半醛向高丝氨酸的转变。选择了不同生物的不同hom基因用于该实验。可以使用谷氨酸棒杆菌的hom基因,也可以使用来自其他原核生物如大肠杆菌或枯草芽孢杆菌的hom基因,或者真核生物如酿酒酵母、粟酒裂殖酵母、棉桃阿舒氏囊霉菌或海藻、高等植物或动物的hom基因。Hom基因可以对任何天冬氨酸家族的氨基酸如天冬氨酸、赖氨酸、甲硫氨酸、苏氨酸的生物合成途径中任何代谢物的反馈抑制不敏感。这些代谢物如天冬氨酸、赖氨酸、甲硫氨酸、苏氨酸、天冬氨酸磷酸、天冬氨酸半醛、高丝氨酸、胱硫醚、高半胱氨酸或任何该生物合成途径中的其他代谢物。除了这些代谢物外,高丝氨酸脱氢酶可能对所有这些的类似物的抑制不敏感,甚至对参与该代谢的其他化合物的抑制也不敏感,因为还有其他氨基酸如半胱氨酸或辅因子如维生素B12和其所有衍生物和S腺苷甲硫氨酸和其代谢物和衍生物和类似物。高丝氨酸脱氢酶对所有这些不敏感,这些化合物的一部分或一种可以是其天然趋向,或者可以是使用化学试剂或辐射或其他诱变剂的一种或多种经典突变和选择的结果。突变可以使用基因技术例如导入位点特异点突变或者任何上述MP或MP编码DNA序列适用的方法导入到hom基因中。
当hom基因与metZ和metA基因组合并导入到过量产生赖氨酸的谷氨酸棒杆菌中时,赖氨酸积累减少了,高半胱氨酸和甲硫氨酸积累增加了。如果使用过量产生谷氨酸棒杆菌的菌株并且在用含有hom基因和metZ和metA基因组合的DNA转化之前,破坏其中ddh基因或lysA基因,则高半胱氨酸和甲硫氨酸积累还可增加。使用不同的硫源均可实现高半胱氨酸和甲硫氨酸的过量产生。硫酸盐、硫代硫酸盐、亚硫酸盐和更加还原的硫源如硫化氢和硫醚和其衍生物也可以使用。有机硫源如甲基硫醇、硫代羟乙酸、硫代氰酸盐、硫脲、含硫氨基酸如半胱氨酸和其他含硫化合物也可用来实现高半胱氨酸和甲硫氨酸过量生产。
在另一个实施方案中,metC基因使用前述方法被导入到谷氨酸棒杆菌中。metC基因可以与其他基因如metB、metA和metA一起转化到菌株中。也可加入hom基因。当hom基因、metC、metB和metA基因被组合到一个载体上时并导入到谷氨酸棒杆菌中时,就实现了高半胱氨酸和甲硫氨酸的过量生产。硫酸盐、硫代硫酸盐、亚硫酸盐和更加还原的硫源如硫化氢和硫醚和其衍生物也可以使用。有机硫源如甲基硫醇、硫代羟乙酸、硫代氰酸盐、硫脲、含硫氨基酸如半胱氨酸和其他含硫化合物也可用来实现高半胱氨酸和甲硫氨酸过量生产。
本发明进一步由以下实例阐明,这些实例不应该被解释为仅局限于此。本申请中所引用的所有参考文献、专利申请、专利、发表的专利申请、表和序列列表中的内容,全部引入作为参考。
实施例
实施例1:谷氨酸棒杆菌ATCC 13032全部基因组DNA的制备
谷氨酸棒杆菌(ATCC 13032)培养物在BHI培养基(Difco)中,30℃剧烈振荡培养过夜。离心收集细胞,弃上清,细胞重新悬浮在5ml缓冲液I(培养物原体积的5%-所有指出的体积都是对于100ml培养物体积计算的)中。缓冲液I的组成:140.34g/l蔗糖,2.46g/l MgSO4x 7H2O,10ml/l KH2PO4溶液(100g/l,KOH调节至PH6.7),50g/l M12浓缩物(10g/l(NH4)2SO4,1g/l NaCl,2g/l MgSO4x 7H2O,0.2g/l CaCl2,0.5g/l酵母提取物(Difco)),10ml/l微量元素混合物(200mg/l FeSO4xH2O,10mg/l ZnSO4x7H2O,3mg/l MnCl2x4H2O,30mg/l H3BO3,20mg/lCoCl2x6H2O,1mg/l NiCl2x6H2O,3mg/l Na2MoO4x2H2O),500mg/l络合剂(EDTA或者柠檬酸),100ml/l维生素混合物(0.2mg/l生物素,0.2mg/l叶酸,20mg/l p-氨基安息香酸,20mg/l核黄素,40mg/lpanthothenate,140mg/l烟酸,40mg/l盐酸吡多醛,200mg/l肌醇)。悬浮液中加入溶菌酶至终浓度2.5mg/ml。37℃孵育大约4小时之后,细胞壁被降解,得到的原生质体用离心收集。沉淀用5ml缓冲液I洗一次,用5ml TE缓冲液(10mM Tris-HCl,1ml EDTA,pH8)洗一次。沉淀用4ml TE缓冲液重悬,并加入0.5ml SDS溶液(10%)和0.5ml NaCl溶液(5M)。加入蛋白酶K至终浓度200μg/ml,悬浮液在37℃孵育约18小时。DNA用苯酚、苯酚-氯仿-异戊醇、氯仿-异戊醇按照标准程序提取纯化。然后,加入1/50体积的3M乙酸钠和2倍体积的乙醇,在-20℃孵育30分钟,用使用SS34转头(Sorvall)的高速离心机12,000rpm离心30分钟,沉淀DNA。把DNA溶解在含有20μg/ml RNaseA的1ml TE缓冲液中,在1000ml TE缓冲液中4℃透析至少3小时。这段时间中,更换缓冲液3次。每0.4ml透析的DNA溶液中,加入0.4ml 2M LiCl和0.8ml乙醇。在-20℃孵育30分钟后,离心(13,000rpm,Biofuge Fresco,Heraeus,Hanau,Germany)收集DNA。DNA沉淀融解在TE缓冲液中。按该程序制备的DNA可以用于所有目的,包括southern杂交和基因组文库的构建。
实施例2:在大肠杆菌中谷氨酸棒杆菌ATCC13032的基因组文库的构建
使用如在实施例1中所描述制备的DNA,按照已知的和充分建立的方法(参见,例如Sambrook,J.et al.(1989)“Molecular Cloning:ALaboratory Manual”Cold Spring Harbor Laboratory,Cold Spring HarborLaboratory Press,或者Ausubel,F.M.et al.(1994)“Current Protocols inMolecular Bilogy”,John Wiley&Sons.),可以构建粘粒文库和质粒文库。
可以使用任何质粒和粘粒。质粒pBR322(Sutcliffe,J.G.(1979)Proc.Natl.Acad.Sci.USA,75:3737-3741);pACY177(Change&Cohen(1978)J.Bacteriol 134:1141-1156),pBS系列质粒(pBSSK+,pBSSK-和其他质粒;Stratagene,LaJolla,USA),粘粒SuperCos1(Stratagene,LaJolla,USA)或者Lorist6(Gibson,T.J.,Rosenthal A.and Waterson,R.H.(1987)Gene53:283-286)可以用于特殊用途。专门在谷氨酸棒杆菌中使用的基因文库可以用质粒pSL109(Lee,H.-S.and A.J.Sinskey(1994)J.Microbiol.Biotechnol.4:256-263)构建。
为了分离metC克隆,大肠杆菌JE6839细胞用文库DNA转化并在含有氨苄青霉素和合适添加物的M9基本培养基上铺平板。平板在37℃温育5天。分离集落并筛选质粒内含物。分离的metC基因的完整核苷酸序列通过本领域普通技术人员熟知的方法确定。
实施例3:DNA测序和计算机功能分析
按照标准方法,使用如在实施例2中所描述基因组文库,可以进行DNA测序,特别是用使用ABI377测序仪的链终止方法(参见,例如Fleischman,R.D.et al.(1995)“Whole-genome Random Sequencing andAssembly of Haemophilus Influenzae Rd.,Science,269:496-512)。使用具有以下核苷酸序列的测序引物:5’-GGAAACAGTATGACCATG-3’(SEQID NO:123)或者5’-GTAAA CGACGGCCAGT-3’(SEQ ID NO:124)。
实施例4:体内诱变
可以通过大肠杆菌或者其他微生物(例如,芽孢杆菌某些菌或者像是酿酒酵母的酵母)的质粒(或者其他载体)DNA的传代,进行谷氨酸棒杆菌的体内诱变,其中这些微生物保持其遗传信息整体性的能力已被损伤。典型的突变株,在DNA修复系统的基因中有突变(例如,mutHLS,mutD,mutT等;参考文献参见Rupp,W.D.(1996)DNA repair mechanisms,in:Escherichia coli and Salmonella,p.2277-2294,ASM:Washington.)。这些菌株对于技术熟练的人来说是熟知的。这些菌株的使用阐述在,例如Greener,A.and Callahan,M.(1994)Strategies 7:32-34中。
实施例5:在大肠杆菌和谷氨酸棒杆菌之间传递的DNA
棒杆菌和短杆菌菌种含有能自发复制的内源质粒(像是例如,pHM1519或者pBL1)(评论参见,例如,Martin,J.F.et al.(1987)Biotechnology,5:137-146)。大肠杆菌和谷氨酸棒杆菌的穿梭载体,可以使用大肠杆菌的标准载体容易的构建(Sambrook,J.et al.(1989)“Molecular Cloning:A Laboratory Manual”Cold Spring Harbor Laboratory,Cold Spring Harbor Laboratory Press或者Ausubel,F.M.et al.(1994)“Current Protocols in Molecular Bilogy”,John Wiley&Sons.),即在其中加入谷氨酸棒杆菌的复制叉起始点和合适的标记。这种复制起始点,优选是从棒杆菌和短杆菌菌种中分离的内源质粒获得的。用作这些菌种转化标记这一特殊用途的是卡那霉素抗性基因(例如来自Tn5或者Tn903转座子的那些卡那霉素抗性基因)或者氯霉素抗性基因(Winnacker,E.L.(1987)“From Genes to Clones-Introduction to Gene Technology,VCH,Weinheim)。在构建各种野生型穿梭载体的文献中有许多实例,这些穿梭载体可以在大肠杆菌和谷氨酸棒杆菌中复制,并且可以用于各种目的,其中包括基因过量表达(参考文献参见,例如,Yoshihama,M.et al.(1985)J.Bacteriol.162:591-597,Martin J.F.et al.(1987)Biotechnology,5:137-146和Eikmanns,B.J.et a;.(1991)Gene,102:93-98)。
使用标准方法可以把感兴趣的基因克隆到上述穿梭载体中,并且可以把该杂交载引入谷氨酸棒杆菌菌株中。谷氨酸棒杆菌的转化可以通过原生质体转化(Kastsumata,R.et al.(1984)J.Bacteriol.159306-311),电传孔(Liebl,E.et al.(1989)FEMSMicrobiol.Letters,53:399-303)实现,当使用特殊的载体时,也可以通过结合作用(例如在,A et al.(1990)J.Bacteriol.172:1663-1666)实现。也可以通过从谷氨酸棒杆菌制备质粒DNA(使用技术上已知的标准方法)并将其转化到大肠杆菌中,而把穿梭载体从谷氨酸棒杆菌转移到大肠杆菌。这一转化步骤可以使用标准方法进行,但是使用Mcr缺陷型大肠杆菌菌株,例如NM522(Gough&Murray(1983)J.Mol.Biol.166:1-19)是有利的。
使用含有pCG1(U.S.Patent No.4,617,267)或者其片段的质粒,并且可以选择来自TN903的卡那霉素抗性基因(Grindley,N.D.andJoyce,C.M.(1980)Proc.Natl.Acad Sci.USA 77(12):7176-7180),就可以在谷氨酸棒杆菌中过量表达基因。另外,使用质粒pSL109(Lee,H.-S.and A.J.Sinskey(1994)J.Microbiol.Biotechnol.4:256-263)也可以在谷氨酸棒杆菌中过量表达基因。
除了使用可复制质粒以外,也可以通过基因组整合而实现基因的过量表达。谷氨酸棒杆菌或者其他棒杆菌或者短杆菌菌种的基因组整合,可以通过熟知的方法实现,例如基因组区域的同源重组,限制性核酸内切酶介导的整合(REMI)(参见例如,DE Patent 19823834),或者通过使用转座子。也可以通过修饰调节区域(例如,启动子、阻抑物和/或增强子),通过使用定向位点方法(例如同源重组)或者基于随机事件方法(例如转座子诱变或者REMI)的序列修饰、插入或者缺失,来调节感兴趣基因的活性。用作转录终止子的核酸序列,也可以被插入到本发明一个或者多个基因编码区域的3’;这样的终止子在技术上是熟知的,并且描述在例如Winnacker,E.L.1987)From Genes to Clones-Introduction to Gene Technology.VCH:Weinheim中。
实施例6:突变蛋白质表达的估算
被转化宿主细胞中突变蛋白质活性的观测,依赖于这一事实,即突变蛋白质以与野生型蛋白质相似的方式和相似的数量表达。确定突变基因转录水平(用于基因产物翻译的mRNA的数量指标)的一种有用的方法是进行Northern杂交(参考文献参见,例如,Ausubel et al.(1988)CurrentProtocols in Molecular Biology,Wiley:New York),其中设计的用于结合感兴趣基因的引物标记有可探测的标记(通常是放射性的或者化学发光的),从而,当生物体培养物的全部RNA被提取出,跑凝胶电泳,转移到稳定介质上并与该探针孵育,结合探针的结合和数量便指示了该基因mRNA的存在和数量。该信息是突变基因转录程度的证据。可以使用几种方法从谷氨酸棒杆菌中制备全部细胞RNA,这在技术上是熟知的,例如描述在Bormann,E.R.et al.(1992)Mol.Microbiol.6:317-326中的。
为了估算由该mRNA翻译的蛋白质的存在和相对数量,可以使用标准技术例如SDS聚丙烯酰胺凝胶电泳。在谷氨酸棒杆菌中metC和metZ与metA组合的过量生产通过该方法得到证实。Western印迹也可使用(参见,例如,Ausubel et al.(1988)Current Protocols in Molecular Biology,Wiley:New York)。在该方法中,提取全部细胞蛋白质,通过凝胶电泳分离,转移到像是硝酸纤维素这样的介质上,和与所需蛋白质特异结合的探针共孵育,例如抗体。该探针通常标记有易于检测的化学发光的或者比色的标记。观测到的标记的存在和数量,指示了出现在细胞中的所需突变蛋白质的存在和数量。
实施例7:大肠杆菌和遗传修饰的谷氨酸棒杆菌的生长-培养基和培养条件
大肠杆菌菌株按常规分别在MB和LB培养液中培养(Follettie,M.T.,et al.(1993)J.Bacteriol.175,4096-4103)。大肠杆菌基本培养基是M9和改进的MCGC(Yoshihama,M.,et al.(1985)J.Bacteriol.162,591-507)。加入葡萄糖至终浓度1%。按以下量加入抗体(每毫升微克数):氨苄青霉素50;卡那霉素25;萘啶酮酸,25。氨基酸、维生素和其他添加物按以下量加入:甲硫氨酸9.3mM;精氨酸9.3mM;组氨酸9.3mM;硫胺素0.05mM。大肠杆菌细胞按常规在37℃下培养。
遗传修饰的谷氨酸棒杆菌可以培养在合成或者天然生长培养基中。用于谷氨酸棒杆菌的各种不同的生长培养基是已知的并且是易于得到的(Lieb,et al.(1989)Appl.Microbiol.Biotechno.,32:205-210;von der Ostenet al.(1998)Biotechnology Letters,11:11-16;Patent DE 4,120,867;Liebl(1992)“The Genus Corynebacterium,in:Procaryotes,Volume II,Balows,A.et al.,eds.Springer-Verlag)。这些培养基含有一种或者多种碳源、氮源、无机盐、维生素和微量元素。优选的碳源是糖类,例如单糖、二糖或者多糖。例如,葡萄糖、果糖、甘露糖、半乳糖、核糖、山梨糖、核酮糖、乳糖、麦芽糖、蔗糖,棉子糖,淀粉或者纤维素,都可用作很好的碳源。也可以通过复杂化合物向培养基提供糖类,例如糖蜜或者其他糖类精炼的副产物。提高不同碳源的混合物也是有利的。其他可用的碳源有酒精和有机酸,例如甲醇、乙醇、乙酸或者乳酸。氮源通常是有机或者无机的氮化合物,或者含有这些化合物的物质。代表性的氮源包括氨气或者铵盐,例如NH4Cl或者(NH4)2SO4、NH4OH、硝酸盐、尿素、氨基酸或者复杂的氮源,例如玉米浸泡液、大豆粉、大豆蛋白、酵母提取物、肉类提取物或者其他。
含硫氨基酸如高半胱氨酸和甲硫氨酸的过量生产可以使用不同的硫源实现。硫酸盐、硫代硫酸盐、亚硫酸盐和更加还原的硫源如硫化氢和硫醚和其衍生物均可使用。有机硫源如甲基硫醇、硫代羟乙酸、硫代氰酸盐、硫脲、含硫氨基酸如半胱氨酸和其他含硫化合物也可用来实现高半胱氨酸和甲硫氨酸过量生产。
可以包含在培养基中的无机盐化合物,包括盐酸盐、磷酸盐或者硫酸盐的钙、镁、钠、钴、钼、钾、锰、锌、铜或者铁。螯合剂可以加到培养基中,以维持溶液中的金属离子。特别有用的螯合剂包括二羟基苯酚,像是儿茶酚和原儿茶酸,或者有机酸,像是柠檬酸。培养基典型的也含有生长因子,例如维生素和生长促进剂,它们的实例包括生物素、核黄素、硫胺、叶酸、烟酸、泛酸盐和吡多醇。生长因子和盐经常来自复杂的培养基成分,例如酵母提取物、糖蜜、玉米浸泡液和其他成分。培养基化合物的确切组成强烈的依赖于直接实验,而且对于每一个具体情况具体决定。关于培养基最优化的信息通过在教科书“AppliedMicrobiol.Physiology,A Practical Approach”(eds.P.M.Rhodes,P.F.Stanbury,IRL Press(1997)pp.53-73,ISBN 0199635773)”中。也可以从商业供应商那里选择生长培养基,像是standard 1(Merck)或者BHI(grain heart infusion,DIFCO)或者其他的。
所有培养基组分都要通过加热(1.5bar,120℃,20分钟)或者无菌过滤灭菌。组分可以一起灭菌,或者如果必要的话分开单独灭菌。所有的培养基组分可以在生长的开始就加入,或者可以选择连续性或者分批加入。
培养条件对每个实验分别确定。温度应该在15℃到45℃范围内。温度可以保持恒定,或者在实验中改变。培养基的pH在5到8.5范围内,优选的在大约7.0,并且可以通过培养基中缓冲液的添加来维持。针对这一目的有代表性的缓冲液是磷酸钾缓冲液。合成缓冲液,例如MOPS、HEPES、ACES以及其他的,也可以代替使用或者同时使用。也可以在生长过程中通过添加NaOH或者NH4OH,以维持稳定的培养pH。如果使用像是酵母提取物这样的复杂培养基组分,可以减少添加缓冲液的必要性,这是因为许多复杂化合物具有很强的缓冲能力这一事实。如果使用发酵罐培养微生物,也可以使用氨气控制pH。
孵育时间通常在几小时到几天范围内。这一时间的选取是为了允许在液体培养基中积累最大量的产物。公布的生长实验可是在各种容器中进行,例如微量滴定板、玻璃试管、玻璃摇瓶或者不同大小的玻璃或者金属的发酵罐。为了筛选大量的克隆,微生物应该培养在有挡板或者没有挡板的微量滴定板、玻璃试管或者摇瓶中。优选的使用100ml摇瓶,加入10%(体积)的所需培养基。摇瓶应该放在摇床上摇动(振幅25毫米),速度范围100-300rpm。可以通过保持湿润的空气减少蒸发损失;或者,对蒸发损失进行数学修正。
如果要检测遗传修饰的克隆,那么也应该检测未经修饰的对照克隆或者含有基本质粒但没有任何插入的对照克隆。使用生长在琼脂板上30℃孵育的细胞,例如CM平板(10g/l葡萄糖,2.5g/l NaCl,2g/l尿素,10g/l多胨,5g/l酵母提取物,5g/l肉汁提取物,22g/l琼脂,2M NaOH调至pH 6.8),接种培养基至OD600值为0.5-1.5。培养基的接种可以通过引入来自CM平板的谷氨酸棒杆菌细胞的盐悬浮液,或者通过加入该细菌的液体预培养物实现。
实施例8:突变蛋白质功能的体外分析
酶的活性和动力学参数的测定在技术上是已经很好建立了的。任何对给定的经过改变的酶的活性测定实验,必须适合野生型酶的特殊活性,这完全在技术熟练者的能力之内。关于酶的概括评论,以及关于结构、动力学、原理、方法、应用和确定许多酶活性实例的明确细节,可以在例如以下参考文献中找到:Dixon,M.,and Webb,E.C.,(1979)Enzymes.Longmans:London;Fersht,(1985)Enzyme Structure and Mechanism.Freeman:NewYork;Walsh,(1979)Enzymatic Reaction Mechanisms.Freeman:San Francisco;Price,N.C.,Stevens,L.(1982)Fundamentals of Enzymology.Oxford Univ.Press:Oxford;Boyer,P.D.,ed.(1983)The Enzymes,3rd ed.Academic Press:New York;Bisswanger,H.,(1994)Enzymkinetik,2nd ed.VCH:Weinheim(ISBN 3527300325);Bergmeyer,H.U.,Bergmeyer,J.,Graβ1,M.,eds.(1983-1986)Methods of EnzymaticAhalysis,3rd ed.,vol.I-XII,Verlag Chemie:Weinheim;and Ullmann’s Encyclopediaof Industrial Chemistry(1987)vol.A9,“Enzymes”.VCH:Weinheim,p.352-363。
谷氨酸棒杆菌细胞提取物按照以前所述制备(Park,S.-D.,et al.(1998)Mol.Cells 8,286-294)。胱硫醚β裂合酶按照以下分析。分析混合物含有100mM Tris-HCl(pH8.5),0.1mM NADH,1mML-胱硫醚,5单位L-乳酸脱氢酶和适量的粗提取物。在340纳米监测光度变化。S-(□-氨基乙基)-半胱氨酸(AEC)抗性分析按照Rossol,I.and Pühler,A.(1992)J.Bacteriol.174,2968-77所述进行。不同谷氨酸棒杆菌菌株提取物的胱硫醚β裂合酶分析和同一菌株的AEC抗性分析见以下表5。
表5.胱硫醚β裂合酶的表达a
a酶通过在含有1%葡萄糖的基本培养基中生长至稳定期诱导。收获细胞、破碎并如材料和方法部分分析活性。
b使用MCGC基本培养基。在平板上监测生长。
o细胞在含有40mM S-(□-氨基乙基)-半胱氨酸(AEC)的平板上生长5天。
d本研究中制备的突变体。
e未测定。
metC克隆表达胱硫醚β裂合酶的能力通过酶促实验测定。分析由携带质粒pSL173的谷氨酸棒杆菌ASO19E12细胞制备的粗提取物。携带该质粒的细胞与携带空载体pMT1的细胞相比,胱硫醚β裂合酶活性增加5倍(表5),显然具有基因剂量效应。粗提取物的SDS-PAGE分析显示了Mr约为41,000的推定的胱硫醚β裂合酶带。各个推定的胱硫醚β裂合酶带的强度与互补和酶实验数据(表5)一致。如上所述,metC的一个区域似乎与原来报导的aceD基因几乎一致。因为aecD基因是根据其赋予对赖氨酸毒性类似物S-(□-氨基乙基)-半胱氨酸(AEC)的抗性分离的,我们测定了metC蛋白产物的该活性存在。如表5所示,过量表达胱硫醚β裂合酶的细胞对AEC的抗性增加。携带有突变metC基因(见后文)的菌株完全失去了对AEC的抗性表型。
对O-乙酰高丝氨酸硫化氢解酶的分析如下进行(Belfaiza,J.,et al.(1998)J.Bacteriol.180,250-255;Ravanel,S.,M.Droux,and R.Douce(1995)Arch.Biochem.Biophys.316,572-584;Foglino,M.(1995)Microbiology 141,431-439)。0.1毫升分析混合物含有20mM MOPS-NaOH(pH7.5),10mM O-乙酰高丝氨酸,50mM NaOH中的2mM Na2S,和适量的酶。最后加入Na2S后,用50微升矿物油覆盖混合物。30℃温育30分钟,煮沸混合物3分钟终止反应。反应中产生的高半胱氨酸如前所述进行定量(Yamagata,S.(1987)Method Enzymol.143,478-483.)。取出0j.1毫升反应混合物,与0.1ml H2O,0.6ml饱和NaCl,0.1ml含有67mMKCN的1.5M Na2CO3和0.1ml 2%硝普盐。室温温育1分钟后,520纳米测定光密度。携带多余拷贝的metZ基因如含有metZ基因的质粒的棒杆菌较没有多余metZ基因的棒杆菌细胞,metZ酶活性明显要高。
结合DNA的蛋白质的活性可以通过几种技术上已知的方法测定,例如DNA条带移位分析(也称作凝胶阻滞分析)。这些蛋白质对其他分子表达的作用,可以用报告基因分析测定(例如描述在Kolmar,H.et al.(1995)EMBO J.14:3895-3904中的,及其引用的参考文献)。报告基因测试系统是已知的,并且在原核和真核细胞中的应用都已建立,使用像是beta-半乳糖苷酶、绿色荧光蛋白和几种其他蛋白质这样的酶。
膜转运蛋白质活性的测定可以根据例如描述在Gennis,R.B.(1989)“Pores,Channels and Transporters”,in Biomembrane,Molecular Structureand Function,Springer:Heidelberg,p.85-137;199-234;and 270-322中的那些技术进行。
实施例9:突变蛋白质对于所需产物生产的效果的分析
谷氨酸棒杆菌遗传修饰对于所需化合物(例如氨基酸)生产的作用,可以这样估计,即通过合适条件下(例如以上描述的那些)生长已修饰的微生物,并且分析增加所需产物(例如,氨基酸)生产的培养基和/或细胞组分。这些分析技术对于熟练常规技术者来说是熟知的,包括光谱分析、薄层层析、各种染色方法、酶促方法和微生物方法,以及像是高效液相色谱(Ullman,Encyclopedia of Industrial Chemistry-vol.A2,p.89-90and p.443-613,VCH:Weinheim(1985);Fallon,A.et al.,(1987)“Applications of HPLC in Biochemistry”in:Laboratory Techniques inBiochemistry and Molecular Biology,vol.17;Rehm et al.(1993)Biotechnology,vol.3,Chapter III:“Product recovery and purification”,page 469-714,VCH:Weinheim;Belter,P.A.et al.(1988)Bioseparations:downstream processing for biotechnology,John Wiley and Sons;Kennedy,J.F.and Cabral,J.M.S.(1992)Recovery processes for biological materials,John Wiley and Sons;Shaeiwitz,J.A.and Henry,J.D.(1988)Biochemicalseparations,in:Ulmann’s Encyclopedia of Industrial Chemistry,vol.B3,Chapter 11,page 1-27,VCH:Weinheim;and Dechow,F.J.(1989)Separationand purification techniques in biotechnology,Noyes Publications)这样的分析层析。
除了对最终发酵产物的测定,也可以对用于所需化合物生产的代谢途径的其他组分进行分析,例如中间体和副产物,以确定化合物的总生产效率。分析方法包括培养基中营养物水平(例如,糖类、烃、氮源、磷酸以及其他离子)的测定,生物量组成和生长的测定,生物合成途径常见代谢产物的生产的分析,以及对发酵中产生气体的测定。这些测定的标准方法略述在Applied Microbial Physiology,A Practical Approach,P.M.Rhodes and P.F.Stanbury,eds.,IRL Press,p.103-163;and 165-192(ISBN:0199635773)及其引用的参考文献中。
实施例10:谷氨酸棒杆菌培养物中所需产物的纯化
从谷氨酸棒杆菌细胞中或者上述培养基的上清中回收所需产物,可以通过技术上已知的各种方法进行。如果所需产物不是细胞分泌的,那么可以通过低速离心从培养基中收集细胞,用标准技术裂解细胞,例如机械力或者超声波。离心除去细胞碎片,保留含有可溶蛋白的上清部分用于进一步纯化所需化合物。如果产物是从谷氨酸棒杆菌细胞分泌的,那么用低速离心从培养基中除去细胞,保留上清部分用于进一步纯化。
任何一种纯化方法得到的上清部分,用合适的树脂进行层析,所需分子被层析树脂保留,而样品中的很多杂质不被保留,或者杂质被树脂保留,而样品不被保留。使用相同或者不同的层析树脂,可以根据需要重复这一层析步骤。本领域技术人员可以非常熟练的选择合适的层析树脂,并且熟知这些树脂对于待纯化特定分子最有效的应用。纯化的产物可以用过滤或者超滤浓缩,并且贮存在产物稳定性最大的温度下。
技术上已知的纯化方法非常多,前述的纯化方法并不意味着仅仅局限于此。这些纯化方法描述在,例如Bailey,J.E.& Ollis,D.F.BiochmicalEngineering Fundamentals,McGraw-Hill:New York(1986)中。
分离化合物的特性和纯度,可以技术上的标准技术估计。这包括高效液相色谱(HPLC)、分光方法、染色方法、薄层层析、NIRS、酶促方法或者微生物方法。这些分析方法在以下文献中有评论:Patek et al.(1994)Appl.Environ.Microbiol.60:133-140;Malakhova et al.(1996)Biotekhnologiya 11:27-32;and Schmidt et al.(1998)Bioprocess Engineer.19:67-70.Ulmann’s Encyclopedia of Industrial Chemistry,(1996)vol.A27,VCH:Weinheim,p.89-90,p.521-540,p.540-547,p.559-566,575-581andp.581-587;Michal,G.(1999)Biochemical Pathways:An Atlas ofBiochemistry and Molecular Biology,John Wiley and Sons;Fallon,A.et al.(1987)Applications of HPLC in Biochemistry in:Laboratory Techniques inBiochemistry and Molecular Biology,vol.17。
实施例11:本发明基因序列的分析
序列比较和两条序列之间同源性百分比的测定,是技术上已知的技术,可以使用数学运算法则完成,例如Karlin and Altschul(1990)Proc.Natl.Acad.Sci.USA 87:2264-68中的运算法则,该运算法则在Karlin andAltschul(1993)Proc.Natl.Acad.Sci.USA 90:5873-77中有修改。该运算法则被整合在Altschul,et al.(1990)J.Mol.Biol.215:403-10中的NBLAST和XBLAST程序(2.0版)中。BLAST核苷酸搜寻可以用NBLAST程序进行,score=100,wordlength=12,可以得到与本发明MP核酸分子同源的核苷酸序列。BLAST蛋白质搜寻可以用XBLAST程序进行,score=50,wordlength=3,可以得到与本发明MP蛋白质分子同源的氨基酸序列。出于比较的目的,为了获得有间隙的序列对比,可以使用描述在Altschul et al.,(1997)Nucleic Acids Res.25(17):3389-3402中的GappedBLAST。当使用BLAST和Gapped BLAST程序时,本领域技术人员知道对于特定的待分析序列如何优化程序(例如,XBLAST和NBLAST)的参数。
用于序列比较的另一个数学运算法则实例是,Meyers和Miller运算法则((1998)Comput.Appl.Biosci.4:11-17)。该运算法则被整合在ALIGN程序(2.0版)中,该程序是GCG序列序列对比软件包的一部分。当使用ALIGN程序比较氨基酸序列时,可以使用PAM120重量残基表、间隙长度处罚12、间隙处罚4。其他的序列分析运算法则在技术上也是已知的,包括ADVANCE和ADAM,叙述在Torelli and Robotti(1994)Comput.Appl.Biosci.10:3-5中;和FASTA,叙述在Pearson and Lipman(1998)P.N.A.S.85:2444-8中。
两条氨基酸序列之间的百分比同源性也可以使用GCG软件包(http://www.gcg.com有提供)中的GAP程序实现,使用Blosum 62矩阵或者PAM 250矩阵,间隙分量12、10、8、6或者4,长度分量2、3或者4。两条核酸序列之间的百分比同源性可以使用GCG软件包中的GAP程序实现,使用标准参数,例如间隙分量50和长度分量3。
本发明基因序列与Genbank中序列之间的比较分析,可以使用技术上已知的技术进行(参见,例如,Bexevanis and Ouellette,eds.(1998)Bioinformatics:A Practical Guide to the Analysis of Genes and Proteins.John Wiley and Sons:New York)。本发明基因序列,通过三个步骤的方法与Genbank中的序列进行比较。在第一步中,对本发明的每一条序列相对Genbank中的核苷酸序列进行BLASTN分析(例如,本地序列对比分析),保留最高的500个匹配作进一步分析。然后对这500个匹配作FASTA搜寻(例如,本地与全世界的组合序列对比分析,在其中对限定的序列区域进行序列对比)。接下来,对本发明的每条基因序列与FASTA的三个最高匹配,使用GCG软件包中的GAP程序(使用标准参数)进行全世界序列对比。为了得到正确结果,从Genbank选出的序列长度,使用技术上熟知的方法调节为查询序列的长度。该分析的结果列在表4中。虽然这样得到的结果,与对本发明每条基因相对于Genbank每条对照所进行的单独GAP(全世界)分析得到的结果,是一致的,但是相对于大数据库的GAP(全世界)分析来说,所需的计算时间大大减少。没有得到截止值以上序列对比的本发明序列,在表4中表明,缺少序列对比信息。本领域技术人员能够深一层的理解,在表4中列出的标题“%homology(GPA)”下的GAP序列对比同源性百分比,是以欧洲数字格式列出的,其中‘,’代表十进制点。例如,该列中的值“40,345”代表“40.345%”。
实施例12:DNA微阵列的构建和操作
本发明的序列还可以用于DNA微阵列(DNA阵列的设计、方法和应用技术上是熟知的,描述在,例如,Schena,M.te al.(1995)Science 270:467-470;Wodicka,L.et al.(1997)Nature Biotechnology 15:1359-1367;DeSaizieu,A.et al.(1998)Nature Biotechnology 16:45-48;and DeRisi,J.L.et al.(1997)Science 278:680-686)的构建和应用。
DNA微阵列使用固体或者可弯曲的支持物,包括硝酸纤维素、尼龙、玻璃、硅或者其他材料。核酸分子可以以有序的方式连接在表面。合适标记之后,其他核酸或者核酸混合物可以与固定的核酸分子杂交,标记可以用于监控和测量确定区域杂交分子的单独的信号强度。本方法允许同时定量适用的核酸样品或者混合物中的全部或者所选择核酸的相对或者绝对数量。因此,DNA微阵列允许多种(多至6800或者更多)类似核酸表达的分析(参见例如,Schena,M.(1996)BioEssays 18(5):427-431)。
本发明序列可以用于设计寡聚核苷酸引物,这些引物可以通过像聚合酶链式反应这样的核酸扩增反应扩增一条或者多条谷氨酸棒杆菌基因的确定区域。5’或者3’寡聚核苷酸引物或者合适连接体的选择和设计,允许得到的PCR产物共价连接到上述支持介质的表面(也描述在,例如,Schena,M.et al.(1995)Science 270:467-470)。
核酸微阵列也可以通过如在Wodicka,L.et al.(1997)NatureBiotechnology 15:1359-1367中描述的原位寡聚核苷酸合成构建。通过照相平板方法,可将矩阵中精确确定的区域暴露在光线中。保护基团是光不稳定的,从而被激活并经受核苷酸添加,但是被掩饰而见不到光的区域不进行任何修饰。接下来的保护和光激活循环,允许在确定位置不同寡聚核苷酸的合成。本发明确定的小区域可以在微阵列上通过固相寡聚核苷酸合成而合成。
出现在样品或者核苷酸混合物中的本发明核酸分子,可以与微阵列杂交。可以根据标准方法标记这些核酸分子。简单的说,核酸分子(例如,mRNA分子或者DNA分子)可以通过与同位素或者荧光标记的核苷酸结合而被标记,例如,在逆转录或者DNA合成中。标记核酸与微阵列的杂交有描述(例如在Schena,M.et al.(1995)supra;Wodicka,L.et al.(1997),supra;and DeSaizieu A.et al.(1998),supra中)。杂交分子的检测和定量要适合特定的结合标记。放射性标记可被探测,例如,在Schena,M.et al.(1995)supra中描述的,荧光标记也可以探测,例如使用Shalon etal.(1996)Gemone Research 6:639-645的方法。
如上所述,本发明序列在DNA微阵列中的应用,允许不同的谷氨酸棒杆菌菌株或者其他棒杆菌的比较分析。例如,通过核酸阵列方法,可以促进基于个别转录分部图的菌株内改变的研究,以及促进对特定和/或所需的像是致病性、生产能力和压力承受能力这样的菌株性质重要的基因的鉴定。同样,使用核酸阵列技术,也可以比较发酵反应过程中本发明基因表达的分部图。
实施例13:细胞蛋白质群体动力学的分析(蛋白质组学)
本发明的基因、组成和方法,可以用于研究蛋白质群体的相互作用和动力学,称作“蛋白质组学”。感兴趣的蛋白质群体包括,但是不局限于,谷氨酸棒杆菌的全部蛋白质群体(例如,和其他生物体的蛋白质群体比较起来),在特殊环境或者代谢条件下(例如,发酵中、高温或者低温、或者高pH或低pH)有活性的那些蛋白质,或者在特定生长或者发育阶段有活性的那些蛋白质。
可以用各种熟知的技术分析蛋白质群体,例如凝胶电泳。细胞蛋白质可以通过例如裂解或者提取获得,也可以使用各种电泳技术彼此分离。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分离蛋白质,很大程度上基于它们的分子重量。等电聚焦聚丙烯酰胺凝胶电泳(IEF-PAGE)通过等点点(这不仅反映了氨基酸序列,而且放映了蛋白质的翻译后修饰)分离蛋白质。另一种更加优选的蛋白质分析方法是,IEF-PAGE和SDS-PAGE的连续结合,称为2-D-凝胶电泳(在例如Hermann et al.(1998)Electrophoresis 19:3217-3221;Fountoulakis et al.(1998)Electrophoresis 19:1193-1202;Langen etal.(1997)Electrophoresis 18:1184-1192;Antelmann et al.(1997)Electrophoresis18:1451-1463中有描述)。
用这些方法分离的蛋白质可以通过标准技术显现,例如通过染色或者标记。合适的染色在技术上是已知的,包括考马斯亮蓝、银染或者荧光染料,例如Sypro Ruby(Molecular Probes)。谷氨酸棒杆菌培养基中包含有放射性标记的氨基酸或者其他蛋白质前体(例如,35S-甲硫氨酸,35S-半胱氨酸,14C-标记氨基酸,15N-氨基酸,15NO3或者15NH4 +或者13C-标记氨基酸),可以使得这些细胞在其蛋白质分离之前就标记蛋白质。类似的,也可以使用荧光标记。根据前述技术可以提取、隔离和分离这些标记蛋白质。
用这些技术显现的蛋白质,可以通过测量所用的染料或者标记作进一步分析。特定蛋白质的数量可以使用例如光学方法,进行定量确定,并且可以与在同一块凝胶上或者其他凝胶上的其他蛋白质的数量进行比较。可以通过例如光学比较、分光分析、凝胶图象分析和扫描,或者通过使用照相胶片或者显示器,对凝胶上的蛋白质进行比较。这些技术在技术上是熟知的。
为了确定特定蛋白质的特性,可以使用直接序列测定或者其他标准技术。例如,可以使用N-和/或C-末端氨基酸测序(例如Edman降解),以及质谱分析(特别是MALDI或者ESI技术(参见例如,Langen et al.(1997)Electrophoresis 18:1184-1192))。此处提供的蛋白质序列,可以用作通过这些技术进行的谷氨酸棒杆菌蛋白质鉴定。
通过这些技术得到的信息,可以用于比较蛋白质存在、活性、不同生物条件下(例如,在其他条件中的不同生物体、发酵时间点、培养基条件、或者生物环境)不同样品间修饰的各种模式。这些试验得到的数据,可以单独的,或者与其他技术相结合的用于各种应用,例如比较特定情况下(例如代谢情况)各种生物体的行为,增加生产精细化学物质的菌株的生产能力,或者增加精细化学物质生产的效率。
实施例14:利用聚合酶反应(PCR)克隆基因
可以使用含有与谷氨酸棒杆菌或其他菌株序列同源的核苷酸序列或本领域熟知的限制酶识别位点的特异性寡核苷酸扩增基因(例如可参见Sambrook,J.,Fritsh,E.F.,and Maniatis,T.Molecular Cloning:ALaboratory Manual.2nd,ed,Cold Spring Harbor Laboratoy,Cold SpringHarbor Laboratory Press,Cold Spring Harbor,NY,1989)。这些可以用来扩增含有上述菌株部分基因组的特异性DNA片段,使用以下DNA聚合酶如水生栖热菌DNA聚合酶、P.furiosus DNA聚合酶或P.woesei DNA聚合酶,在适量的缓冲溶液中含有dNTP,按照厂商说明进行操作。
基因片段如包括基因编码区中不存在的合适上游和下游区域的RXA00657编码序列可以使用上述技术扩增。而且,这些片段可以由未整合的寡核苷酸和核苷酸中纯化。DNA限制酶可以用来产生突出末端,用来连接DNA片段至用互补性酶或匹配的酶消化的载体上,所述酶产生可以用来连接DNA至Sinskey等美国专利4649119中所述的载体中的末端,使用谷氨酸棒杆菌和相关的短杆菌(如乳发酵短杆菌)遗传操作技术见以下文献(Yoshihama et al,J.Bacteriol.162:591-597(1985);Katsumata et al.,J.Bacteriol.159:306-311(1984);and Santamaria et al.,J.Gen.Microbiol.130:2237-2246(1984)。用于扩增RXA00657上游DNA序列、编码区序列和下游区域的引物如下:
TCGGGTATCCGCGCTACACTTAGA(SEQ ID NO:121);
GGAAACCGGGGCATCGAAACTTA (SEQ ID NO:122)
200ng谷氨酸棒杆菌染色体DNA用作模板,反应体积100μl,含有2,5UPfu Turbo-PolymeraseTM(StratageneTM),和200μM dNTP-核苷酸。在PCR-CyclerTM(Perkin Elmer 2400TM)上进行PCR,使用以下温度/时间循环:
1个循环:94℃:2min,;
20个循环:94℃:1min.;
52℃:1min,72℃:1.5min.,
1个循环:72℃:5min.
从获得的扩增DNA片段中除去引物,所得的片段克隆进pBS KS(StratageneTM)平端EcoRV位点。用限制酶BamHI/XhoI消化切下片段,并连接到BamHI SalI消化的载体pB(SEQ ID NO.:125)中。所得载体称为pBRXA00657。
所得重组载体使用标准技术分析,具体可参见Sambrook,J.,Fritsh,E.F.,and Maniatis,T.Molecular Cloning:ALaboratory Manual.2nd,ed.,ColdSpring Harbor Laboratory,Cold Spring Harbor Laboratory Press,Cold SpringHarbor,NY,1989),也可以使用上述技术转移进谷氨酸棒杆菌中。
棒杆菌菌株(ATCC 13286)如上所述转化处理。棒杆菌菌株转化可以通过原生质体转化(Kastsumata,R.et al.(1984)J.Bacteriol.159306-311),电穿孔(Liebl,E.et al.(1989)FEMSMicrobiol.Letters,53:399-303)实现,在使用特殊载体时,也可以通过辍合(conjugation)实现(见例如A.et al.(1990)J.Bacteriol.172:1663-1666)。也可以将穿梭载体由谷氨酸棒杆菌转化到大肠杆菌中,由谷氨酸棒杆菌中制备质粒DNA(使用本领域熟知的标准方法)并转化进大肠杆菌中。该转化步骤可以通过标准方法实现,使用Mcr-缺陷大肠杆菌菌株如NM522(Gough&Murray(1983)J.Mol.Biol.166:1-19)更为有利。
细菌菌株如谷氨酸棒杆菌菌株(ATCC 13286)的转化使用含有前述RXA00657DNA区域的(SEQ ID NO.:6)质粒pB或不含插入核酸的载体pB(SEQ ID NO.:)进行。
所得菌株铺平板并由CM培养基中分离,培养基含有10g/l葡萄糖,2,5g/l NaCl,2,0g/l尿素,10g/l细菌用蛋白胨(Difco/Becton Dicinson/SparksUSATM),5g/l酵母提取物(Difco/Becton Dicinson/Sparks USATM),5g/l肉提取物(Difco/Becton Dicinson/Sparks USATM),22g/l琼脂(Difco/BectonDickinson/Sparks USATM)和15μg/ml硫酸卡那霉素(Serva,Germany),用NaOH调pH至6.8。
由上述琼脂培养基中分离的菌株以10毫升接种于没有覆盖(baffles)的100ml烧瓶中的液体培养基中,培养基中含有100g/蔗糖,50g/l(NH4)2SO4,2,5g/NaCl,2,0g/l尿素,10g/l细菌用蛋白胨(Difco/BectonDickinson/Sparks USA),5g/酵母提取物(Difco/Becton Dickinson/SparksUSA),5g/l肉提取物(Difco/Becton Dickinson/Sparks USA),和25g/l CaCO3(Riedel de Haen,Germany)。用NaOH调pH至6.8。
菌株30℃温育48h。EppendorfTM离心机中12,000rpm离心20′制备上清。稀释液体上清并进行氨基酸分析(这些测定的标准方法见AppliedMicrobial Physiology,A Practical Approach,P.M.Rhodes and P.F.Stanbury,eds.,IRL Press,p.103-129;131-163;and 165-192(ISBN:0199635773)及其中引用的文献)。
结果见以下表6。
表6
菌株ATCC13286 | 含有的质粒 | pB | pB RXA00657 |
产生的赖氨酸(g/l) | 13.5 | 14.93 | |
选择性(mol赖氨酸/mol消耗的糖) | 0.235 | 0.25 |
等同声明
本领域技术人员可以认识到,或者能够确定仅仅使用常规实验,此处描述的本发明的特定实施方案有很多等价物。下面的权利要求意图包含这些等价物。
表3:可用于本发明实施的棒杆菌和短杆菌菌株
属 | 种 | ATCC | EERM | NRRL | CECT | NCIMB | CRS | NCTC | DSMZ | |
短杆菌 | 产氨短杆菌 | 21054 | ||||||||
短杆菌 | 产氨短杆菌 | 19350 | ||||||||
短杆菌 | 产氨短杆菌 | 19351 | ||||||||
短杆菌 | 产氨短杆菌 | 19352 | ||||||||
短杆菌 | 产氨短杆菌 | 19353 | ||||||||
短杆菌 | 产氨短杆菌 | 19354 | ||||||||
短杆菌 | 产氨短杆菌 | 19355 | ||||||||
短杆菌 | 产氨短杆菌 | 19356 | ||||||||
短杆菌 | 产氨短杆菌 | 21055 | ||||||||
短杆菌 | 产氨短杆菌 | 21077 | ||||||||
短杆菌 | 产氨短杆菌 | 21553 | ||||||||
短杆菌 | 产氨短杆菌 | 21580 | ||||||||
短杆菌 | 产氨短杆菌 | 39101 | ||||||||
短杆菌 | butanicum | 21196 | ||||||||
短杆菌 | 分歧短杆菌 | 21792 | P928 | |||||||
短杆菌 | 黄色短杆菌 | 21474 | ||||||||
短杆菌 | 黄色短杆菌 | 21129 | ||||||||
短杆菌 | 黄色短杆菌 | 21518 | ||||||||
短杆菌 | 黄色短杆菌 | B11474 | ||||||||
短杆菌 | 黄色短杆菌 | B11472 | ||||||||
短杆菌 | 黄色短杆菌 | 21127 | ||||||||
短杆菌 | 黄色短杆菌 | 21128 | ||||||||
短杆菌 | 黄色短杆菌 | 21427 | ||||||||
短杆菌 | 黄色短杆菌 | 21475 | ||||||||
短杆菌 | 黄色短杆菌 | 21517 | ||||||||
短杆菌 | 黄色短杆菌 | 21528 | ||||||||
短杆菌 | 黄色短杆菌 | 21529 | ||||||||
短杆菌 | 黄色短杆菌 | B11477 | ||||||||
短杆菌 | 黄色短杆菌 | B11478 | ||||||||
短杆菌 | 黄色短杆菌 | 21127 | ||||||||
短杆菌 | 黄色短杆菌 | B11474 | ||||||||
短杆菌 | 希氏短杆菌 | 15527 | ||||||||
短杆菌 | 酮戊二酸短杆菌 | 21004 | ||||||||
短杆菌 | 酮戊二酸短杆菌 | 21089 | ||||||||
短杆菌 | ketosoreductum | 21914 | ||||||||
短杆菌 | 乳发酵短杆菌 | 70 | ||||||||
短杆菌 | 乳发酵短杆菌 | 74 | ||||||||
短杆菌 | 乳发酵短杆菌 | 77 | ||||||||
短杆菌 | 乳发酵短杆菌 | 21798 | ||||||||
短杆菌 | 乳发酵短杆菌 | 21799 | ||||||||
短杆菌 | 乳发酵短杆菌 | 21800 | ||||||||
短杆菌 | 乳发酵短杆菌 | 21801 | ||||||||
短杆菌 | 乳发酵短杆菌 | B11470 | ||||||||
短杆菌 | 乳发酵短杆菌 | B11471 |
属 | 种 | ATCC | FERM | NRRL | CECT | NCIMB | CBS | NCTC | DSMZ | |
短杆菌 | 乳发酵短杆菌 | 21086 | ||||||||
短杆菌 | 乳发酵短杆菌 | 21420 | ||||||||
短杆菌 | 乳发酵短杆菌 | 21086 | ||||||||
短杆菌 | 乳发酵短杆菌 | 31269 | ||||||||
短杆菌 | 扩展短杆菌 | 9174 | ||||||||
短杆菌 | 扩展短杆菌 | 19391 | ||||||||
短杆菌 | 扩展短杆菌 | 8377 | ||||||||
短杆菌 | paraffinolyticum | 11160 | ||||||||
短杆菌 | 短杆菌属种. | 717.73 | ||||||||
短杆菌 | 短杆菌属种. | 717.73 | ||||||||
短杆菌 | 短杆菌属种. | 14604 | ||||||||
短杆菌 | 短杆菌属种. | 21860 | ||||||||
短杆菌 | 短杆菌属种. | 21864 | ||||||||
短杆菌 | 短杆菌属种. | 21865 | ||||||||
短杆菌 | 短杆菌属种. | 21866 | ||||||||
短杆菌 | 短杆菌属种. | 19240 | ||||||||
棒杆菌 | 嗜乙酰乙酸棒杆菌 | 21476 | ||||||||
棒杆菌 | 嗜乙酰乙酸棒杆菌 | 13870 | ||||||||
棒杆菌 | 乙酰谷氨酸棒杆菌 | B11473 | ||||||||
棒杆菌 | 乙酰谷氨酸棒杆菌 | B11475 | ||||||||
棒杆菌 | 乙酰谷氨酸棒杆菌 | 15806 | ||||||||
棒杆菌 | 乙酰谷氨酸棒杆菌 | 21491 | ||||||||
棒杆菌 | 乙酰谷氨酸棒杆菌 | 31270 | ||||||||
棒杆菌 | 嗜乙酰棒杆菌 | B3671 | ||||||||
棒杆菌 | 产氨棒杆菌 | 6872 | 2399 | |||||||
棒杆菌 | 产氨棒杆菌 | 15511 | ||||||||
棒杆菌 | fujiokense | 21496 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 14067 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 39137 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21254 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21255 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 31830 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13032 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 14305 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 15455 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13058 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13059 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13060 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21492 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21513 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21526 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21543 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13287 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21851 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21253 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21514 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21516 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21299 |
属 | 种 | ATCC | FERM | NRRI | CECT | NCIMB | CBS | NCTC | DSMZ | |
棒杆菌 | 谷氨酸棒杆菌 | 21300 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 39684 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21488 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21649 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21650 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19223 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13869 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21157 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21158 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21159 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21355 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 31808 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21674 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21562 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21563 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21564 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21565 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21566 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21567 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21568 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21569 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21570 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21571 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21572 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21573 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21579 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19049 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19050 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19051 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19052 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19053 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19054 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19055 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19056 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19057 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19058 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19059 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19060 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 19185 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 13286 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21515 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21527 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21544 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21492 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | B8183 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | B8182 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | B12416 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | B12417 |
属 | 种 | ATCC | FERM | NRRI | CECT | NCIMB | CBS | NCTC | DSMZ | Otherorigin |
棒杆菌 | 谷氨酸棒杆菌 | B12418 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | B11476 | ||||||||
棒杆菌 | 谷氨酸棒杆菌 | 21608 | ||||||||
棒杆菌 | 百合花棒杆菌 | P973 | ||||||||
棒杆菌 | nitrilophilus | 21419 | 11594 | |||||||
棒杆菌 | 棒杆菌属种 | P4445 | ||||||||
棒杆菌 | 棒杆菌属种 | P4446 | ||||||||
棒杆菌 | 棒杆菌属种 | 31088 | ||||||||
棒杆菌 | 棒杆菌属种 | 31089 | ||||||||
棒杆菌 | 棒杆菌属种 | 31090 | ||||||||
棒杆菌 | 棒杆菌属种 | 31090 | ||||||||
棒杆菌 | 棒杆菌属种 | 31090 | ||||||||
棒杆菌 | 棒杆菌属种 | 15954 | 20145 | |||||||
棒杆菌 | 棒杆菌属种 | 21857 | ||||||||
棒杆菌 | 棒杆菌属种 | 21862 | ||||||||
棒杆菌 | 棒杆菌属种 | 21863 | ||||||||
棒杆菌 | 谷氨酸棒杆菌* | ASO19 | ||||||||
棒杆菌 | 谷氨酸棒杆菌** | ASO19E12 | ||||||||
棒杆菌 | 谷氨酸棒杆菌*** | HL457 | ||||||||
棒杆菌 | 谷氨酸棒杆菌**** | HL459 |
ATCC:美国典型培养物保藏中心,Rockville,MD,USA
FERM:发酵研究所,Chiba,Japan
NRRL;ARS培养物保藏中心,北方区域研究实验室,Peoria,IL,USA
CECT:西班牙典型培养物保藏中心,Valencia,Spain
NCIMB:国立工业和海洋微生物保藏有限公司,Aberdeen,UK
CBS:真菌菌种保藏中心,Baarn,NL
NCTC:国立典型培养物保藏中心,London,UK
DSMZ:德意志微生物保藏中心,Braunschweig,Germany
具体可参见Sugawara,H.et al.(1993)World directory ofcollections ofcultures of microorganisms:Bacteria,fungi and yeasts(4th edn),World federation for culture collections world data center on microorganisms,Saimata,Japen.
*谷氨酸棒杆菌ATCC13059d的自发利福平抗性突变体Yoshihama et al.,1985
**ASO19的限制缺陷变体Follettie et al.,1993
***ASO19E12metC-破坏突变体 本研究
****ASO19E12metC-破坏突变体 本研究
序列表
<110>BASF公司(BASF Aktiengesellschaft)
<120>编码代谢途径蛋白的谷氨酸棒杆菌基因
<130>BGI-121CP2PC
<140>
<141>
<150>09/606740
<151>2000-06-23
<150>60/187970
<151>2000-03-09
<160>125
<170>PatentIn Vers.2.0
<210>1
<211>1840
<212>DNA
<213>谷氨酸棒杆菌(Corynebacterium glutamicum)
<220>
<221>CDS
<222>(363)..(1676)
<400>1
cagaaactgt gtgcagaaat gcatgcagaa aaaggaaagt tcgggccaag atgggtgttt 60
ctgtatgccg atgatcggat ctttgacagc tgggtatgcg acaaatcacc gagagttgtt 120
aattcttaac aatggaaaag taacattgag agatgattta taccatcctg caccatttag 180
agtggggcta gtcatacccc cataacccta gctgtacgca atcgatttca aatcagttgg 240
aaaaagtcaa gaaaattacc cgagaattaa tttataccac acagtctatt gcaatagacc 300
aagctgttca gtagggtgca tgggagaaga atttcctaat aaaaactctt aaggacctcc 360
aa atg cca aag tac gac aat tcc aat gct gac cag tgg ggc ttt gaa 407
Met Pro Lys Tyr Asp Asn Ser Asn Ala Asp Gln Trp Gly Phe Glu
1 5 10 15
acc cgc tcc att cac gca ggc cag tca gta gac gca cag acc agc gca 455
Thr Arg Ser Ile His Ala Gly Gln Ser Val Asp Ala Gln Thr Ser Ala
20 25 30
cga aac ctt ccg atc tac caa tcc acc gct ttc gtg ttc gac tcc gct 503
Arg Asn Leu Pro Ile Tyr Gln Ser Thr Ala Phe Val Phe Asp Ser Ala
35 40 45
gag cac gcc aag cag cgt ttc gca ctt gag gat cta ggc cct gtt tac 551
Glu His Ala Lys Gln Arg Phe Ala Leu Glu Asp Leu Gly Pro Val Tyr
50 55 60
tcc cgc ctc acc aac cca acc gtt gag gct ttg gaa aac cgc atc gct 599
Ser Arg Leu Thr Asn Pro Thr Va1 Glu Ala Leu Glu Asn Arg Ile Ala
65 70 75
tcc ctc gaa ggt ggc gtc cac gct gta gcg ttc tcc tcc gga cag gcc 647
Ser Leu Glu Gly Gly Val His Ala Val Ala Phe Ser Ser Gly Gln Ala
80 85 90 95
gca acc acc aac gcc att ttg aac ctg gca gga gcg ggc gac cac atc 695
Ala Thr Thr Asn Ala Ile Leu Asn Leu Ala Gly Ala Gly Asp His Ile
100 105 110
gtc acc tcc cca cgc ctc tac ggt ggc acc gag act cta ttc ctt atc 743
Val Thr Ser Pro Arg Leu Tyr Gly Gly Thr Glu Thr Leu Phe Leu Ile
115 120 125
act ctt aac cgc ctg ggt atc gat gtt tcc ttc gtg gaa aac ccc gac 791
Thr Leu Asn Arg Leu Gly Ile Asp Val Ser Phe Val Glu Asn Pro Asp
130 135 140
gac cct gag tcc tgg cag gca gcc gtt cag cca aac acc aaa gca ttc 839
Asp Pro Glu Ser Trp Gln Ala Ala Val Gln Pro Asn Thr Lys Ala Phe
145 150 155
ttc ggc gag act ttc gcc aac cca cag gca gac gtc ctg gat att cct 887
Phe Gly Glu Thr Phe Ala Asn Pro Gln Ala Asp Val Leu Asp Ile Pro
160 165 170 175
gcg gtg gct gaa gtt gcg cac cgc aac agc gtt cca ctg atc atc gac 935
Ala Val Ala Glu Val Ala His Arg Asn Ser Val Pro Leu Ile Ile Asp
180 185 190
aac acc atc gct acc gca gcg ctc gtg cgc ccg ctc gag ctc ggc gca 983
Asn Thr Ile Ala Thr Ala Ala Leu Val Arg Pro Leu Glu Leu Gly Ala
195 200 205
gac gtt gtc gtc gct tcc ctc acc aag ttc tac acc ggc aac ggc tcc 1031
Asp Val Val Val Ala Ser Leu Thr Lys Phe Tyr Thr Gly Asn Gly Ser
210 215 220
gga ctg ggc ggc gtg ctt atc gac ggc gga aag ttc gat tgg act gtc 1079
Gly Leu Gly Gly Val Leu Ile Asp Gly Gly Lys Phe Asp Trp Thr Val
225 230 235
gaa aag gat gga aag cca gta ttc ccc tac ttc gtc act cca gat gct 1127
Glu Lys Asp Gly Lys Pro Val Phe Pro Tyr Phe Val Thr Pro Asp Ala
240 245 250 255
gct tac cac gga ttg aag tac gca gac ctt ggt gca cca gcc ttc ggc 1175
Ala Tyr His Gly Leu Lys Tyr Ala Asp Leu Gly Ala Pro Ala Phe Gly
260 265 270
ctc aag gtt cgc gtt ggc ctt cta cgc gac acc ggc tcc acc ctc tcc 1223
Leu Lys Val Arg Val Gly Leu Leu Arg Asp Thr Gly Ser Thr Leu Ser
275 280 285
gca ttc aac gca tgg gct gca gtc cag ggc atc gac acc ctt tcc ctg 1271
Ala Phe Asn Ala Trp Ala Ala Val Gln Gly Ile Asp Thr Leu Ser Leu
290 295 300
cgc ctg gag cgc cac aac gaa aac gcc atc aag gtt gca gaa ttc ctc 1319
Arg Leu Glu Arg His Asn Glu Asn Ala Ile Lys Val Ala Glu Phe Leu
305 310 315
aac aac cac gag aag gtg gaa aag gtt aac ttc gca ggc ctg aag gat 1367
Asn Asn His Glu Lys Val Glu Lys Val Asn Phe Ala Gly Leu Lys Asp
320 325 330 335
tcc cct tgg tac gca acc aag gaa aag ctt ggc ctg aag tac acc ggc 1415
Ser Pro Trp Tyr Ala Thr Lys Glu Lys Leu Gly Leu Lys Tyr Thr Gly
340 345 350
tcc gtt ctc acc ttc gag atc aag ggc ggc aag gat gag gct tgg gca 1463
Ser Val Leu Thr Phe Glu Ile Lys Gly Gly Lys Asp Glu Ala Trp Ala
355 360 365
ttt atc gac gcc ctg aag cta cac tcc aac ctt gca aac atc ggc gat 1511
Phe Ile Asp Ala Leu Lys Leu His Ser Asn Leu Ala Asn Ile Gly Asp
370 375 380
gtt cgc tcc ctc gtt gtt cac cca gca acc acc acc cat tca cag tcc 1559
Val Arg Ser Leu Val Val His Pro Ala Thr Thr Thr His Ser Gln Ser
385 390 395
gac gaa gct ggc ctg gca cgc gcg ggc gtt acc cag tcc acc gtc cgc 1607
Asp Glu Ala Gly Leu Ala Arg Ala Gly Val Thr Gln Ser Thr Val Arg
400 405 410 415
ctg tcc gtt ggc atc gag acc att gat gat atc atc gct gac ctc gaa 1655
Leu Ser Val Gly Ile Glu Thr Ile Asp Asp Ile Ile Ala Asp Leu Glu
420 425 430
ggc ggc ttt gct gca atc tag ctttaaatag actcacccca gtgcttaaag 1706
Gly Gly Phe Ala Ala Ile
435
cgctgggttt ttctttttca gactcgtgag aatgcaaact agactagaca gagctgtcca 1766
tatacactgg acgaagtttt agtcttgtcc acccagaaca ggcggttatt ttcatgccca 1826
ccctcgcgcc ttca 1840
<210>2
<211>437
<212>PRT
<213>谷氨酸棒杆菌
<400>2
Met Pro Lys Tyr Asp Asn Ser Asn Ala Asp Gln Trp Gly Phe Glu Thr
1 5 10 15
Arg Ser Ile His Ala Gly Gln Ser Val Asp Ala Gln Thr Ser Ala Arg
20 25 30
Asn Leu Pro Ile Tyr Gln Ser Thr Ala Phe Val Phe Asp Ser Ala Glu
35 40 45
His Ala Lys Gln Arg Phe Ala Leu Glu Asp Leu Gly Pro Va1 Tyr Ser
50 55 60
Arg Leu Thr Asn Pro Thr Val Glu Ala Leu Glu Asn Arg Ile Ala Ser
65 70 75 80
Leu Glu Gly Gly Val His Ala Val Ala Phe Ser Ser Gly Gln Ala Ala
85 90 95
Thr Thr Asn Ala Ile Leu Asn Leu Ala Gly Ala Gly Asp His Ile Val
100 105 110
Thr Ser Pro Arg Leu Tyr Gly Gly Thr Glu Thr Leu Phe Leu Ile Thr
115 120 125
Leu Asn Arg Leu Gly Ile Asp Val Ser Phe Val Glu Asn Pro Asp Asp
130 135 140
Pro Glu Ser Trp Gln Ala Ala Val Gln Pro Asn Thr Lys Ala Phe Phe
145 150 155 160
Gly Glu Thr Phe Ala Asn Pro Gln Ala Asp Val Leu Asp Ile Pro Ala
165 170 175
Val Ala Glu Val Ala His Arg Asn Ser Val Pro Leu Ile Ile Asp Asn
180 185 190
Thr Ile Ala Thr Ala Ala Leu Val Arg Pro Leu Glu Leu Gly Ala Asp
195 200 205
Val Val Val Ala Ser Leu Thr Lys Phe Tyr Thr Gly Asn Gly Ser Gly
210 215 220
Leu Gly Gly Val Leu Ile Asp Gly Gly Lys Phe Asp Trp Thr Val Glu
225 230 235 240
Lys Asp Gly Lys Pro Val Phe Pro Tyr Phe Val Thr Pro Asp Ala Ala
245 250 255
Tyr His Gly Leu Lys Tyr Ala Asp Leu Gly Ala Pro Ala Phe Gly Leu
260 265 270
Lys Val Arg Val Gly Leu Leu Arg Asp Thr Gly Ser Thr Leu Ser Ala
275 280 285
Phe Asn Ala Trp Ala Ala Val Gln Gly Ile Asp Thr Leu Ser Leu Arg
290 295 300
Leu Glu Arg His Asn Glu Asn Ala Ile Lys Val Ala Glu Phe Leu Asn
305 310 315 320
Asn His Glu Lys Val Glu Lys Val Asn Phe Ala Gly Leu Lys Asp Ser
325 330 335
Pro Trp Tyr Ala Thr Lys Glu Lys Leu Gly Leu Lys Tyr Thr Gly Ser
340 345 350
Val Leu Thr Phe Glu Ile Lys Gly Gly Lys Asp Glu Ala Trp Ala Phe
355 360 365
Ile Asp Ala Leu Lys Leu His Ser Asn Leu Ala Asn Ile Gly Asp Val
370 375 380
Arg Ser Leu Val Val His Pro Ala Thr Thr Thr His Ser Gln Ser Asp
385 390 395 400
Glu Ala Gly Leu Ala Arg Ala Gly Val Thr Gln Ser Thr Val Arg Leu
405 410 415
Ser Val Gly Ile Glu Thr Ile Asp Asp Ile Ile Ala Asp Leu Glu Gly
420 425 430
Gly Phe Ala Ala Ile
435
<210>3
<211>1495
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(287)..(1264)
<400>3
ccatggtttc ctcagcggaa acggcttggc tatcagcact ttcacccgaa cagcctgcaa 60
gaagtgcgac ggctaacagg gctgggattg tcctcaactt cacttcgggc tccttcttag 120
taataggttc gtagaaaagt ttactagcct agagagtatg cgatttcctg aactcgaaga 180
attgaagaat cgccggacct tgaaatggac ccggtttcca gaagacgtgc ttcctttgtg 240
ggttgcggaa agtgattttg gcacctgccc gcagttgaag gaagct atg gca gat 295
Met Ala Asp
1
gcc gtt gag cgc gag gtc ttc gga tac cca cca gat gct act ggg ttg 343
Ala Val Glu Arg Glu Val Phe Gly Tyr Pro Pro Asp Ala Thr Gly Leu
5 10 15
aat gat gcg ttg act gga ttc tac gag cgt cgc tat ggg ttt ggc cca 391
Asn Asp Ala Leu Thr Gly Phe Tyr Glu Arg Arg Tyr Gly Phe Gly Pro
20 25 30 35
aat ccg gaa agt gtt ttc gcc att ccg gat gtg gtt cgt ggc ctg aag 439
Asn Pro Glu Ser Val Phe Ala Ile Pro Asp Va1 Val Arg Gly Leu Lys
40 45 50
ctt gcc att gag cat ttc act aag cct ggt tcg gcg atc att gtg ccg 487
Leu Ala Ile Glu His Phe Thr Lys Pro Gly Ser Ala Ile Ile Val Pro
55 60 65
ttg cct gca tac cct cct ttc att gag ttg cct aag gtg act ggt cgt 535
Leu Pro Ala Tyr Pro Pro Phe Ile Glu Leu Pro Lys Val Thr Gly Arg
70 75 80
cag gcg atc tac att gat gcg cat gag tac gat ttg aag gaa att gag 583
Gln Ala Ile Tyr Ile Asp Ala His Glu Tyr Asp Leu Lys Glu Ile Glu
85 90 95
aag gcc ttc gct gac ggt gcg gga tca ctg ttg ttc tgc aat cca cac 631
Lys Ala Phe Ala Asp Gly Ala Gly Ser Leu Leu Phe Cys Asn Pro His
100 105 110 115
aac cca ctg ggc acg gtc ttt tct gaa gag tac atc cgc gag ctc acc 679
Asn Pro Leu Gly Thr Val Phe Ser Glu Glu Tyr Ile Arg Glu Leu Thr
120 125 130
gat att gcg gcg aag tac gat gcc cgc atc atc gtc gat gag atc cac 727
Asp Ile Ala Ala Lys Tyr Asp Ala Arg Ile Ile Val Asp Glu Ile His
135 140 145
gcg cca ctg gtt tat gaa ggc acc cat gtg gtt gct gct ggt gtt tct 775
Ala Pro Leu Val Tyr Glu Gly Thr His Val Val Ala Ala Gly Val Ser
150 155 160
gag aac gct gca aac act tgc atc acc atc acc gca act tct aag gcg 823
Glu Asn Ala Ala Asn Thr Cys Ile Thr Ile Thr Ala Thr Ser Lys Ala
165 170 175
tgg aac act gct ggt ttg aag tgt gct cag atc ttc ttc agt aat gaa 871
Trp Asn Thr Ala Gly Leu Lys Cys Ala Gln Ile Phe Phe Ser Asn Glu
180 185 190 195
gcc gat gtg aag gcc tgg aag aat ttg tcg gat att acc cgt gac ggt 919
Ala Asp Val Lys Ala Trp Lys Asn Leu Ser Asp Ile Thr Arg Asp Gly
200 205 210
gtg tcc atc ctt gga ttg atc gct gcg gag aca gtg tac aac gag ggc 967
Val Ser Ile Leu Gly Leu Ile Ala Ala Glu Thr Val Tyr Asn Glu Gly
215 220 225
gaa gaa ttc ctt gat gag tca att cag att ctc aag gac aac cgt gac 1015
Glu Glu Phe Leu Asp Glu Ser Ile Gln Ile Leu Lys Asp Asn Arg Asp
230 235 240
ttt gcg gct gct gaa ctg gaa aag ctt ggc gtg aag gtc tac gca ccg 1063
Phe Ala Ala Ala Glu Leu Glu Lys Leu Gly Val Lys Val Tyr Ala Pro
245 250 255
gac tcc act tat ttg atg tgg ttg gac ttc gct ggc acc aag atc gaa 1111
Asp Ser Thr Tyr Leu Met Trp Leu Asp Phe Ala Gly Thr Lys Ile Glu
260 265 270 275
gag gcg cct tct aaa att ctt cgt gag gag ggt aag gtc atg ctg aat 1159
Glu Ala Pro Ser Lys Ile Leu Arg Glu Glu Gly Lys Val Met Leu Asn
280 285 290
gat ggc gca gct ttt ggt ggt ttc acc acc tgc gct cgt ctt aat ttt 1207
Asp Gly Ala Ala Phe Gly Gly Phe Thr Thr Cys Ala Arg Leu Asn Phe
295 300 305
gcg tgt tcc aga gag acc ctt gag gag ggg ctg cgc cgt atc gcc agc 1255
Ala Cys Ser Arg Glu Thr Leu Glu Glu Gly Leu Arg Arg Ile Ala Ser
310 315 320
gtg ttg taa ataatgagta aaaagtctgt cctgattact tctttgatgc 1304
Val Leu
325
tgttttccat gttcttcgga gctggaaacc tcatcttccc gccgatgctt ggattgtcgg 1364
caggaaccaa ctatctacca gctatcttag gatttctagc aacgagtgtt ctgctcccgg 1424
tgctggcgat tatcgcggtg gtgttgtcgg gagaaaatgt caaggacatg gcttctcgtg 1484
gcggtaagat c 1495
<210>4
<211>325
<212>PRT
<213>谷氨酸棒杆菌
<400>4
Met Ala Asp Ala Val Glu Arg Glu Val Phe Gly Tyr Pro Pro Asp Ala
l 5 10 15
Thr Gly Leu Asn Asp Ala Leu Thr Gly Phe Tyr Glu Arg Arg Tyr Gly
20 25 30
Phe Gly Pro Asn Pro Glu Ser Val Phe Ala Ile Pro Asp Val Val Arg
35 40 45
Gly Leu Lys Leu Ala Ile Glu His Phe Thr Lys Pro Gly Ser Ala Ile
50 55 60
Ile Val Pro Leu Pro Ala Tyr Pro Pro Phe Ile Glu Leu Pro Lys Val
65 70 75 80
Thr Gly Arg Gln Ala Ile Tyr Ile Asp Ala His Glu Tyr Asp Leu Lys
85 90 95
Glu Ile Glu Lys Ala Phe Ala Asp Gly Ala Gly Ser Leu Leu Phe Cys
100 105 110
Asn Pro His Asn Pro Leu Gly Thr Val Phe Ser Glu Glu Tyr Ile Arg
115 120 125
Glu Leu Thr Asp Ile Ala Ala Lys Tyr Asp Ala Arg Ile Ile Val Asp
130 135 140
Glu Ile His Ala Pro Leu Val Tyr Glu Gly Thr His Val Val Ala Ala
145 150 155 160
Gly Val Ser Glu Asn Ala Ala Asn Thr Cys Ile Thr Ile Thr Ala Thr
165 170 175
Ser Lys Ala Trp Asn Thr Ala Gly Leu Lys Cys Ala Gln Ile Phe Phe
180 185 190
Ser Asn Glu Ala Asp Val Lys Ala Trp Lys Asn Leu Ser Asp Ile Thr
195 200 205
Arg Asp Gly Val Ser Ile Leu Gly Leu Ile Ala Ala Glu Thr Val Tyr
210 215 220
Asn Glu Gly Glu Glu Phe Leu Asp Glu Ser Ile Gln Ile Leu Lys Asp
225 230 235 240
Asn Arg Asp Phe Ala Ala Ala Glu Leu Glu Lys Leu Gly Val Lys Val
245 250 255
Tyr Ala Pro Asp Ser Thr Tyr Leu Met Trp Leu Asp Phe Ala Gly Thr
260 265 270
Lys Ile Glu Glu Ala Pro Ser Lys Ile Leu Arg Glu Glu Gly Lys Val
275 280 285
Met Leu Asn Asp Gly Ala Ala Phe Gly Gly Phe Thr Thr Cys Ala Arg
290 295 300
Leu Asn Phe Ala Cys Ser Arg Glu Thr Leu Glu Glu Gly Leu Arg Arg
305 310 315 320
Ile Ala Ser Val Leu
325
<210>5
<211>1033
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1006)
<400>5
gtgcggatcg ggtatccgcg ctacacttag aggtgttaga gatcatgagt ttccacgaac 60
tgtaacgcag gattcaccaa tcaatgaaag gtcgaccgac atg agc act gaa gac 115
Met Ser Thr Glu Asp
1 5
att gtc gtc gta gca gta gat ggc tcg gac gcc tca aaa caa gct gtt 163
Ile Val Val Val Ala Val Asp Gly Ser Asp Ala Ser Lys Gln Ala Val
10 15 20
cgg tgg gct gca aat acc gcc aac aaa cgt ggc att cca ctt cgc ttg 211
Arg Trp Ala Ala Asn Thr Ala Asn Lys Arg Gly Ile Pro Leu Arg Leu
25 30 35
gct tcc agc tac acc atg cct cag ttc ctc tac gca gag gga atg gtt 259
Ala Ser Ser Tyr Thr Met Pro Gln Phe Leu Tyr Ala Glu Gly Met Val
40 45 50
cca cca caa gag ctt ttc gat gac ctc cag gcc gaa gcc ctg gaa aag 307
Pro Pro Gln Glu Leu Phe Asp Asp Leu Gln Ala Glu Ala Leu Glu Lys
55 60 65
att aac gaa gcc cgt gac atc gcc cat gag gta gcg cca gaa atc aag 355
Ile Asn Glu Ala Arg Asp Ile Ala His Glu Val Ala Pro Glu Ile Lys
70 75 80 85
atc ggg cac acc atc gct gaa ggc agt ccc atc gac atg ctg ttg gaa 403
Ile Gly His Thr Ile Ala Glu Gly Ser Pro Ile Asp Met Leu Leu Glu
90 95 100
atg tct ccc gat gcc aca atg atc gtc atg ggt tcc cgc gga ctc ggc 451
Met Ser Pro Asp Ala Thr Met Ile Val Met Gly Ser Arg Gly Leu Gly
105 110 115
gga ctc tcc gga atg gtc atg ggc tcc gtc tcc ggt gca gtg gtc agc 499
Gly Leu Ser Gly Met Val Met Gly Ser Val Ser Gly Ala Val Val Ser
120 125 130
cac gca aag tgt cca gtc gtt gtt gtc cgt gaa gac agc gca gtc aac 547
His Ala Lys Cys Pro Val Val Val Val Arg Glu Asp Ser Ala Val Asn
135 140 145
gaa gac agc aag tac ggc cca gtc gtc gtc ggt gtg gat ggc tcc gaa 595
Glu Asp Ser Lys Tyr Gly Pro Val Val Val Gly Val Asp Gly Ser Glu
150 155 160 165
gtc tcc caa cag gca acc gaa tac gca ttt gcg gaa gct gaa gct cgt 643
Val Ser Gln Gln Ala Thr Glu Tyr Ala Phe Ala Glu Ala Glu Ala Arg
170 175 180
ggc gcc gaa ctc gtt gca gtt cac acc tgg atg gac atg cag gta cag 691
Gly Ala Glu Leu Val Ala Val His Thr Trp Met Asp Met Gln Val Gln
185 190 195
gca tca ctt gca ggt ctt gca gct gct caa cag cag tgg gat gaa gtg 739
Ala Ser Leu Ala Gly Leu Ala Ala Ala Gln Gln Gln Trp Asp Glu Val
200 205 210
gaa cgt cag caa acc gac atg ctg atc gaa cgc ctc gca cca ctg gtg 787
Glu Arg Gln Gln Thr Asp Met Leu Ile Glu Arg Leu Ala Pro Leu Val
215 220 225
gaa aag tac cca agt gta acc gtc aag aag atc atc acc cgt gac cgc 835
Glu Lys Tyr Pro Ser Val Thr Val Lys Lys Ile Ile Thr Arg Asp Arg
230 235 240 245
cca gtt cgc gca ctt gca gaa gca tct gaa aac gcg cag ctc cta gtc 883
Pro Val Arg Ala Leu Ala Glu Ala Ser Glu Asn Ala Gln Leu Leu Val
250 255 260
gtt ggt tcc cat ggt cgt ggc gga ttt aag ggc atg ctc ctt ggc tcc 931
Val Gly Ser His Gly Arg Gly Gly Phe Lys Gly Met Leu Leu Gly Ser
265 270 275
acc tcc cgc gca ctg ctg caa tcc gca ccg tgc cca atg atg gtg gtt 979
Thr Ser Arg Ala Leu Leu Gln Ser Ala Pro Cys Pro Met Met Val Val
280 285 290
cgc cca cct gag aag att aag aag tag tttcttttaa gtttcgatgc cccggtt 1033
Arg Pro Pro Glu Lys Ile Lys Lys
295 300
<210>6
<211>301
<212>PRT
<213>谷氨酸棒杆菌
<400>6
Met Ser Thr Glu Asp Ile Val Val Val Ala Val Asp Gly Ser Asp Ala
1 5 10 15
Ser Lys Gln Ala Val Arg Trp Ala Ala Asn Thr Ala Asn Lys Arg Gly
20 25 30
Ile Pro Leu Arg Leu Ala Ser Ser Tyr Thr Met Pro Gln Phe Leu Tyr
35 40 45
Ala Glu Gly Met Val Pro Pro Gln Glu Leu Phe Asp Asp Lau Gln Ala
50 55 60
Glu Ala Leu Glu Lys Ile Asn Glu Ala Arg Asp Ile Ala His Glu Val
65 70 75 80
Ala Pro Glu Ile Lys Ile Gly His Thr Ile Ala Glu Gly Ser Pro Ile
85 90 95
Asp Met Leu Leu Glu Met Ser Pro Asp Ala Thr Met Ile Val Met Gly
100 105 110
Ser Arg Gly Leu Gly Gly Leu Ser Gly Met Val Met Gly Ser Val Ser
115 120 125
Gly Ala Val Val Ser His Ala Lys Cys Pro Val Val Val Val Arg Glu
130 135 140
Asp Ser Ala Val Asn Glu Asp Ser Lys Tyr Gly Pro Val Val Val Gly
145 150 155 160
Val Asp Gly Ser Glu Val Ser Gln Gln Ala Thr Glu Tyr Ala Phe Ala
165 170 175
Glu Ala Glu Ala Arg Gly Ala Glu Leu Val Ala Val His Thr Trp Met
180 185 190
Asp Met Gln Val Gln Ala Ser Leu Ala Gly Leu Ala Ala Ala Gln Gln
195 200 205
Gln Trp Asp Glu Val Glu Arg Gln Gln Thr Asp Met Leu Ile Glu Arg
210 215 220
Leu Ala Pro Leu Val Glu Lys Tyr Pro Ser Val Thr Val Lys Lys Ile
225 230 235 240
Ile Thr Arg Asp Arg Pro Val Arg Ala Leu Ala Glu Ala Ser Glu Asn
245 250 255
Ala Gln Leu Leu Val Val Gly Ser His Gly Arg Gly Gly Phe Lys Gly
260 265 270
Met Leu Leu Gly Ser Thr Ser Arg Ala Leu Leu Gln Ser Ala Pro Cys
275 280 285
Pro Met Met Val Val Arg Pro Pro Glu Lys Ile Lys Lys
290 295 300
<210>7
<211>948
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(925)
<223>RXA02229
<400>7
gctggttcaa cagagaccac cgcgtgtcct gggtcgacgc ctctggcgat cccaccgcac 60
aagccttgga gattttgggt ctacaatagc gagggtgaat ttg acc atc ccc ttt 115
Leu Thr Ile Pro Phe
1 5
gcc aaa ggc cac gcc acc gaa aac gac ttc atc atc atc ccc gat gag 163
Ala Lys Gly His Ala Thr Glu Asn Asp Phe Ile Ile Ile Pro Asp Glu
10 15 20
gat gcg cgc cta gat tta act cca gaa atg gtg gtc acg ctg tgt gac 211
Asp Ala Arg Leu Asp Leu Thr Pro Glu Met Val Val Thr Leu Cys Asp
25 30 35
cgc cgc gcc ggg atc ggt gct gat ggt atc ctc cgc gtg gtt aaa gct 259
Arg Arg Ala Gly Ile Gly Ala Asp Gly Ile Leu Arg Val Val Lys Ala
40 45 50
gca gac gta gaa ggc tcc acg gtc gac cca tcg ctg tgg ttc atg gat 307
Ala Asp Val Glu Gly Ser Thr Val Asp Pro Ser Leu Trp Phe Met Asp
55 60 65
tac cgc aac gcc gat gga tct ttg gct gaa atg tgc ggc aat ggt gtg 355
Tyr Arg Asn Ala Asp Gly Ser Leu Ala Glu Met Cys Gly Asn Gly Val
70 75 80 85
cgc ctg ttc gcg cac tgg ctg tac tcc cgc ggt ctt gtt gat aat acg 403
Arg Leu Phe Ala His Trp Leu Tyr Ser Arg Gly Leu Val Asp Asn Thr
90 95 100
agc ttt gat atc ggt acc cgc gcc ggt gtc cgc cac gtt gat att ttg 451
Ser Phe Asp Ile Gly Thr Arg Ala Gly Val Arg His Val Asp Ile Leu
105 110 115
cag gca gat caa cat tct gcg cag gtc cgc gtt gat atg ggc atc cct 499
Gln Ala Asp Gln His Ser Ala Gln Val Arg Val Asp Met Gly Ile Pro
120 125 130
gac gtc acg gga tta tcc acc tgc gac atc aac ggc caa gta ttc gct 547
Asp Val Thr Gly Leu Ser Thr Cys Asp Ile Asn Gly Gln Val Phe Ala
135 140 145
ggc ctt ggc gtt gat atg ggt aac cca cac cta gcg tgc gtt gtg ccg 595
Gly Leu Gly Val Asp Met Gly Asn Pro His Leu Ala Cys Val Val Pro
150 155 160 165
ggc tta agt gcg tcg gct ctt gcc gat atg gaa ctg cgc gca cct acg 643
Gly Leu Ser Ala Ser Ala Leu Ala Asp Met Glu Leu Arg Ala Pro Thr
170 175 180
ttt gat cag gaa ttc ttc ccc cac ggt gtg aac gta gaa atc gtc aca 691
Phe Asp Gln Glu Phe Phe Pro His Gly Val Asn Val Glu Ile Val Thr
185 190 195
gaa tta gaa gat gac gca gta tcg atg cgc gtg tgg gaa cgc gga gtg 739
Glu Leu Glu Asp Asp Ala Val Ser Met Arg Val Trp Glu Arg Gly Val
200 205 210
ggc gaa acc cgc tcc tgt ggc acg gga acc gtt gct gca gcg tgt gct 787
Gly Glu Thr Arg Ser Cys Gly Thr Gly Thr Val Ala Ala Ala Cys Ala
215 220 225
gct tta gct gat gct gga ttg gga gaa ggc aca gct aaa gtg tgc gtt 835
Ala Leu Ala Asp Ala Gly Leu Gly Glu Gly Thr Ala Lys Val Cys Val
230 235 240 245
cca cgt ggg gaa gta gaa gtc cag atc ttt gac gac ggc tcc aca ctc 883
Pro Arg Gly Glu Val Glu Val Gln Ile Phe Asp Asp Gly Ser Thr Leu
250 255 260
acc ggc cca agc gcc atc atc gca ctc ggt gag gtg cag atc 925
Thr Gly Pro Ser Ala Ile Ile Ala Leu Gly Glu Val Gln Ile
265 270 275
taagattcgc gattgtagtt cgg 948
<210>8
<211>275
<212>PRT
<213>谷氨酸棒杆菌
<400>8
Leu Thr Ile Pro Phe Ala Lys Gly His Ala Thr Glu Asn Asp Phe Ile
1 5 10 15
Ile Ile Pro Asp Glu Asp Ala Arg Leu Asp Leu Thr Pro Glu Met Val
20 25 30
Val Thr Leu Cys Asp Arg Arg Ala Gly Ile Gly Ala Asp Gly Ile Leu
35 40 45
Arg Val Val Lys Ala Ala Asp Val Glu Gly Ser Thr Val Asp Pro Ser
50 55 60
Leu Trp Phe Met Asp Tyr Arg Asn Ala Asp Gly Ser Leu Ala Glu Met
65 70 75 80
Cys Gly Asn Gly Val Arg Leu Phe Ala His Trp Leu Tyr Ser Arg Gly
85 90 95
Leu Val Asp Asn Thr Ser Phe Asp Ile Gly Thr Arg Ala Gly Val Arg
100 105 110
His Val Asp Ile Leu Gln Ala Asp Gln His Ser Ala Gln Val Arg Val
115 120 125
Asp Met Gly Ile Pro Asp Val Thr Gly Leu Ser Thr Cys Asp Ile Asn
130 135 140
Gly Gln Val Phe Ala Gly Leu Gly Val Asp Met Gly Asn Pro His Leu
145 150 155 160
Ala Cys Val Val Pro Gly Leu Ser Ala Ser Ala Leu Ala Asp Met Glu
165 170 175
Leu Arg Ala Pro Thr Phe Asp Gln Glu Phe Phe Pro His Gly Val Asn
180 185 190
Val Glu Ile Val Thr Glu Leu Glu Asp Asp Ala Val Ser Met Arg Val
195 200 205
Trp Glu Arg Gly Val Gly Glu Thr Arg Ser Cys Gly Thr Gly Thr Val
210 215 220
Ala Ala Ala Cys Ala Ala Leu Ala Asp Ala Gly Leu Gly Glu Gly Thr
225 230 235 240
Ala Lys Val Cys Val Pro Arg Gly Glu Val Glu Val Gln Ile Phe Asp
245 250 255
Asp Gly Ser Thr Leu Thr Gly Pro Ser Ala Ile Ile Ala Leu Gly Glu
260 265 270
Val Gln Ile
275
<210>9
<211>1491
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1468)
<223>RXS02970
<400>9
aaccgacaaa acagccgttc acgtgctaaa gcagctcggc ttgatctagg gtgaggtgag 60
ttatttaaag acttcataat attttgggga gtgaactggt ttg gca ttg aag ggt 115
Leu Ala Leu Lys Gly
1 5
tac acc aac ttt gac ggt gaa ttc atc gaa ttc gga tct gtg caa gca 163
Tyr Thr Asn Phe Asp Gly Glu Phe Ile Glu Phe Gly Ser Val Gln Ala
10 15 20
aaa gaa gag gaa aaa cgg gca ttc gac aac gat cgc gcg cac gtt ttc 211
Lys Glu Glu Glu Lys Arg Ala Phe Asp Asn Asp Arg Ala His Val Phe
25 30 35
cac tcc tgg tcc gcg cag gac aaa atc agc ccc aaa gta tgg gca gct 259
His Ser Trp Ser Ala Gln Asp Lys Ile Ser Pro Lys Val Trp Ala Ala
40 45 50
gcc gaa ggt tcc acg ctg tac gac ttc gac ggc aac gcc ttc atc gac 307
Ala Glu Gly Ser Thr Leu Tyr Asp Phe Asp Gly Asn Ala Phe Ile Asp
55 60 65
atg ggt tcc caa ctt gtc tcg gca aac tta ggc cac aac aac cct cga 355
Met Gly Ser Gln Leu Val Ser Ala Asn Leu Gly His Asn Asn Pro Arg
70 75 80 85
tta gtt gag gcg atc cag cgc caa gca gcc cgg ttg acc aac atc aac 403
Leu Val Glu Ala Ile Gln Arg Gln Ala Ala Arg Leu Thr Asn Ile Asn
90 95 100
ccg gcc ttc ggc aat gat gtg cgc tct gat gtt gct gca aag atc gtg 451
Pro Ala Phe Gly Asn Asp Val Arg Ser Asp Val Ala Ala Lys Ile Val
105 110 115
tcg atg gcc cgt ggc gaa ttc tcc cac gtg ttt ttc acc aac ggc ggc 499
Ser Met Ala Arg Gly Glu Phe Ser His Val Phe Phe Thr Asn Gly Gly
120 125 130
gcc gac gcc atc gag cac tcc atc cgc atg gct cgc ctg cac acc gga 547
Ala Asp Ala Ile Glu His Ser Ile Arg Met Ala Arg Leu His Thr Gly
135 140 145
cgc aac aaa att ctg tcc gca tac cgc agc tac cac ggc gca acc gga 595
Arg Asn Lys Ile Leu Ser Ala Tyr Arg Ser Tyr His Gly Ala Thr Gly
150 155 160 165
tcc gcg atg atg ctc acc ggc gaa cac cgc cgc ctg ggc aac ccc acc 643
Ser Ala Met Met Leu Thr Gly Glu His Arg Arg Leu Gly Asn Pro Thr
170 175 180
acc gac cca gat atc tac cac ttc tgg gca cca ttc ctg cac cac tcc 691
Thr Asp Pro Asp Ile Tyr His Phe Trp Ala Pro Phe Leu His His Ser
185 190 195
tca ttc ttt gcc acc acc caa gaa gaa gaa tgc gaa cgc gca ctc aag 739
Ser Phe Phe Ala Thr Thr Gln Glu Glu Glu Cys Glu Arg Ala Leu Lys
200 205 210
cac ttg gaa gat gtc atc gcg ttt gaa ggt gct ggc atg atc gca gcg 787
His Leu Glu Asp Val Ile Ala Phe Glu Gly Ala Gly Met Ile Ala Ala
215 220 225
atc gtc ctg gag cca gtg gtg gga tca tca gga atc atc ctg cca cca 835
Ile Val Leu Glu Pro Val Val Gly Ser Ser Gly Ile Ile Leu Pro Pro
230 235 240 245
gca ggt tac tta aat ggc gtg cgc gaa ctt tgc aac aag cac ggc atc 883
Ala Gly Tyr Leu Asn Gly Val Arg Glu Leu Cys Asn Lys His Gly Ile
250 255 260
ctc ttc atc gcc gac gaa gtc atg gtc gga ttc gga cgc acc gga aaa 931
Leu Phe Ile Ala Asp Glu Val Met Val Gly Phe Gly Arg Thr Gly Lys
265 270 275
ctg ttt gct tac gag cat gct ggc gac gat ttc cag cca gac atg atc 979
Leu Phe Ala Tyr Glu His Ala Gly Asp Asp Phe Gln Pro Asp Met Ile
280 285 290
acc ttc gcc aag ggt gtt aac gca ggt tac gcc cca ctc ggt ggc atc 1027
Thr Phe Ala Lys Gly Val Asn Ala Gly Tyr Ala Pro Leu Gly Gly Ile
295 300 305
gtg atg acc caa tca atc cgc gat acc ttc gga tca gag gca tac tcc 1075
Val Met Thr Gln Ser Ile Arg Asp Thr Phe Gly Ser Glu Ala Tyr Ser
310 315 320 325
ggc gga ctc acc tac tcc gga cac cca ctt gca gta gca ccc gcc aag 1123
Gly Gly Leu Thr Tyr Ser Gly His Pro Leu Ala Val Ala Pro Ala Lys
330 335 340
gca gcg ctg gag att tac gcg gaa gga gag atc att cca cgc gta gct 1171
Ala Ala Leu Glu Ile Tyr Ala Glu Gly Glu Ile Ile Pro Arg Val Ala
345 350 355
cga ctt ggc gct gaa ctg atc gaa cct cgc ctt cgt gaa cta gcg gaa 1219
Arg Leu Gly Ala Glu Leu Ile Glu Pro Arg Leu Arg Glu Leu Ala Glu
360 365 370
gaa aac gta gcg atc gct gac gtg cgg ggc atc gga ttc ttc tgg gca 1267
Glu Asn Val Ala Ile Ala Asp Val Arg Gly Ile Gly Phe Phe Trp Ala
375 380 385
gtg gag ttc aat gca gac gcc act gcc atg gct gcc ggt gct gca gaa 1315
Val Glu Phe Asn Ala Asp Ala Thr Ala Met Ala Ala Gly Ala Ala Glu
390 395 400 405
ttc aag gaa cgc ggc gtg tgg ccg atg atc tcc ggc aac cga ttc cac 1363
Phe Lys Glu Arg Gly Val Trp Pro Met Ile Ser Gly Asn Arg Phe His
410 415 420
atc gcg ccg ccg ctg acc acc act gat gac gaa ttg gta gca ctg ctg 1411
Ile Ala Pro Pro Leu Thr Thr Thr Asp Asp Glu Leu Val Ala Leu Leu
425 430 435
gac gcg gtg gaa gct gca gcc caa gct gtc gag ctg acc ttc gct ggg 1459
Asp Ala Val Glu Ala Ala Ala Gln Ala Val Glu Leu Thr Phe Ala Gly
440 445 450
gcg ttg ttc taagttttct agataacaag gcc 1491
Ala Leu Phe
455
<210>10
<211>456
<212>PRT
<213>谷氨酸棒杆菌
<400>10
Leu Ala Leu Lys Gly Tyr Thr Asn Phe Asp Gly Glu Phe Ile Glu Phe
1 5 10 15
Gly Ser Val Gln Ala Lys Glu Glu Glu Lys Arg Ala Phe Asp Asn Asp
20 25 30
Arg Ala His Val Phe His Ser Trp Ser Ala Gln Asp Lys Ile Ser Pro
35 40 45
Lys Val Trp Ala Ala Ala Glu Gly Ser Thr Leu Tyr Asp Phe Asp Gly
50 55 60
Asn Ala Phe Ile Asp Met Gly Ser Gln Leu Val Ser Ala Asn Leu Gly
65 70 75 80
His Asn Asn Pro Arg Leu Val Glu Ala Ile Gln Arg Gln Ala Ala Arg
85 90 95
Leu Thr Asn Ile Asn Pro Ala Phe Gly Asn Asp Val Arg Ser Asp Val
100 105 110
Ala Ala Lys Ile Val Ser Met Ala Arg Gly Glu Phe Ser His Val Phe
115 120 125
Phe Thr Asn Gly Gly Ala Asp Ala Ile Glu His Ser Ile Arg Met Ala
130 135 140
Arg Leu His Thr Gly Arg Asn Lys Ile Leu Ser Ala Tyr Arg Ser Tyr
145 150 155 160
His Gly Ala Thr Gly Ser Ala Met Met Leu Thr Gly Glu His Arg Arg
165 170 175
Leu Gly Asn Pro Thr Thr Asp Pro Asp Ile Tyr His Phe Trp Ala Pro
180 185 190
Phe Leu His His Ser Ser Phe Phe Ala Thr Thr Gln Glu Glu Glu Cys
195 200 205
Glu Arg Ala Leu Lys His Leu Glu Asp Val Ile Ala Phe Glu Gly Ala
210 215 220
Gly Met Ile Ala Ala Ile Val Leu Glu Pro Val Val Gly Ser Ser Gly
225 230 235 240
Ile Ile Leu Pro Pro Ala Gly Tyr Leu Asn Gly Val Arg Glu Leu Cys
245 250 255
Asn Lys His Gly Ile Leu Phe Ile Ala Asp Glu Val Met Val Gly Phe
260 265 270
Gly Arg Thr Gly Lys Leu Phe Ala Tyr Glu His Ala Gly Asp Asp Phe
275 280 285
Gln Pro Asp Met Ile Thr Phe Ala Lys Gly Val Asn Ala Gly Tyr Ala
290 295 300
Pro Leu Gly Gly Ile Val Met Thr Gln Ser Ile Arg Asp Thr Phe Gly
305 310 315 320
Ser Glu Ala Tyr Ser Gly Gly Leu Thr Tyr Ser Gly His Pro Leu Ala
325 330 335
Val Ala Pro Ala Lys Ala Ala Leu Glu Ile Tyr Ala Glu Gly Glu Ile
340 345 350
Ile Pro Arg Val Ala Arg Leu Gly Ala Glu Leu Ile Glu Pro Arg Leu
355 360 365
Arg Glu Leu Ala Glu Glu Asn Val Ala Ile Ala Asp Val Arg Gly Ile
370 375 380
Gly Phe Phe Trp Ala Val Glu Phe Asn Ala Asp Ala Thr Ala Met Ala
385 390 395 400
Ala Gly Ala Ala Glu Phe Lys Glu Arg Gly Val Trp Pro Met Ile Ser
405 410 415
Gly Asn Arg Phe His Ile Ala Pro Pro Leu Thr Thr Thr Asp Asp Glu
420 425 430
Leu Val Ala Leu Leu Asp Ala Val Glu Ala Ala Ala Gln Ala Val Glu
435 440 445
Leu Thr Phe Ala Gly Ala Leu Phe
450 455
<210>11
<211>1330
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1330)
<223>FRXA01009
<400>11
aaccgacaaa acagccgttc acgtgctaaa gcagctcggc ttgatctagg gtgaggtgag 60
ttatttaaag acttcataat attttgggga gtgaactggt ttg gca ttg aag ggt 115
Leu Ala Leu Lys Gly
1 5
tac acc aac ttt gac ggt gaa ttc atc gaa ttc gga tct gtg caa gca 163
Tyr Thr Asn Phe Asp Gly Glu Phe Ile Glu Phe Gly Ser Val Gln Ala
10 15 20
aaa gaa gag gaa aaa cgg gca ttc gac aac gat cgc gcg cac gtt ttc 211
Lys Glu Glu Glu Lys Arg Ala Phe Asp Asn Asp Arg Ala His Val Phe
25 30 35
cac tcc tgg tcc gcg cag gac aaa atc agc ccc aaa gta tgg gca gct 259
His Ser Trp Ser Ala Gln Asp Lys Ile Ser Pro Lys Val Trp Ala Ala
40 45 50
gcc gaa ggt tcc acg ctg tac gac ttc gac ggc aac gcc ttc atc gac 307
Ala Glu Gly Ser Thr Leu Tyr Asp Phe Asp Gly Asn Ala Phe Ile Asp
55 60 65
atg ggt tcc caa ctt gtc tcg gca aac tta ggc cac aac aac cct cga 355
Met Gly Ser Gln Leu Val Ser Ala Asn Leu Gly His Asn Asn Pro Arg
70 75 80 85
tta gtt gag gcg atc cag cgc caa gca gcc cgg ttg acc aac atc aac 403
Leu Val Glu Ala Ile Gln Arg Gln Ala Ala Arg Leu Thr Asn Ile Asn
90 95 100
ccg gcc ttc ggc aat gat gtg cgc tct gat gtt gct gca aag atc gtg 451
Pro Ala Phe Gly Asn Asp Val Arg Ser Asp Val Ala Ala Lys Ile Val
105 110 115
tcg atg gcc cgt ggc gaa ttc tcc cac gtg ttt ttc acc aac ggc ggc 499
Ser Met Ala Arg Gly Glu Phe Ser His Val Phe Phe Thr Asn Gly Gly
120 125 130
gcc gac gcc atc gag cac tcc atc cgc atg gct cgc ctg cac acc gga 547
Ala Asp Ala Ile Glu His Ser Ile Arg Met Ala Arg Leu His Thr Gly
135 140 145
cgc aac aaa att ctg tcc gca tac cgc agc tac cac ggc gca acc gga 595
Arg Asn Lys Ile Leu Ser Ala Tyr Arg Ser Tyr His Gly Ala Thr Gly
150 155 160 165
tcc gcg atg atg ctc acc ggc gaa cac cgc cgc ctg ggc aac ccc acc 643
Ser Ala Met Met Leu Thr Gly Glu His Arg Arg Leu Gly Asn Pro Thr
170 175 180
acc gac cca gat atc tac cac ttc tgg gca cca ttc ctg cac cac tcc 691
Thr Asp Pro Asp Ile Tyr His Phe Trp Ala Pro Phe Leu His His Ser
185 190 195
tca ttc ttt gcc acc acc caa gaa gaa gaa tgc gaa cgc gca ctc aag 739
Ser Phe Phe Ala Thr Thr Gln Glu Glu Glu Cys Glu Arg Ala Leu Lys
200 205 210
cac ttg gaa gat gtc atc gcg ttt gaa ggt gct ggc atg atc gca gcg 787
His Leu Glu Asp Val Ile Ala Phe Glu Gly Ala Gly Met Ile Ala Ala
215 220 225
atc gtc ctg gag cca gtg gtg gga tca tca gga atc atc ctg cca cca 835
Ile Val Leu Glu Pro Val Val Gly Ser Ser Gly Ile Ile Leu Pro Pro
230 235 240 245
gca ggt tac tta aat ggc gtg cgc gaa ctt tgc aac aag cac ggc atc 883
Ala Gly Tyr Leu Asn Gly Val Arg Glu Leu Cys Asn Lys His Gly Ile
250 255 260
ctc ttc atc gcc gac gaa gtc atg gtc gga ttc gga cgc acc gga aaa 931
Leu Phe Ile Ala Asp Glu Val Met Val Gly Phe Gly Arg Thr Gly Lys
265 270 275
ctg ttt gct tac gag cat gct ggc gac gat ttc cag cca gac atg atc 979
Leu Phe Ala Tyr Glu His Ala Gly Asp Asp Phe Gln Pro Asp Met Ile
280 285 290
acc ttc gcc aag ggt gtt aac gca ggt tac gcc cca ctc ggt ggc atc 1027
Thr Phe Ala Lys Gly Val Asn Ala Gly Tyr Ala Pro Leu Gly Gly Ile
295 300 305
gtg atg acc caa tca atc cgc gat acc ttc gga tca gag gca tac tcc 1075
Val Met Thr Gln Ser Ile Arg Asp Thr Phe Gly Ser Glu Ala Tyr Ser
310 315 320 325
ggc gga ctc acc tac tcc gga cac cca ctt gca gta gca ccc gcc aag 1123
Gly Gly Leu Thr Tyr Ser Gly His Pro Leu Ala Val Ala Pro Ala Lys
330 335 340
gca gcg ctg gag att tac gcg gaa gga gag atc att cca cgc gta gct 1171
Ala Ala Leu Glu Ile Tyr Ala Glu Gly Glu Ile Ile Pro Arg Val Ala
345 350 355
cga ctt ggc gct gaa ctg atc gaa cct cgc ctt cgt gaa cta gcg gaa 1219
Arg Leu Gly Ala Glu Leu Ile Glu Pro Arg Leu Arg Glu Leu Ala Glu
360 365 370
gaa aac gta gcg atc gct gac gtg cgg ggc atc gga ttc ttc tgg gca 1267
Glu Asn Val Ala Ile Ala Asp Val Arg Gly Ile Gly Phe Phe Trp Ala
375 380 385
gtg gag ttc aat gca gac gcc act gcc atg gct gcc ggt gct gca gaa 1315
Val Glu Phe Asn Ala Asp Ala Thr Ala Met Ala Ala Gly Ala Ala Glu
390 395 400 405
ttc aag gaa cgc ggc 1330
Phe Lys Glu Arg Gly
410
<210>12
<211>410
<212>PRT
<213>谷氨酸棒杆菌
<400>12
Leu Ala Leu Lys Gly Tyr Thr Asn Phe Asp Gly Glu Phe Ile Glu Phe
1 5 10 15
Gly Ser Val Gln Ala Lys Glu Glu Glu Lys Arg Ala Phe Asp Asn Asp
20 25 30
Arg Ala His Val Phe His Ser Trp Ser Ala Gln Asp Lys Ile Ser Pro
35 40 45
Lys Val Trp Ala Ala Ala Glu Gly Ser Thr Leu Tyr Asp Phe Asp Gly
50 55 60
Asn Ala Phe Ile Asp Met Gly Ser Gln Leu Val Ser Ala Asn Leu Gly
65 70 75 80
His Asn Asn Pro Arg Leu Val Glu AlaI le Gln Arg Gln Ala Ala Arg
85 90 95
Leu Thr Asn Ile Asn Pro Ala Phe Gly Asn Asp Val Arg Ser Asp Val
100 105 110
Ala Ala Lys Ile Val Ser Met Ala Arg Gly Glu Phe Ser His Val Phe
115 120 125
Phe Thr Asn Gly Gly Ala Asp Ala Ile Glu His Ser Ile Arg Met Ala
130 135 140
Arg Leu His Thr Gly Arg Asn Lys Ile Leu Ser Ala Tyr Arg Ser Tyr
145 150 155 160
His Gly Ala Thr Gly Ser Ala Met Met Leu Thr Gly Glu His Arg Arg
165 170 175
Leu Gly Asn Pro Thr Thr Asp Pro Asp Ile Tyr His Phe Trp Ala Pro
180 185 190
Phe Leu His His Ser Ser Phe Phe Ala Thr Thr Gln Glu Glu Glu Cys
195 200 205
Glu Arg Ala Leu Lys His Leu Glu Asp Val Ile Ala Phe Glu Gly Ala
210 215 220
Gly Met Ile Ala Ala Ile Val Leu Glu Pro Val Val Gly Ser Ser Gly
225 230 235 240
Ile Ile Leu Pro Pro Ala Gly Tyr Leu Asn Gly Val Arg Glu Leu Cys
245 250 255
Asn Lys His Gly Ile Leu Phe Ile Ala Asp Glu Val Met Val Gly Phe
260 265 270
Gly Arg Thr Gly Lys Leu Phe Ala Tyr Glu His Ala Gly Asp Asp Phe
275 280 285
Gln Pro Asp Met Ile Thr Phe Ala Lys Gly Val Asn Ala Gly Tyr Ala
290 295 300
Pro Leu Gly Gly Ile Val Met Thr Gln Ser Ile Arg Asp Thr Phe Gly
305 310 315 320
Ser Glu Ala Tyr Ser Gly Gly Leu Thr Tyr Ser Gly His Pro Leu Ala
325 330 335
Val Ala Pro Ala Lys Ala Ala Leu Glu Ile Tyr Ala Glu Gly Glu Ile
340 345 350
Ile Pro Arg Val Ala Arg Leu Gly Ala Glu Leu Ile Glu Pro Arg Leu
355 360 365
Arg Glu Leu Ala Glu Glu Asn Val Ala Ile Ala Asp Val Arg Gly Ile
370 375 380
Gly Phe Phe Trp Ala Val Glu Phe Asn Ala Asp Ala Thr Ala Met Ala
385 390 395 400
Ala Gly Ala Ala Glu Phe Lys Glu Arg Gly
405 410
<210>13
<211>792
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(769)
<223>RXC02390
<400>13
gctggtggtg ctgacccata cgctggaact ccaactgctg ttgataccgc caagatgttt 60
ggccgcgagg atctcgtagc tcgcttcgag tcataggccg gtg gag tgg acc gct 115
Val Glu Trp Thr Ala
1 5
ttt ggc acc ctg att ctg ctc aat ttg gtg ggc agt tta tcc ccg ggg 163
Phe Gly Thr Leu Ile Leu Leu Asn Leu Val Gly Ser Leu Ser Pro Gly
10 15 20
cct gat acc ttt ttc ctc ctc cgc tta gcc acc cgc tcc aga gcg cac 211
Pro Asp Thr Phe Phe Leu Leu Arg Leu Ala Thr Arg Ser Arg Ala His
25 30 35
gcg atc gct ggc gtc gcc ggc atc gtc acc gga ctc acg gtg tgg gtg 259
Ala Ile Ala Gly Val Ala Gly Ile Val Thr Gly Leu Thr Val Trp Val
40 45 50
acg ctg acg gtc gtg gga gca gcg gcg ctg ctc acc act tat ccg tcg 307
Thr Leu Thr Val Val Gly Ala Ala Ala Leu Leu Thr Thr Tyr Pro Ser
55 60 65
att ctc gga atc atc cag ctc gtc ggc ggc acg tac cta agc ttc att 355
Ile Leu Gly Ile Ile Gln Leu Val Gly Gly Thr Tyr Leu Ser Phe Ile
70 75 80 85
ggg tac aag ttg ctg cgc tcg gcg tcg aga gag ctt atc gac gcc cgc 403
Gly Tyr Lys Leu Leu Arg Ser Ala Ser Arg Glu Leu Ile Asp Ala Arg
90 95 100
cag ttc cgt ttc aac gcc gat gcc cga cct atc ccg gat gcg gta gaa 451
Gln Phe Arg Phe Asn Ala Asp Ala Arg Pro Ile Pro Asp Ala Val Glu
105 110 115
gca ctg gga acc cgc act cag gta tat cga caa ggt ttg gcc acc aac 499
Ala Leu Gly Thr Arg Thr Gln Val Tyr Arg Gln Gly Leu Ala Thr Asn
120 125 130
ctg tca aac cct aaa gtt gtc atg tac ttc gcg gca att ctg gct ccg 547
Leu Ser Asn Pro Lys Val Val Met Tyr Phe Ala Ala Ile Leu Ala Pro
135 140 145
ttg atg cca gcg cac cca tca ccg gtg ctg gcg ttc tct atc atc gtg 595
Leu Met Pro Ala His Pro Ser Pro Val Leu Ala Phe Ser Ile Ile Val
150 155 160 165
gcg att tta gtg cag acc ttt gtt acc ttc tct gct gtg tgc ctc att 643
Ala Ile Leu Val Gln Thr Phe Val Thr Phe Ser Ala Val Cys Leu Ile
170 175 180
gtc tct acg gag cgt gtg cgc aaa gca atg ctg cgt gca ggt ccc tgg 691
Val Ser Thr Glu Arg Val Arg Lys Ala Met Leu Arg Ala Gly Pro Trp
185 190 195
ttt gac ctg ctt gct ggc gtt gtc ttc ctc gtt gtg ggt gtg act ctg 739
Phe Asp Leu Leu Ala Gly Val Val Phe Leu Val Val Gly Val Thr Leu
200 205 210
ctg tat gaa ggc ctg acc ggt tta ctc ggg taaaggcata aaaaatggct 789
Leu Tyr Glu Gly Leu Thr Gly Leu Leu Gly
215 220
tcc 792
<210>14
<211>223
<212>PRT
<213>谷氨酸棒杆菌
<400>14
Val Glu Trp Thr Ala Phe Gly Thr Leu Ile Leu Leu Asn Leu Val Gly
1 5 10 15
Ser Leu Ser Pro Gly Pro Asp Thr Phe Phe Leu Leu Arg Leu Ala Thr
20 25 30
Arg Ser Arg Ala His Ala Ile Ala Gly Val Ala Gly Ile Val Thr Gly
35 40 45
Leu Thr Val Trp Val Thr Leu Thr Val Val Gly Ala Ala Ala Leu Leu
50 55 60
Thr Thr Tyr Pro Ser Ile Leu Gly Ile Ile Gln Leu Val Gly Gly Thr
65 70 75 80
Tyr Leu Ser Phe Ile Gly Tyr Lys Leu Leu Arg Ser Ala Ser Arg Glu
85 90 95
Leu Ile Asp Ala Arg Gln Phe Arg Phe Asn Ala Asp Ala Arg Pro Ile
100 105 110
Pro Asp Ala Val Glu Ala Leu Gly Thr Arg Thr Gln Val Tyr Arg Gln
115 120 125
Gly Leu Ala Thr Asn Leu Ser Asn Pro Lys Val Val Met Tyr Phe Ala
130 135 140
Ala Ile Leu Ala Pro Leu Met Pro Ala His Pro Ser Pro Val Leu Ala
145 150 155 160
Phe Ser Ile Ile Val Ala Ile Leu Val Gln Thr Phe Val Thr Phe Ser
165 170 175
Ala Val Cys Leu Ile Val Ser Thr Glu Arg Val Arg Lys Ala Met Leu
180 185 190
Arg Ala Gly Pro Trp Phe Asp Leu Leu Ala Gly Val Val Phe Leu Val
195 200 205
Val Gly Val Thr Leu Leu Tyr Glu Gly Leu Thr Gly Leu Leu Gly
210 215 220
<210>15
<211>897
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(874)
<223>RXC01796
<400>15
atgtaactcg atcaggtgga aatgcccgca aaagtggcgg cggtggccga gggatggccg 60
ttggtgcggc atcggtggcc tgctactagt cgggctcttc ttg ctc ctt ggc ggt 115
Leu Leu Leu Gly Gly
1 5
aac cct gcc gag atc gac cag gtt tta ggt ggc gat caa acc cag atc 163
Asn Pro Ala Glu Ile Asp Gln Val Leu Gly Gly Asp Gln Thr Gln Ile
10 15 20
gag tct gga gag tcc acc gga gcc ggc gac ttt gat cac tgc caa acc 211
Glu Ser Gly Glu Ser Thr Gly Ala Gly Asp Phe Asp His Cys Gln Thr
25 30 35
ggc gca gat gcc aac gcc agt gat gat tgt cgc ctt tac tac acc tca 259
Gly Ala Asp Ala Asn Ala Ser Asp Asp Cys Arg Leu Tyr Tyr Thr Ser
40 45 50
ttc tcc gtc aat gaa atg tgg cag act ttg ctt cca gct cag gct ggt 307
Phe Ser Val Asn Glu Met Trp Gln Thr Leu Leu Pro Ala Gln Ala Gly
55 60 65
atc gaa tac acc gag ccg aca ttg act ctt ttc aaa aac tcc acc caa 355
Ile Glu Tyr Thr Glu Pro Thr Leu Thr Leu Phe Lys Asn Ser Thr Gln
70 75 80 85
acc ggc tgc ggt ttc gct tct gcg tcc act ggg ccg ttt tac tgt ccg 403
Thr Gly Cys Gly Phe Ala Ser Ala Ser Thr Gly Pro Phe Tyr Cys Pro
90 95 100
tca gac caa gat gct tat ttt gac ttg act ttc ttc gat cag atg cgt 451
Ser Asp Gln Asp Ala Tyr Phe Asp Leu Thr Phe Phe Asp Gln Met Arg
105 110 115
cag ttc ggt gca gaa aac gcc ccg ctt gcc cag atg tac atc gtg gcg 499
Gln Phe Gly Ala Glu Asn Ala Pro Leu Ala Gln Met Tyr Ile Val Ala
120 125 130
cac gag tac ggc cac cac gtc caa aac ctc gag ggc aca ctc gga ctg 547
His Glu Tyr Gly His His Val Gln Asn Leu Glu Gly Thr Leu Gly Leu
135 140 145
tcc aat tac aac gat ccg ggc gct gat tcc aac gcc gtc aag atc gag 595
Ser Asn Tyr Asn Asp Pro Gly Ala Asp Ser Asn Ala Val Lys Ile Glu
150 155 160 165
ttg cag gcc gat tgc tac gca ggc att tgg gct aat cac tcc agc gaa 643
Leu Gln Ala Asp Cys Tyr Ala Gly Ile Trp Ala Asn His Ser Ser Glu
170 175 180
ggc ccg gat ccg cta ctc caa ccc atc acc gaa tct gag cta gat tcc 691
Gly Pro Asp Pro Leu Leu Gln Pro Ile Thr Glu Ser Glu Leu Asp Ser
185 190 195
gct ctc ctt gct gca agc gcc gtg ggc gac gac aat atc cag caa cga 739
Ala Leu Leu Ala Ala Ser Ala Val Gly Asp Asp Asn Ile Gln Gln Arg
200 205 210
tcc ggt ggc gat gtc aat cct gaa agc tgg act cac ggc tca tcg cag 787
Ser Gly Gly Asp Val Asn Pro Glu Ser Trp Thr His Gly Ser Ser Gln
215 220 225
cag cgc aaa gac gcg ttc ctc gcc ggc tac aac acc ggc cag atg agc 835
Gln Arg Lys Asp Ala Phe Leu Ala Gly Tyr Asn Thr Gly Gln Met Ser
230 235 240 245
gcc tgc gac ttc ctc ggc cgg ggc gtc tac aac gac gct taaagcattg 884
Ala Cys Asp Phe Leu Gly Arg Gly Val Tyr Asn Asp Ala
250 255
cttttcgacg tct 897
<210>16
<211>258
<212>PRT
<213>谷氨酸棒杆菌
<400>16
Leu Leu Leu Gly Gly Asn Pro Ala Glu Ile Asp Gln Val Leu Gly Gly
1 5 10 15
Asp Gln Thr Gln Ile Glu Ser Gly Glu Ser Thr Gly Ala Gly Asp Phe
20 25 30
Asp His Cys Gln Thr Gly Ala Asp Ala Asn Ala Ser Asp Asp Cys Arg
35 40 45
Leu Tyr Tyr Thr Ser Phe Ser Val Asn Glu Met Trp Gln Thr Leu Leu
50 55 60
Pro Ala Gln Ala Gly Ile Glu Tyr Thr Glu Pro Thr Leu Thr Leu Phe
65 70 75 80
Lys Asn Ser Thr Gln Thr Gly Cys Gly Phe Ala Ser Ala Ser Thr Gly
85 90 95
Pro Phe Tyr Cys Pro Ser Asp Gln Asp Ala Tyr Phe Asp Leu Thr Phe
100 105 110
Phe Asp Gln Met Arg Gln Phe Gly Ala Glu Asn Ala Pro Leu Ala Gln
115 120 125
Met Tyr Ile Val Ala His Glu Tyr Gly His His Val Gln Asn Leu Glu
130 135 140
Gly Thr Leu Gly Leu Ser Asn Tyr Asn Asp Pro Gly Ala Asp Ser Asn
145 150 155 160
Ala Val Lys Ile Glu Leu Gln Ala Asp Cys Tyr Ala Gly Ile Trp Ala
165 170 175
Asn His Ser Ser Glu Gly Pro Asp Pro Leu Leu Gln Pro Ile Thr Glu
180 185 190
Ser Glu Leu Asp Ser Ala Leu Leu Ala Ala Ser Ala Val Gly Asp Asp
195 200 205
Asn Ile Gln Gln Arg Ser Gly Gly Asp Val Asn Pro Glu Ser Trp Thr
210 215 220
His Gly Ser Ser Gln Gln Arg Lys Asp Ala Phe Leu Ala Gly Tyr Asn
225 230 235 240
Thr Gly Gln Met Ser Ala Cys Asp Phe Leu Gly Arg Gly Val Tyr Asn
245 250 255
Asp Ala
<210>17
<211>771
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(748)
<223>RXC01207
<400>17
cttcatgatc tcaccggcag agcgcgtttt gttacagcgc gtaaactgtg actttgaaaa 60
atttttgaac aatccgtaca ccaacttcag gagaaaaaca gtg agc aga atc tat 115
Val Ser Arg Ile Tyr
1 5
gac tgt gcc gac caa gac tcc cgt gca gca ggc cta aag gcg gct gtc 163
Asp Cys Ala Asp Gln Asp Ser Arg Ala Ala Gly Leu Lys Ala Ala Val
10 15 20
gat gca gtc aaa gcc ggt cag ctc gtt gtc ctt ccc acg gat acc ctt 211
Asp Ala Val Lys Ala Gly Gln Leu Val Val Leu Pro Thr Asp Thr Leu
25 30 35
tat gga ctc ggc tgc gac gct ttc aac aac gag gca gta gcc aac ctt 259
Tyr Gly Leu Gly Cys Asp Ala Phe Asn Asn Glu Ala Val Ala Asn Leu
40 45 50
ctg gcc acc aaa cac cgt ggc ccc gat atg ccc gtt cca gtg ctc gtc 307
Leu Ala Thr Lys His Arg Gly Pro Asp Met Pro Val Pro Val Leu Val
55 60 65
ggc agc tgg gac acc att caa gga ctt gtg cac tcc tat tct gcg cag 355
Gly Ser Trp Asp Thr Ile Gln Gly Leu Val His Ser Tyr Ser Ala Gln
70 75 80 85
gca aaa gcg ctt gtg gag gcg ttc tgg cct ggt gga ctg tcc atc atc 403
Ala Lys Ala Leu Val Glu Ala Phe Trp Pro Gly Gly Leu Ser Ile Ile
90 95 100
gtt ccg cag gca cca agc ctt ccg tgg aac ctt ggc gat acc cgt ggc 451
Val Pro Gln Ala Pro Ser Leu Pro Trp Asn Leu Gly Asp Thr Arg Gly
105 110 115
acc gta atg ctg cgc atg cca ctg cac cca gtt gcc att gaa ttg ctg 499
Thr Val Met Leu Arg Met Pro Leu His Pro Val Ala Ile Glu Leu Leu
120 125 130
cgc caa acc gga cca atg gct gtc tcc tcc gcc aac atc tcc gga cat 547
Arg Gln Thr Gly Pro Met Ala Val Ser Ser Ala Asn Ile Ser Gly His
135 140 145
act cct cca acc acc gtg ctg gag gct cgt cag cag ctc aac caa aat 595
Thr Pro Pro Thr Thr Val Leu Glu Ala Arg Gln Gln Leu Asn Gln Asn
150 155 160 165
gtc gct gtc tac ctc gat ggt ggc gaa tgc gcg ctg gcc acc cct tca 643
Val Ala Val Tyr Leu Asp Gly Gly Glu Cys Ala Leu Ala Thr Pro Ser
170 175 180
acc atc gtg gat att tca ggc ccc gca cca aag att ttg cgt gag ggt 691
Thr Ile Val Asp Ile Ser Gly Pro Ala Pro Lys Ile Leu Arg Glu Gly
185 190 195
gcc atc agc gca gaa cgc gtt ggc gaa gta ctt gga gtg tcg gca gaa 739
Ala Ile Ser Ala Glu Arg Val Gly Glu Val Leu Gly Val Ser Ala Glu
200 205 210
agc ctg cgc taaatgggag tcggtttcgc ggg 771
Ser Leu Arg
215
<210>18
<211>216
<212>PRT
<213>谷氨酸棒杆菌
<400>18
Val Ser Arg Ile Tyr Asp Cys Ala Asp Gln Asp Ser Arg Ala Ala Gly
1 5 10 15
Leu Lys Ala Ala Val Asp Ala Val Lys Ala Gly Gln Leu Val Val Leu
20 25 30
Pro Thr Asp Thr Leu Tyr Gly Leu Gly Cys Asp Ala Phe Asn Asn Glu
35 40 45
Ala Val Ala Asn Leu Leu Ala Thr Lys His Arg Gly Pro Asp Met Pro
50 55 60
Val Pro Val Leu Val Gly Ser Trp Asp Thr Ile Gln Gly Leu Val His
65 70 75 80
Ser Tyr Ser Ala Gln Ala Lys Ala Leu Val Glu Ala Phe Trp Pro Gly
85 90 95
Gly Leu Ser Ile Ile Val Pro Gln Ala Pro Ser Leu Pro Trp Asn Leu
100 105 110
Gly Asp Thr Arg Gly Thr Val Met Leu Arg Met Pro Leu His Pro Val
115 120 125
Ala Ile Glu Leu Leu Arg Gln Thr Gly Pro Met Ala Val Ser Ser Ala
130 135 140
Asn Ile Ser Gly His Thr Pro Pro Thr Thr Val Leu Glu Ala Arg Gln
145 150 155 160
Gln Leu Asn Gln Asn Val Ala Val Tyr Leu Asp Gly Gly Glu Cys Ala
165 170 175
Leu Ala Thr Pro Ser Thr Ile Val Asp Ile Ser Gly Pro Ala Pro Lys
180 185 190
Ile Leu Arg Glu Gly Ala Ile Ser Ala Glu Arg Val Gly Glu Val Leu
195 200 205
Gly Val Ser Ala Glu Ser Leu Arg
210 215
<210>19
<211>1026
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1003)
<223>RXC00657
<400>19
gtgcggatcg ggtatccgcg ctacacttag aggtgttaga gatcatgagt ttccacgaac 60
tgtaacgcag gattcaccaa tcaatgaaag gtcgaccgac atg agc act gaa gac 115
Met Ser Thr Glu Asp
1 5
att gtc gtc gta gca gta gat ggc tcg gac gcc tca aaa caa gct gtt 163
Ile Val Val Val Ala Val Asp Gly Ser Asp Ala Ser Lys Gln Ala Val
10 15 20
cgg tgg gct gca aat acc gcc aac aaa cgt ggc att cca ctt cgc ttg 211
Arg Trp Ala Ala Asn Thr Ala Asn Lys Arg Gly Ile Pro Leu Arg Leu
25 30 35
gct tcc agc tac acc atg cct cag ttc ctc tac gca gag gga atg gtt 259
Ala Ser Ser Tyr Thr Met Pro Gln Phe Leu Tyr Ala Glu Gly Met Val
40 45 50
cca cca caa gag ctt ttc gat gac ctc cag gcc gaa gcc ctg gaa aag 307
Pro Pro Gln Glu Leu Phe Asp Asp Leu Gln Ala Glu Ala Leu Glu Lys
55 60 65
att aac gaa gcc cgt gac atc gcc cat gag gta gcg cca gaa atc aag 355
Ile Asn Glu Ala Arg Asp Ile Ala His Glu Val Ala Pro Glu Ile Lys
70 75 80 85
atc ggg cac acc atc gct gaa ggc agt ccc atc gac atg ctg ttg gaa 403
Ile Gly His Thr Ile Ala Glu Gly Ser Pro Ile Asp Met Leu Leu Glu
90 95 100
atg tct ccc gat gcc aca atg atc gtc atg ggt tcc cgc gga ctc ggc 451
Met Ser Pro Asp Ala Thr Met Ile Val Met Gly Ser Arg Gly Leu Gly
105 110 115
gga ctc tcc gga atg gtc atg ggc tcc gtc tcc ggt gca gtg gtc agc 499
Gly Leu Ser Gly Met Val Met Gly Ser Val Ser Gly Ala Val Val Ser
120 125 130
cac gca aag tgt cca gtc gtt gtt gtc cgt gaa gac agc gca gtc aac 547
His Ala Lys Cys Pro Val Val Val Val Arg Glu Asp Ser Ala Val Asn
135 140 145
gaa gac agc aag tac ggc cca gtc gtc gtc ggt gtg gat ggc tcc gaa 595
Glu Asp Ser Lys Tyr Gly Pro Val Val Val Gly Val Asp Gly Ser Glu
150 155 160 165
gtc tcc caa cag gca acc gaa tac gca ttt gcg gaa gct gaa gct cgt 643
Val Ser Gln Gln Ala Thr Glu Tyr Ala Phe Ala Glu Ala Glu Ala Arg
170 175 180
ggc gcc gaa ctc gtt gca gtt cac acc tgg atg gac atg cag gta cag 691
Gly Ala Glu Leu Val Ala Val His Thr Trp Met Asp Met Gln Val Gln
185 190 195
gca tca ctt gca ggt ctt gca gct gct caa cag cag tgg gat gaa gtg 739
Ala Ser Leu Ala Gly Leu Ala Ala Ala Gln Gln Gln Trp Asp Glu Val
200 205 210
gaa cgt cag caa acc gac atg ctg atc gaa cgc ctc gca cca ctg gtg 787
Glu Arg Gln Gln Thr Asp Met Leu Ile Glu Arg Leu Ala Pro Leu Val
215 220 225
gaa aag tac cca agt gta acc gtc aag aag atc atc acc cgt gac cgc 835
Glu Lys Tyr Pro Ser Val Thr Val Lys Lys Ile Ile Thr Arg Asp Arg
230 235 240 245
cca gtt cgc gca ctt gca gaa gca tct gaa aac gcg cag ctc cta gtc 883
Pro Val Arg Ala Leu Ala Glu Ala Ser Glu Asn Ala Gln Leu Leu Val
250 255 260
gtt ggt tcc cat ggt cgt ggc gga ttt aag ggc atg ctc ctt ggc tcc 931
Val Gly Ser His Gly Arg Gly Gly Phe Lys Gly Met Leu Leu Gly Ser
265 270 275
acc tcc cgc gca ctg ctg caa tcc gca ccg tgc cca atg atg gtg gtt 979
Thr Ser Arg Ala Leu Leu Gln Ser Ala Pro Cys Pro Met Met Val Val
280 285 290
cgc cca cct gag aag att aag aag tagtttcttt taagtttcga tgc 1026
Arg Pro Pro Glu Lys Ile Lys Lys
295 300
<210>20
<211>301
<212>PRT
<213>谷氨酸棒杆菌
<400>20
Met Ser Thr Glu Asp Ile Val Val Val Ala Val Asp Gly Ser Asp Ala
1 5 10 15
Ser Lys Gln Ala Val Arg Trp Ala Ala Asn Thr Ala Asn Lys Arg Gly
20 25 30
Ile Pro Leu Arg Leu Ala Ser Ser Tyr Thr Met Pro Gln Phe Leu Tyr
35 40 45
Ala Glu Gly Met Val Pro Pro Gln Glu Leu Phe Asp Asp Leu Gln Ala
50 55 60
Glu Ala Leu Glu Lys Ile Asn Glu Ala Arg Asp Ile Ala His Glu Val
65 70 75 80
Ala Pro Glu Ile Lys Ile Gly His Thr Ile Ala Glu Gly Ser Pro Ile
85 90 95
Asp Met Leu Leu Glu Met Ser Pro Asp Ala Thr Met Ile Val Met Gly
100 105 110
Ser Arg Gly Leu Gly Gly Leu Ser Gly Met Val Met Gly Ser Val Ser
115 120 125
Gly Ala Val Val Ser His Ala Lys Cys Pro Val Val Val Val Arg Glu
130 135 140
Asp Ser Ala Val Asn Glu Asp Ser Lys Tyr Gly Pro Val Val Val Gly
145 150 155 160
Val Asp Gly Ser Glu Val Ser Gln Gln Ala Thr Glu Tyr Ala Phe Ala
165 170 175
Glu Ala Glu Ala Arg Gly Ala Glu Leu Val Ala Val His Thr Trp Met
180 185 190
Asp Met Gln Val Gln Ala Ser Leu Ala Gly Leu Ala Ala Ala Gln Gln
195 200 205
Gln Trp Asp Glu Val Glu Arg Gln Gln Thr Asp Met Leu Ile Glu Arg
210 215 220
Leu Ala Pro Leu Val Glu Lys Tyr Pro Ser Val Thr Val Lys Lys Ile
225 230 235 240
Ile Thr Arg Asp Arg Pro Val Arg Ala Leu Ala Glu Ala Ser Glu Asn
245 250 255
Ala Gln Leu Leu Val Val Gly Ser His Gly Arg Gly Gly Phe Lys Gly
260 265 270
Met Leu Leu Gly Ser Thr Ser Arg Ala Leu Leu Gln Ser Ala Pro Cys
275 280 285
Pro Met Met Val Val Arg Pro Pro Glu Lys Ile Lys Lys
290 295 300
<210>21
<211>1059
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1036)
<223>RXC00552
<400>21
ccgccaacaa ggcagcaaag ctcgatccaa ttgacgcctt gcgttatgag taaaagcctc 60
gtttttaagg tagccacaca tcgcactaga ctgaagaact gtg gct acc tca aaa 115
Val Ala Thr Ser Lys
1 5
att ctt ctt tat tac gca ttc acc ccg ctc tct gac cct aaa gcg gtt 163
Ile Leu Leu Tyr Tyr Ala Phe Thr Pro Leu Ser Asp Pro Lys Ala Val
10 15 20
cag ctg tgg cag cgt gag ctc tgc gag tca ctg aat ctt cgt ggc cgc 211
Gln Leu Trp Gln Arg Glu Leu Cys Glu Ser Leu Asn Leu Arg Gly Arg
25 30 35
atc ctg atc tcc act cac ggc atc aat gga acc gtg ggc gga gat att 259
Ile Leu Ile Ser Thr His Gly Ile Asn Gly Thr Val Gly Gly Asp Ile
40 45 50
gat gat tgc aag gcg tac att aaa aag acc cgc gag tac cca ggt ttc 307
Asp Asp Cys Lys Ala Tyr Ile Lys Lys Thr Arg Glu Tyr Pro Gly Phe
55 60 65
aac cgc atg cag ttt aag tgg tcc gag ggt ggc gct gag gat ttc cca 355
Asn Arg Met Gln Phe Lys Trp Ser Glu Gly Gly Ala Glu Asp Phe Pro
70 75 80 85
aag ctc agt gtc aaa gtc cgc gat gag atc gtt gcc ttc ggc gct cca 403
Lys Leu Ser Val Lys Val Arg Asp Glu Ile Val Ala Phe Gly Ala Pro
90 95 100
gat gag ctc aaa gtg gat gaa aac ggc gtc gtc ggt ggc ggc gtt cac 451
Asp Glu Leu Lys Val Asp Glu Asn Gly Val Val Gly Gly Gly Val His
105 110 115
ctg aaa cca cag cag gtc aat gag ctt gtg gaa gcc cgt ggc gat gaa 499
Leu Lys Pro Gln Gln Val Asn Glu Leu Val Glu Ala Arg Gly Asp Glu
120 125 130
gtt gtg ttc ttt gac ggc cgc aac gca atg gaa gcc cag atc ggc aag 547
Val Val Phe Phe Asp Gly Arg Asn Ala Met Glu Ala Gln Ile Gly Lys
135 140 145
ttc aag gac gct gtt gtc cct gac gta gaa acc act cat gat ttc atc 595
Phe Lys Asp Ala Val Val Pro Asp Val Glu Thr Thr His Asp Phe Ile
150 155 160 165
gca gaa att gag tct gga aaa tac gac gat ctc aaa gac aag cct gtg 643
Ala Glu Ile Glu Ser Gly Lys Tyr Asp Asp Leu Lys Asp Lys Pro Val
170 175 180
gtc acc tac tgc acc ggc gga att cgt tgt gag atc ctg agt tca ctc 691
Val Thr Tyr Cys Thr Gly Gly Ile Arg Cys Glu Ile Leu Ser Ser Leu
185 190 195
atg atc aac cgt ggt ttc aaa gag gtc tac caa atc gat ggc ggc atc 739
Met Ile Asn Arg Gly Phe Lys Glu Val Tyr Gln Ile Asp Gly Gly Ile
200 205 210
gtt cgc tac ggc gag cag ttt ggc aac aag ggc ctg tgg gaa ggc tcc 787
Val Arg Tyr Gly Glu Gln Phe Gly Asn Lys Gly Leu Trp Glu Gly Ser
215 220 225
ctc tac gtt ttc gat aag cgc atg cat atg gaa ttc ggc gag gat tac 835
Leu Tyr Val Phe Asp Lys Arg Met His Met Glu Phe Gly Glu Asp Tyr
230 235 240 245
aaa gag gtc gga cac tgc atc cat tgc gat act ccc acc aac aaa ttt 883
Lys Glu Val Gly His Cys Ile His Cys Asp Thr Pro Thr Asn Lys Phe
250 255 260
gag cac tgc ctc aac gaa gat gat tgc cgc gag ctc gtg ttg atg tgc 931
Glu His Cys Leu Asn Glu Asp Asp Cys Arg Glu Leu Val Leu Met Cys
265 270 275
cct gat tgc ttc gcc aat gtt gag acc cgt cat tgc aag cgc gaa cgc 979
Pro Asp Cys Phe Ala Asn Val Glu Thr Arg His Cys Lys Arg Glu Arg
280 285 290
tgt gca gca att gct gcg gat ttc gct gag caa gga att gat ccg ctc 1027
Cys Ala Ala Ile Ala Ala Asp Phe Ala Glu Gln Gly Ile Asp Pro Leu
295 300 305
gtt act tct taaaaagggt atggtggctg ggt 1059
Val Thr Ser
310
<210>22
<211>312
<212>PRT
<213>谷氨酸棒杆菌
<400>22
Val Ala Thr Ser Lys Ile Leu Leu Tyr Tyr Ala Phe Thr Pro Leu Ser
1 5 10 15
Asp Pro Lys Ala Val Gln Leu Trp Gln Arg Glu Leu Cys Glu Ser Leu
20 25 30
Asn Leu Arg Gly Arg Ile Leu Ile Ser Thr His Gly Ile Asn Gly Thr
35 40 45
Val Gly Gly Asp Ile Asp Asp Cys Lys Ala Tyr Ile Lys Lys Thr Arg
50 55 60
Glu Tyr Pro Gly Phe Asn Arg Met Gln Phe Lys Trp Ser Glu Gly Gly
65 70 75 80
Ala Glu Asp Phe Pro Lys Leu Ser Val Lys Val Arg Asp Glu Ile Val
85 90 95
Ala Phe Gly Ala Pro Asp Glu Leu Lys Val Asp Glu Asn Gly Val Val
100 105 110
Gly Gly Gly Val His Leu Lys Pro Gln Gln Val Asn Glu Leu Val Glu
115 120 125
Ala Arg Gly Asp Glu Val Val Phe Phe Asp Gly Arg Asn Ala Met Glu
130 135 140
Ala Gln Ile Gly Lys Phe Lys Asp Ala Val Val Pro Asp Val Glu Thr
145 150 155 160
Thr His Asp Phe Ile Ala Glu Ile Glu Ser Gly Lys Tyr Asp Asp Leu
165 170 175
Lys Asp Lys Pro Val Val Thr Tyr Cys Thr Gly Gly Ile Arg Cys Glu
180 185 190
Ile Leu Ser Ser Leu Met Ile Asn Arg Gly Phe Lys Glu Val Tyr Gln
195 200 205
Ile Asp Gly Gly Ile Val Arg Tyr Gly Glu Gln Phe Gly Asn Lys Gly
210 215 220
Leu Trp Glu Gly Ser Leu Tyr Val Phe Asp Lys Arg Met His Met Glu
225 230 235 240
Phe Gly Glu Asp Tyr Lys Glu Val Gly His Cys Ile His Cys Asp Thr
245 250 255
Pro Thr Asn Lys Phe Glu His Cys Leu Asn Glu Asp Asp Cys Arg Glu
260 265 270
Leu Val Leu Met Cys Pro Asp Cys Phe Ala Asn Val Glu Thr Arg His
275 280 285
Cys Lys Arg Glu Arg Cys Ala Ala Ile Ala Ala Asp Phe Ala Glu Gln
290 295 300
Gly Ile Asp Pro Leu Val Thr Ser
305 310
<210>23
<211>1386
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1363)
<223>RXA00534
<400>23
ctgtgcagaa agaaaacact cctctggcta ggtagacaca gtttataaag gtagagttga 60
gcgggtaact gtcagcacgt agatcgaaag gtgcacaaag gtg gcc ctg gtc gta 115
Val Ala Leu Val Val
1 5
cag aaa tat ggc ggt tcc tcg ctt gag agt gcg gaa cgc att aga aac 163
Gln Lys Tyr Gly Gly Ser Ser Leu Glu Ser Ala Glu Arg Ile Arg Asn
10 15 20
gtc gct gaa cgg atc gtt gcc acc aag aag gct gga aat gat gtc gtg 211
Val Ala Glu Arg Ile Val Ala Thr Lys Lys Ala Gly Asn Asp Val Val
25 30 35
gtt gtc tgc tcc gca atg gga gac acc acg gat gaa ctt cta gaa ctt 259
Val Val Cys Ser Ala Met Gly Asp Thr Thr Asp Glu Leu Leu Glu Leu
40 45 50
gca gcg gca gtg aat ccc gtt ccg cca gct cgt gaa atg gat atg ctc 307
Ala Ala Ala Val Asn Pro Val Pro Pro Ala Arg Glu Met Asp Met Leu
55 60 65
ctg act gct ggt gag cgt att tct aac gct ctc gtc gcc atg gct att 355
Leu Thr Ala Gly Glu Arg Ile Ser Asn Ala Leu Val Ala Met Ala Ile
70 75 80 85
gag tcc ctt ggc gca gaa gcc caa tct ttc acg ggc tct cag gct ggt 403
Glu Ser Leu Gly Ala Glu Ala Gln Ser Phe Thr Gly Ser Gln Ala Gly
90 95 100
gtg ctc acc acc gag cgc cac gga aac gca cgc att gtt gat gtc act 451
Val Leu Thr Thr Glu Arg His Gly Asn Ala Arg Ile Val Asp Val Thr
105 110 115
cca ggt cgt gtg cgt gaa gca ctc gat gag ggc aag atc tgc att gtt 499
Pro Gly Arg Val Arg Glu Ala Leu Asp Glu Gly Lys Ile Cys Ile Val
120 125 130
gct ggt ttc cag ggt gtt aat aaa gaa acc cgc gat gtc acc acg ttg 547
Ala Gly Phe Gln Gly Val Asn Lys Glu Thr Arg Asp Val Thr Thr Leu
135 140 145
ggt cgt ggt ggt tct gac acc act gca gtt gcg ttg gca gct gct ttg 595
Gly Arg Gly Gly Ser Asp Thr Thr Ala Val Ala Leu Ala Ala Ala Leu
150 155 160 165
aac gct gat gtg tgt gag att tac tcg gac gtt gac ggt gtg tat acc 643
Asn Ala Asp Val Cys Glu Ile Tyr Ser Asp Val Asp Gly Val Tyr Thr
170 175 180
gct gac ccg cgc atc gtt cct aat gca cag aag ctg gaa aag ctc agc 691
Ala Asp Pro Arg Ile Val Pro Asn Ala Gln Lys Leu Glu Lys Leu Ser
185 190 195
ttc gaa gaa atg ctg gaa ctt gct gct gtt ggc tcc aag att ttg gtg 739
Phe Glu Glu Met Leu Glu Leu Ala Ala Val Gly Ser Lys Ile Leu Val
200 205 210
ctg cgc agt gtt gaa tac gct cgt gca ttc aat gtg cca ctt cgc gta 787
Leu Arg Ser Val Glu Tyr Ala Arg Ala Phe Asn Val Pro Leu Arg Val
215 220 225
cgc tcg tct tat agt aat gat ccc ggc act ttg att gcc ggc tct atg 835
Arg Ser Ser Tyr Ser Asn Asp Pro Gly Thr Leu Ile Ala Gly Ser Met
230 235 240 245
gag gat att cct gtg gaa gaa gca gtc ctt acc ggt gtc gca acc gac 883
Glu Asp Ile Pro Val Glu Glu Ala Val Leu Thr Gly Val Ala Thr Asp
250 255 260
aag tcc gaa gcc aaa gta acc gtt ctg ggt att tcc gat aag cca ggc 931
Lys Ser Glu Ala Lys Val Thr Val Leu Gly Ile Ser Asp Lys Pro Gly
265 270 275
gag gct gcg aag gtt ttc cgt gcg ttg gct gat gca gaa atc aac att 979
Glu Ala Ala Lys Val Phe Arg Ala Leu Ala Asp Ala Glu Ile Asn Ile
280 285 290
gac atg gtt ctg cag aac gtc tct tct gta gaa gac ggc acc acc gac 1027
Asp Met Val Leu Gln Asn Val Ser Ser Val Glu Asp Gly Thr Thr Asp
295 300 305
atc acc ttc acc tgc cct cgt tcc gac ggc cgc cgc gcg atg gag atc 1075
Ile Thr Phe Thr Cys Pro Arg Ser Asp Gly Arg Arg Ala Met Glu Ile
310 315 320 325
ttg aag aag ctt cag gtt cag ggc aac tgg acc aat gtg ctt tac gac 1123
Leu Lys Lys Leu Gln Val Gln Gly Asn Trp Thr Asn Val Leu Tyr Asp
330 335 340
gac cag gtc ggc aaa gtc tcc ctc gtg ggt gct ggc atg aag tct cac 1171
Asp Gln Val Gly Lys Val Ser Leu Val Gly Ala Gly Met Lys Ser His
345 350 355
cca ggt gtt acc gca gag ttc atg gaa gct ctg cgc gat gtc aac gtg 1219
Pro Gly Val Thr Ala Glu Phe Met Glu Ala Leu Arg Asp Val Asn Val
360 365 370
aac atc gaa ttg att tcc acc tct gag att cgt att tcc gtg ctg atc 1267
Asn Ile Glu Leu Ile Ser Thr Ser Glu Ile Arg Ile Ser Val Leu Ile
375 380 385
cgt gaa gat gat ctg gat gct gct gca cgt gca ttg cat gag cag ttc 1315
Arg Glu Asp Asp Leu Asp Ala Ala Ala Arg Ala Leu His Glu Gln Phe
390 395 400 405
cag ctg ggc ggc gaa gac gaa gcc gtc gtt tat gca ggc acc gga cgc 1363
Gln Leu Gly Gly Glu Asp Glu Ala Val Val Tyr Ala Gly Thr Gly Arg
410 415 420
taaagtttta aaggagtagt ttt 1386
<210>24
<211>421
<212>PRT
<213>谷氨酸棒杆菌
<400>24
Val Ala Leu Val Val Gln Lys Tyr Gly Gly Ser Ser Leu Glu Ser Ala
1 5 10 15
Glu Arg Ile Arg Asn Val Ala Glu Arg Ile Val Ala Thr Lys Lys Ala
20 25 30
Gly Asn Asp Val Val Val Val Cys Ser Ala Met Gly Asp Thr Thr Asp
35 40 45
Glu Leu Leu Glu Leu Ala Ala Ala Val Asn Pro Val Pro Pro Ala Arg
50 55 60
Glu Met Asp Met Leu Leu Thr Ala Gly Glu Arg Ile Ser Asn Ala Leu
65 70 75 80
Val Ala Met Ala Ile Glu Ser Leu Gly Ala Glu Ala Gln Ser Phe Thr
85 90 95
Gly Ser Gln Ala Gly Val Leu Thr Thr Glu Arg His Gly Asn Ala Arg
100 105 110
Ile Val Asp Val Thr Pro Gly Arg Val Arg Glu Ala Leu Asp Glu Gly
115 120 125
Lys Ile Cys Ile Val Ala Gly Phe Gln Gly Val Asn Lys Glu Thr Arg
130 135 140
Asp Val Thr Thr Leu Gly Arg Gly Gly Ser Asp Thr Thr Ala Val Ala
145 150 155 160
Leu Ala Ala Ala Leu Asn Ala Asp Val Cys Glu Ile Tyr Ser Asp Val
165 170 175
Asp Gly Val Tyr Thr Ala Asp Pro Arg Ile Val Pro Asn Ala Gln Lys
180 185 190
Leu Glu Lys Leu Ser Phe Glu Glu Met Leu Glu Leu Ala Ala Val Gly
195 200 205
Ser Lys Ile Leu Val Leu Arg Ser Val Glu Tyr Ala Arg Ala Phe Asn
210 215 220
Val Pro Leu Arg Val Arg Ser Ser Tyr Ser Asn Asp Pro Gly Thr Leu
225 230 235 240
Ile Ala Gly Ser Met Glu Asp Ile Pro Val Glu Glu Ala Val Leu Thr
245 250 255
Gly Val Ala Thr Asp Lys Ser Glu Ala Lys Val Thr Val Leu Gly Ile
260 265 270
Ser Asp Lys Pro Gly Glu Ala Ala Lys Val Phe Arg Ala Leu Ala Asp
275 280 285
Ala Glu Ile Asn Ile Asp Met Val Leu Gln Asn Val Ser Ser Val Glu
290 295 300
Asp Gly Thr Thr Asp Ile Thr Phe Thr Cys Pro Arg Ser Asp Gly Arg
305 310 315 320
Arg Ala Met Glu Ile Leu Lys Lys Leu Gln Val Gln Gly Asn Trp Thr
325 330 335
Asn Val Leu Tyr Asp Asp Gln Val Gly Lys Val Ser Leu Val Gly Ala
340 345 350
Gly Met Lys Ser His Pro Gly Val Thr Ala Glu Phe Met Glu Ala Leu
355 360 365
Arg Asp Val Asn Val Asn Ile Glu Leu Ile Ser Thr Ser Glu Ile Arg
370 375 380
Ile Ser Val Leu Ile Arg Glu Asp Asp Leu Asp Ala Ala Ala Arg Ala
385 390 395 400
Leu His Glu Gln Phe Gln Leu Gly Gly Glu Asp Glu Ala Val Val Tyr
405 410 415
Ala Gly Thr Gly Arg
420
<210>25
<211>1155
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1132)
<223>RXA00533
<400>25
ctgcacgtgc attgcatgag cagttccagc tgggcggcga agacgaagcc gtcgtttatg 60
caggcaccgg acgctaaagt tttaaaggag tagttttaca atg acc acc atc gca 115
Met Thr Thr Ile Ala
1 5
gtt gtt ggt gca acc ggc cag gtc ggc cag gtt atg cgc acc ctt ttg 163
Val Val Gly Ala Thr Gly Gln Val Gly Gln Val Met Arg Thr Leu Leu
10 15 20
gaa gag cgc aat ttc cca gct gac act gtt cgt ttc ttt gct tcc cca 211
Glu Glu Arg Asn Phe Pro Ala Asp Thr Val Arg Phe Phe Ala Ser Pro
25 30 35
cgt tcc gca ggc cgt aag att gaa ttc cgt ggc acg gaa atc gag gta 259
Arg Ser Ala Gly Arg Lys Ile Glu Phe Arg Gly Thr Glu Ile Glu Val
40 45 50
gaa gac att act cag gca acc gag gag tcc ctc aag gac atc gac gtt 307
Glu Asp Ile Thr Gln Ala Thr Glu Glu Ser Leu Lys Asp Ile Asp Val
55 60 65
gcg ttg ttc tcc gct gga ggc acc gct tcc aag cag tac gct cca ctg 355
Ala Leu Phe Ser Ala Gly Gly Thr Ala Ser Lys Gln Tyr Ala Pro Leu
70 75 80 85
ttc gct gct gca ggc gcg act gtt gtg gat aac tct tct gct tgg cgc 403
Phe Ala Ala Ala Gly Ala Thr Val Val Asp Asn Ser Ser Ala Trp Arg
90 95 100
aag gac gac gag gtt cca cta atc gtc tct gag gtg aac cct tcc gac 451
Lys Asp Asp Glu Val Pro Leu Ile Val Ser Glu Val Asn Pro Ser Asp
105 110 115
aag gat tcc ctg gtc aag ggc att att gcg aac cct aac tgc acc acc 499
Lys Asp Ser Leu Val Lys Gly Ile Ile Ala Asn Pro Asn Cys Thr Thr
120 125 130
atg gct gcg atg cca gtg ctg aag cca ctt cac gat gcc gct ggt ctt 547
Met Ala Ala Met Pro Val Leu Lys Pro Leu His Asp Ala Ala Gly Leu
135 140 145
gta aag ctt cac gtt tcc tct tac cag gct gtt tcc ggt tct ggt ctt 595
Val Lys Leu His Val Ser Ser Tyr Gln Ala Val Ser Gly Ser Gly Leu
150 155 160 165
gca ggt gtg gaa acc ttg gca aag cag gtt gct gca gtt gga gac cac 643
Ala Gly Val Glu Thr Leu Ala Lys Gln Val Ala Ala Val Gly Asp His
170 175 180
aac gtt gag ttc gtc cat gat gga cag gct gct gac gca ggc gat gtc 691
Asn Val Glu Phe Val His Asp Gly Gln Ala Ala Asp Ala Gly Asp Val
185 190 195
gga cct tat gtt tca cca atc gct tac aac gtg ctg cca ttc gcc gga 739
Gly Pro Tyr Val Ser Pro Ile Ala Tyr Asn Val Leu Pro Phe Ala Gly
200 205 210
aac ctc gtc gat gac ggc acc ttc gaa acc gat gaa gag cag aag ctg 787
Asn Leu Val Asp Asp Gly Thr Phe Glu Thr Asp Glu Glu Gln Lys Leu
215 220 225
cgc aac gaa tcc cgc aag att ctc ggt ctc cca gac ctc aag gtc tca 835
Arg Asn Glu Ser Arg Lys Ile Leu Gly Leu Pro Asp Leu Lys Val Ser
230 235 240 245
ggc acc tgc gtc cgc gtg ccg gtt ttc acc ggc cac acg ctg acc att 883
Gly Thr Cys Val Arg Val Pro Val Phe Thr Gly His Thr Leu Thr Ile
250 255 260
cac gcc gaa ttc gac aag gca atc acc gtg gac cag gcg cag gag atc 931
His Ala Glu Phe Asp Lys Ala Ile Thr Val Asp Gln Ala Gln Glu Ile
265 270 275
ttg ggt gcc gct tca ggc gtc aag ctt gtc gac gtc cca acc cca ctt 979
Leu Gly Ala Ala Ser Gly Val Lys Leu Val Asp Val Pro Thr Pro Leu
280 285 290
gca gct gcc ggc att gac gaa tcc ctc gtt gga cgc atc cgt cag gac 1027
Ala Ala Ala Gly Ile Asp Glu Ser Leu Val Gly Arg Ile Arg Gln Asp
295 300 305
tcc act gtc gac gat aac cgc ggt ctg gtt ctc gtc gta tct ggc gac 1075
Ser Thr Val Asp Asp Asn Arg Gly Leu Val Leu Val Val Ser Gly Asp
310 315 320 325
aac ctc cgc aag ggt gct gcg cta aac acc atc cag atc gct gag ctg 1123
Asn Leu Arg Lys Gly Ala Ala Leu Asn Thr Ile Gln Ile Ala Glu Lau
330 335 340
ctg gtt aag taaaaacccg ccattaaaaa ctc 1155
Leu Val Lys
<210>26
<211>344
<212>PRT
<213>谷氨酸棒杆菌
<400>26
Met Thr Thr Ile Ala Val Val Gly Ala Thr Gly Gln Val Gly Gln Val
1 5 10 15
Met Arg Thr Leu Leu Glu Glu Arg Asn Phe Pro Ala Asp Thr Val Arg
20 25 30
Phe Phe Ala Ser Pro Arg Ser Ala Gly Arg Lys Ile Glu Phe Arg Gly
35 40 45
Thr Glu Ile Glu Val Glu Asp Ile Thr Gln Ala Thr Glu Glu Ser Leu
50 55 60
Lys Asp Ile Asp Val Ala Leu Phe Ser Ala Gly Gly Thr Ala Ser Lys
65 70 75 80
Gln Tyr Ala Pro Leu Phe Ala Ala Ala Gly Ala Thr Val Val Asp Asn
85 90 95
Ser Ser Ala Trp Arg Lys Asp Asp Glu Val Pro Leu Ile Val Ser Glu
100 105 110
Val Asn Pro Ser Asp Lys Asp Ser Leu Val Lys Gly Ile Ile Ala Asn
115 120 125
Pro Asn Cys Thr Thr Met Ala Ala Met Pro Val Leu Lys Pro Leu His
130 135 140
Asp Ala Ala Gly Leu Val Lys Leu His Val Ser Ser Tyr Gln Ala Val
145 150 155 160
Ser Gly Ser Gly Leu Ala Gly Val Glu Thr Leu Ala Lys Gln Val Ala
165 170 175
Ala Val Gly Asp His Asn Val Glu Phe Val His Asp Gly Gln Ala Ala
180 185 190
Asp Ala Gly Asp Val Gly Pro Tyr Val Ser Pro Ile Ala Tyr Asn Val
195 200 205
Leu Pro Phe Ala Gly Asn Leu Val Asp Asp Gly Thr Phe Glu Thr Asp
210 215 220
Glu Glu Gln Lys Leu Arg Asn Glu Ser Arg Lys Ile Leu Gly Leu Pro
225 230 235 240
Asp Leu Lys Val Ser Gly Thr Cys Val Arg Val Pro Val Phe Thr Gly
245 250 255
His Thr Leu Thr Ile His Ala Glu Phe Asp Lys Ala Ile Thr Val Asp
260 265 270
Gln Ala Gln Glu Ile Leu Gly Ala Ala Ser Gly Val Lys Leu Val Asp
275 280 285
Val Pro Thr Pro Leu Ala Ala Ala Gly Ile Asp Glu Ser Leu Val Gly
290 295 300
Arg Ile Arg Gln Asp Ser Thr Val Asp Asp Asn Arg Gly Leu Val Leu
305 310 315 320
Val Val Ser Gly Asp Asn Leu Arg Lys Gly Ala Ala Leu Asn Thr Ile
325 330 335
Gln Ile Ala Glu Leu Leu Val Lys
340
<210>27
<211>608
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(69)..(608)
<223>RXA02843
<400>27
cccattgcgc ggaggtcgca ccccttccga cttgaactga taggccgata gaaattattc 60
tggacgtc atg act act gct tcc gca acc gga att gca aca ctg acc tcc 110
Met Thr Thr Ala Ser Ala Thr Gly Ile Ala Thr Leu Thr Ser
1 5 10
acc ggc gac gtc ctg gac gtg tgg tat cca gaa atc ggg tcc acc gac 158
Thr Gly Asp Val Leu Asp Val Trp Tyr Pro Glu Ile Gly Ser Thr Asp
15 20 25 30
cag tcc gcg ctc aca cct cta gaa ggc gtc gat gaa gat cga aac gtc 206
Gln Ser Ala Leu Thr Pro Leu Glu Gly Val Asp Glu Asp Arg Asn Val
35 40 45
acc cgc aaa atc gtg acg aca act atc gac acc gac gca gcc ccc acc 254
Thr Arg Lys Ile Val Thr Thr Thr Ile Asp Thr Asp Ala Ala Pro Thr
50 55 60
gac acc tac gat gca tgg ctg cgc ctt cac ctc ctc tcc cac cgc gtt 302
Asp Thr Tyr Asp Ala Trp Leu Arg Leu His Leu Leu Ser His Arg Val
65 70 75
ttc cgc cct cac acc atc aac cta gac ggc att ttc ggc ctc ctc aac 350
Phe Arg Pro His Thr Ile Asn Leu Asp Gly Ile Phe Gly Leu Leu Asn
80 85 90
aat gtc gtg tgg acc aac ttc gga ccg tgc gca gtt gac ggt ttc gca 398
Asn Val Val Trp Thr Asn Phe Gly Pro Cys Ala Val Asp Gly Phe Ala
95 100 105 110
ctc acc cgc gcg cgc ctg tca cgc cga ggc caa gtt acg gtt tat agc 446
Leu Thr Arg Ala Arg Leu Ser Arg Arg Gly Gln Val Thr Val Tyr Ser
115 120 125
gtc gac aag ttc cca cgc atg gtc gac tat gtg gtt ccc tcg ggc gtg 494
Val Asp Lys Phe Pro Arg Met Val Asp Tyr Val Val Pro Ser Gly Val
130 135 140
cgc atc ggt gac gcc gac cgc gtc cga ctt ggc gcg tac ctg gca gat 542
Arg Ile Gly Asp Ala Asp Arg Val Arg Leu Gly Ala Tyr Leu Ala Asp
145 150 155
ggc acc acc gtg atg cat gag ggc ttc gtg aac ttc aac gct ggc acg 590
Gly Thr Thr Val Met His Glu Gly Phe Val Asn Phe Asn Ala Gly Thr
160 165 170
ctc ggc gct tcc atg gtt 608
Leu Gly Ala Ser Met Val
175 180
<210>28
<211>180
<212>PRT
<213>谷氨酸棒杆菌
<400>28
Met Thr Thr Ala Ser Ala Thr Gly Ile Ala Thr Leu Thr Ser Thr Gly
1 5 10 15
Asp Val Leu Asp Val Trp Tyr Pro Glu Ile Gly Ser Thr Asp Gln Ser
20 25 30
Ala Leu Thr Pro Leu Glu Gly Val Asp Glu Asp Arg Asn Val Thr Arg
35 40 45
Lys Ile Val Thr Thr Thr Ile Asp Thr Asp Ala Ala Pro Thr Asp Thr
50 55 60
Tyr Asp Ala Trp Leu Arg Leu His Leu Leu Ser His Arg Val Phe Arg
65 70 75 80
Pro His Thr Ile Asn Leu Asp Gly Ile Phe Gly Leu Leu Asn Asn Val
85 90 95
Val Trp Thr Asn Phe Gly Pro Cys Ala Val Asp Gly Phe Ala Leu Thr
100 105 110
Arg Ala Arg Leu Ser Arg Arg Gly Gln Val Thr Val Tyr Ser Val Asp
115 120 125
Lys Phe Pro Arg Met Val Asp Tyr Val Val Pro Ser Gly Val Arg Ile
130 135 140
Gly Asp Ala Asp Arg Val Arg Leu Gly Ala Tyr Leu Ala Asp Gly Thr
145 150 155 160
Thr Val Met His Glu Gly Phe Val Asn Phe Asn Ala Gly Thr Leu Gly
165 170 175
Ala Ser Met Val
180
<210>29
<211>1230
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1207)
<223>RXA02022
<400>29
tatttgcgat tccaactgct tgggctccgc gaatgttttc actcattttt taatcgaccg 60
cttccatcat gttttaacta aggtttgtag gcttaaacct gtg aac tct gaa ctc 115
Val Asn Ser Glu Leu
1 5
aaa cca gga tta gat ctc ctc ggc gac cca att gtc ctt act caa cgt 163
Lys Pro Gly Leu Asp Leu Leu Gly Asp Pro Ile Val Leu Thr Gln Arg
10 15 20
ttg gta gat ata ccg agt ccg tcg ggt cag gaa aag cag att gct gat 211
Leu Val Asp Ile Pro Ser Pro Ser Gly Gln Glu Lys Gln Ile Ala Asp
25 30 35
gaa att gaa gat gcc ctt cgg aac ctt aat cta cct ggt gta gag gtc 259
Glu Ile Glu Asp Ala Leu Arg Asn Leu Asn Leu Pro Gly Val Glu Val
40 45 50
ttc cgc ttc aac aac aac gtt ctt gct cgc acg aac agg gga ttg gcc 307
Phe Arg Phe Asn Asn Asn Val Leu Ala Arg Thr Asn Arg Gly Leu Ala
55 60 65
tcg agg gtc atg ctt gct ggt cat atc gat aca gtg ccg atc gcg gac 355
Ser Arg Val Met Leu Ala Gly His Ile Asp Thr Val Pro Ile Ala Asp
70 75 80 85
aat ctg cca agc cgt gtg gaa gac ggc atc atg tat ggc tgt ggc acc 403
Asn Leu Pro Ser Arg Val Glu Asp Gly Ile Met Tyr Gly Cys Gly Thr
90 95 100
gtc gat atg aaa tct ggg ttg gcg gtg tat ttg cat act ttt gcc acc 451
Val Asp Met Lys Ser Gly Leu Ala Val Tyr Leu His Thr Phe Ala Thr
105 110 115
ttg gcc acg tcg act gag ctt aaa cat gat ctg acg ctg att gcg tat 499
Leu Ala Thr Ser Thr Glu Leu Lys His Asp Leu Thr Leu Ile Ala Tyr
120 125 130
gag tgc gag gaa gtt gct gat cac ctc aat ggt ttg ggc cac att cgc 547
Glu Cys Glu Glu Val Ala Asp His Leu Asn Gly Leu Gly His Ile Arg
135 140 145
gat gag cat ccg gag tgg ttg gcg gct gat ttg gcg ttg ttg ggt gag 595
Asp Glu His Pro Glu Trp Leu Ala Ala Asp Leu Ala Leu Leu Gly Glu
150 155 160 165
cct act ggc ggc tgg att gag gcg ggc tgc cag ggc aat ctg cgc atc 643
Pro Thr Gly Gly Trp Ile Glu Ala Gly Cys Gln Gly Asn Leu Arg Ile
170 175 180
aag gtg acg gcg cat ggt gtg cgt gcc cat tcg gcg aga agc tgg ttg 691
Lys Val Thr Ala His Gly Val Arg Ala His Ser Ala Arg Ser Trp Leu
185 190 195
ggt gat aat gcg atg cat aag ttg tcg ccg atc att tcg aag gtt gct 739
Gly Asp Asn Ala Met His Lys Leu Ser Pro Ile Ile Ser Lys Val Ala
200 205 210
gcg tat aag gcc gca gaa gtc aac att gat ggc ttg acc tac cgt gaa 787
Ala Tyr Lys Ala Ala Glu Val Asn Ile Asp Gly Leu Thr Tyr Arg Glu
215 220 225
ggc ctc aac atc gtt ttc tgc gaa tcg ggc gtg gca aac aac gtc att 835
Gly Leu Asn Ile Val Phe Cys Glu Ser Gly Val Ala Asn Asn Val Ile
230 235 240 245
cca gac ctc gcg tgg atg aac ctc aac ttc cgt ttc gcg ccg aat cgc 883
Pro Asp Leu Ala Trp Met Asn Leu Asn Phe Arg Phe Ala Pro Asn Arg
250 255 260
gat ctc aac gag gcg atc gag cat gtc gtc gaa acg ctt gag ctt gac 931
Asp Leu Asn Glu Ala Ile Glu His Val Val Glu Thr Leu Glu Leu Asp
265 270 275
ggt caa gac ggc atc gaa tgg gcc gta gaa gac ggg gca ggc ggt gcc 979
Gly Gln Asp Gly Ile Glu Trp Ala Val Glu Asp Gly Ala Gly Gly Ala
280 285 290
ctt cca ggc ttg ggg cag cag gtg aca agc ggg ctt atc gac gcc gtc 1027
Leu Pro Gly Leu Gly Gln Gln Val Thr Ser Gly Leu Ile Asp Ala Val
295 300 305
ggc cgc gaa aaa atc cgc gca aaa ttc ggc tgg acc gat gtc tca cgt 1075
Gly Arg Glu Lys Ile Arg Ala Lys Phe Gly Trp Thr Asp Val Ser Arg
310 315 320 325
ttt tca gcc atg gga att cca gcc cta aac ttt ggc gct ggt gat cca 1123
Phe Ser Ala Met Gly Ile Pro Ala Leu Asn Phe Gly Ala Gly Asp Pro
330 335 340
agt ttc gcg cat aaa cgc gac gag cag tgc cca gtg gag caa atc acg 1171
Ser Phe Ala His Lys Arg Asp Glu Gln Cys Pro Val Glu Gln Ile Thr
345 350 355
gat gtg gca gca att ttg aag cag tac ctg agc gag taaccgcatt 1217
Asp Val Ala Ala Ile Leu Lys Gln Tyr Leu Ser Glu
360 365
cggggttatc gtg 1230
<210>30
<211>369
<212>PRT
<213>谷氨酸棒杆菌
<400>30
Val Asn Ser Glu Leu Lys Pro Gly Leu Asp Leu Leu Gly Asp Pro Ile
1 5 10 15
Val Leu Thr Gln Arg Leu Val Asp Ile Pro Ser Pro Ser Gly Gln Glu
20 25 30
Lys Gln Ile Ala Asp Glu Ile Glu Asp Ala Leu Arg Asn Leu Asn Leu
35 40 45
Pro Gly Val Glu Val Phe Arg Phe Asn Asn Asn Val Leu Ala Arg Thr
50 55 60
Asn Arg Gly Leu Ala Ser Arg Val Met Leu Ala Gly His Ile Asp Thr
65 70 75 80
Val Pro Ile Ala Asp Asn Leu Pro Ser Arg Val Glu Asp Gly Ile Met
85 90 95
Tyr Gly Cys Gly Thr Val Asp Met Lys Ser Gly Leu Ala Val Tyr Leu
100 105 110
His Thr Phe Ala Thr Leu Ala Thr Ser Thr Glu Leu Lys His Asp Leu
115 120 125
Thr Leu Ile Ala Tyr Glu Cys Glu Glu Val Ala Asp His Leu Asn Gly
130 135 140
Leu Gly His Ile Arg Asp Glu His Pro Glu Trp Leu Ala Ala Asp Leu
145 150 155 160
Ala Leu Leu Gly Glu Pro Thr Gly Gly Trp Ile Glu Ala Gly Cys Gln
165 170 175
Gly Asn Leu Arg Ile Lys Val Thr Ala His Gly Val Arg Ala His Ser
180 185 190
Ala Arg Ser Trp Leu Gly Asp Asn Ala Met His Lys Leu Ser Pro Ile
195 200 205
Ile Ser Lys Val Ala Ala Tyr Lys Ala Ala Glu Val Asn Ile Asp Gly
210 215 220
Leu Thr Tyr Arg Glu Gly Leu Asn Ile Val Phe Cys Glu Ser Gly Val
225 230 235 240
Ala Asn Asn Val Ile Pro Asp Leu Ala Trp Met Asn Leu Asn Phe Arg
245 250 255
Phe Ala Pro Asn Arg Asp Leu Asn Glu Ala Ile Glu His Val Val Glu
260 265 270
Thr Leu Glu Leu Asp Gly Gln Asp Gly Ile Glu Trp Ala Val Glu Asp
275 280 285
Gly Ala Gly Gly Ala Leu Pro Gly Leu Gly Gln Gln Val Thr Ser Gly
290 295 300
Leu Ile Asp Ala Val Gly Arg Glu Lys Ile Arg Ala Lys Phe Gly Trp
305 310 315 320
Thr Asp Val Ser Arg Phe Ser Ala Met Gly Ile Pro Ala Leu Asn Phe
325 330 335
Gly Ala Gly Asp Pro Ser Phe Ala His Lys Arg Asp Glu Gln Cys Pro
340 345 350
Val Glu Gln Ile Thr Asp Val Ala Ala Ile Leu Lys Gln Tyr Leu Ser
355 360 365
Glu
<210>31
<211>1059
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1036)
<223>RXA00044
<400>31
attacctcag ccttccaagc tgatgatgca ttacttaaaa actgcagaca cttgaaaaac 60
ttctcacccg cactcgttcc ctcaacccac aaggagcacc atg gct tcc gca act 115
Met Ala Ser Ala Thr
1 5
ttc acc ggc gtg atc cca ccc gta atg acc cca ctc cac gcc gac ggc 163
Phe Thr Gly Val Ile Pro Pro Val Met Thr Pro Leu His Ala Asp Gly
10 15 20
agt gtg gat gta gaa agc ctc cgc aag ctc gtt gac cac ctc atc aat 211
Ser Val Asp Val Glu Ser Leu Arg Lys Leu Val Asp His Leu Ile Asn
25 30 35
ggt ggc gtc gac gga ctt ttc gca ctg ggc tcc tca ggc gaa gcg gca 259
Gly Gly Val Asp Gly Leu Phe Ala Leu Gly Ser Ser Gly Glu Ala Ala
40 45 50
ttc ctc acc cgc gcc cag cgc aaa ctc gca ctg acc acc atc atc gag 307
Phe Leu Thr Arg Ala Gln Arg Lys Leu Ala Leu Thr Thr Ile Ile Glu
55 60 65
cac acc gca ggc cgc gtt ccc gta act gct ggt gtc att gaa acc acc 355
His Thr Ala Gly Arg Val Pro Val Thr Ala Gly Val Ile Glu Thr Thr
70 75 80 85
act gct cgc gtg att gag ctc gtg gaa gat gcc ctg gag gct ggt gcc 403
Thr Ala Arg Val Ile Glu Leu Val Glu Asp Ala Leu Glu Ala Gly Ala
90 95 100
gaa ggc ctc gtt gcc act gca cct ttc tac acc cgc acc cac gat gtg 451
Glu Gly Leu Val Ala Thr Ala Pro Phe Tyr Thr Arg Thr His Asp Val
105 110 115
gaa att gaa gaa cac ttc cgc aag atc cac gcc gcc gct cca gag ctt 499
Glu Ile Glu Glu His Phe Arg Lys Ile His Ala Ala Ala Pro Glu Leu
120 125 130
cca ctg ttt gcc tac aac atc cca gtg tcg gtg cac tcc aac ctc aac 547
Pro Leu Phe Ala Tyr Asn Ile Pro Val Ser Val His Ser Asn Leu Asn
135 140 145
cca gtc atg ctt ttg acg ctg gcc aag gat ggc gtt ctt gca ggc acc 595
Pro Val Met Leu Leu Thr Leu Ala Lys Asp Gly Val Leu Ala Gly Thr
150 155 160 165
aag gat tcc agt ggc aat gat ggc gca atc cgc tca ctg atc gaa gct 643
Lys Asp Ser Ser Gly Asn Asp Gly Ala Ile Arg Ser Leu Ile Glu Ala
170 175 180
cgt gat gat gct gga ctc act gag cag ttc aag atc ctc acc ggc agc 691
Arg Asp Asp Ala Gly Leu Thr Glu Gln Phe Lys Ile Leu Thr Gly Ser
185 190 195
gaa acc acc gtt gat ttc gcc tac ctt gcg ggt gcc gat gga gtt gtc 739
Glu Thr Thr Val Asp Phe Ala Tyr Leu Ala Gly Ala Asp Gly Val Val
200 205 210
cca ggc ctg ggc aat gtt gat cct gca gca tac gca gct tta gca aaa 787
Pro Gly Leu Gly Asn Val Asp Pro Ala Ala Tyr Ala Ala Leu Ala Lys
215 220 225
ctc tgc ctc gat gga aag tgg gca gaa gct gct gct ttg cag aag cgc 835
Leu Cys Leu Asp Gly Lys Trp Ala Glu Ala Ala Ala Leu Gln Lys Arg
230 235 240 245
atc aac cac ctc ttc cac atc gtc ttc gtg gga gac acc tcc cat atg 883
Ile Asn His Leu Phe His Ile Val Phe Val Gly Asp Thr Ser His Met
250 255 260
tcc gga tcc agc gct ggt ttg ggc ggt ttc aag aca gca ctc gca cac 931
Ser Gly Ser Ser Ala Gly Leu Gly Gly Phe Lys Thr Ala Leu Ala His
265 270 275
ctt ggc att att gaa tcc aat gcg atg gca gtt cct cac cag agc ctc 979
Leu Gly Ile Ile Glu Ser Asn Ala Met Ala Val Pro His Gln Ser Leu
280 285 290
agc gac gaa gaa act gct cgc att cac gcc att gtt gat gaa ttc ctg 1027
Ser Asp Glu Glu Thr Ala Arg Ile His Ala Ile Val Asp Glu Phe Leu
295 300 305
tac acc gct taaggcccac acctcatgac tga 1059
Tyr Thr Ala
310
<210>32
<211>312
<212>PRT
<213>谷氨酸棒杆菌
<400>32
Met Ala Ser Ala Thr Phe Thr Gly Val Ile Pro Pro Val Met Thr Pro
1 5 10 15
Leu His Ala Asp Gly Ser Val Asp Val Glu Ser Leu Arg Lys Leu Val
20 25 30
Asp His Leu Ile Asn Gly Gly Val Asp Gly Leu Phe Ala Leu Gly Ser
35 40 45
Ser Gly Glu Ala Ala Phe Leu Thr Arg Ala Gln Arg Lys Leu Ala Leu
50 55 60
Thr Thr Ile Ile Glu His Thr Ala Gly Arg Val Pro Val Thr Ala Gly
65 70 75 80
Val Ile Glu Thr Thr Thr Ala Arg Val Ile Glu Leu Val Glu Asp Ala
85 90 95
Leu Glu Ala Gly Ala Glu Gly Leu Val Ala Thr Ala Pro Phe Tyr Thr
100 105 110
Arg Thr His Asp Val Glu Ile Glu Glu His Phe Arg Lys Ile His Ala
115 120 125
Ala Ala Pro Glu Leu Pro Leu Phe Ala Tyr Asn Ile Pro Val Ser Val
130 135 140
His Ser Asn Leu Asn Pro Val Met Leu Leu Thr Leu Ala Lys Asp Gly
145 150 155 160
Val Leu Ala Gly Thr Lys Asp Ser Ser Gly Asn Asp Gly Ala Ile Arg
165 170 175
Ser Leu Ile Glu Ala Arg Asp Asp Ala Gly Leu Thr Glu Gln Phe Lys
180 185 190
Ile Leu Thr Gly Ser Glu Thr Thr Val Asp Phe Ala Tyr Leu Ala Gly
195 200 205
Ala Asp Gly Val Val Pro Gly Leu Gly Asn Val Asp Pro Ala Ala Tyr
210 215 220
Ala Ala Leu Ala Lys Leu Cys Leu Asp Gly Lys Trp Ala Glu Ala Ala
225 230 235 240
Ala Leu Gln Lys Arg Ile Asn His Leu Phe His Ile Val Phe Val Gly
245 250 255
Asp Thr Ser His Met Ser Gly Ser Ser Ala Gly Leu Gly Gly Phe Lys
260 265 270
Thr Ala Leu Ala His Leu Gly Ile Ile Glu Ser Asn Ala Met Ala Val
275 280 285
Pro His Gln Ser Leu Ser Asp Glu Glu Thr Ala Arg Ile His Ala Ile
290 295 300
Val Asp Glu Phe Leu Tyr Thr Ala
305 310
<210>33
<211>867
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(844)
<223>RXA00863
<400>33
aacggtcagt taggtatgga tatcagcacc ttctgaacgg gtacgtctag actggtgggc 60
gtttgaaaaa ctcttcgccc cacgaaaatg aaggagcata atg gga atc aag gtt 115
Met Gly Ile Lys Val
1 5
ggc gtt ctc gga gcc aaa ggc cgt gtt ggt caa act att gtg gca gca 163
Gly Val Leu Gly Ala Lys Gly Arg Val Gly Gln Thr Ile Val Ala Ala
10 15 20
gtc aat gag tcc gac gat ctg gag ctt gtt gca gag atc ggc gtc gac 211
Val Asn Glu Ser Asp Asp Leu Glu Leu Val Ala Glu Ile Gly Val Asp
25 30 35
gat gat ttg agc ctt ctg gta gac aac ggc gct gaa gtt gtc gtt gac 259
Asp Asp Leu Ser Leu Leu Val Asp Asn Gly Ala Glu Val Val Val Asp
40 45 50
ttc acc act cct aac gct gtg atg ggc aac ctg gag ttc tgc atc aac 307
Phe Thr Thr Pro Asn Ala Val Met Gly Asn Leu Glu Phe Cys Ile Asn
55 60 65
aac ggc att tct gcg gtt gtt gga acc acg ggc ttc gat gat gct cgt 355
Asn Gly Ile Ser Ala Val Val Gly Thr Thr Gly Phe Asp Asp Ala Arg
70 75 80 85
ttg gag cag gtt cgc gac tgg ctt gaa gga aaa gac aat gtc ggt gtt 403
Leu Glu Gln Val Arg Asp Trp Leu Glu Gly Lys Asp Asn Val Gly Val
90 95 100
ctg atc gca cct aac ttt gct atc tct gcg gtg ttg acc atg gtc ttt 451
Leu Ile Ala Pro Asn Phe Ala Ile Ser Ala Val Leu Thr Met Val Phe
105 110 115
tcc aag cag gct gcc cgc ttc ttc gaa tca gct gaa gtt att gag ctg 499
Ser Lys Gln Ala Ala Arg Phe Phe Glu Ser Ala Glu Val Ile Glu Leu
120 125 130
cac cac ccc aac aag ctg gat gca cct tca ggc acc gcg atc cac act 547
His His Pro Asn Lys Leu Asp Ala Pro Ser Gly Thr Ala Ile His Thr
135 140 145
gct cag ggc att gct gcg gca cgc aaa gaa gca ggc atg gac gca cag 595
Ala Gln Gly Ile Ala Ala Ala Arg Lys Glu Ala Gly Met Asp Ala Gln
150 155 160 165
cca gat gcg acc gag cag gca ctt gag ggt tcc cgt ggc gca agc gta 643
Pro Asp Ala Thr Glu Gln Ala Leu Glu Gly Ser Arg Gly Ala Ser Val
170 175 180
gat gga atc ccg gtt cat gca gtc cgc atg tcc ggc atg gtt gct cac 691
Asp Gly Ile Pro Val His Ala Val Arg Met Ser Gly Met Val Ala His
185 190 195
gag caa gtt atc ttt ggc acc cag ggt cag acc ttg acc atc aag cag 739
Glu Gln Val Ile Phe Gly Thr Gln Gly Gln Thr Leu Thr Ile Lys Gln
200 205 210
gac tcc tat gat cgc aac tca ttt gca cca ggt gtc ttg gtg ggt gtg 787
Asp Ser Tyr Asp Arg Asn Ser Phe Ala Pro Gly Val Leu Val Gly Val
215 220 225
cgc aac att gca cag cac cca ggc cta gtc gta gga ctt gag cat tac 835
Arg Asn Ile Ala Gln His Pro Gly Leu Val Val Gly Leu Glu His Tyr
230 235 240 245
cta ggc ctg taaaggctca tttcagcagc ggg 867
Leu Gly Leu
<210>34
<211>248
<212>PRT
<213>谷氨酸棒杆菌
<400>34
Met Gly Ile Lys Val Gly Val Leu Gly Ala Lys Gly Arg Val Gly Gln
1 5 10 15
Thr Ile Val Ala Ala Val Asn Glu Ser Asp Asp Leu Glu Leu Val Ala
20 25 30
Glu Ile Gly Val Asp Asp Asp Leu Ser Leu Leu Val Asp Asn Gly Ala
35 40 45
Glu Val Val Val Asp Phe Thr Thr Pro Asn Ala Val Met Gly Asn Leu
50 55 60
Glu Phe Cys Ile Asn Asn Gly Ile Ser Ala Val Val Gly Thr Thr Gly
65 70 75 80
Phe Asp Asp Ala Arg Leu Glu Gln Val Arg Asp Trp Leu Glu Gly Lys
85 90 95
Asp Asn Val Gly Val Leu Ile Ala Pro Asn Phe Ala Ile Ser Ala Val
100 105 110
Leu Thr Met Val Phe Ser Lys Gln Ala Ala Arg Phe Phe Glu Ser Ala
115 120 125
Glu Val Ile Glu Leu His His Pro Asn Lys Leu Asp Ala Pro Ser Gly
130 135 140
Thr Ala Ile His Thr Ala Gln Gly Ile Ala Ala Ala Arg Lys Glu Ala
145 150 155 160
Gly Met Asp Ala Gln Pro Asp Ala Thr Glu Gln Ala Leu Glu Gly Ser
165 170 175
Arg Gly Ala Ser Val Asp Gly Ile Pro Val His Ala Val Arg Met Ser
180 185 190
Gly Met Val Ala His Glu Gln Val Ile Phe Gly Thr Gln Gly Gln Thr
195 200 205
Leu Thr Ile Lys Gln Asp Ser Tyr Asp Arg Asn Ser Phe Ala Pro Gly
210 215 220
Val Leu Val Gly Val Arg Asn Ile Ala Gln His Pro Gly Leu Val Val
225 230 235 240
Gly Leu Glu His Tyr Leu Gly Leu
245
<210>35
<211>873
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(850)
<223>RXA00864
<400>35
acagcaccca ggcctagtcg taggacttga gcattaccta ggcctgtaaa ggctcatttc 60
agcagcgggt ggaatttttt aaaaggagcg tttaaaggct gtg gcc gaa caa gtt 115
Val Ala Glu Gln Val
1 5
aaa ttg agc gtg gag ttg ata gcg tgc agt tct ttt act cca ccc gct 163
Lys Leu Ser Val Glu Leu Ile Ala Cys Ser Ser Phe Thr Pro Pro Ala
10 15 20
gat gtt gag tgg tca act gat gtt gag ggc gcg gaa gca ctc gtc gag 211
Asp Val Glu Trp Ser Thr Asp Val Glu Gly Ala Glu Ala Leu Val Glu
25 30 35
ttt gcg ggt cgt gcc tgc tac gaa act ttt gat aag ccg aac cct cga 259
Phe Ala Gly Arg Ala Cys Tyr Glu Thr Phe Asp Lys Pro Asn Pro Arg
40 45 50
act gct tcc aat gct gcg tat ctg cgc cac atc atg gaa gtg ggg cac 307
Thr Ala Ser Asn Ala Ala Tyr Leu Arg His Ile Met Glu Val Gly His
55 60 65
act gct ttg ctt gag cat gcc aat gcc acg atg tat atc cga ggc att 355
Thr Ala Leu Leu Glu His Ala Asn Ala Thr Met Tyr Ile Arg Gly Ile
70 75 80 85
tct cgg tcc gcg acc cat gaa ttg gtc cga cac cgc cat ttt tcc ttc 403
Ser Arg Ser Ala Thr His Glu Leu Val Arg His Arg His Phe Ser Phe
90 95 100
tct caa ctg tct cag cgt ttc gtg cac agc gga gaa tcg gaa gta gtg 451
Ser Gln Leu Ser Gln Arg Phe Val His Ser Gly Glu Ser Glu Val Val
105 110 115
gtg ccc act ctc atc gat gaa gat ccg cag ttg cgt gaa ctt ttc atg 499
Val Pro Thr Leu Ile Asp Glu Asp Pro Gln Leu Arg Glu Leu Phe Met
120 125 130
cac gcc atg gat gag tct cgg ttc gct ttc aat gag ctg ctt aat gcg 547
His Ala Met Asp Glu Ser Arg Phe Ala Phe Asn Glu Leu Leu Asn Ala
135 140 145
ctg gaa gaa aaa ctt ggc gat gaa ccg aat gca ctt tta agg aaa aag 595
Leu Glu Glu Lys Leu Gly Asp Glu Pro Asn Ala Leu Leu Arg Lys Lys
150 155 160 165
cag gct cgt caa gca gct cgc gct gtg ctg ccc aac gct aca gag tcc 643
Gln Ala Arg Gln Ala Ala Arg Ala Val Leu Pro Asn Ala Thr Glu Ser
170 175 180
aga atc gtg gtg tct gga aac ttc cgc acc tgg agg cat ttc att ggc 691
Arg Ile Val Val Ser Gly Asn Phe Arg Thr Trp Arg His Phe Ile Gly
185 190 195
atg cga gcc agt gaa cat gca gac gtc gaa atc cgc gaa gta gcg gta 739
Met Arg Ala Ser Glu His Ala Asp Val Glu Ile Arg Glu Val Ala Val
200 205 210
gaa tgt tta aga aag ctg cag gta gca gcg cca act gtt ttc ggt gat 787
Glu Cys Leu Arg Lys Leu Gln Val Ala Ala Pro Thr Val Phe Gly Asp
215 220 225
ttt gag att gaa act ttg gca gac gga tcg caa atg gca aca agc ccg 835
Phe Glu Ile Glu Thr Leu Ala Asp Gly Ser Gln Met Ala Thr Ser Pro
230 235 240 245
tat gtc atg gac ttt taacgcaaag ctcacaccca cga 873
Tyr Val Met Asp Phe
250
<210>36
<211>250
<212>PRT
<213>谷氨酸棒杆菌
<400>36
Val Ala Glu Gln Val Lys Leu Ser Val Glu Leu Ile Ala Cys Ser Ser
1 5 10 15
Phe Thr Pro Pro Ala Asp Val Glu Trp Ser Thr Asp Val Glu Gly Ala
20 25 30
Glu Ala Leu Val Glu Phe Ala Gly Arg Ala Cys Tyr Glu Thr Phe Asp
35 40 45
Lys Pro Asn Pro Arg Thr Ala Ser Asn Ala Ala Tyr Leu Arg His Ile
50 55 60
Met Glu Val Gly His Thr Ala Leu Leu Glu His Ala Asn Ala Thr Met
65 70 75 80
Tyr Ile Arg Gly Ile Ser Arg Ser Ala Thr His Glu Leu Val Arg His
85 90 95
Arg His Phe Ser Phe Ser Gln Leu Ser Gln Arg Phe Val His Ser Gly
100 105 110
Glu Ser Glu Val Val Val Pro Thr Leu Ile Asp Glu Asp Pro Gln Leu
115 120 125
Arg Glu Leu Phe Met His Ala Met Asp Glu Ser Arg Phe Ala Phe Asn
130 135 140
Glu Leu Leu Asn Ala Leu Glu Glu Lys Leu Gly Asp Glu Pro Asn Ala
145 150 155 160
Leu Leu Arg Lys Lys Gln Ala Arg Gln Ala Ala Arg Ala Val Leu Pro
165 170 175
Asn Ala Thr Glu Ser Arg Ile Val Val Ser Gly Asn Phe Arg Thr Trp
180 185 190
Arg His Phe Ile Gly Met Arg Ala Ser Glu His Ala Asp Val Glu Ile
195 200 205
Arg Glu Val Ala Val Glu Cys Leu Arg Lys Leu Gln Val Ala Ala Pro
210 215 220
Thr Val Phe Gly Asp Phe Glu Ile Glu Thr Leu Ala Asp Gly Ser Gln
225 230 235 240
Met Ala Thr Ser Pro Tyr Val Met Asp Phe
245 250
<210>37
<211>608
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(69)..(608)
<223>RXA02843
<400>37
cccattgcgc ggaggtcgca ccccttccga cttgaactga taggccgata gaaattattc 60
tggacgtc atg act act gct tcc gca acc gga att gca aca ctg acc tcc 110
Met Thr Thr Ala Ser Ala Thr Gly Ile Ala Thr Leu Thr Ser
1 5 10
acc ggc gac gtc ctg gac gtg tgg tat cca gaa atc ggg tcc acc gac 158
Thr Gly Asp Val Leu Asp Val Trp Tyr Pro Glu Ile Gly Ser Thr Asp
15 20 25 30
cag tcc gcg ctc aca cct cta gaa ggc gtc gat gaa gat cga aac gtc 206
Gln Ser Ala Leu Thr Pro Leu Glu Gly Val Asp Glu Asp Arg Asn Val
35 40 45
acc cgc aaa atc gtg acg aca act atc gac acc gac gca gcc ccc acc 254
Thr Arg Lys Ile Val Thr Thr Thr Ile Asp Thr Asp Ala Ala Pro Thr
50 55 60
gac acc tac gat gca tgg ctg cgc ctt cac ctc ctc tcc cac cgc gtt 302
Asp Thr Tyr Asp Ala Trp Leu Arg Leu His Leu Leu Ser His Arg Val
65 70 75
ttc cgc cct cac acc atc aac cta gac ggc att ttc ggc ctc ctc aac 350
Phe Arg Pro His Thr Ile Asn Leu Asp Gly Ile Phe Gly Leu Leu Asn
80 85 90
aat gtc gtg tgg acc aac ttc gga ccg tgc gca gtt gac ggt ttc gca 398
Asn Val Val Trp Thr Asn Phe Gly Pro Cys Ala Val Asp Gly Phe Ala
95 100 105 110
ctc acc cgc gcg cgc ctg tca cgc cga ggc caa gtt acg gtt tat agc 446
Leu Thr Arg Ala Arg Leu Ser Arg Arg Gly Gln Val Thr Val Tyr Ser
115 120 125
gtc gac aag ttc cca cgc atg gtc gac tat gtg gtt ccc tcg ggc gtg 494
Val Asp Lys Phe Pro Arg Met Val Asp Tyr Val Val Pro Ser Gly Val
130 135 140
cgc atc ggt gac gcc gac cgc gtc cga ctt ggc gcg tac ctg gca gat 542
Arg Ile Gly Asp Ala Asp Arg Val Arg Leu Gly Ala Tyr Leu Ala Asp
145 150 155
ggc acc acc gtg atg cat gag ggc ttc gtg aac ttc aac gct ggc acg 590
Gly Thr Thr Val Met His Glu Gly Phe Val Asn Phe Asn Ala Gly Thr
160 165 170
ctc ggc gct tcc atg gtt 608
Leu Gly Ala Ser Met Val
175 180
<210>38
<211>180
<212>PRT
<213>谷氨酸棒杆菌
<400>38
Met Thr Thr Ala Ser Ala Thr Gly Ile Ala Thr Leu Thr Ser Thr Gly
1 5 10 15
Asp Val Leu Asp Val Trp Tyr Pro Glu Ile Gly Ser Thr Asp Gln Ser
20 25 30
Ala Leu Thr Pro Leu Glu Gly Val Asp Glu Asp Arg Asn Val Thr Arg
35 40 45
Lys Ile Val Thr Thr Thr Ile Asp Thr Asp Ala Ala Pro Thr Asp Thr
50 55 60
Tyr Asp Ala Trp Leu Arg Leu His Leu Leu Ser His Arg Val Phe Arg
65 70 75 80
Pro His Thr Ile Asn Leu Asp Gly Ile Phe Gly Leu Leu Asn Asn Val
85 90 95
Val Trp Thr Asn Phe Gly Pro Cys Ala Val Asp Gly Phe Ala Leu Thr
100 105 110
Arg Ala Arg Leu Ser Arg Arg Gly Gln Val Thr Val Tyr Ser Val Asp
115 120 125
Lys Phe Pro Arg Met Val Asp Tyr Val Val Pro Ser Gly Val Arg Ile
130 135 140
Gly Asp Ala Asp Arg Val Arg Leu Gly Ala Tyr Leu Ala Asp Gly Thr
145 150 155 160
Thr Val Met His Glu Gly Phe Val Asn Phe Asn Ala Gly Thr Leu Gly
165 170 175
Ala Ser Met Val
180
<210>39
<211>1143
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1120)
<223>RXN00355
<400>39
aatagatcag cgcatccgtg gtggaaccaa aaggctcaac aatacgaaac gttcgctttc 60
ggtcctgatg aaagagatgt ccctgaatca tcatctaagt atg cat ctc ggt aag 115
Met His Leu Gly Lys
1 5
ctc gac cag gac agt gcc acc aca att ttg gag gat tac aag aac atg 163
Leu Asp Gln Asp Ser Ala Thr Thr Ile Leu Glu Asp Tyr Lys Asn Met
10 15 20
acc aac atc cgc gta gct atc gtg ggc tac gga aac ctg gga cgc agc 211
Thr Asn Ile Arg Val Ala Ile Val Gly Tyr Gly Asn Leu Gly Arg Ser
25 30 35
gtc gaa aag ctt att gcc aag cag ccc gac atg gac ctt gta gga atc 259
Val Glu Lys Leu Ile Ala Lys Gln Pro Asp Met Asp Leu Val Gly Ile
40 45 50
ttc tcg cgc cgg gcc acc ctc gac aca aag acg cca gtc ttt gat gtc 307
Phe Ser Arg Arg Ala Thr Leu Asp Thr Lys Thr Pro Val Phe Asp Val
55 60 65
gcc gac gtg gac aag cac gcc gac gac gtg gac gtg ctg ttc ctg tgc 355
Ala Asp Val Asp Lys His Ala Asp Asp Val Asp Val Leu Phe Leu Cys
70 75 80 85
atg ggc tcc gcc acc gac atc cct gag cag gca cca aag ttc gcg cag 403
Met Gly Ser Ala Thr Asp Ile Pro Glu Gln Ala Pro Lys Phe Ala Gln
90 95 100
ttc gcc tgc acc gta gac acc tac gac aac cac cgc gac atc cca cgc 451
Phe Ala Cys Thr Val Asp Thr Tyr Asp Asn His Arg Asp Ile Pro Arg
105 110 115
cac cgc cag gtc atg aac gaa gcc gcc acc gca gcc ggc aac gtt gca 499
His Arg Gln Val Met Asn Glu Ala Ala Thr Ala Ala Gly Asn Val Ala
120 125 130
ctg gtc tct acc ggc tgg gat cca gga atg ttc tcc atc aac cgc gtc 547
Leu Val Ser Thr Gly Trp Asp Pro Gly Met Phe Ser Ile Asn Arg Val
135 140 145
tac gca gcg gca gtc tta gcc gag cac cag cag cac acc ttc tgg ggc 595
Tyr Ala Ala Ala Val Leu Ala Glu His Gln Gln His Thr Phe Trp Gly
150 155 160 165
cca ggt ttg tca cag ggc cac tcc gat gct ttg cga cgc atc cct ggc 643
Pro Gly Leu Ser Gln Gly His Ser Asp Ala Leu Arg Arg Ile Pro Gly
170 175 180
gtt caa aag gca gtc cag tac acc ctc cca tcc gaa gac gcc ctg gaa 691
Val Gln Lys Ala Val Gln Tyr Thr Leu Pro Ser Glu Asp Ala Leu Glu
185 190 195
aag gcc cgc cgc ggc gaa gcc ggc gac ctt acc gga aag caa acc cac 739
Lys Ala Arg Arg Gly Glu Ala Gly Asp Leu Thr Gly Lys Gln Thr His
200 205 210
aag cgc caa tgc ttc gtg gtt gcc gac gcg gcc gat cac gag cgc atc 787
Lys Arg Gln Cys Phe Val Val Ala Asp Ala Ala Asp His Glu Arg Ile
215 220 225
gaa aac gac atc cgc acc atg cct gat tac ttc gtt ggc tac gaa gtc 835
Glu Asn Asp Ile Arg Thr Met Pro Asp Tyr Phe Val Gly Tyr Glu Val
230 235 240 245
gaa gtc aac ttc atc gac gaa gca acc ttc gac tcc gag cac acc ggc 883
Glu Val Asn Phe Ile Asp Glu Ala Thr Phe Asp Ser Glu His Thr Gly
250 255 260
atg cca cac ggt ggc cac gtg att acc acc ggc gac acc ggt ggc ttc 931
Met Pro His Gly Gly His Val Ile Thr Thr Gly Asp Thr Gly Gly Phe
265 270 275
aac cac acc gtg gaa tac atc ctc aag ctg gac cga aac cca gat ttc 979
Asn His Thr Val Glu Tyr Ile Leu Lys Leu Asp Arg Asn Pro Asp Phe
280 285 290
acc gct tcc tca cag atc gct ttc ggt cgc gca gct cac cgc atg aag 1027
Thr Ala Ser Ser Gln Ile Ala Phe Gly Arg Ala Ala His Arg Met Lys
295 300 305
cag cag ggc caa agc gga gct ttc acc gtc ctc gaa gtt gct cca tac 1075
Gln Gln Gly Gln Ser Gly Ala Phe Thr Val Leu Glu Val Ala Pro Tyr
310 315 320 325
ctg ctc tcc cca gag aac ttg gac gat ctg atc gca cgc gac gtc 1120
Leu Leu Ser Pro Glu Asn Leu Asp Asp Leu Ile Ala Arg Asp Val
330 335 340
taatttagct cgaggggcaa gga 1143
<210>40
<211>340
<212>PRT
<213>谷氨酸棒杆菌
<400>40
Met His Leu Gly Lys Leu Asp Gln Asp Ser Ala Thr Thr Ile Leu Glu
1 5 10 15
Asp Tyr Lys Asn Met Thr Asn Ile Arg Val Ala Ile Val Gly Tyr Gly
20 25 30
Asn Leu Gly Arg Ser Val Glu Lys Leu Ile Ala Lys Gln Pro Asp Met
35 40 45
Asp Leu Val Gly Ile Phe Ser Arg Arg Ala Thr Leu Asp Thr Lys Thr
50 55 60
Pro Val Phe Asp Val Ala Asp Val Asp Lys His Ala Asp Asp Val Asp
65 70 75 80
Val Leu Phe Leu Cys Met Gly Ser Ala Thr Asp Ile Pro Glu Gln Ala
85 90 95
Pro Lys Phe Ala Gln Phe Ala Cys Thr Val Asp Thr Tyr Asp Asn His
100 105 110
Arg Asp Ile Pro Arg His Arg Gln Val Met Asn Glu Ala Ala Thr Ala
115 120 125
Ala Gly Asn Val Ala Leu Val Ser Thr Gly Trp Asp Pro Gly Met Phe
130 135 140
Ser Ile Asn Arg Val Tyr Ala Ala Ala Val Leu Ala Glu His Gln Gln
145 150 155 160
His Thr Phe Trp Gly Pro Gly Leu Ser Gln Gly His Ser Asp Ala Leu
165 170 175
Arg Arg Ile Pro Gly Val Gln Lys Ala Val Gln Tyr Thr Leu Pro Ser
180 185 190
Glu Asp Ala Leu Glu Lys Ala Arg Arg Gly Glu Ala Gly Asp Leu Thr
195 200 205
Gly Lys Gln Thr His Lys Arg Gln Cys Phe Val Val Ala Asp Ala Ala
210 215 220
Asp His Glu Arg Ile Glu Asn Asp Ile Arg Thr Met Pro Asp Tyr Phe
225 230 235 240
Val Gly Tyr Glu Val Glu Val Asn Phe Ile Asp Glu Ala Thr Phe Asp
245 250 255
Ser Glu His Thr Gly Met Pro His Gly Gly His Val Ile Thr Thr Gly
260 265 270
Asp Thr Gly Gly Phe Asn His Thr Val Glu Tyr Ile Leu Lys Leu Asp
275 280 285
Arg Asn Pro Asp Phe Thr Ala Ser Ser Gln Ile Ala Phe Gly Arg Ala
290 295 300
Ala His Arg Met Lys Gln Gln Gly Gln Ser Gly Ala Phe Thr Val Leu
305 310 315 320
Glu Val Ala Pro Tyr Leu Leu Ser Pro Glu Asn Leu Asp Asp Leu Ile
325 330 335
Ala Arg Asp Val
340
<210>41
<211>958
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(958)
<223>FRXA00352
<400>41
aatagatcag cgcatccgtg gtggaaccaa aaggctcaac aatacgaaac gttcgctttc 60
ggtcctgatg aaagagatgt ccctgaatca tcatctaagt atg cat ctc ggt aag 115
Met His Leu Gly Lys
1 5
ctc gac cag gac agt gcc acc aca att ttg gag gat tac aag aac atg 163
Leu Asp Gln Asp Ser Ala Thr Thr Ile Leu Glu Asp Tyr Lys Asn Met
10 15 20
acc aac atc cgc gta gct atc gtg ggc tac gga aac ctg gga cgc agc 211
Thr Asn Ile Arg Val Ala Ile Val Gly Tyr Gly Asn Leu Gly Arg Ser
25 30 35
gtc gaa aag ctt att gcc aag cag ccc gac atg gac ctt gta gga atc 259
Val Glu Lys Leu Ile Ala Lys Gln Pro Asp Met Asp Leu Val Gly Ile
40 45 50
ttc tcg cgc cgg gcc acc ctc gac aca aag acg cca gtc ttt gat gtc 307
Phe Ser Arg Arg Ala Thr Leu Asp Thr Lys Thr Pro Val Phe Asp Val
55 60 65
gcc gac gtg gac aag cac gcc gac gac gtg gac gtg ctg ttc ctg tgc 355
Ala Asp Val Asp Lys His Ala Asp Asp Val Asp Val Leu Phe Leu Cys
70 75 80 85
atg ggc tcc gcc acc gac atc cct gag cag gca cca aag ttc gcg cag 403
Met Gly Ser Ala Thr Asp Ile Pro Glu Gln Ala Pro Lys Phe Ala Gln
90 95 100
ttc gcc tgc acc gta gac acc tac gac aac cac cgc gac atc cca cgc 451
Phe Ala Cys Thr Val Asp Thr Tyr Asp Asn His Arg Asp Ile Pro Arg
105 110 115
cac cgc cag gtc atg aac gaa gcc gcc acc gca gcc ggc aac gtt gca 499
His Arg Gln Val Met Asn Glu Ala Ala Thr Ala Ala Gly Asn Val Ala
120 125 130
ctg gtc tct acc ggc tgg gat cca gga atg ttc tcc atc aac cgc gtc 547
Leu Val Ser Thr Gly Trp Asp Pro Gly Met Phe Ser Ile Asn Arg Val
135 140 145
tac gca gcg gca gtc tta gcc gag cac cag cag cac acc ttc tgg ggc 595
Tyr Ala Ala Ala Val Leu Ala Glu His Gln Gln His Thr Phe Trp Gly
150 155 160 165
cca ggt ttg tca cag ggc cac tcc gat gct ttg cga cgc atc cct ggc 643
Pro Gly Leu Ser Gln Gly His Ser Asp Ala Leu Arg Arg Ile Pro Gly
170 175 180
gtt caa aag gca gtc cag tac acc ctc cca tcc gaa gac gcc ctg gaa 691
Val Gln Lys Ala Val Gln Tyr Thr Leu Pro Ser Glu Asp Ala Leu Glu
185 190 195
aag gcc cgc cgc ggc gaa gcc ggc gac ctt acc gga aag caa acc cac 739
Lys Ala Arg Arg Gly Glu Ala Gly Asp Leu Thr Gly Lys Gln Thr His
200 205 210
aag cgc caa tgc ttc gtg gtt gcc gac gcg gcc gat cac gag cgc atc 787
Lys Arg Gln Cys Phe Val Val Ala Asp Ala Ala Asp His Glu Arg Ile
215 220 225
gaa aac gac atc cgc acc atg cct gat tac ttc gtt ggc tac gaa gtc 835
Glu Asn Asp Ile Arg Thr Met Pro Asp Tyr Phe Val Gly Tyr Glu Val
230 235 240 245
gaa gtc aac ttc atc gac gaa gca acc ttc gac tcc gag cac acc ggc 883
Glu Val Asn Phe Ile Asp Glu Ala Thr Phe Asp Ser Glu His Thr Gly
250 255 260
atg cca cac ggt ggc cac gtg att acc acc ggc gac acc ggt ggc ttc 931
Met Pro His Gly Gly His Val Ile Thr Thr Gly Asp Thr Gly Gly Phe
265 270 275
aac cac acc gtg gaa tac atc ctc aag 958
Asn His Thr Val Glu Tyr Ile Leu Lys
280 285
<210>42
<211>286
<212>PRT
<213>谷氨酸棒杆菌
<400>42
Met His Leu Gly Lys Leu Asp Gln Asp Ser Ala Thr Thr Ile Leu Glu
1 5 10 15
Asp Tyr Lys Asn Met Thr Asn Ile Arg Val Ala Ile Val Gly Tyr Gly
20 25 30
Asn Leu Gly Arg Ser Val Glu Lys Leu Ile Ala Lys Gln Pro Asp Met
35 40 45
Asp Leu Val Gly Ile Phe Ser Arg Arg Ala Thr Leu Asp Thr Lys Thr
50 55 60
Pro Val Phe Asp Val Ala Asp Val Asp Lys His Ala Asp Asp Val Asp
65 70 75 80
Val Leu Phe Leu Cys Met Gly Ser Ala Thr Asp Ile Pro Glu Gln Ala
85 90 95
Pro Lys Phe Ala Gln Phe Ala Cys Thr Val Asp Thr Tyr Asp Asn His
100 105 110
Arg Asp Ile Pro Arg His Arg Gln Val Met Asn Glu Ala Ala Thr Ala
115 120 125
Ala Gly Asn Val Ala Leu Val Ser Thr Gly Trp Asp Pro Gly Met Phe
130 135 140
Ser Ile Asn Arg Val Tyr Ala Ala Ala Val Leu Ala Glu His Gln Gln
145 150 155 160
His Thr Phe Trp Gly Pro Gly Leu Ser Gln Gly His Ser Asp Ala Leu
165 170 175
Arg Arg Ile Pro Gly Val Gln Lys Ala Val Gln Tyr Thr Leu Pro Ser
180 185 190
Glu Asp Ala Leu Glu Lys Ala Arg Arg Gly Glu Ala Gly Asp Leu Thr
195 200 205
Gly Lys Gln Thr His Lys Arg Gln Cys Phe Val Val Ala Asp Ala Ala
210 215 220
Asp His Glu Arg Ile Glu Asn Asp Ile Arg Thr Met Pro Asp Tyr Phe
225 230 235 240
Val Gly Tyr Glu Val Glu Val Asn Phe Ile Asp Glu Ala Thr Phe Asp
245 250 255
Ser Glu His Thr Gly Met Pro His Gly Gly His Val Ile Thr Thr Gly
260 265 270
Asp Thr Gly Gly Phe Asn His Thr Val Glu Tyr Ile Leu Lys
275 280 285
<210>43
<211>1400
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(1377)
<223>RXA00972
<400>43
cct gca cct ggt tgg cgt ttc cgc acc gga gaa gat gta aca atg gct 48
Pro Ala Pro Gly Trp Arg Phe Arg Thr Gly Glu Asp Val Thr Met Ala
1 5 10 15
aca gtt gaa aat ttc aat gaa ctt ccc gca cac gta tgg cca cgc aat 96
Thr Val Glu Asn Phe Asn Glu Leu Pro Ala His Val Trp Pro Arg Asn
20 25 30
gcc gtg cgc caa gaa gac ggc gtt gtc acc gtc gct ggt gtg cct ctg 144
Ala Val Arg Gln Glu Asp Gly Val Val Thr Val Ala Gly Val Pro Leu
35 40 45
cct gac ctc gct gaa gaa tac gga acc cca ctg ttc gta gtc gac gag 192
Pro Asp Leu Ala Glu Glu Tyr Gly Thr Pro Leu Phe Val Val Asp Glu
50 55 60
gac gat ttc cgt tcc cgc tgt cgc gac atg gct acc gca ttc ggt gga 240
Asp Asp Phe Arg Ser Arg Cys Arg Asp Met Ala Thr Ala Phe Gly Gly
65 70 75 80
cca ggc aat gtg cac tac gca tct aaa gcg ttc ctg acc aag acc att 288
Pro Gly Asn Val His Tyr Ala Ser Lys Ala Phe Leu Thr Lys Thr Ile
85 90 95
gca cgt tgg gtt gat gaa gag ggg ctg gca ctg gac att gca tcc atc 336
Ala Arg Trp Val Asp Glu Glu Gly Leu Ala Leu Asp Ile Ala Ser Ile
100 105 110
aac gaa ctg ggc att gcc ctg gcc gct ggt ttc ccc gcc agc cgt atc 384
Asn Glu Leu Gly Ile Ala Leu Ala Ala Gly Phe Pro Ala Ser Arg Ile
115 120 125
acc gcg cac ggc aac aac aaa ggc gta gag ttc ctg cgc gcg ttg gtt 432
Thr Ala His Gly Asn Asn Lys Gly Val Glu Phe Leu Arg Ala Leu Val
130 135 140
caa aac ggt gtg gga cac gtg gtg ctg gac tcc gca cag gaa cta gaa 480
Gln Asn Gly Val Gly His Val Val Leu Asp Ser Ala Gln Glu Leu Glu
1451 50 155 160
ctg ttg gat tac gtt gcc gct ggt gaa ggc aag att cag gac gtg ttg 528
Leu Leu Asp Tyr Val Ala Ala Gly Glu Gly Lys Ile Gln Asp Val Leu
165 170 175
atc cgc gta aag cca ggc atc gaa gca cac acc cac gag ttc atc gcc 576
Ile Arg Val Lys Pro Gly Ile Glu Ala His Thr His Glu Phe Ile Ala
180 185 190
act agc cac gaa gac cag aag ttc gga ttc tcc ctg gca tcc ggt tcc 624
Thr Ser His Glu Asp Gln Lys Phe Gly Phe Ser Leu Ala Ser Gly Ser
195 200 205
gca ttc gaa gca gca aaa gcc gcc aac aac gca gaa aac ctg aac ctg 672
Ala Phe Glu Ala Ala Lys Ala Ala Asn Asn Ala Glu Asn Leu Asn Leu
210 215 220
gtt ggc ctg cac tgc cac gtt ggt tcc cag gtg ttc gac gcc gaa ggc 720
Val Gly Leu His Cys His Val Gly Ser Gln Val Phe Asp Ala Glu Gly
225 230 235 240
ttc aag ctg gca gca gaa cgc gtg ttg ggc ctg tac tca cag atc cac 768
Phe Lys Leu Ala Ala Glu Arg Val Leu Gly Leu Tyr Ser Gln Ile His
245 250 255
agc gaa ctg ggc gtt gcc ctt cct gaa ctg gat ctc ggt ggc gga tac 816
Ser Glu Leu Gly Val Ala Leu Pro Glu Leu Asp Leu Gly Gly Gly Tyr
260 265 270
ggc att gcc tat acc gca gct gaa gaa cca ctc aac gtc gca gaa gtt 864
Gly Ile Ala Tyr Thr Ala Ala Glu Glu Pro Leu Asn Val Ala Glu Val
275 280 285
gcc tcc gac ctg ctc acc gca gtc gga aaa atg gca gcg gaa cta ggc 912
Ala Ser Asp Leu Leu Thr Ala Val Gly Lys Met Ala Ala Glu Leu Gly
290 295 300
atc gac gca cca acc gtg ctt gtt gag ccc ggc cgc gct atc gca ggc 960
Ile Asp Ala Pro Thr Val Leu Val Glu Pro Gly Arg Ala Ile Ala Gly
305 310 315 320
ccc tcc acc gtg acc atc tac gaa gtc ggc acc acc aaa gac gtc cac 1008
Pro Ser Thr Val Thr Ile Tyr Glu Val Gly Thr Thr Lys Asp Val His
325 330 335
gta gac gac gac aaa acc cgc cgt tac atc gcc gtg gac gga ggc atg 1056
Val Asp Asp Asp Lys Thr Arg Arg Tyr Ile Ala Val Asp Gly Gly Met
340 345 350
tcc gac aac atc cgc cca gca ctc tac ggg tcc gaa tac gac gcc cgc 1104
Ser Asp Asn Ile Arg Pro Ala Leu Tyr Gly Ser Glu Tyr Asp Ala Arg
355 360 365
gta gta tcc cgc ttc gcc gaa gga gac cca gta agc acc cgc atc gtg 1152
Val Val Ser Arg Phe Ala Glu Gly Asp Pro Val Ser Thr Arg Ile Val
370 375 380
ggc tcc cac tgc gaa tcc ggc gat atc ctg atc aac gat gaa atc tac 1200
Gly Ser His Cys Glu Ser Gly Asp Ile Leu Ile Asn Asp Glu Ile Tyr
385 390 395 400
cca tct gac atc acc agc ggc gac ttc ctt gca ctc gca gcc acc ggc 1248
Pro Ser Asp Ile Thr Ser Gly Asp Phe Leu Ala Leu Ala Ala Thr Gly
405 410 415
gca tac tgc tac gcc atg agc tcc cgc tac aac gcc ttc aca cgg ccc 1296
Ala Tyr Cys Tyr Ala Met Ser Ser Arg Tyr Asn Ala Phe Thr Arg Pro
420 425 430
gcc gtc gtg tcc gtc cgc gct ggc agc tcc cgc ctc atg ctg cgc cgc 1344
Ala Val Val Ser Val Arg Ala Gly Ser Ser Arg Leu Met Leu Arg Arg
435 440 445
gaa acg ctc gac gac atc ctc tca cta gag gca taacgctttt cgacgcctga 1397
Glu Thr Leu Asp Asp Ile Leu Ser Leu Glu Ala
450 455
ccc 1400
<210>44
<211>459
<212>PRT
<213>谷氨酸棒杆菌
<400>44
Pro Ala Pro Gly Trp Arg Phe Arg Thr Gly Glu Asp Val Thr Met Ala
1 5 10 15
Thr Val Glu Asn Phe Asn Glu Leu Pro Ala His Val Trp Pro Arg Asn
20 25 30
Ala Val Arg Gln Glu Asp Gly Val Val Thr Val Ala Gly Val Pro Leu
35 40 45
Pro Asp Leu Ala Glu Glu Tyr Gly Thr Pro Leu Phe Val Val Asp Glu
50 55 60
Asp Asp Phe Arg Ser Arg Cys Arg Asp Met Ala Thr Ala Phe Gly Gly
65 70 75 80
Pro Gly Asn Val His Tyr Ala Ser Lys Ala Phe Leu Thr Lys Thr Ile
85 90 95
Ala Arg Trp Val Asp Glu Glu Gly Leu Ala Leu Asp Ile Ala Ser Ile
100 105 110
Asn Glu Leu Gly Ile Ala Leu Ala Ala Gly Phe Pro Ala Ser Arg Ile
115 120 125
Thr Ala His Gly Asn Asn Lys Gly Val Glu Phe Leu Arg Ala Leu Val
130 135 140
Gln Asn Gly Val Gly His Val Val Leu Asp Ser Ala Gln Glu Leu Glu
145 150 155 160
Leu Leu Asp Tyr Val Ala Ala Gly Glu Gly Lys Ile Gln Asp Val Leu
165 170 175
Ile Arg Val Lys Pro Gly Ile Glu Ala His Thr His Glu Phe Ile Ala
180 185 190
Thr Ser His Glu Asp Gln Lys Phe Gly Phe Ser Leu Ala Ser Gly Ser
195 200 205
Ala Phe Glu Ala Ala Lys Ala Ala Asn Asn Ala Glu Asn Leu Asn Leu
210 215 220
Val Gly Leu His Cys His Val Gly Ser Gln Val Phe Asp Ala Glu Gly
225 230 235 240
Phe Lys Leu Ala Ala Glu Arg Val Leu Gly Leu Tyr Ser Gln Ile His
245 250 255
Ser Glu Leu Gly Val Ala Leu Pro Glu Leu Asp Leu Gly Gly Gly Tyr
260 265 270
Gly Ile Ala Tyr Thr Ala Ala Glu Glu Pro Leu Asn Val Ala Glu Val
275 280 285
Ala Ser Asp Leu Leu Thr Ala Val Gly Lys Met Ala Ala Glu Leu Gly
290 295 300
Ile Asp Ala Pro Thr Val Leu Val Glu Pro Gly Arg Ala Ile Ala Gly
305 310 315 320
Pro Ser Thr Val Thr Ile Tyr Glu Val Gly Thr Thr Lys Asp Val His
325 330 335
Val Asp Asp Asp Lys Thr Arg Arg Tyr Ile Ala Val Asp Gly Gly Met
340 345 350
Ser Asp Asn Ile Arg Pro Ala Leu Tyr Gly Ser Glu Tyr Asp Ala Arg
355 360 365
Val Val Ser Arg Phe Ala Glu Gly Asp Pro Val Ser Thr Arg Ile Val
370 375 380
Gly Ser His Cys Glu Ser Gly Asp Ile Leu Ile Asn Asp Glu Ile Tyr
385 390 395 400
Pro Ser Asp Ile Thr Ser Gly Asp Phe Leu Ala Leu Ala Ala Thr Gly
405 410 415
Ala Tyr Cys Tyr Ala Met Ser Ser Arg Tyr Asn Ala Phe Thr Arg Pro
420 425 430
Ala Val Val Ser Val Arg Ala Gly Ser Ser Arg Leu Met Leu Arg Arg
435 440 445
Glu Thr Leu Asp Asp Ile Leu Ser Leu Glu Ala
450 455
<210>45
<211>2121
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(2098)
<223>RXA02653
<400>45
agacagagtg ttagtgcgtg gggcagctct cactttcatc gacatcactc gagtatgctc 60
accggccgta ttcattccaa taacccgcac agggaaacta atg ata ccg aag ccc 115
Met Ile Pro Lys Pro
1 5
gac gtg acc gac tta tat tta gag gac ctc tta aat gag ggt tcg gaa 163
Asp Val Thr Asp Leu Tyr Leu Glu Asp Leu Leu Asn Glu Gly Ser Glu
10 15 20
aag att cgg tcc gcc aag gat ctt tcc gaa ctt agg aca gtt cta aaa 211
Lys Ile Arg Ser Ala Lys Asp Leu Ser Glu Leu Arg Thr Val Leu Lys
25 30 35
gag gtt tcc tcc caa att cag gaa cga gct ggg aaa aaa gat gaa gaa 259
Glu Val Ser Ser Gln Ile Gln Glu Arg Ala Gly Lys Lys Asp Glu Glu
40 45 50
tgg gga atg ggg gcc act tgg cgg gag ctg tac ccc agc atc gtg gaa 307
Trp Gly Met Gly Ala Thr Trp Arg Glu Leu Tyr Pro Ser Ile Val Glu
55 60 65
cgc gct tcc tac gaa ggg cgt gac agc cta atc gga ttt gat cac tta 355
Arg Ala Ser Tyr Glu Gly Arg Asp Ser Leu Ile Gly Phe Asp His Leu
70 75 80 85
gcc cgg gaa atg gaa aga tta gcc ttc ggc cca cca tcc gaa agt ttt 403
Ala Arg Glu Met Glu Arg Leu Ala Phe Gly Pro Pro Ser Glu Ser Phe
90 95 100
gaa tac ctc caa gaa ctc gta aaa tcc gga gtg gta gac atc act cac 451
Glu Tyr Leu Gln Glu Leu Val Lys Ser Gly Val Val Asp Ile Thr His
105 110 115
ctg cat cgt ggc cgg gaa cca ctg aca gat tta gtt cgt gaa ctt gaa 499
Leu His Arg Gly Arg Glu Pro Leu Thr Asp Leu Val Arg Glu Leu Glu
120 125 130
ata act gtg gtg ata gac gct gtt ctt ccc ccg ccg gga gta gtg cca 547
Ile Thr Val Val Ile Asp Ala Val Leu Pro Pro Pro Gly Val Val Pro
135 140 145
ggc aca ttg gtg cac aat ttg gta aaa gag gga tat gcc aga atg cgt 595
Gly Thr Leu Val His Asn Leu Val Lys Glu Gly Tyr Ala Arg Met Arg
150 155 160 165
cct ggg act cgg ggg tta gat gta gcg gct gac ggc acc gtt caa ggg 643
Pro Gly Thr Arg Gly Leu Asp Val Ala Ala Asp Gly Thr Val Gln Gly
170 175 180
caa cga cat ttg gct gca gtc gga cgg atg acg gaa gat gtg gtt ttg 691
Gln Arg His Leu Ala Ala Val Gly Arg Met Thr Glu Asp Val Val Leu
185 190 195
ggt aat gac aca ttg tcg cga tca tta cat gac ata atc ccg aag tgg 739
Gly Asn Asp Thr Leu Ser Arg Ser Leu His Asp Ile Ile Pro Lys Trp
200 205 210
gct cgt cga gtt atc cgc gac gcg agc acg tat ccc gat agg gta cat 787
Ala Arg Arg Val Ile Arg Asp Ala Ser Thr Tyr Pro Asp Arg Val His
215 220 225
ggt act cca ccg ctt ccg gca cgg ttg gaa ccc tgg gcg gaa aag ctc 835
Gly Thr Pro Pro Leu Pro Ala Arg Leu Glu Pro Trp Ala Glu Lys Leu
230 235 240 245
act tca gat ccg gcc aca tgc cgc cac ctg att gaa gaa ttc ggg agt 883
Thr Ser Asp Pro Ala Thr Cys Arg His Leu Ile Glu Glu Phe Gly Ser
250 255 260
cct gtg aat gta ctc cat tca ggt tct atg cct cgt aat ata aat gag 931
Pro Val Asn Val Leu His Ser Gly Ser Met Pro Arg Asn Ile Asn Glu
265 270 275
ttg gtt gac gcc ggc att cag atg ggg gtg gat act cga ata ttt ttt 979
Leu Val Asp Ala Gly Ile Gln Met Gly Val Asp Thr Arg Ile Phe Phe
280 285 290
gcc cgc aaa gcg aat aag ggt ctt acc ttc gtt gat gcc gtt aaa gac 1027
Ala Arg Lys Ala Asn Lys Gly Leu Thr Phe Val Asp Ala Val Lys Asp
295 300 305
acc ggt cat ggt gta gat gta gcc agt gaa cga gag tta tct cag gtg 1075
Thr Gly His Gly Val Asp Val Ala Ser Glu Arg Glu Leu Ser Gln Val
310 315 320 325
ctt aat cgt gga gtc cca gga gag cgg atc att cta tcc gca gct atc 1123
Leu Asn Arg Gly Val Pro Gly Glu Arg Ile Ile Leu Ser Ala Ala Ile
330 335 340
aaa ccg gac aga cta ttg gca tta gcg atc gaa aat ggc gtg atc atc 1171
Lys Pro Asp Arg Leu Leu Ala Leu Ala Ile Glu Asn Gly Val Ile Ile
345 350 355
tct gtg gat tcg cgt gat gaa tta gat cgc att tcg gct ttg gtt ggt 1219
Ser Val Asp Ser Arg Asp Glu Leu Asp Arg Ile Ser Ala Leu Val Gly
360 365 370
gac cgc gtt gca cga gtt gcg cct aga gta gct cca gat cct gca gtc 1267
Asp Arg Val Ala Arg Val Ala Pro Arg Val Ala Pro Asp Pro Ala Val
375 380 385
tta cct cca act aga ttt ggt gag cgt gct gca gac tgg ggt aat cgg 1315
Leu Pro Pro Thr Arg Phe Gly Glu Arg Ala Ala Asp Trp Gly Asn Arg
390 395 400 405
ctt acc gag gtg ata ccc ggc gtg gat att gtg ggt ctt cac gtt cac 1363
Leu Thr Glu Val Ile Pro Gly Val Asp Ile Val Gly Leu His Val His
410 415 420
ctc cat ggc tat gct gca aaa gac cgt gct ctg gct ctg cag gaa tgt 1411
Leu His Gly Tyr Ala Ala Lys Asp Arg Ala Leu Ala Leu Gln Glu Cys
425 430 435
tgc caa ctc gtc gat tct ctc aga gaa tgc ggg cat tcc cca cag ttt 1459
Cys Gln Leu Val Asp Ser Leu Arg Glu Cys Gly His Ser Pro Gln Phe
440 445 450
att gac ctt gga gga ggg gtg cct atg agc tac att gaa tct gag gaa 1507
Ile Asp Leu Gly Gly Gly Val Pro Met Ser Tyr Ile Glu Ser Glu Glu
455 460 465
gat tgg atc cgt tat caa tcc gct aaa tct gcg act tca gcc ggg tat 1555
Asp Trp Ile Arg Tyr Gln Ser Ala Lys Ser Ala Thr Ser Ala Gly Tyr
470 475 480 485
gcc gaa tcc ttt acg tgg aaa gac gat ccg tta tct aat acg tac ccg 1603
Ala Glu Ser Phe Thr Trp Lys Asp Asp Pro Leu Ser Asn Thr Tyr Pro
490 495 500
ttc tat cag acc cca gtg cgc ggt aat tgg ttg aaa gac gtg ctt tct 1651
Phe Tyr Gln Thr Pro Val Arg Gly Asn Trp Leu Lys Asp Val Leu Ser
505 510 515
aag ggg gta gct cag atg ctc att gac cgg gga ttg cgg tta cac ata 1699
Lys Gly Val Ala Gln Met Leu Ile Asp Arg Gly Leu Arg Leu His Ile
520 525 530
gag cct ggt cga agt tta cta gat ggg tgt ggc gtc act ctt gcc gaa 1747
Glu Pro Gly Arg Ser Leu Leu Asp Gly Cys Gly Val Thr Leu Ala Glu
535 540 545
gtt gct ttt gtg aaa acc cga agt gac ggg ttg cct cta gtg gga ctg 1795
Val Ala Phe Val Lys Thr Arg Ser Asp Gly Leu Pro Leu Val Gly Leu
550 555 560 565
gct atg aac cga acg cag tgc cgg act aca tcc gat gat ttt ctc att 1843
Ala Met Asn Arg Thr Gln Cys Arg Thr Thr Ser Asp Asp Phe Leu Ile
570 575 580
gat ccc ctg cat atc act gac ggt gat gta ggc gag gaa atc gaa gca 1891
Asp Pro Leu His Ile Thr Asp Gly Asp Val Gly Glu Glu Ile Glu Ala
585 590 595
tat cta gtg ggt gcc tac tgc atc gaa gat gag ctg att tta cgc cgg 1939
Tyr Leu Val Gly Ala Tyr Cys Ile Glu Asp Glu Leu Ile Leu Arg Arg
600 605 610
cga atc cgc ttc ccg aga gga gtc aaa cca gga gat atc atc gga att 1987
Arg Ile Arg Phe Pro Arg Gly Val Lys Pro Gly Asp Ile Ile Gly Ile
615 620 625
cct aac acc gca gga tac ttc atg cat atc ttg gaa agt gca tcg cac 2035
Pro Asn Thr Ala Gly Tyr Phe Met His Ile Leu Glu Ser Ala Ser His
630 635 640 645
caa atc ccg ttg gcg aaa aat gta gtg tgg ccg gag ggg cag tta gac 2083
Gln Ile Pro Leu Ala Lys Asn Val Val Trp Pro Glu Gly Gln Leu Asp
650 655 660
gat atc gat gcg gat taagacataa ccattcgcta atc 2121
Asp Ile Asp Ala Asp
665
<210>46
<211>666
<212>PRT
<213>谷氨酸棒杆菌
<400>46
Met Ile Pro Lys Pro Asp Val Thr Asp Leu Tyr Leu Glu Asp Leu Leu
1 5 10 15
Asn Glu Gly Ser Glu Lys Ile Arg Ser Ala Lys Asp Leu Ser Glu Leu
20 25 30
Arg Thr Val Leu Lys Glu Val Ser Ser Gln Ile Gln Glu Arg Ala Gly
35 40 45
Lys Lys Asp Glu Glu Trp Gly Met Gly Ala Thr Trp Arg Glu Leu Tyr
50 55 60
Pro Ser Ile Val Glu Arg Ala Ser Tyr Glu Gly Arg Asp Ser Leu Ile
65 70 75 80
Gly Phe Asp His Leu Ala Arg Glu Met Glu Arg Leu Ala Phe Gly Pro
85 90 95
Pro Ser Glu Ser Phe Glu Tyr Leu Gln Glu Leu Val Lys Ser Gly Val
100 105 110
Val Asp Ile Thr His Leu His Arg Gly Arg Glu Pro Leu Thr Asp Leu
115 120 125
Val Arg Glu Leu Glu Ile Thr Val Val Ile Asp Ala Val Leu Pro Pro
130 135 140
Pro Gly Val Val Pro Gly Thr Leu Val His Asn Leu Val Lys Glu Gly
145 150 155 160
Tyr Ala Arg Met Arg Pro Gly Thr Arg Gly Leu Asp Val Ala Ala Asp
165 170 175
Gly Thr Val Gln Gly Gln Arg His Leu Ala Ala Val Gly Arg Met Thr
180 185 190
Glu Asp Val Val Leu Gly Asn Asp Thr Leu Ser Arg Ser Leu His Asp
195 200 205
Ile Ile Pro Lys Trp Ala Arg Arg Val Ile Arg Asp Ala Ser Thr Tyr
210 215 220
Pro Asp Arg Val His Gly Thr Pro Pro Leu Pro Ala Arg Leu Glu Pro
225 230 235 240
Trp Ala Glu Lys Leu Thr Ser Asp Pro Ala Thr Cys Arg His Leu Ile
245 250 255
Glu Glu Phe Gly Ser Pro Val Asn Val Leu His Ser Gly Ser Met Pro
260 265 270
Arg Asn Ile Asn Glu Leu Val Asp Ala Gly Ile Gln Met Gly Val Asp
275 280 285
Thr Arg Ile Phe Phe Ala Arg Lys Ala Asn Lys Gly Leu Thr Phe Val
290 295 300
Asp Ala Val Lys Asp Thr Gly His Gly Val Asp Val Ala Ser Glu Arg
305 310 315 320
Glu Leu Ser Gln Val Leu Asn Arg Gly Val Pro Gly Glu Arg Ile Ile
325 330 335
Leu Ser Ala Ala Ile Lys Pro Asp Arg Leu Leu Ala Leu Ala Ile Glu
340 345 350
Asn Gly Val Ile Ile Ser Val Asp Ser Arg Asp Glu Leu Asp Arg Ile
355 360 365
Ser Ala Leu Val Gly Asp Arg Val Ala Arg Val Ala Pro Arg Val Ala
370 375 380
Pro Asp Pro Ala Val Leu Pro Pro Thr Arg Phe Gly Glu Arg Ala Ala
385 390 395 400
Asp Trp Gly Asn Arg Leu Thr Glu Val Ile Pro Gly Val Asp Ile Val
405 410 415
Gly Leu His Val His Leu His Gly Tyr Ala Ala Lys Asp Arg Ala Leu
420 425 430
Ala Leu Gln Glu Cys Cys Gln Leu Val Asp Ser Leu Arg Glu Cys Gly
435 440 445
His Ser Pro Gln Phe Ile Asp Leu Gly Gly Gly Val Pro Met Ser Tyr
450 455 460
Ile Glu Ser Glu Glu Asp Trp Ile Arg Tyr Gln Ser Ala Lys Ser Ala
465 470 475 480
Thr Ser Ala Gly Tyr Ala Glu Ser Phe Thr Trp Lys Asp Asp Pro Leu
485 490 495
Ser Asn Thr Tyr Pro Phe Tyr Gln Thr Pro Val Arg Gly Asn Trp Leu
500 505 510
Lys Asp Val Leu Ser Lys Gly Val Ala Gln Met Leu Ile Asp Arg Gly
515 520 525
Leu Arg Leu His Ile Glu Pro Gly Arg Ser Leu Leu Asp Gly Cys Gly
530 535 540
Val Thr Leu Ala Glu Val Ala Phe Val Lys Thr Arg Ser Asp Gly Leu
545 550 555 560
Pro Leu Val Gly Leu Ala Met Asn Arg Thr Gln Cys Arg Thr Thr Ser
565 570 575
Asp Asp Phe Leu Ile Asp Pro Leu His Ile Thr Asp Gly Asp Val Gly
580 585 590
Glu Glu Ile Glu Ala Tyr Leu Val Gly Ala Tyr Cys Ile Glu Asp Glu
595 600 605
Leu Ile Leu Arg Arg Arg Ile Arg Phe Pro Arg Gly Val Lys Pro Gly
610 615 620
Asp Ile Ile Gly Ile Pro Asn Thr Ala Gly Tyr Phe Met His Ile Leu
625 630 635 640
Glu Ser Ala Ser His Gln Ile Pro Leu Ala Lys Asn Val Val Trp Pro
645 650 655
Glu Gly Gln Leu Asp Asp Ile Asp Ala Asp
660 665
<210>47
<211>993
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(970)
<223>RXA01393
<400>47
caaaagcaga cctgtaatga agatttccat gatcaccatc gtgacctatg gaagtactta 60
agtaaaatga ttggttctta acatggttta atatagcttc atg aac ccc att caa 115
Met Asn Pro Ile Gln
1 5
ctg gac act ttg ctc tca atc att gat gaa ggc agc ttc gaa ggc gcc 163
Leu Asp Thr Leu Leu Ser Ile Ile Asp Glu Gly Ser Phe Glu Gly Ala
10 15 20
tcc tta gcc ctt tcc att tcc ccc tcg gcg gtg agt cag cgc gtt aaa 211
Ser Leu Ala Leu Ser Ile Ser Pro Ser Ala Val Ser Gln Arg Val Lys
25 30 35
gct ctc gag cat cac gtg ggt cga gtg ttg gta tcg cgc acc caa ccg 259
Ala Leu Glu His His Val Gly Arg Val Leu Val Ser Arg Thr Gln Pro
40 45 50
gcc aaa gca acc gaa gcg ggt gaa gtc ctt gtg caa gca gcg cgg aaa 307
Ala Lys Ala Thr Glu Ala Gly Glu Val Leu Val Gln Ala Ala Arg Lys
55 60 65
atg gtg ttg ctg caa gca gaa act aaa gcg caa cta tct gga cgc ctt 355
Met Val Leu Leu Gln Ala Glu Thr Lys Ala Gln Leu Ser Gly Arg Leu
70 75 80 85
gct gaa atc ccg tta acc atc gcc atc aac gca gat tcg cta tcc aca 403
Ala Glu Ile Pro Leu Thr Ile Ala Ile Asn Ala Asp Ser Leu Ser Thr
90 95 100
tgg ttt cct ccc gtg ttc aac gag gta gct tct tgg ggt gga gca acg 451
Trp Phe Pro Pro Val Phe Asn Glu Val Ala Ser Trp Gly Gly Ala Thr
105 110 115
ctc acg ctg cgc ttg gaa gat gaa gcg cac aca tta tcc ttg ctg cgg 499
Leu Thr Leu Arg Leu Glu Asp Glu Ala His Thr Leu Ser Leu Leu Arg
120 125 130
cgt gga gat gtt tta gga gcg gta acc cgt gaa gct aat ccc gtg gcg 547
Arg Gly Asp Val Leu Gly Ala Val Thr Arg Glu Ala Asn Pro Val Ala
135 140 145
gga tgt gaa gta gta gaa ctt gga acc atg cgc cac ttg gcc att gca 595
Gly Cys Glu Val Val Glu Leu Gly Thr Met Arg His Leu Ala Ile Ala
150 155 160 165
acc ccc tca ttg cgg gat gcc tac atg gtt gat ggg aaa cta gat tgg 643
Thr Pro Ser Leu Arg Asp Ala Tyr Met Val Asp Gly Lys Leu Asp Trp
170 175 180
gct gcg atg ccc gtc tta cgc ttc ggt ccc aaa gat gtg ctt caa gac 691
Ala Ala Met Pro Val Leu Arg Phe Gly Pro Lys Asp Val Leu Gln Asp
185 190 195
cgt gac ctg gac ggg cgc gtc gat ggt cct gtg ggg cgc agg cgc gta 739
Arg Asp Leu Asp Gly Arg Val Asp Gly Pro Val Gly Arg Arg Arg Val
200 205 210
tcc att gtc ccg tcg gcg gaa ggt ttt ggt gag gca att cgc cga ggc 787
Ser Ile Val Pro Ser Ala Glu Gly Phe Gly Glu Ala Ile Arg Arg Gly
215 220 225
ctt ggt tgg gga ctt ctt ccc gaa acc caa gct gct ccc atg cta aaa 835
Leu Gly Trp Gly Leu Leu Pro Glu Thr Gln Ala Ala Pro Met Leu Lys
230 235 240 245
gca gga gaa gtg atc ctc ctc gat gag ata ccc att gac aca ccg atg 883
Ala Gly Glu Val Ile Leu Leu Asp Glu Ile Pro Ile Asp Thr Pro Met
250 255 260
tat tgg caa cga tgg cgc ctg gaa tct aga tct cta gct aga ctc aca 931
Tyr Trp Gln Arg Trp Arg Leu Glu Ser Arg Ser Leu Ala Arg Leu Thr
265 270 275
gac gcc gtc gtt gat gca gca atc gag gga ttg cgg cct tagttacttc 980
Asp Ala Val Val Asp Ala Ala Ile Glu Gly Leu Arg Pro
280 285 290
tgaaaaggtt cag 993
<210>48
<211>290
<212>PRT
<213>谷氨酸棒杆菌
<400>48
Met Asn Pro Ile Gln Leu Asp Thr Leu Leu Ser Ile Ile Asp Glu Gly
1 5 10 15
Ser Phe Glu Gly Ala Ser Leu Ala Leu Ser Ile Ser Pro Ser Ala Val
20 25 30
Ser Gln Arg Val Lys Ala Leu Glu His His Val Gly Arg Val Leu Val
35 40 45
Ser Arg Thr Gln Pro Ala Lys Ala Thr Glu Ala Gly Glu Val Leu Val
50 55 60
Gln Ala Ala Arg Lys Met Val Leu Leu Gln Ala Glu Thr Lys Ala Gln
65 70 75 80
Leu Ser Gly Arg Leu Ala Glu Ile Pro Leu Thr Ile Ala Ile Asn Ala
85 90 95
Asp Ser Leu Ser Thr Trp Phe Pro Pro Val Phe Asn Glu Val Ala Ser
100 105 110
Trp Gly Gly Ala Thr Leu Thr Leu Arg Leu Glu Asp Glu Ala His Thr
115 120 125
Leu Ser Leu Leu Arg Arg Gly Asp Val Leu Gly Ala Val Thr Arg Glu
130 135 140
Ala Asn Pro Val Ala Gly Cys Glu Val Val Glu Leu Gly Thr Met Arg
145 150 155 160
His Leu Ala Ile Ala Thr Pro Ser Leu Arg Asp Ala Tyr Met Val Asp
165 170 175
Gly Lys Leu Asp Trp Ala Ala Met Pro Val Leu Arg Phe Gly Pro Lys
180 185 190
Asp Val Leu Gln Asp Arg Asp Leu Asp Gly Arg Val Asp Gly Pro Val
195 200 205
Gly Arg Arg Arg Val Ser Ile Val Pro Ser Ala Glu Gly Phe Gly Glu
210 215 220
Ala Ile Arg Arg Gly Leu Gly Trp Gly Leu Leu Pro Glu Thr Gln Ala
225 230 235 240
Ala Pro Met Leu Lys Ala Gly Glu Val Ile Leu Leu Asp Glu Ile Pro
245 250 255
Ile Asp Thr Pro Met Tyr Trp Gln Arg Trp Arg Leu Glu Ser Arg Ser
260 265 270
Leu Ala Arg Leu Thr Asp Ala Val Val Asp Ala Ala Ile Glu Gly Leu
275 280 285
Arg Pro
290
<210>49
<211>1626
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1603)
<223>RXA00241
<400>49
ggtctccagc ctttctaaac aattcatctg cacttgatta attggcccca agattacgcg 60
aagtttagcg acttcgccgt acgtcaacta cgttaaatga gtg aat act caa tca 115
Val Asn Thr Gln Ser
1 5
gat tct gcg ggg tct caa ggt gca gcg gcc aca agt cgt act gta tct 163
Asp Ser Ala Gly Ser Gln Gly Ala Ala Ala Thr Ser Arg Thr Val Ser
10 15 20
att aga acc ctc atc gcg ctg atc atc gga tcg acc gtc ggc gcg gga 211
Ile Arg Thr Leu Ile Ala Leu Ile Ile Gly Ser Thr Val Gly Ala Gly
25 30 35
att ttc tcc atc cct caa aac atc ggc tca gtc gca ggt ccc ggc gcg 259
Ile Phe Ser Ile Pro Gln Asn Ile Gly Ser Val Ala Gly Pro Gly Ala
40 45 50
atg ctc atc ggc tgg ctg atc gcc ggt gtg ggc atg ttg tcc gta gcg 307
Met Leu Ile Gly Trp Leu Ile Ala Gly Val Gly Met Leu Ser Val Ala
55 60 65
ttc gtg ttc cat gtt ctt gcc cgc cgt aaa cct cac ctc gat tct ggc 355
Phe Val Phe His Val Leu Ala Arg Arg Lys Pro His Leu Asp Ser Gly
70 75 80 85
gtc tac gca tat gcg cgt gtt gga ttg ggc gat tat gta ggt ttc tcc 403
Val Tyr Ala Tyr Ala Arg Val Gly Leu Gly Asp Tyr Val Gly Phe Ser
90 95 100
tcc gct tgg ggt tat tgg ctg ggt tca gtc atc gcc caa gtt ggc tac 45l
Ser Ala Trp Gly Tyr Trp Leu Gly Ser Val Ile Ala Gln Val Gly Tyr
105 110 115
gca acg tta ttt ttc tcc acg ttg ggc cac tac gta ccg ctg ttt tcc 499
Ala Thr Leu Phe Phe Ser Thr Leu Gly His Tyr Val Pro Leu Phe Ser
120 125 130
caa gat cat cca ttt gtg tca gcg ttg gca gtt agc gct ttg acc tgg 547
Gln Asp His Pro Phe Val Ser Ala Leu Ala Val Ser Ala Leu Thr Trp
135 140 145
ctg gtg ttt gga gtt gtt tcc cga gga att agc caa gct gct ttc ttg 595
Leu Val Phe Gly Val Val Ser Arg Gly Ile Ser Gln Ala Ala Phe Leu
150 155 160 165
aca acg gtc acc acc gtg gcc aaa att ctg cct ctg ttg tgc ttc atc 643
Thr Thr Val Thr Thr Val Ala Lys Ile Leu Pro Leu Leu Cys Phe Ile
170 175 180
atc ctt gtt gca ttc ttg ggc ttt agc tgg gag aag ttc act gtt gat 691
Ile Leu Val Ala Phe Leu Gly Phe Ser Trp Glu Lys Phe Thr Val Asp
185 190 195
tta tgg gcg cgt gat ggt ggc gtg ggc agc att ttt gat cag gtg cgc 739
Leu Trp Ala Arg Asp Gly Gly Val Gly Ser Ile Phe Asp Gln Val Arg
200 205 210
ggc atc atg gtg tac acc gtg tgg gtg ttc atc ggt atc gaa ggt gca 787
Gly Ile Met Val Tyr Thr Val Trp Val Phe Ile Gly Ile Glu Gly Ala
215 220 225
tcg gta tat tcc cgc cag gca cgc tca cgc agt gat gtc agc cga gct 835
Ser Val Tyr Ser Arg Gln Ala Arg Ser Arg Ser Asp Val Ser Arg Ala
230 235 240 245
acc gtg att ggt ttt gtg gct gtt ctc ctt ttg ctg gtg tcg att tct 883
Thr Val Ile Gly Phe Val Ala Val Leu Leu Leu Leu Val Ser Ile Ser
250 255 260
tcg ctg agc ttc ggt gta ctg acc caa caa gag ctc gct gcg tta cca 931
Ser Leu Ser Phe Gly Val Leu Thr Gln Gln Glu Leu Ala Ala Leu Pro
265 270 275
gat aat tcc atg gcg tcg gtg ctc gaa gct gtt gtt ggt cca tgg ggt 979
Asp Asn Ser Met Ala Ser Val Leu Glu Ala Val Val Gly Pro Trp Gly
280 285 290
gcc gca ttg att tcg ttg ggt ctg tgt ctt tcg gtt ctt ggg gcc tat 1027
Ala Ala Leu Ile Ser Leu Gly Leu Cys Leu Ser Val Leu Gly Ala Tyr
295 300 305
gtg tcc tgg cag atg ctc tgc gca gaa cca ctg gcg ttg atg gca atg 1075
Val Ser Trp Gln Met Leu Cys Ala Glu Pro Leu Ala Leu Met Ala Met
310 315 320 325
gat ggc ctc att cca agc aaa atc ggg gcc atc aac agc cgc ggt gct 1123
Asp Gly Leu Ile Pro Ser Lys Ile Gly Ala Ile Asn Ser Arg Gly Ala
330 335 340
gcc tgg atg gct cag ctg atc tcc acc atc gtg att cag att ttc atc 1171
Ala Trp Met Ala Gln Leu Ile Ser Thr Ile Val Ile Gln Ile Phe Ile
345 350 355
atc att ttc ttc ctc aac gag acc acc tac gtc tcc atg gtg caa ttg 1219
Ile Ile Phe Phe Leu Asn Glu Thr Thr Tyr Val Ser Met Val Gln Leu
360 365 370
gct acc aac cta tac ttg gtg cct tac ctg ttc tct gcc ttt tat ctg 1267
Ala Thr Asn Leu Tyr Leu Val Pro Tyr Leu Phe Ser Ala Phe Tyr Leu
375 380 385
gtc atg ctg gca aca cgt gga aaa gga atc acc cac cca cat gcc ggc 1315
Val Met Leu Ala Thr Arg Gly Lys Gly Ile Thr His Pro His Ala Gly
390 395 400 405
aca cgt ttt gat gat tcc ggt cca gag ata tcc cgc cga gaa aac cgc 1363
Thr Arg Phe Asp Asp Ser Gly Pro Glu Ile Ser Arg Arg Glu Asn Arg
410 415 420
aaa cac ctc atc gtc ggt tta gta gca acg gtg tat tca gtg tgg ctg 1411
Lys His Leu Ile Val Gly Leu Val Ala Thr Val Tyr Ser Val Trp Leu
425 430 435
ttt tac gct gca gaa ccg cag ttt gtc ctc ttc gga gcc atg gcg atg 1459
Phe Tyr Ala Ala Glu Pro Gln Phe Val Leu Phe Gly Ala Met Ala Met
440 445 450
ctt ccc ggc tta atc ccc tat gtg tgg aca agg att tat cgt ggc gaa 1507
Leu Pro Gly Leu Ile Pro Tyr Val Trp Thr Arg Ile Tyr Arg Gly Glu
455 460 465
cag gtg ttt aac cgc ttt gaa atc ggc gtg gtt gtt gtc ctg gtc gtt 1555
Gln Val Phe Asn Arg Phe Glu Ile Gly Val Val Val Val Leu Val Val
470 475 480 485
gct gcc agc gcg ggc gtt att ggt ttg gtc aac gga tca cta tcg ctt 1603
Ala Ala Ser Ala Gly Val Ile Gly Leu Val Asn Gly Ser Leu Ser Leu
490 495 500
taaacaccga aaccttcctg cta 1626
<210>50
<211>501
<212>PRT
<213>谷氨酸棒杆菌
<400>50
Val Asn Thr Gln Ser Asp Ser Ala Gly Ser Gln Gly Ala Ala Ala Thr
1 5 10 15
Ser Arg Thr Val Ser Ile Arg Thr Leu Ile Ala Leu Ile Ile Gly Ser
20 25 30
Thr Val Gly Ala Gly Ile Phe Ser Ile Pro Gln Asn Ile Gly Ser Val
35 40 45
Ala Gly Pro Gly Ala Met Leu Ile Gly Trp Leu Ile Ala Gly Val Gly
50 55 60
Met Leu Ser Val Ala Phe Val Phe His Val Leu Ala Arg Arg Lys Pro
65 70 75 80
His Leu Asp Ser Gly Val Tyr Ala Tyr Ala Arg Val Gly Leu Gly Asp
85 90 95
Tyr Val Gly Phe Ser Ser Ala Trp Gly Tyr Trp Leu Gly Ser Val Ile
100 105 110
Ala Gln Val Gly Tyr Ala Thr Leu Phe Phe Ser Thr Leu Gly His Tyr
115 120 125
Val Pro Leu Phe Ser Gln Asp His Pro Phe Val Ser Ala Leu Ala Val
130 135 140
Ser Ala Leu Thr Trp Leu Val Phe Gly Val Val Ser Arg Gly Ile Ser
145 150 155 160
Gln Ala Ala Phe Leu Thr Thr Val Thr Thr Val Ala Lys Ile Leu Pro
165 170 175
Leu Leu Cys Phe Ile Ile Leu Val Ala Phe Leu Gly Phe Ser Trp Glu
180 185 190
Lys Phe Thr Val Asp Leu Trp Ala Arg Asp Gly Gly Val Gly Ser Ile
195 200 205
Phe Asp Gln Val Arg Gly Ile Met Val Tyr Thr Val Trp Val Phe Ile
210 215 220
Gly Ile Glu Gly Ala Ser Val Tyr Ser Arg Gln Ala Arg Ser Arg Ser
225 230 235 240
Asp Val Ser Arg Ala Thr Val Ile Gly Phe Val Ala Val Leu Leu Leu
245 250 255
Leu Val Ser Ile Ser Ser Leu Ser Phe Gly Val Leu Thr Gln Gln Glu
260 265 270
Leu Ala Ala Leu Pro Asp Asn Ser Met Ala Ser Val Leu Glu Ala Val
275 280 285
Val Gly Pro Trp Gly Ala Ala Leu Ile Ser Leu Gly Leu Cys Leu Ser
290 295 300
Val Leu Gly Ala Tyr Val Ser Trp Gln Met Leu Cys Ala Glu Pro Leu
305 310 315 320
Ala Leu Met Ala Met Asp Gly Leu Ile Pro Ser Lys Ile Gly Ala Ile
325 330 335
Asn Ser Arg Gly Ala Ala Trp Met Ala Gln Leu Ile Ser Thr Ile Val
340 345 350
Ile Gln Ile Phe Ile Ile Ile Phe Phe Leu Asn Glu Thr Thr Tyr Val
355 360 365
Ser Met Val Gln Leu Ala Thr Asn Leu Tyr Leu Val Pro Tyr Leu Phe
370 375 380
Ser Ala Phe Tyr Leu Val Met Leu Ala Thr Arg Gly Lys Gly Ile Thr
385 390 395 400
His Pro His Ala Gly Thr Arg Phe Asp Asp Ser Gly Pro Glu Ile Ser
405 410 415
Arg Arg Glu Asn Arg Lys His Leu Ile Val Gly Leu Val Ala Thr Val
420 425 430
Tyr Ser Val Trp Leu Phe Tyr Ala Ala Glu Pro Gln Phe Val Leu Phe
435 440 445
Gly Ala Met Ala Met Leu Pro Gly Leu Ile Pro Tyr Val Trp Thr Arg
450 455 460
Ile Tyr Arg Gly Glu Gln Val Phe Asn Arg Phe Glu Ile Gly Val Val
465 470 475 480
Val Val Leu Val Val Ala Ala Ser Ala Gly Val Ile Gly Leu Val Asn
485 490 495
Gly Ser Leu Ser Leu
500
<210>51
<211>822
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(799)
<223>RXA01394
<400>51
gagcaaagtg tccagttgaa tggggttcat gaagctatat taaaccatgt taagaaccaa 60
tcattttact taagtacttc cataggtcac gatggtgatc atg gaa atc ttc att 115
Met Glu Ile Phe Ile
1 5
aca ggt ctg ctt ttg ggg gcc agt ctt tta ctg tcc atc gga ccg cag 163
Thr Gly Leu Leu Leu Gly Ala Ser Leu Leu Leu Ser Ile Gly Pro Gln
10 15 20
aat gta ctg gtg att aaa caa gga att aag cgc gaa gga ctc att gcg 211
Asn Val Leu Val Ile Lys Gln Gly Ile Lys Arg Glu Gly Leu Ile Ala
25 30 35
gtt ctt ctc gtg tgt tta att tct gac gtc ttt ttg ttc atc gcc ggc 259
Val Leu Leu Val Cys Leu Ile Ser Asp Val Phe Leu Phe Ile Ala Gly
40 45 50
acc ttg ggc gtt gat ctt ttg tcc aat gcc gcg ccg atc gtg ctc gat 307
Thr Leu Gly Val Asp Leu Leu Ser Asn Ala Ala Pro Ile Val Leu Asp
55 60 65
att atg cgc tgg ggt ggc atc gct tac ctg tta tgg ttt gcc gtc atg 355
Ile Met Arg Trp Gly Gly Ile Ala Tyr Leu Leu Trp Phe Ala Val Met
70 75 80 85
gca gcg aaa gac gcc atg aca aac aag gtg gaa gcg cca cag atc att 403
Ala Ala Lys Asp Ala Met Thr Asn Lys Val Glu Ala Pro Gln Ile Ile
90 95 100
gaa gaa aca gaa cca acc gtg ccc gat gac acg cct ttg ggc ggt tcg 451
Glu Glu Thr Glu Pro Thr Val Pro Asp Asp Thr Pro Leu Gly Gly Ser
105 110 115
gcg gtg gcc act gac acg cgc aac cgg gtg cgg gtg gag gtg agc gtc 499
Ala Val Ala Thr Asp Thr Arg Asn Arg Val Arg Val Glu Val Ser Val
120 125 130
gat aag cag cgg gtt tgg gta aag ccc atg ttg atg gca atc gtg ctg 547
Asp Lys Gln Arg Val Trp Val Lys Pro Met Leu Met Ala Ile Val Leu
135 140 145
acc tgg ttg aac ccg aat gcg tat ttg gac gcg ttt gtg ttt atc ggc 595
Thr Trp Leu Asn Pro Asn Ala Tyr Leu Asp Ala Phe Val Phe Ile Gly
150 155 160 165
ggc gtc ggc gcg caa tac ggc gac acc gga cgg tgg att ttc gcc gct 643
Gly Val Gly Ala Gln Tyr Gly Asp Thr Gly Arg Trp Ile Phe Ala Ala
170 175 180
ggc gcg ttc gcg gca agc ctg atc tgg ttc ccg ctg gtg ggt ttc ggc 691
Gly Ala Phe Ala Ala Ser Leu Ile Trp Phe Pro Leu Val Gly Phe Gly
185 190 195
gca gca gca ttg tca cgc ccg ctg tcc agc ccc aag gtg tgg cgc tgg 739
Ala Ala Ala Leu Ser Arg Pro Leu Ser Ser Pro Lys Val Trp Arg Trp
200 205 210
atc aac gtc gtc gtg gca gtt gtg atg acc gca ttg gcc atc aaa ctg 787
Ile Asn Val Val Val Ala Val Val Met Thr Ala Leu Ala Ile Lys Leu
215 220 225
atg ttg atg ggt tagttttcgc gggttttgga atc 822
Met Leu Met Gly
230
<210>52
<211>233
<212>PRT
<213>谷氨酸棒杆菌
<400>52
Met Glu Ile Phe Ile Thr Gly Leu Leu Leu Gly Ala Ser Leu Leu Leu
1 5 10 15
Ser Ile Gly Pro Gln Asn Val Leu Val Ile Lys Gln Gly Ile Lys Arg
20 25 30
Glu Gly Leu Ile Ala Val Leu Leu Val Cys Leu Ile Ser Asp Val Phe
35 40 45
Leu Phe Ile Ala Gly Thr Leu Gly Val Asp Leu Leu Ser Asn Ala Ala
50 55 60
Pro Ile Val Leu Asp Ile Met Arg Trp Gly Gly Ile Ala Tyr Leu Leu
65 70 75 80
Trp Phe Ala Val Met Ala Ala Lys Asp Ala Met Thr Asn Lys Val Glu
85 90 95
Ala Pro Gln Ile Ile Glu Glu Thr Glu Pro Thr Val Pro Asp Asp Thr
100 105 110
Pro Leu Gly Gly Ser Ala Val Ala Thr Asp Thr Arg Asn Arg Val Arg
115 120 125
Val Glu Val Ser Val Asp Lys Gln Arg Val Trp Val Lys Pro Met Leu
130 135 140
Met Ala Ile Val Leu Thr Trp Leu Asn Pro Asn Ala Tyr Leu Asp Ala
145 150 155 160
Phe Val Phe Ile Gly Gly Val Gly Ala Gln Tyr Gly Asp Thr Gly Arg
165 170 175
Trp Ile Phe Ala Ala Gly Ala Phe Ala Ala Ser Leu Ile Trp Phe Pro
180 185 190
Leu Val Gly Phe Gly Ala Ala Ala Leu Ser Arg Pro Leu Ser Ser Pro
195 200 205
Lys Val Trp Arg Trp Ile Asn Val Val Val Ala Val Val Met Thr Ala
210 215 220
Leu Ala Ile Lys Leu Met Leu Met Gly
225 230
<210>53
<211>1026
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1003)
<223>RXA00865
<400>53
ttatcggaat gtggcttggg cgattgttat gcaaaagttg ttaggttttt tgcggggttg 60
tttaaccccc aaatgaggga agaaggtaac cttgaactct atg agc aca ggt tta 115
Met Ser Thr Gly Leu
1 5
aca gct aag acc gga gta gag cac ttc ggc acc gtt gga gta gca atg 163
Thr Ala Lys Thr Gly Val Glu His Phe Gly Thr Val Gly Val Ala Met
10 15 20
gtt act cca ttc acg gaa tcc gga gac atc gat atc gct gct ggc cgc 211
Val Thr Pro Phe Thr Glu Ser Gly Asp Ile Asp Ile Ala Ala Gly Arg
25 30 35
gaa gtc gcg gct tat ttg gtt gat aag ggc ttg gat tct ttg gtt ctc 259
Glu Val Ala Ala Tyr Leu Val Asp Lys Gly Leu Asp Ser Leu Val Leu
40 45 50
gcg ggc acc act ggt gaa tcc cca acg aca acc gcc gct gaa aaa cta 307
Ala Gly Thr Thr Gly Glu Ser Pro Thr Thr Thr Ala Ala Glu Lys Leu
55 60 65
gaa ctg ctc aag gcc gtt cgt gag gaa gtt ggg gat cgg gcg aag ctc 355
Glu Leu Leu Lys Ala Val Arg Glu Glu Val Gly Asp Arg Ala Lys Leu
70 75 80 85
atc gcc ggt gtc gga acc aac aac acg cgg aca tct gtg gaa ctt gcg 403
Ile Ala Gly Val Gly Thr Asn Asn Thr Arg Thr Ser Val Glu Leu Ala
90 95 100
gaa gct gct gct tct gct ggc gca gac ggc ctt tta gtt gta act cct 451
Glu Ala Ala Ala Ser Ala Gly Ala Asp Gly Leu Leu Val Val Thr Pro
105 110 115
tat tac tcc aag ccg agc caa gag gga ttg ctg gcg cac ttc ggt gca 499
Tyr Tyr Ser Lys Pro Ser Gln Glu Gly Leu Leu Ala His Phe Gly Ala
120 125 130
att gct gca gca aca gag gtt cca att tgt ctc tat gac att cct ggt 547
Ile Ala Ala Ala Thr Glu Val Pro Ile Cys Leu Tyr Asp Ile Pro Gly
135 140 145
cgg tca ggt att cca att gag tct gat acc atg aga cgc ctg agt gaa 595
Arg Ser Gly Ile Pro Ile Glu Ser Asp Thr Met Arg Arg Leu Ser Glu
150 155 160 165
tta cct acg att ttg gcg gtc aag gac gcc aag ggt gac ctc gtt gca 643
Leu Pro Thr Ile Leu Ala Val Lys Asp Ala Lys Gly Asp Leu Val Ala
170 175 180
gcc acg tca ttg atc aaa gaa acg gga ctt gcc tgg tat tca ggc gat 691
Ala Thr Ser Leu Ile Lys Glu Thr Gly Leu Ala Trp Tyr Ser Gly Asp
185 190 195
gac cca cta aac ctt gtt tgg ctt gct ttg ggc gga tca ggt ttc att 739
Asp Pro Leu Asn Leu Val Trp Leu Ala Leu Gly Gly Ser Gly Phe Ile
200 205 210
tcc gta att gga cat gca gcc ccc aca gca tta cgt gag ttg tac aca 787
Ser Val Ile Gly His Ala Ala Pro Thr Ala Leu Arg Glu Leu Tyr Thr
215 220 225
agc ttc gag gaa ggc gac ctc gtc cgt gcg cgg gaa atc aac gcc aaa 835
Ser Phe Glu Glu Gly Asp Leu Val Arg Ala Arg Glu Ile Asn Ala Lys
230 235 240 245
cta tca ccg ctg gta gct gcc caa ggt cgc ttg ggt gga gtc agc ttg 883
Leu Ser Pro Leu Val Ala Ala Gln Gly Arg Leu Gly Gly Val Ser Leu
250 255 260
gca aaa gct gct ctg cgt ctg cag ggc atc aac gta gga gat cct cga 931
Ala Lys Ala Ala Leu Arg Leu Gln Gly Ile Asn Val Gly Asp Pro Arg
265 270 275
ctt cca att atg gct cca aat gag cag gaa ctt gag gct ctc cga gaa 979
Leu Pro Ile Met Ala Pro Asn Glu Gln Glu Leu Glu Ala Leu Arg Glu
280 285 290
gac atg aaa aaa gct gga gtt cta taaatatgaa tgattcccga aat 1026
Asp Met Lys Lys Ala Gly Val Leu
295 300
<210>54
<211>301
<212>PRT
<213>谷氨酸棒杆菌
<400>54
Met Ser Thr Gly Leu Thr Ala Lys Thr Gly Val Glu His Phe Gly Thr
1 5 10 15
Val Gly Val Ala Met Val Thr Pro Phe Thr Glu Sar Gly Asp Ile Asp
20 25 30
Ile Ala Ala Gly Arg Glu Val Ala Ala Tyr Leu Val Asp Lys Gly Leu
35 40 45
Asp Ser Leu Val Leu Ala Gly Thr Thr Gly Glu Ser Pro Thr Thr Thr
50 55 60
Ala Ala Glu Lys Leu lu Leu Leu Lys Ala Val Arg Glu Glu Val Gly
65 70 75 80
Asp Arg Ala Lys Leu Ile Ala Gly Val Gly Thr Asn Asn Thr Arg Thr
85 90 95
Ser Val Glu Leu Ala Glu Ala Ala Ala Ser Ala Gly Ala Asp Gly Leu
100 105 110
Leu Val Val Thr Pro Tyr Tyr Ser Lys Pro Ser Gln Glu Gly Leu Leu
115 120 125
Ala His Phe Gly Ala Ile Ala Ala Ala Thr Glu Val Pro Ile Cys Leu
130 135 140
Tyr Asp Ile Pro Gly Arg Ser Gly Ile Pro Ile Glu Ser Asp Thr Met
145 150 155 160
Arg Arg Leu Ser Glu Leu Pro Thr Ile Leu Ala Val Lys Asp Ala Lys
165 170 175
Gly Asp Leu Val Ala Ala Thr Ser Leu Ile Lys Glu Thr Gly Leu Ala
180 185 190
Trp Tyr Ser Gly Asp Asp Pro Leu Asn Leu Val Trp Leu Ala Leu Gly
195 200 205
Gly Ser Gly Phe Ile Ser Val Ile Gly His Ala Ala Pro Thr Ala Leu
210 215 220
Arg Glu Leu Tyr Thr Ser Phe Glu Glu Gly Asp Leu Val Arg Ala Arg
225 230 235 240
Glu Ile Asn Ala Lys Leu Ser Pro Leu Val Ala Ala Gln Gly Arg Leu
245 250 255
Gly Gly Val Ser Leu Ala Lys Ala Ala Leu Arg Leu Gln Gly Ile Asn
260 265 270
Val Gly Asp Pro Arg Leu Pro Ile Met Ala Pro Asn Glu Gln Glu Leu
275 280 285
Glu Ala Leu Arg Glu Asp Met Lys Lys Ala Gly Val Leu
290 295 300
<210>55
<211>1071
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1048)
<223>RXS02021
<400>55
ttgggtcgcc gaggagatct aatcctggtt tgagttcaga gttcacaggt ttaagcctac 60
aaaccttagt taaaacatga tggaagcggt cgattaaaaa atg agt gaa aac att 115
Met Ser Glu Asn Ile
1 5
cgc gga gcc caa gca gtt gga atc gca aat atc gcc atg gac ggg acc 163
Arg Gly Ala Gln Ala Val Gly Ile Ala Asn Ile Ala Met Asp Gly Thr
10 15 20
atc ctg gac acg tgg tac cca gaa ccc caa att ttc aac ccg gat cag 211
Ile Leu Asp Thr Trp Tyr Pro Glu Pro Gln Ile Phe Asn Pro Asp Gln
25 30 35
tgg gct gaa cgc tac cca ttg gaa gtg ggc acc aca cgc ctc gga gca 259
Trp Ala Glu Arg Tyr Pro Leu Glu Val Gly Thr Thr Arg Leu Gly Ala
40 45 50
aac gaa ctc acc cca cgg atg ctg cag ttg gta aaa ctg gac caa gat 307
Asn Glu Leu Thr Pro Arg Met Leu Gln Leu Val Lys Leu Asp Gln Asp
55 60 65
cgc ctc gtc gaa cag gta gca gtc cgc acc gtt atc ccc gat ctg tct 355
Arg Leu Val Glu Gln Val Ala Val Arg Thr Val Ile Pro Asp Leu Ser
70 75 80 85
caa cct cca gta gac gcg cac gat gtt tac ctg cgc ctc cac ctg ctt 403
Gln Pro Pro Val Asp Ala His Asp Val Tyr Leu Arg Leu His Leu Leu
90 95 100
tcc cac cgg ctg gtc cgc ccc cac gaa atg cac atg caa aac acc ttg 451
Ser His Arg Leu Val Arg Pro His Glu Met His Met Gln Asn Thr Leu
105 110 115
gag ctg ctg tcc gac gtg gtg tgg aca aac aag ggc cct tgc ctt cct 499
Glu Leu Leu Ser Asp Val Val Trp Thr Asn Lys Gly Pro Cys Leu Pro
120 125 130
gaa aac ttt gag tgg gtg cgt ggt gct ctg cgg tcc cgc gga ctc atc 547
Glu Asn Phe Glu Trp Val Arg Gly Ala Leu Arg Ser Arg Gly Leu Ile
135 140 145
cac gtc tac tgt gtg gac cgt ctt ccc cgc atg gtc gac tat gtg gtt 595
His Val Tyr Cys Val Asp Arg Leu Pro Arg Met Val Asp Tyr Val Val
150 155 160 165
ccc cct gga gtc cgc atc tcc gaa gca gaa cgc gtg cgc cta ggt gca 643
Pro Pro Gly Val Arg Ile Ser Glu Ala Glu Arg Val Arg Leu Gly Ala
170 175 180
tac ctt gct ccg ggt acc tct gtg ctg cgt gaa ggt ttc gtg tct ttc 691
Tyr Leu Ala Pro Gly Thr Ser Val Leu Arg Glu Gly Phe Val Ser Phe
185 190 195
aac tcc ggc acc ttg ggt gcc gca aag gtg gaa ggc cgc ctg agt tcc 739
Asn Ser Gly Thr Leu Gly Ala Ala Lys Val Glu Gly Arg Leu Ser Ser
200 205 210
ggt gtg gtc atc ggt gaa ggt tcc gag att gga ctg tct tct act att 787
Gly Val Val Ile Gly Glu Gly Ser Glu Ile Gly Leu Ser Ser Thr Ile
215 220 225
cag tcc ccg aga gat gaa cag cgc cgc cgt ttg ccg ttg agc atc ggc 835
Gln Ser Pro Arg Asp Glu Gln Arg Arg Arg Leu Pro Leu Ser Ile Gly
230 235 240 245
caa aac tgc aac ttt ggt gtc agc tcc gga atc atc gga gtc agt ctg 883
Gln Asn Cys Asn Phe Gly Val Ser Ser Gly Ile Ile Gly Val Ser Leu
250 255 260
gga gac aat tgc gac atc gga aat aac att gtc ttg gat gga gat acc 931
Gly Asp Asn Cys Asp Ile Gly Asn Asn Ile Val Leu Asp Gly Asp Thr
265 270 275
ccc att tgg ttc gca gcc gat gag gag tta cgc act atc gac tcc atc 979
Pro Ile Trp Phe Ala Ala Asp Glu Glu Leu Arg Thr Ile Asp Ser Ile
280 285 290
gaa ggc caa gca aat tgg tca atc aag cgt gaa tcc ggc ttc cat gag 1027
Glu Gly Gln Ala Asn Trp Ser Ile Lys Arg Glu Ser Gly Phe His Glu
295 300 305
cca gtt gcc cgc ctc aaa gct tgacccattt tcataaccag tgc 1071
Pro Val Ala Arg Leu Lys Ala
310 315
<210>56
<211>316
<212>PRT
<213>谷氨酸棒杆菌
<400>56
Met Ser Glu Asn Ile Arg Gly Ala Gln Ala Val Gly Ile Ala Asn Ile
1 5 10 15
Ala Met Asp Gly Thr Ile Leu Asp Thr Trp Tyr Pro Glu Pro Gln Ile
20 25 30
Phe Asn Pro Asp Gln Trp Ala Glu Arg Tyr Pro Leu Glu Val Gly Thr
35 40 45
Thr Arg Leu Gly Ala Asn Glu Leu Thr Pro Arg Met Leu Gln Leu Val
50 55 60
Lys Leu Asp Gln Asp Arg Leu Val Glu Gln Val Ala Val Arg Thr Val
65 70 75 80
Ile Pro Asp Leu Ser Gln Pro Pro Val Asp Ala His Asp Val Tyr Leu
85 90 95
Arg Leu His Leu Leu Ser His Arg Leu Val Arg Pro His Glu Met His
100 105 110
Met Gln Asn Thr Leu Glu Leu Leu Ser Asp Val Val Trp Thr Asn Lys
115 120 125
Gly Pro Cys Leu Pro Glu Asn Phe Glu Trp Val Arg Gly Ala Leu Arg
130 135 140
Ser Arg Gly Leu Ile His Val Tyr Cys Val Asp Arg Leu Pro Arg Met
145 150 155 160
Val Asp Tyr Val Val Pro Pro Gly Val Arg Ile Ser Glu Ala Glu Arg
165 170 175
Val Arg Leu Gly Ala Tyr Leu Ala Pro Gly Thr Ser Val Leu Arg Glu
180 185 190
Gly Phe Val Ser Phe Asn Ser Gly Thr Leu Gly Ala Ala Lys Val Glu
195 200 205
Gly Arg Leu Ser Ser Gly Val Val Ile Gly Glu Gly Ser Glu Ile Gly
210 215 220
Leu Ser Ser Thr Ile Gln Ser Pro Arg Asp Glu Gln Arg Arg Arg Leu
225 230 235 240
Pro Leu Ser Ile Gly Gln Asn Cys Asn Phe Gly Val Ser Ser Gly Ile
245 250 255
Ile Gly Val Ser Leu Gly Asp Asn Cys Asp Ile Gly Asn Asn Ile Val
260 265 270
Leu Asp Gly Asp Thr Pro Ile Trp Phe Ala Ala Asp Glu Glu Leu Arg
275 280 285
Thr Ile Asp Ser Ile Glu Gly Gln Ala Asn Trp Ser Ile Lys Arg Glu
290 295 300
Ser Gly Phe His Glu Pro Val Ala Arg Leu Lys Ala
305 310 315
<210>57
<211>1296
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1273)
<223>RXS02157
<400>57
gggtggaatt ggcacgatgg tgctgccgga tgtttttgat cgggagaatt atcctgaagg 60
caccgttttt agaaaagacg acaaggatgg ggaactgtaa atg agc acg ctg gaa 115
Met Ser Thr Leu Glu
1 5
act tgg cca cag gtc att att aat acg tac ggc acc cca cca gtt gag 163
Thr Trp Pro Gln Val Ile Ile Asn Thr Tyr Gly Thr Pro Pro Val Glu
10 15 20
ctg gtg tcc ggc aag ggc gca acc gtc act gat gac cag ggc aat gtc 211
Leu Val Ser Gly Lys Gly Ala Thr Val Thr Asp Asp Gln Gly Asn Val
25 30 35
tac atc gac ttg ctc gcg ggc atc gca gtc aac gcg ttg ggc cac gcc 259
Tyr Ile Asp Leu Leu Ala Gly Ile Ala Val Asn Ala Leu Gly His Ala
40 45 50
cac ccg gcg atc atc gag gcg gtc acc aac cag atc ggc caa ctt ggt 307
His Pro Ala Ile Ile Glu Ala Val Thr Asn Gln Ile Gly Gln Leu Gly
55 60 65
cac gtc tca aac ttg ttc gca tcc agg ccc gtc gtc gag gtc gcc gag 355
His Val Ser Asn Leu Phe Ala Ser Arg Pro Val Val Glu Val Ala Glu
70 75 80 85
gag ctc atc aag cgt ttt tcg ctt gac gac gcc acc ctc gcc gcg caa 403
Glu Leu Ile Lys Arg Phe Ser Leu Asp Asp Ala Thr Leu Ala Ala Gln
90 95 100
acc cgg gtt ttc ttc tgc aac tcg ggc gcc gaa gca aac gag gct gct 451
Thr Arg Val Phe Phe Cys Asn Ser Gly Ala Glu Ala Asn Glu Ala Ala
105 110 115
ttc aag att gca cgc ttg act ggt cgt tcc cgg att ctg gct gca gtt 499
Phe Lys Ile Ala Arg Leu Thr Gly Arg Ser Arg Ile Leu Ala Ala Val
120 125 130
cat ggt ttc cac ggc cgc acc atg ggt tcc ctc gcg ctg act ggc cag 547
His Gly Phe His Gly Arg Thr Met Gly Ser Leu Ala Leu Thr Gly Gln
135 140 145
cca gac aag cgt gaa gcg ttc ctg cca atg cca agc ggt gtg gag ttc 595
Pro Asp Lys Arg Glu Ala Phe Leu Pro Met Pro Ser Gly Val Glu Phe
150 155 160 165
tac cct tac ggc gac acc gat tac ttg cgc aaa atg gta gaa acc aac 643
Tyr Pro Tyr Gly Asp Thr Asp Tyr Leu Arg Lys Met Val Glu Thr Asn
170 175 180
cca acg gat gtg gct gct atc ttc ctc gag cca atc cag ggt gaa acg 691
Pro Thr Asp Val Ala Ala Ile Phe Leu Glu Pro Ile Gln Gly Glu Thr
185 190 195
ggc gtt gtt cca gca cct gaa gga ttc ctc aag gca gtg cgc gag ctg 739
Gly Val Val Pro Ala Pro Glu Gly Phe Leu Lys Ala Val Arg Glu Leu
200 205 210
tgc gat gag tac ggc atc ttg atg atc acc gat gaa gtc cag act ggc 787
Cys Asp Glu Tyr Gly Ile Leu Met Ile Thr Asp Glu Val Gln Thr Gly
215 220 225
gtt ggc cgt acc ggc gat ttc ttt gca cat cag cac gat ggc gtt gtt 835
Val Gly Arg Thr Gly Asp Phe Phe Ala His Gln His Asp Gly Val Val
230 235 240 245
ccc gat gtg gtg acc atg gcc aag gga ctt ggc ggc ggt ctt ccc atc 883
Pro Asp Val Val Thr Met Ala Lys Gly Leu Gly Gly Gly Leu Pro Ile
250 255 260
ggt gct tgt ttg gcc act ggc cgt gca gct gaa ttg atg acc cca ggc 931
Gly Ala Cys Leu Ala Thr Gly Arg Ala Ala Glu Leu Met Thr Pro Gly
265 270 275
aag cac ggc acc act ttc ggt ggc aac cca gtt gct tgt gca gct gcc 979
Lys His Gly Thr Thr Phe Gly Gly Asn Pro Val Ala Cys Ala Ala Ala
280 285 290
aag gca gtg ctg tct gtt gtc gat gac gct ttc tgc gca gaa gtt gcc 1027
Lys Ala Val Leu Ser Val Val Asp Asp Ala Phe Cys Ala Glu Val Ala
295 300 305
cgc aag ggc gag ctg ttc aag gaa ctt ctt gcc aag gtt gac ggc gtt 1075
Arg Lys Gly Glu Leu Phe Lys Glu Leu Leu Ala Lys Val Asp Gly Val
310 315 320 325
gta gac gtc cgt ggc agg ggc ttg atg ttg ggc gtg gtg ctg gag cgc 1123
Val Asp Val Arg Gly Arg Gly Leu Met Leu Gly Val Val Leu Glu Arg
330 335 340
gac gtc gca aag caa gct gtt ctt gat ggt ttt aag cac ggc gtt att 1171
Asp Val Ala Lys Gln Ala Val Leu Asp Gly Phe Lys His Gly Val Ile
345 350 355
ttg aat gca ccg gcg gac aac att atc cgt ttg acc ccg ccg ctg gtg 1219
Leu Asn Ala Pro Ala Asp Asn Ile Ile Arg Leu Thr Pro Pro Leu Val
360 365 370
atc acc gac gaa gaa atc gca gac gca gtc aag gct att gcc gag aca 1267
Ile Thr Asp Glu Glu Ile Ala Asp Ala Val Lys Ala Ile Ala Glu Thr
375 380 385
atc gca taaaggactc aaacttatga ctt 1296
Ile Ala
390
<210>58
<211>391
<212>PRT
<213>谷氨酸棒杆菌
<400>58
Met Ser Thr Leu Glu Thr Trp Pro Gln Val Ile Ile Asn Thr Tyr Gly
1 5 10 15
Thr Pro Pro Val Glu Leu Val Ser Gly Lys Gly Ala Thr Val Thr Asp
20 25 30
Asp Gln Gly Asn Val Tyr Ile Asp Leu Leu Ala Gly Ile Ala Val Asn
35 40 45
Ala Leu Gly His Ala His Pro Ala Ile Ile Glu Ala Val Thr Asn Gln
50 55 60
Ile Gly Gln Leu Gly His Val Ser Asn Leu Phe Ala Ser Arg Pro Val
65 70 75 80
Val Glu Val Ala Glu Glu Leu Ile Lys Arg Phe Ser Leu Asp Asp Ala
85 90 95
Thr Leu Ala Ala Gln Thr Arg Val Phe Phe Cys Asn Ser Gly Ala Glu
100 105 110
Ala Asn Glu Ala Ala Phe Lys Ile Ala Arg Leu Thr Gly Arg Ser Arg
115 120 125
Ile Leu Ala Ala Val His Gly Phe His Gly Arg Thr Met Gly Ser Leu
130 135 140
Ala Leu Thr Gly Gln Pro Asp Lys Arg Glu Ala Phe Leu Pro Met Pro
145 150 155 160
Ser Gly Val Glu Phe Tyr Pro Tyr Gly Asp Thr Asp Tyr Leu Arg Lys
165 170 175
Met Val Glu Thr Asn Pro Thr Asp Val Ala Ala Ile Phe Leu Glu Pro
180 185 190
Ile Gln Gly Glu Thr Gly Val Val Pro Ala Pro Glu Gly Phe Leu Lys
195 200 205
Ala Val Arg Glu Leu Cys Asp Glu Tyr Gly Ile Leu Met Ile Thr Asp
210 215 220
Glu Val Gln Thr Gly Val Gly Arg Thr Gly Asp Phe Phe Ala His Gln
225 230 235 240
His Asp Gly Val Val Pro Asp Val Val Thr Met Ala Lys Gly Leu Gly
245 250 255
Gly Gly Leu Pro Ile Gly Ala Cys Leu Ala Thr Gly Arg Ala Ala Glu
260 265 270
Leu Met Thr Pro Gly Lys His Gly Thr Thr Phe Gly Gly Asn Pro Val
275 280 285
Ala Cys Ala Ala Ala Lys Ala Val Leu Ser Val Val Asp Asp Ala Phe
290 295 300
Cys Ala Glu Val Ala Arg Lys Gly Glu Leu Phe Lys Glu Leu Leu Ala
305 310 315 320
Lys Val Asp Gly Val Val Asp Val Arg Gly Arg Gly Leu Met Leu Gly
325 330 335
Val Val Leu Glu Arg Asp Val Ala Lys Gln Ala Val Leu Asp Gly Phe
340 345 350
Lys His Gly Val Ile Leu Asn Ala Pro Ala Asp Asn Ile Ile Arg Leu
355 360 365
Thr Pro Pro Leu Val Ile Thr Asp Glu Glu Ile Ala Asp Ala Val Lys
370 375 380
Ala Ile Ala Glu Thr Ile Ala
385 390
<210>59
<211>1008
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(985)
<223>RXC00733
<400>59
acggcgaggt tgtcggtatt ggaacgcaca cgaatttgct gaacacgtgc ggtacctacc 60
gtgaaattgt tgaatcccaa gagactgcgc aggcgcaatc atg agt aat act gca 115
Met Ser Asn Thr Ala
1 5
ggc ccc cgc ggg cgt tcc cat cag gca gac gcc gcg ccg aat caa aag 163
Gly Pro Arg Gly Arg Ser His Gln Ala Asp Ala Ala Pro Asn Gln Lys
10 15 20
gca cag aat ttc gga cca tct gcc aaa agg ctt ttc gga att cta ggc 211
Ala Gln Asn Phe Gly Pro Ser Ala Lys Arg Leu Phe Gly Ile Leu Gly
25 30 35
cat gac cgt aac acc tta att ttt gtt atc ttc cta gcc gtc ctg agc 259
His Asp Arg Asn Thr Leu Ile Phe Val Ile Phe Leu Ala Val Leu Ser
40 45 50
gtt gga ctt acc gtc ttg ggc cca tgg ttg ctg ggt aaa gcc acc aac 307
Val Gly Leu Thr Val Leu Gly Pro Trp Leu Leu Gly Lys Ala Thr Asn
55 60 65
gtg gtg ttt gaa gga ttc cta tct aag cgc atg ccg gct ggt gcg tca 355
Val Val Phe Glu Gly Phe Leu Ser Lys Arg Met Pro Ala Gly Ala Ser
70 75 80 85
aag gaa gat atc atc gcg cag ttg cag gct gca ggt aaa cat aat cag 403
Lys Glu Asp Ile Ile Ala Gln Leu Gln Ala Ala Gly Lys His Asn Gln
90 95 100
gct tcc atg atg gaa gac atg aac ctt gtt cca ggc tca ggc att gat 451
Ala Ser Met Met Glu Asp Met Asn Leu Val Pro Gly Ser Gly Ile Asp
105 110 115
ttt gaa aaa tta gcc atg atc ctc gga ctg gtg atc ggt gct tat ctc 499
Phe Glu Lys Leu Ala Met Ile Leu Gly Leu Val Ile Gly Ala Tyr Leu
120 125 130
atc ggt agc ctg ttg tcg ttg ttc cag gcg cgg atg ctc aac cgc atc 547
Ile Gly Ser Leu Leu Ser Leu Phe Gln Ala Arg Met Leu Asn Arg Ile
135 140 145
gtg caa agt gcc atg cac cgg ctg cgc atg gag gtg gag gaa aaa atc 595
Val Gln Ser Ala Met His Arg Leu Arg Met Glu Val Glu Glu Lys Ile
150 155 160 165
cac cgc cta ccg ctg agc tat ttc gat tcc atc aaa cgt ggt gat ctg 643
His Arg Leu Pro Leu Ser Tyr Phe Asp Ser Ile Lys Arg Gly Asp Leu
170 175 180
ctt agc cgt gtg acc aac gat gtg gat aat atc ggt caa tcc ctg caa 691
Leu Ser Arg Val Thr Asn Asp Val Asp Asn Ile Gly Gln Ser Leu Gln
185 190 195
caa acc ttg tca cag gcg atc act tcc cta ctg acc gtc atc ggt gtg 739
Gln Thr Leu Ser Gln Ala Ile Thr Ser Leu Leu Thr Val Ile Gly Val
200 205 210
ttg gtg atg atg ttt atc atc tcc cca ctg ctc gca ctc gtg gcg ctg 787
Leu Val Met Met Phe Ile Ile Ser Pro Leu Leu Ala Leu Val Ala Leu
215 220 225
gta tcc att ccg gtc acc atc gtg gtc act gtg gtg gtt gcg agc cgt 835
Val Ser Ile Pro Val Thr Ile Val Val Thr Val Val Val Ala Ser Arg
230 235 240 245
tcc cag aaa ctc ttt gcg gaa cag tgg aag cag acc ggt att ttg aat 883
Ser Gln Lys Leu Phe Ala Glu Gln Trp Lys Gln Thr Gly Ile Leu Asn
250 255 260
gcg cgc ctg gag gaa acc tac tct ggc cac gcc gtg gtt aag gtt ttc 931
Ala Arg Leu Glu Glu Thr Tyr Ser Gly His Ala Val Val Lys Val Phe
265 270 275
gga cac caa aag gat gtt caa gaa gca ttc gag gaa gaa aat caa gct 979
Gly His Gln Lys Asp Val Gln Glu Ala Phe Glu Glu Glu Asn Gln Ala
280 285 290
tgt gta taaggccagc tttggtgccc agt 1008
Cys Val
295
<210>60
<211>295
<212>PRT
<213>谷氨酸棒杆菌
<400>60
Met Ser Asn Thr Ala Gly Pro Arg Gly Arg Ser His Gln Ala Asp Ala
1 5 10 15
Ala Pro Asn Gln Lys Ala Gln Asn Phe Gly Pro Ser Ala Lys Arg Leu
20 25 30
Phe Gly Ile Leu Gly His Asp Arg Asn Thr Leu Ile Phe Val Ile Phe
35 40 45
Leu Ala Val Leu Ser Val Gly Leu Thr Val Leu Gly Pro Trp Leu Leu
50 55 60
Gly Lys Ala Thr Asn Val Val Phe Glu Gly Phe Leu Ser Lys Arg Met
65 70 75 80
Pro Ala Gly Ala Ser Lys Glu Asp Ile Ile Ala Gln Leu Gln Ala Ala
85 90 95
Gly Lys His Asn Gln Ala Ser Met Met Glu Asp Met Asn Leu Val Pro
100 105 110
Gly Ser Gly Ile Asp Phe Glu Lys Leu Ala Met Ile Leu Gly Leu Val
115 120 125
Ile Gly Ala Tyr Leu Ile Gly Ser Leu Leu Ser Leu Phe Gln Ala Arg
130 135 140
Met Leu Asn Arg Ile Val Gln Ser Ala Met His Arg Leu Arg Met Glu
145 150 155 160
Val Glu Glu Lys Ile His Arg Leu Pro Leu Ser Tyr Phe Asp Ser Ile
165 170 175
Lys Arg Gly Asp Leu Leu Ser Arg Val Thr Asn Asp Val Asp Asn Ile
180 185 190
Gly Gln Ser Leu Gln Gln Thr Leu Ser Gln Ala Ile Thr Ser Leu Leu
195 200 205
Thr Val Ile Gly Val Leu Val Met Met Phe Ile Ile Ser Pro Leu Leu
210 215 220
Ala Leu Val Ala Leu Val Ser Ile Pro Val Thr Ile Val Val Thr Val
225 230 235 240
Val Val Ala Ser Arg Ser Gln Lys Leu Phe Ala Glu Gln Trp Lys Gln
245 250 255
Thr Gly Ile Leu Asn Ala Arg Leu Glu Glu Thr Tyr Ser Gly His Ala
260 265 270
Val Val Lys Val Phe Gly His Gln Lys Asp Val Gln Glu Ala Phe Glu
275 280 285
Glu Glu Asn Gln Ala Cys Val
290 295
<210>61
<211>426
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(426)
<223>RXC00861
<400>61
atg gct cct cac aag gtc atg ctg att acc act ggt act cag ggt gag 48
Met Ala Pro His Lys Val Met Leu Ile Thr Thr Gly Thr Gln Gly Glu
1 5 10 15
cct atg gct gcg ctg tct cgc atg gcg cgt cgt gag cac cga cag atc 96
Pro Met Ala Ala Leu Ser Arg Met Ala Arg Arg Glu His Arg Gln Ile
20 25 30
act gtc cgt gat gga gac ttg att atc ctt tct tcc tcc ctg gtt cca 144
Thr Val Arg Asp Gly Asp Leu Ile Ile Leu Ser Ser Ser Leu Val Pro
35 40 45
ggt aac gaa gaa gca gtg ttc ggt gtc atc aac atg ctg gct cag atc 192
Gly Asn Glu Glu Ala Val Phe Gly Val Ile Asn Met Leu Ala Gln Ile
50 55 60
ggt gca act gtt gtt acc ggt cgc gac gcc aag gtg cac acc tcg ggc 240
Gly Ala Thr Val Val Thr Gly Arg Asp Ala Lys Val His Thr Ser Gly
65 70 75 80
cac ggc tac tcc gga gag ctg ttg ttc ttg tac aac gcc gct cgt ccg 288
His Gly Tyr Ser Gly Glu Leu Leu Phe Leu Tyr Asn Ala Ala Arg Pro
85 90 95
aag aac gct atg cct gtc cac ggc gag tgg cgc cac ctg cgc gcc aac 336
Lys Asn Ala Met Pro Val His Gly Glu Trp Arg His Leu Arg Ala Asn
100 105 110
aag gaa ctg gct atc tcc act ggt gtt aac cgc gac aac gtt gtg ctt 384
Lys Glu Leu Ala Ile Ser Thr Gly Val Asn Arg Asp Asn Val Val Leu
115 120 125
gca caa aac ggt gtt gtg gtt gat atg gtc aac ggt cgc gca 426
Ala Gln Asn Gly Val Val Val Asp Met Val Asn Gly Arg Ala
130 135 140
<210>62
<211>142
<212>PRT
<213>谷氨酸棒杆菌
<400>62
Met Ala Pro His Lys Val Met Leu Ile Thr Thr Gly Thr Gln Gly Glu
1 5 10 15
Pro Met Ala Ala Leu Ser Arg Met Ala Arg Arg Glu His Arg Gln Ile
20 25 30
Thr Val Arg Asp Gly Asp Leu Ile Ile Leu Ser Ser Ser Leu Val Pro
35 40 45
Gly Asn Glu Glu Ala Val Phe Gly Val Ile Asn Met Leu Ala Gln Ile
50 55 60
Gly Ala Thr Val Val Thr Gly Arg Asp Ala Lys Val His Thr Ser Gly
65 70 75 80
His Gly Tyr Ser Gly Glu Leu Leu Phe Leu Tyr Asn Ala Ala Arg Pro
85 90 95
Lys Asn Ala Met Pro Val His Gly Glu Trp Arg His Leu Arg Ala Asn
100 105 110
Lys Glu Leu Ala Ile Ser Thr Gly Val Asn Arg Asp Asn Val Val Leu
115 120 125
Ala Gln Asn Gly Val Val Val Asp Met Val Asn Gly Arg Ala
130 135 140
<210>63
<211>1066
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1066)
<223>RXC00866
<400>63
gcatcaacgt aggagatcct cgacttccaa ttatggctcc aaatgagcag gaacttgagg 60
ctctccgaga agacatgaaa aaagctggag ttctataaat atg aat gat tcc cga 115
Met Asn Asp Ser Arg
1 5
aat cgc ggc cgg aag gtt acc cgc aag gcg ggc cca cca gaa gct ggt 163
Asn Arg Gly Arg Lys Val Thr Arg Lys Ala Gly Pro Pro Glu Ala Gly
10 15 20
cag gaa aac cat ctg gat acc cct gtc ttt cag gca cca gat gct tcc 211
Gln Glu Asn His Leu Asp Thr Pro Val Phe Gln Ala Pro Asp Ala Ser
25 30 35
tct aac cag agc gct gta aaa gct gag acc gcc gga aac gac aat cgg 259
Ser Asn Gln Ser Ala Val Lys Ala Glu Thr Ala Gly Asn Asp Asn Arg
40 45 50
gat gct gcg caa ggt gct caa gga tcc caa gat tct cag ggt tcc cag 307
Asp Ala Ala Gln Gly Ala Gln Gly Ser Gln Asp Ser Gln Gly Ser Gln
55 60 65
aac gct caa ggt tcc cag aac cgc gag tcc gga aac aac aac cgc aac 355
Asn Ala Gln Gly Ser Gln Asn Arg Glu Ser Gly Asn Asn Asn Arg Asn
70 75 80 85
cgt tcc aac aac aac cgt cgc ggt ggt cgt gga cgt cgt gga tcc gga 403
Arg Ser Asn Asn Asn Arg Arg Gly Gly Arg Gly Arg Arg Gly Ser Gly
90 95 100
aac gcc aat gag ggc gcg aac aac aac agc ggt aac cag aac cgt cag 451
Asn Ala Asn Glu Gly Ala Asn Asn Asn Ser Gly Asn Gln Asn Arg Gln
105 110 115
ggc gga aac cgt ggc aac cgc ggt ggc gga cgc cga aac gtt gtt aag 499
Gly Gly Asn Arg Gly Asn Arg Gly Gly Gly Arg Arg Asn Val Val Lys
120 125 130
tcg atg cag ggt gcg gat ctg acc cag cgc ctg cca gag cca cca aag 547
Ser Met Gln Gly Ala Asp Leu Thr Gln Arg Leu Pro Glu Pro Pro Lys
135 140 145
gca ccg gca aac ggt ctg cgt att tac gca ctt ggt ggc att tcc gaa 595
Ala Pro Ala Asn Gly Leu Arg Ile Tyr Ala Leu Gly Gly Ile Ser Glu
150 155 160 165
atc ggt cgc aac atg acc gtg ttt gag tac aac aac cgt ctg ctc atc 643
Ile Gly Arg Asn Met Thr Val Phe Glu Tyr Asn Asn Arg Leu Leu Ile
170 175 180
gtg gac tgt ggt gtg ctc ttc cca tct tca ggt gag cca ggc gtt gac 691
Val Asp Cys Gly Val Leu Phe Pro Ser Ser Gly Glu Pro Gly Val Asp
185 190 195
ctg att ctt cct gac ttc ggc cca att gag gat cac ctg cac cgc gtc 739
Leu Ile Leu Pro Asp Phe Gly Pro Ile Glu Asp His Leu His Arg Val
200 205 210
gat gca ttg gtg gtt act cac gga cac gaa gac cac att ggt gct att 787
Asp Ala Leu Val Val Thr His Gly His Glu Asp His Ile Gly Ala Ile
215 220 225
ccc tgg ctg ctg aag ctg cgc aac gat atc cca atc ttg gca tcc cgt 835
Pro Trp Leu Leu Lys Leu Arg Asn Asp Ile Pro Ile Leu Ala Ser Arg
230 235 240 245
ttc acc ttg gct ctg att gca gct aag tgt aag gaa cac cgt cag cgt 883
Phe Thr Leu Ala Leu Ile Ala Ala Lys Cys Lys Glu His Arg Gln Arg
250 255 260
ccg aag ctg atc gag gtc aac gag cag tcc aat gag gac cgc gga ccg 931
Pro Lys Leu Ile Glu Val Asn Glu Gln Ser Asn Glu Asp Arg Gly Pro
265 270 275
ttc aac att cgc ttc tgg gct gtt aac cac tcc atc cca gac tgc ctt 979
Phe Asn Ile Arg Phe Trp Ala Val Asn His Ser Ile Pro Asp Cys Leu
280 285 290
ggt ctt gct atc aag act cct gct ggt ttg gtc atc cac acc ggt gac 1027
Gly Leu Ala Ile Lys Thr Pro Ala Gly Leu Val Ile His Thr Gly Asp
295 300 305
atc aag ctg gat cag act cct cct gat gga cgc cca act 1066
Ile Lys Leu Asp Gln Thr Pro Pro Asp Gly Arg Pro Thr
310 315 320
<210>64
<211>322
<212>PRT
<213>谷氨酸棒杆菌
<400>64
Met Asn Asp Ser Arg Asn Arg Gly Arg Lys Val Thr Arg Lys Ala Gly
1 5 10 15
Pro Pro Glu Ala Gly Gln Glu Asn His Leu Asp Thr Pro Val Phe Gln
20 25 30
Ala Pro Asp Ala Ser Ser Asn Gln Ser Ala Val Lys Ala Glu Thr Ala
35 40 45
Gly Asn Asp Asn Arg Asp Ala Ala Gln Gly Ala Gln Gly Ser Gln Asp
50 55 60
Ser Gln Gly Ser Gln Asn Ala Gln Gly Ser Gln Asn Arg Glu Ser Gly
65 70 75 80
Asn Asn Asn Arg Asn Arg Ser Asn Asn Asn Arg Arg Gly Gly Arg Gly
85 90 95
Arg Arg Gly Ser Gly Asn Ala Asn Glu Gly Ala Asn Asn Asn Ser Gly
100 105 110
Asn Gln Asn Arg Gln Gly Gly Asn Arg Gly Asn Arg Gly Gly Gly Arg
115 120 125
Arg Asn Val Val Lys Ser Met Gln Gly Ala Asp Leu Thr Gln Arg Leu
130 135 140
Pro Glu Pro Pro Lys Ala Pro Ala Asn Gly Leu Arg Ile Tyr Ala Leu
145 150 155 160
Gly Gly Ile Ser Glu Ile Gly Arg Asn Met Thr Val Phe Glu Tyr Asn
165 170 175
Asn Arg Leu Leu Ile Val Asp Cys Gly Val Leu Phe Pro Ser Ser Gly
180 185 190
Glu Pro Gly Val Asp Leu Ile Leu Pro Asp Phe Gly Pro Ile Glu Asp
195 200 205
His Leu His Arg Val Asp Ala Leu Val Val Thr His Gly His Glu Asp
210 215 220
His Ile Gly Ala Ile Pro Trp Leu Leu Lys Leu Arg Asn Asp Ile Pro
225 230 235 240
Ile Leu Ala Ser Arg Phe Thr Leu Ala Leu Ile Ala Ala Lys Cys Lys
245 250 255
Glu His Arg Gln Arg Pro Lys Leu Ile Glu Val Asn Glu Gln Ser Asn
260 265 270
Glu Asp Arg Gly Pro Phe Asn Ile Arg Phe Trp Ala Val Asn His Ser
275 280 285
Ile Pro Asp Cys Leu Gly Leu Ala Ile Lys Thr Pro Ala Gly Leu Val
290 295 300
Ile His Thr Gly Asp Ile Lys Leu Asp Gln Thr Pro Pro Asp Gly Arg
305 310 315 320
Pro Thr
<210>65
<211>1527
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1504)
<223>RXC02095
<400>65
ctctcttggt cctctcccca cccattttta agtactcaag acccttccaa cagaaaggat 60
tactccccca acaggctcaa aaatactgaa aggctcacgc atg aaa act gag caa 115
Met Lys Thr Glu Gln
1 5
tcc caa aaa gca caa tta gcc cct aag aaa gca cct gaa aag cca caa 163
Ser Gln Lys Ala Gln Leu Ala Pro Lys Lys Ala Pro Glu Lys Pro Gln
10 15 20
cgc atc cgc caa ctt att tcc gtg gcg tgg cag cga cct tgg ctc acc 211
Arg Ile Arg Gln Leu Ile Ser Val Ala Trp Gln Arg Pro Trp Leu Thr
25 30 35
tca ttc acc gta atc agc gct tta gct gca acg ttg ttt gaa ctt aca 259
Ser Phe Thr Val Ile Ser Ala Leu Ala Ala Thr Leu Phe Glu Leu Thr
40 45 50
ctt cct ctt ttg acc ggt ggc gcc atc gat atc gcg ctc gga aat acc 307
Leu Pro Leu Leu Thr Gly Gly Ala Ile Asp Ile Ala Leu Gly Asn Thr
55 60 65
gga gat act tta acc act gac ctg ctg gac cgg ttc act ccg agt gga 355
Gly Asp Thr Leu Thr Thr Asp Leu Leu Asp Arg Phe Thr Pro Ser Gly
70 75 80 85
tta agc gtg ttg acc agc gtc att gcc ctt atc gtg ctt ctc gcg ttg 403
Leu Ser Val Leu Thr Ser Val Ile Ala Leu Ile Val Leu Leu Ala Leu
90 95 100
ctt cgc tat gcc agt caa ttt gga cgg cga tac acc gca ggc aag ctc 451
Leu Arg Tyr Ala Ser Gln Phe Gly Arg Arg Tyr Thr Ala Gly Lys Leu
105 110 115
agc atg ggg gta cag cat gat gtc cgg ctt aaa acg atg cgc tca ttg 499
Ser Met Gly Val Gln His Asp Val Arg Leu Lys Thr Met Arg Ser Leu
120 125 130
cag aac ctc gat ggg cca ggt cag gac tct att cgc aca ggc caa gta 547
Gln Asn Leu Asp Gly Pro Gly Gln Asp Ser Ile Arg Thr Gly Gln Val
135 140 145
gtc agt cgg tcc att tcg gat atc aac atg gtg caa agc ctt gtg gcg 595
Val Ser Arg Ser Ile Ser Asp Ile Asn Met Val Gln Ser Leu Val Ala
150 155 160 165
atg ttg ccg atg ttg atc gga aat gtg gtc aag ctt gtg ctc act ttg 643
Met Leu Pro Met Leu Ile Gly Asn Val Val Lys Leu Val Leu Thr Leu
170 175 180
gtg atc atg ctg gct att tcc ccg ccg ctg acc atc atc gct gca gtg 691
Val Ile Met Leu Ala Ile Ser Pro Pro Leu Thr Ile Ile Ala Ala Val
185 190 195
ttg gtg cct ttg ctg ttg tgg gcc gtg gcc tat tcg cga aaa gcg ctt 739
Leu Val Pro Leu Leu Leu Trp Ala Val Ala Tyr Ser Arg Lys Ala Leu
200 205 210
ttt gcg tcc acg tgg tcg gcc cag caa aag gct gcg gat ctg acc act 787
Phe Ala Ser Thr Trp Ser Ala Gln Gln Lys Ala Ala Asp Leu Thr Thr
215 220 225
cat gtg gaa gaa act gtc acg ggt atc cgc gtg gtc aag gca ttt gcg 835
His Val Glu Glu Thr Val Thr Gly Ile Arg Val Val Lys Ala Phe Ala
230 235 240 245
cag gaa gac cgc gag acc gac aaa ttg gat ctc acc gca cgt gag tta 883
Gln Glu Asp Arg Glu Thr Asp Lys Leu Asp Leu Thr Ala Arg Glu Leu
250 255 260
ttt gcc cag cgc atg cgc act gca cgt ctg acg gca aag ttc atc ccc 931
Phe Ala Gln Arg Met Arg Thr Ala Arg Leu Thr Ala Lys Phe Ile Pro
265 270 275
atg gtt gag cag ctt ccg cag ctt gct ttg gtg gtc aac att gtt ggc 979
Met Val Glu Gln Leu Pro Gln Leu Ala Leu Val Val Asn Ile Val Gly
280 285 290
ggt ggc tat ttg gcc atg act ggt cac atc acg gtg ggc acg ttt gtg 1027
Gly Gly Tyr Leu Ala Met Thr Gly His Ile Thr Val Gly Thr Phe Val
295 300 305
gcg ttt tct tcc tat ctc act agc ttg tcg gcg gtg gct agg tcc ctg 1075
Ala Phe Ser Ser Tyr Leu Thr Ser Leu Ser Ala Val Ala Arg Ser Leu
310 315 320 325
tcg ggc atg ctc atg cgc gtg cag ttg gcg ctg tct tct gtg gag cgc 1123
Ser Gly Met Leu Met Arg Val Gln Leu Ala Leu Ser Ser Val Glu Arg
330 335 340
atc ttt gaa gtc att gat ctt cag cct gaa cgc acc gat cct gca cac 1171
Ile Phe Glu Val Ile Asp Leu Gln Pro Glu Arg Thr Asp Pro Ala His
345 350 355
ccc ctg tca ctt ccc gac act ccc ctg ggt ctg tcg ttc aac aac gta 1219
Pro Leu Ser Leu Pro Asp Thr Pro Leu Gly Leu Ser Phe Asn Asn Val
360 365 370
gat ttc cgt ggg att ctc aac ggt ttt gag ctg ggt gtt cag gcc ggt 1267
Asp Phe Arg Gly Ile Leu Asn Gly Phe Glu Leu Gly Val Gln Ala Gly
375 380 385
gaa acc gtt gtg ttg gtg ggc cct cca ggt tca ggc aag acc atg gct 1315
Glu Thr Val Val Leu Val Gly Pro Pro Gly Ser Gly Lys Thr Met Ala
390 395 400 405
gtg cag ctt gct gga aac ttt tat caa cca gac agc ggc cac atc gcc 1363
Val Gln Leu Ala Gly Asn Phe Tyr Gln Pro Asp Ser Gly His Ile Ala
410 415 420
ttt gat agc aac ggc cat cgc act cgc ttc gac gac ctc acc cac agc 1411
Phe Asp Ser Asn Gly His Arg Thr Arg Phe Asp Asp Leu Thr His Ser
425 430 435
gat atc cgc agg aat ctc atc gcg gtt ttt gat gag ccg ttc ttg tac 1459
Asp Ile Arg Arg Asn Leu Ile Ala Val Phe Asp Glu Pro Phe Leu Tyr
440 445 450
tcc tcc tcc ata ccg cga gaa cat ctc gat ggg ttt gga tgt cag 1504
Ser Ser Ser Ile Pro Arg Glu His Leu Asp Gly Phe Gly Cys Gln
455 460 465
tgatgagcag atcgaacacg cag 1527
<210>66
<211>468
<212>PRT
<213>谷氨酸棒杆菌
<400>66
Met Lys Thr Glu Gln Ser Gln Lys Ala Gln Leu Ala Pro Lys Lys Ala
1 5 10 15
Pro Glu Lys Pro Gln Arg Ile Arg Gln Leu Ile Ser Val Ala Trp Gln
20 25 30
Arg Pro Trp Leu Thr Ser Phe Thr Val Ile Ser Ala Leu Ala Ala Thr
35 40 45
Leu Phe Glu Leu Thr Leu Pro Leu Leu Thr Gly Gly Ala Ile Asp Ile
50 55 60
Ala Leu Gly Asn Thr Gly Asp Thr Leu Thr Thr Asp Leu Leu Asp Arg
65 70 75 80
Phe Thr Pro Ser Gly Leu Ser Val Leu Thr Ser Val Ile Ala Leu Ile
85 90 95
Val Leu Leu Ala Leu Leu Arg Tyr Ala Ser Gln Phe Gly Arg Arg Tyr
100 105 110
Thr Ala Gly Lys Leu Ser Met Gly Val Gln His Asp Val Arg Leu Lys
115 120 125
Thr Met Arg Ser Leu Gln Asn Leu Asp Gly Pro Gly Gln Asp Ser Ile
130 135 140
Arg Thr Gly Gln Val Val Ser Arg Ser Ile Ser Asp Ile Asn Met Val
145 150 155 160
Gln Ser Leu Val Ala Met Leu Pro Met Leu Ile Gly Asn Val Val Lys
165 170 175
Leu Val Leu Thr Leu Val Ile Met Leu Ala Ile Ser Pro Pro Leu Thr
180 185 190
Ile Ile Ala Ala Val Leu Val Pro Leu Leu Leu Trp Ala Val Ala Tyr
195 200 205
Ser Arg Lys Ala Leu Phe Ala Ser Thr Trp Ser Ala Gln Gln Lys Ala
210 215 220
Ala Asp Leu Thr Thr His Val Glu Glu Thr Val Thr Gly Ile Arg Val
225 230 235 240
Val Lys Ala Phe Ala Gln Glu Asp Arg Glu Thr Asp Lys Leu Asp Leu
245 250 255
Thr Ala Arg Glu Leu Phe Ala Gln Arg Met Arg Thr Ala Arg Leu Thr
260 265 270
Ala Lys Phe Ile Pro Met Val Glu Gln Leu Pro Gln Leu Ala Leu Val
275 280 285
Val Asn Ile Val Gly Gly Gly Tyr Leu Ala Met Thr Gly His Ile Thr
290 295 300
Val Gly Thr Phe Val Ala Phe Ser Ser Tyr Leu Thr Ser Leu Ser Ala
305 310 315 320
Val Ala Arg Ser Leu Ser Gly Met Leu Met Arg Val Gln Leu Ala Leu
325 330 335
Ser Ser Val Glu Arg Ile Phe Glu Val Ile Asp Leu Gln Pro Glu Arg
340 345 350
Thr Asp Pro Ala His Pro Leu Ser Leu Pro Asp Thr Pro Leu Gly Leu
355 360 365
Ser Phe Asn Asn Val Asp Phe Arg Gly Ile Leu Asn Gly Phe Glu Leu
370 375 380
Gly Val Gln Ala Gly Glu Thr Val Val Leu Val Gly Pro Pro Gly Ser
385 390 395 400
Gly Lys Thr Met Ala Val Gln Leu Ala Gly Asn Phe Tyr Gln Pro Asp
405 410 415
Ser Gly His Ile Ala Phe Asp Ser Asn Gly His Arg Thr Arg Phe Asp
420 425 430
Asp Leu Thr His Ser Asp Ile Arg Arg Asn Leu Ile Ala Val Phe Asp
435 440 445
Glu Pro Phe Leu Tyr Ser Ser Ser Ile Pro Arg Glu His Leu Asp Gly
450 455 460
Phe Gly Cys Gln
465
<210>67
<211>295
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(84)..(272)
<223>RXC03185
<400>67
agcgcccaac cgttcagacc agcggtttct ctgaggatgc aaagtccatg atgggtnagg 60
tcactgagct gtccgaaacc acc atg aat gat ctt gca gct gaa ggt gaa aac 113
Met Asn Asp Leu Ala Ala Glu Gly Glu Asn
1 5 10
gat cct tac cgc atg gtt cag cag ctg cgc cgc aag ctc tct cgc ttc 161
Asp Pro Tyr Arg Met Val Gln Gln Leu Arg Arg Lys Leu Ser Arg Phe
15 20 25
gtc gag cag aag tgg aag cgc cag ccg gtc atc atg cca acc gtc att 209
Val Glu Gln Lys Trp Lys Arg Gln Pro Val Ile Met Pro Thr Val Ile
30 35 40
ccg atg act gcg gaa acc acg cac atc ggt gac gat gag gtt cgc gct 257
Pro Met Thr Ala Glu Thr Thr His Ile Gly Asp Asp Glu Val Arg Ala
45 50 55
tca cgc gag tcc ctg taaaagcatt tcgcttttcg acg 295
Ser Arg Glu Ser Leu
60
<210>68
<211>63
<212>PRT
<213>谷氨酸棒杆菌
<400>68
Met Asn Asp Leu Ala Ala Glu Gly Glu Asn Asp Pro Tyr Arg Met Val
1 5 10 15
Gln Gln Leu Arg Arg Lys Leu Ser Arg Phe Val Glu Gln Lys Trp Lys
20 25 30
Arg Gln Pro Val Ile Met Pro Thr Val Ile Pro Met Thr Ala Glu Thr
35 40 45
Thr His Ile Gly Asp Asp Glu Val Arg Ala Ser Arg Glu Ser Leu
50 55 60
<210>69
<211>1170
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1147)
<223>RXA00115
<400>69
tggattctcg agtctgtaca cccttgatca aagcccgagt gttccgtaga ttaactttgt 60
cgtatattgt gacctacacc ccatactgtt aggagttttc atg ctc gac aat agt 115
Met Leu Asp Asn Ser
1 5
ttt tac acc gca gag gtt cag ggc cca tac gaa acc gct tcc att ggc 163
Phe Tyr Thr Ala Glu Val Gln Gly Pro Tyr Glu Thr Ala Ser Ile Gly
10 15 20
cgg ctc gaa ctc gaa gaa ggg ggt gtg att gag gat tgc tgg ttg gct 211
Arg Leu Glu Leu Glu Glu Gly Gly Val Ile Glu Asp Cys Trp Leu Ala
25 30 35
tac gct aca gct gga acg ctc aac gag gac aag tcc aac gcc atc ctc 259
Tyr Ala Thr Ala Gly Thr Leu Asn Glu Asp Lys Ser Asn Ala Ile Leu
40 45 50
att ccg acg tgg tac tcc gga acc cat cag acc tgg ttc cag cag tac 307
Ile Pro Thr Trp Tyr Ser Gly Thr His Gln Thr Trp Phe Gln Gln Tyr
55 60 65
atc ggc act gat cat gcg ctg gat cca tca aag tat ttc atc atc tcc 355
Ile Gly Thr Asp His Ala Leu Asp Pro Ser Lys Tyr Phe Ile Ile Ser
70 75 80 85
atc aac caa atc ggt aat ggt ttg tcg gtc tcc cct gcc aac acg gct 403
Ile Asn Gln Ile Gly Asn Gly Leu Ser Val Ser Pro Ala Asn Thr Ala
90 95 100
gat gac agc atc tcg atg tcc aag ttc ccg aat gtt cgc att ggt gat 451
Asp Asp Ser Ile Ser Met Ser Lys Phe Pro Asn Val Arg Ile Gly Asp
105 110 115
gat gtc gtt gcc cag gac cgg ctc ttg cgc caa gag ttt ggt att acc 499
Asp Val Val Ala Gln Asp Arg Leu Leu Arg Gln Glu Phe Gly Ile Thr
120 125 130
gag ctc ttt gcc gtc gtt ggt ggt tcg atg ggt gcg cag caa acc tat 547
Glu Leu Phe Ala Val Val Gly Gly Ser Met Gly Ala Gln Gln Thr Tyr
135 140 145
gag tgg att gtt cgc ttc cct gac caa gtt cat cga gca gct ccg atc 595
Glu Trp Ile Val Arg Phe Pro Asp Gln Val His Arg Ala Ala Pro Ile
150 155 160 165
gcg ggc act gcg aag aac act cct cat gat ttc atc ttc acc cag act 643
Ala Gly Thr Ala Lys Asn Thr Pro His Asp Phe Ile Phe Thr Gln Thr
170 175 180
ctt aat gag acc gtt gag gcc gat cca ggg ttc aat ggc ggc gaa tac 691
Leu Asn Glu Thr Val Glu Ala Asp Pro Gly Phe Asn Gly Gly Glu Tyr
185 190 195
tcc tcc cat gaa gag gta gct gat gga ctt cgc cgt caa tcg cat ctt 739
Ser Ser His Glu Glu Val Ala Asp Gly Leu Arg Arg Gln Ser His Leu
200 205 210
tgg gct gcc atg gga ttt tcc aca gag ttc tgg aag cag gag gca tgg 787
Trp Ala Ala Met Gly Phe Ser Thr Glu Phe Trp Lys Gln Glu Ala Trp
215 220 225
cgt cgc ctg gga ctt gaa agt aag gag tca gtg ctc gcg gac ttc ctg 835
Arg Arg Leu Gly Leu Glu Ser Lys Glu Ser Val Leu Ala Asp Phe Leu
230 235 240 245
gat ccg ctg ttc atg tcc atg gat cct aat acc ttg ctc aac aac gct 883
Asp Pro Leu Phe Met Ser Met Asp Pro Asn Thr Leu Leu Asn Asn Ala
250 255 260
tgg aag tgg cag cat ggc gat gtc tct cgc cac acc ggc ggc gac ttg 931
Trp Lys Trp Gln His Gly Asp Val Ser Arg His Thr Gly Gly Asp Leu
265 270 275
gca gcg gct ctt ggc cga gtg aag gct aag acc ttc gtt atg ccc atc 979
Ala Ala Ala Leu Gly Arg Val Lys Ala Lys Thr Phe Val Met Pro Ile
280 285 290
agc gag gac atg ttc ttt cct gtt cgt gac tgt gcc gca gaa caa gca 1027
Ser Glu Asp Met Phe Phe Pro Val Arg Asp Cys Ala Ala Glu Gln Ala
295 300 305
ctc atc cca ggc agc gag ctt cga gtg atc gaa gac atc gcc ggt cac 1075
Leu Ile Pro Gly Ser Glu Leu Arg Val Ile Glu Asp Ile Ala Gly His
310 315 320 325
ctt ggg ctt ttt aac gtc tct gag aat tac atc cca cag atc gac aaa 1123
Leu Gly Leu Phe Asn Val Ser Glu Asn Tyr Ile Pro Gln Ile Asp Lys
330 335 340
aat ctg aaa gag ctg ttc gag agc taaacactga tgtcaaagag cct 1170
Asn Leu Lys Glu Leu Phe Glu Ser
345
<210>70
<211>349
<212>PRT
<213>谷氨酸棒杆菌
<400>70
Met Leu Asp Asn Ser Phe Tyr Thr Ala Glu Val Gln Gly Pro Tyr Glu
1 5 10 15
Thr Ala Ser Ile Gly Arg Leu Glu Leu Glu Glu Gly Gly Val Ile Glu
20 25 30
Asp Cys Trp Leu Ala Tyr Ala Thr Ala Gly Thr Leu Asn Glu Asp Lys
35 40 45
Ser Asn Ala Ile Leu Ile Pro Thr Trp Tyr Ser Gly Thr His Gln Thr
50 55 60
Trp Phe Gln Gln Tyr Ile Gly Thr Asp His Ala Leu Asp Pro Ser Lys
65 70 75 80
Tyr Phe Ile Ile Ser Ile Asn Gln Ile Gly Asn Gly Leu Ser Val Ser
85 90 95
Pro ALa Asn Thr Ala Asp Asp Ser Ile Ser Met Ser Lys Phe Pro Asn
100 105 110
Val Arg Ile Gly Asp Asp Val Val Ala Gln Asp Arg Leu Leu Arg Gln
115 120 125
Glu Phe Gly Ile Thr Glu Leu Phe Ala Val Val Gly Gly Ser Met Gly
130 135 140
Ala Gln Gln Thr Tyr Glu Trp Ile Val Arg Phe Pro Asp Gln Val His
145 150 155 160
Arg Ala Ala Pro Ile Ala Gly Thr Ala Lys Asn Thr Pro His Asp Phe
165 170 175
Ile Phe Thr Gln Thr Leu Asn Glu Thr Val Glu Ala Asp Pro Gly Phe
180 185 190
Asn Gly Gly Glu Tyr Ser Ser His Glu Glu Val Ala Asp Gly Leu Arg
195 200 205
Arg Gln Ser His Leu Trp Ala Ala Met Gly Phe Ser Thr Glu Phe Trp
210 215 220
Lys Gln Glu Ala Trp Arg Arg Leu Gly Leu Glu Ser Lys Glu Ser Val
225 230 235 240
Leu Ala Asp Phe Leu Asp Pro Leu Phe Met Ser Met Asp Pro Asn Thr
245 250 255
Leu Leu Asn Asn Ala Trp Lys Trp Gln His Gly Asp Val Ser Arg His
260 265 270
Thr Gly Gly Asp Leu Ala Ala Ala Leu Gly Arg Val Lys Ala Lys Thr
275 280 285
Phe Val Met Pro Ile Ser Glu Asp Met Phe Phe Pro Val Arg Asp Cys
290 295 300
Ala Ala Glu Gln Ala Leu Ile Pro Gly Ser Glu Leu Arg Val Ile Glu
305 310 315 320
Asp Ile Ala Gly His Leu Gly Leu Phe Asn Val Ser Glu Asn Tyr Ile
325 330 335
Pro GlnIle Asp Lys Asn Leu Lys Glu Leu Phe Glu Ser
340 345
<210>71
<211>1254
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1231)
<223>RXN00403
<400>71
tttttcagac tcgtgagaat gcaaactaga ctagacagag ctgtccatat acactggacg 60
aagttttagt cttgtccacc cagaacaggc ggttattttc atg ccc acc ctc gcg 115
Met Pro Thr Leu Ala
1 5
cct tca ggt caa ctt gaa atc caa gcg atc ggt gat gtc tcc acc gaa 163
Pro Ser Gly Gln Leu Glu Ile Gln Ala Ile Gly Asp Val Ser Thr Glu
10 15 20
gcc gga gca atc att aca aac gct gaa atc gcc tat cac cgc tgg ggt 211
Ala Gly Ala Ile Ile Thr Asn Ala Glu Ile Ala Tyr His Arg Trp Gly
25 30 35
gaa tac cgc gta gat aaa gaa gga cgc agc aat gtc gtt ctc atc gaa 259
Glu Tyr Arg Val Asp Lys Glu Gly Arg Ser Asn Val Val Leu Ile Glu
40 45 50
cac gcc ctc act gga gat tcc aac gca gcc gat tgg tgg gct gac ttg 307
His Ala Leu Thr Gly Asp Ser Asn Ala Ala Asp Trp Trp Ala Asp Leu
55 60 65
ctc ggt ccc ggc aaa gcc atc aac act gat att tac tgc gtg atc tgt 355
Leu Gly Pro Gly Lys Ala Ile Asn Thr Asp Ile Tyr Cys Val Ile Cys
70 75 80 85
acc aac gtc atc ggt ggt tgc aac ggt tcc acc gga cct ggc tcc atg 403
Thr Asn Val Ile Gly Gly Cys Asn Gly Ser Thr Gly Pro Gly Ser Met
90 95 100
cat cca gat gga aat ttc tgg ggt aat cgc ttc ccc gcc acg tcc att 451
His Pro Asp Gly Asn Phe Trp Gly Asn Arg Phe Pro Ala Thr Ser Ile
105 110 115
cgt gat cag gta aac gcc gaa aaa caa ttc ctc gac gca ctc ggc atc 499
Arg Asp Gln Val Asn Ala Glu Lys Gln Phe Leu Asp Ala Leu Gly Ile
120 125 130
acc acg gtc gcc gca gta ctt ggt ggt tcc atg ggt ggt gcc cgc acc 547
Thr Thr Val Ala Ala Val Leu Gly Gly Ser Met Gly Gly Ala Arg Thr
135 140 145
cta gag tgg gcc gca atg tac cca gaa act gtt ggc gca gct gct gtt 595
Leu Glu Trp Ala Ala Met Tyr Pro Glu Thr Val Gly Ala Ala Ala Val
150 155 160 165
ctt gca gtt tct gca cgc gcc agc gcc tgg caa atc ggc att caa tcc 643
Leu Ala Val Ser Ala Arg Ala Ser Ala Trp Gln Ile Gly Ile Gln Ser
170 175 180
gcc caa att aag gcg att gaa aac gac cac cac tgg cac gaa ggc aac 691
Ala Gln Ile Lys Ala Ile Glu Asn Asp His His Trp His Glu Gly Asn
185 190 195
tac tac gaa tcc ggc tgc aac cca gcc acc gga ctc ggc gcc gcc cga 739
Tyr Tyr Glu Ser Gly Cys Asn Pro Ala Thr Gly Leu Gly Ala Ala Arg
200 205 210
cgc atc gcc cac ctc acc tac cgt ggc gaa cta gaa atc gac gaa cgc 787
Arg Ile Ala His Leu Thr Tyr Arg Gly Glu Leu Glu Ile Asp Glu Arg
215 220 225
ttc ggc acc aaa gcc caa aag aac gaa aac cca ctc ggt ccc tac cgc 835
Phe Gly Thr Lys Ala Gln Lys Asn Glu Asn Pro Leu Gly Pro Tyr Arg
230 235 240 245
aag ccc gac cag cgc ttc gcc gtg gaa tcc tac ttg gac tac caa gca 883
Lys Pro Asp Gln Arg Phe Ala Val Glu Ser Tyr Leu Asp Tyr Gln Ala
250 255 260
gac aag cta gta cag cgt ttc gac gcc ggc tcc tac gtc ttg ctc acc 931
Asp Lys Leu Val Gln Arg Phe Asp Ala Gly Ser Tyr Val Leu Leu Thr
265 270 275
gac gcc ctc aac cgc cac gac att ggt cgc gac cgc gga ggc ctc aac 979
Asp Ala Leu Asn Arg His Asp Ile Gly Arg Asp Arg Gly Gly Leu Asn
280 285 290
aag gca ctc gaa tcc atc aaa gtt cca gtc ctt gtc gca ggc gta gat 1027
Lys Ala Leu Glu Ser Ile Lys Val Pro Val Leu Val Ala Gly Val Asp
295 300 305
acc gat att ttg tac ccc tac cac cag caa gaa cac ctc tcc aga aac 1075
Thr Asp Ile Leu Tyr Pro Tyr His Gln Gln Glu His Leu Ser Arg Asn
310 315 320 325
ctg gga aat cta ctg gca atg gca aaa atc gta tcc cct gtc ggc cac 1123
Leu Gly Asn Leu Leu Ala Met Ala Lys Ile Val Ser Pro Val Gly His
330 335 340
gat gct ttc ctc acc gaa agc cgc caa atg gat cgc atc gtg agg aac 1171
Asp Ala Phe Leu Thr Glu Ser Arg Gln Met Asp Arg Ile Val Arg Asn
345 350 355
ttc ttc agc ctc atc tcc cca gac gaa gac aac cct tcg acc tac atc 1219
Phe Phe Ser Leu Ile Ser Pro Asp Glu Asp Asn Pro Ser Thr Tyr Ile
360 365 370
gag ttc tac atc taataggtat ttacgacaaa tag 1254
Glu Phe Tyr Ile
375
<210>72
<211>377
<212>PRT
<213>谷氨酸棒杆菌
<400>72
Met Pro Thr Leu Ala Pro Ser Gly Gln Leu Glu Ile Gln Ala Ile Gly
1 5 10 15
Asp Val Ser Thr Glu Ala Gly Ala Ile Ile Thr Asn Ala Glu Ile Ala
20 25 30
Tyr His Arg Trp Gly Glu Tyr Arg Val Asp Lys Glu Gly Arg Ser Asn
35 40 45
Val Val Leu Ile Glu His Ala Leu Thr Gly Asp Ser Asn Ala Ala Asp
50 55 60
Trp Trp Ala Asp Leu Leu Gly Pro Gly Lys Ala Ile Asn Thr Asp Ile
65 70 75 80
Tyr Cys Val Ile Cys Thr Asn Val Ile Gly Gly Cys Asn Gly Ser Thr
85 90 95
Gly Pro Gly Ser Met His Pro Asp Gly Asn Phe Trp Gly Asn Arg Phe
100 105 110
Pro Ala Thr Ser Ile Arg Asp Gln Val Asn Ala Glu Lys Gln Phe Leu
115 120 125
Asp Ala Leu Gly Ile Thr Thr Val Ala Ala Val Leu Gly Gly Ser Met
130 135 140
Gly Gly Ala Arg Thr Leu Glu Trp Ala Ala Met Tyr Pro Glu Thr Val
145 150 155 160
Gly Ala Ala Ala Val Leu Ala Val Ser Ala Arg Ala Ser Ala Trp Gln
165 170 175
Ile Gly Ile Gln Ser Ala Gln Ile Lys Ala Ile Glu Asn Asp His His
180 185 190
Trp His Glu Gly Asn Tyr Tyr Glu Ser Gly Cys Asn Pro Ala Thr Gly
195 200 205
Leu Gly Ala Ala Arg Arg Ile Ala His Leu Thr Tyr Arg Gly Glu Leu
210 215 220
Glu Ile Asp Glu Arg Phe Gly Thr Lys Ala Gln Lys Asn Glu Asn Pro
225 230 235 240
Leu Gly Pro Tyr Arg Lys Pro Asp Gln Arg Phe Ala Val Glu Ser Tyr
245 250 255
Leu Asp Tyr Gln Ala Asp Lys Leu Val Gln Arg Phe Asp Ala Gly Ser
260 265 270
Tyr Val Leu Leu Thr Asp Ala Leu Asn Arg His Asp Ile Gly Arg Asp
275 280 285
Arg Gly Gly Leu Asn Lys Ala Leu Glu Ser Ile Lys Val Pro Val Leu
290 295 300
Val Ala Gly Val Asp Thr Asp Ile Leu Tyr Pro Tyr His Gln Gln Glu
305 310 315 320
His Leu Ser Arg Asn Leu Gly Asn Leu Leu Ala Met Ala Lys Ile Val
325 330 335
Ser Pro Val Gly His Asp Ala Phe Leu Thr Glu Ser Arg Gln Met Asp
340 345 350
Arg Ile Val Arg Asn Phe Phe Ser Leu Ile Ser Pro Asp Glu Asp Asn
355 360 365
Pro Ser Thr Tyr Ile Glu Phe Tyr Ile
370 375
<210>73
<211>1210
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1210)
<223>FRXA00403
<400>73
tttttcagac tcgtgagaat gcaaactaga ctagacagag ctgtccatat acactggacg 60
aagttttagt cttgtccacc cagaacaggc ggttattttc atg ccc acc ctc gcg 115
Met Pro Thr Leu Ala
1 5
cct tca ggt caa ctt gaa atc caa gcg atc ggt gat gtc tcc acc gaa 163
Pro Ser Gly Gln Leu Glu Ile Gln Ala Ile Gly Asp Val Ser Thr Glu
10 15 20
gcc gga gca atc att aca aac gct gaa atc gcc tat cac cgc tgg ggt 211
Ala Gly Ala Ile Ile Thr Asn Ala Glu Ile Ala Tyr His Arg Trp Gly
25 30 35
gaa tac cgc gta gat aaa gaa gga cgc agc aat gtc gtt ctc atc gaa 259
Glu Tyr Arg Val Asp Lys Glu Gly Arg Ser Asn Val Val Leu Ile Glu
40 45 50
cac gcc ctc act gga gat tcc aac gca gcc gat tgg tgg gct gac ttg 307
His Ala Leu Thr Gly Asp Ser Asn Ala Ala Asp Trp Trp Ala Asp Leu
55 60 65
ctc ggt ccc ggc aaa gcc atc aac act gat att tac tgc gtg atc tgt 355
Leu Gly Pro Gly Lys Ala Ile Asn Thr Asp Ile Tyr Cys Val Ile Cys
70 75 80 85
acc aac gtc atc ggt ggt tgc aac ggt tcc acc gga cct ggc tcc atg 403
Thr Asn Val Ile Gly Gly Cys Asn Gly Ser Thr Gly Pro Gly Ser Met
90 95 100
cat cca gat gga aat ttc tgg ggt aat cgc ttc ccc gcc acg tcc att 451
His Pro Asp Gly Asn Phe Trp Gly Asn Arg Phe Pro Ala Thr Ser Ile
105 110 115
cgt gat cag gta aac gcc gaa aaa caa ttc ctc gac gca ctc ggc atc 499
Arg Asp Gln Val Asn Ala Glu Lys Gln Phe Leu Asp Ala Leu Gly Ile
120 125 130
acc acg gtc gcc gca gta ctt ggt ggt tcc atg ggt ggt gcc cgc acc 547
Thr Thr Val Ala Ala Val Leu Gly Gly Ser Met Gly Gly Ala Arg Thr
135 140 145
cta gag tgg gcc gca atg tac cca gaa act gtt ggc gca gct gct gtt 595
Leu Glu Trp Ala Ala Met Tyr Pro Glu Thr Val Gly Ala Ala Ala Val
150 155 160 165
ctt gca gtt tct gca cgc gcc agc gcc tgg caa atc ggc att caa tcc 643
Leu Ala Val Ser Ala Arg Ala Ser Ala Trp Gln Ile Gly Ile Gln Ser
170 175 180
gcc caa att aag gcg att gaa aac gac cac cac tgg cac gaa ggc aac 691
Ala Gln Ile Lys Ala Ile Glu Asn Asp His His Trp His Glu Gly Asn
185 190 195
tac tac gaa tcc ggc tgc aac cca gcc acc gga ctc ggc gcc gcc cga 739
Tyr Tyr Glu Ser Gly Cys Asn Pro Ala Thr Gly Leu Gly Ala Ala Arg
200 205 210
cgc atc gcc cac ctc acc tac cgt ggc gaa cta gaa atc gac gaa cgc 787
Arg Ile Ala His Leu Thr Tyr Arg Gly Glu Leu Glu Ile Asp Glu Arg
215 220 225
ttc ggc acc aaa gcc caa aag aac gaa aac cca ctc ggt ccc tac cgc 835
Phe Gly Thr Lys Ala Gln Lys Asn Glu Asn Pro Leu Gly Pro Tyr Arg
230 235 240 245
aag ccc gac cag cgc ttc gcc gtg gaa tcc tac ttg gac tac caa gca 883
Lys Pro Asp Gln Arg Phe Ala Val Glu Ser Tyr Leu Asp Tyr Gln Ala
250 255 260
gac aag cta gta cag cgt ttc gac gcc ggc tcc tac gtc ttg ctc acc 931
Asp Lys Leu Val Gln Arg Phe Asp Ala Gly Ser Tyr Val Leu Leu Thr
265 270 275
gac gcc ctc aac cgc cac gac att ggt cgc gac cgc gga ggc ctc aac 979
Asp Ala Leu Asn Arg His Asp Ile Gly Arg Asp Arg Gly Gly Leu Asn
280 285 290
aag gca ctc gaa tcc atc aaa gtt cca gtc ctt gtc gca ggc gta gat 1027
Lys Ala Leu Glu Ser Ile Lys Val Pro Val Leu Val Ala Gly Val Asp
295 300 305
acc gat att ttg tac ccc tac cac cag caa gaa cac ctc tcc aga aac 1075
Thr Asp Ile Leu Tyr Pro Tyr His Gln Gln Glu His Leu Ser Arg Asn
310 315 320 325
ctg gga aat cta ctg gca atg gca aaa atc gta tcc cct gtc ggc cac 1123
Leu Gly Asn Leu Leu Ala Met Ala Lys Ile Val Ser Pro Val Gly His
330 335 340
gat gct ttc ctc acc gaa agc cgc caa atg gat cgc atc gtg agg aac 1171
Asp Ala Phe Leu Thr Glu Ser Arg Gln Met Asp Arg Ile Val Arg Asn
345 350 355
ttc ttc agc ctc atc tcc cca gac gaa gac aac cct tcg 1210
Phe Phe Ser Leu Ile Ser Pro Asp Glu Asp Asn Pro Ser
360 365 370
<210>74
<211>370
<212>PRT
<213>谷氨酸棒杆菌
<400>74
Met Pro Thr Leu Ala Pro Ser Gly Gln Leu Glu Ile Gln Ala Ile Gly
1 5 10 15
Asp Val Ser Thr Glu Ala Gly Ala Ile Ile Thr Asn Ala Glu Ile Ala
20 25 30
Tyr His Arg Trp Gly Glu Tyr Arg Val Asp Lys Glu Gly Arg Ser Asn
35 40 45
Val Val Leu Ile Glu His Ala Leu Thr Gly Asp Ser Asn Ala Ala Asp
50 55 60
Trp Trp Ala Asp Leu Leu Gly Pro Gly Lys Ala Ile Asn Thr Asp Ile
65 70 75 80
Tyr Cys Val Ile Cys Thr Asn Val Ile Gly Gly Cys Asn Gly Ser Thr
85 90 95
Gly Pro Gly Ser Met His Pro Asp Gly Asn Phe Trp Gly Asn Arg Phe
100 105 110
Pro Ala Thr Ser Ile Arg Asp Gln Val Asn Ala Glu Lys Gln Phe Leu
115 120 125
Asp Ala Leu Gly Ile Thr Thr Val Ala Ala Val Leu Gly Gly Ser Met
130 135 140
Gly Gly Ala Arg Thr Leu Glu Trp Ala Ala Met Tyr Pro Glu Thr Val
145 150 155 160
Gly Ala Ala Ala Val Leu Ala Val Ser Ala Arg Ala Ser Ala Trp Gln
165 170 175
Ile Gly Ile Gln Ser Ala Gln Ile Lys Ala Ile Glu Asn Asp His His
180 185 190
Trp His Glu Gly Asn Tyr Tyr Glu Ser Gly Cys Asn Pro Ala Thr Gly
195 200 205
Leu Gly Ala Ala Arg Arg Ile Ala His Leu Thr Tyr Arg Gly Glu Leu
210 215 220
Glu Ile Asp Glu Arg Phe Gly Thr Lys Ala Gln Lys Asn Glu Asn Pro
225 230 235 240
Leu Gly Pro Tyr Arg Lys Pro Asp Gln Arg Phe Ala Val Glu Ser Tyr
245 250 255
Leu Asp Tyr Gln Ala Asp Lys Leu Val Gln Arg Phe Asp Ala Gly Ser
260 265 270
Tyr Val Leu Leu Thr Asp Ala Leu Asn Arg His Asp Ile Gly Arg Asp
275 280 285
Arg Gly Gly Leu Asn Lys Ala Leu Glu Ser Ile Lys Val Pro Val Leu
290 295 300
Val Ala Gly Val Asp Thr Asp Ile Leu Tyr Pro Tyr His Gln Gln Glu
305 310 315 320
His Leu Ser Arg Asn Leu Gly Asn Leu Leu Ala Met Ala Lys Ile Val
325 330 335
Ser Pro Val Gly His Asp Ala Phe Leu Thr Glu Ser Arg Gln Met Asp
340 345 350
Arg Ile Val Arg Asn Phe Phe Ser Leu Ile Ser Pro Asp Glu Asp Asn
355 360 365
Pro Ser
370
<210>75
<211>687
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(664)
<223>RXS03158
<400>75
caaagctcac cgaaggcacc aacgccaagt tggttgttga caacaccttg gcatccccat 60
acctgcagca gccactaaaa ctcggcgcac acgcaagtcc ttg cac tcc acc acc 115
Leu His Ser Thr Thr
1 5
aag tac atc gaa gga cac tcc gac gtt gtt ggc ggc ctt gtg ggt acc 163
Lys Tyr Ile Glu Gly His Ser Asp Val Val Gly Gly Leu Val Gly Thr
10 15 20
aac gac cag gaa atg gac gaa gaa ctg ctg ttc atg cag ggc ggc atc 211
Asn Asp Gln Glu Met Asp Glu Glu Leu Leu Phe Met Gln Gly Gly Ile
25 30 35
gga ccg atc cca tca gtt ttc gat gca tac ctg acc gcc cgt ggc ctc 259
Gly Pro Ile Pro Ser Val Phe Asp Ala Tyr Leu Thr Ala Arg Gly Leu
40 45 50
aag acc ctt gca gtg cgc atg gat cgc cac tgc gac aac gca gaa aag 307
Lys Thr Leu Ala Val Arg Met Asp Arg His Cys Asp Asn Ala Glu Lys
55 60 65
atc gcg gaa ttc ctg gac tcc cgc cca gag gtc tcc acc gtg ctc tac 355
Ile Ala Glu Phe Leu Asp Ser Arg Pro Glu Val Ser Thr Val Leu Tyr
70 75 80 85
cca ggt ctg aag aac cac cca ggc cac gaa gtc gca gcg aag cag atg 403
Pro Gly Leu Lys Asn His Pro Gly His Glu Val Ala Ala Lys Gln Met
90 95 100
aag cgc ttc ggc ggc atg atc tcc gtc cgt ttc gca ggc ggc gaa gaa 451
Lys Arg Phe Gly Gly Met Ile Ser Val Arg Phe Ala Gly Gly Glu Glu
105 110 115
gca gct aag aag ttc tgt acc tcc acc aaa ctg atc tgt ctg gcc gag 499
Ala Ala Lys Lys Phe Cys Thr Ser Thr Lys Leu Ile Cys Leu Ala Glu
120 125 130
tcc ctc ggt ggc gtg gaa tcc ctc ctg gag cac cca gca acc atg acc 547
Ser Leu Gly Gly Val Glu Ser Leu Leu Glu His Pro Ala Thr Met Thr
135 140 145
cac cag tca gct gcc ggc tct cag ctc gag gtt ccc cgc gac ctc gtg 595
His Gln Ser Ala Ala Gly Ser Gln Leu Glu Val Pro Arg Asp Leu Val
150 155 160 165
cgc atc tcc att ggt att gaa gac att gaa gac ctg ctc gca gat gtc 643
Arg Ile Ser Ile Gly Ile Glu Asp Ile Glu Asp Leu Leu Ala Asp Val
170 175 180
gag cag gcc ctc aat aac ctt tagaaactat ttggcggcaa gca 687
Glu Gln Ala Leu Asn Asn Leu
185
<210>76
<211>188
<212>PRT
<213>谷氨酸棒杆菌
<400>76
Leu His Ser Thr Thr Lys Tyr Ile Glu Gly His Ser Asp Val Val Gly
1 5 10 15
Gly Leu Val Gly Thr Asn Asp Gln Glu Met Asp Glu Glu Leu Leu Phe
20 25 30
Met Gln Gly Gly Ile Gly Pro Ile Pro Ser Val Phe Asp Ala Tyr Leu
35 40 45
Thr Ala Arg Gly Leu Lys Thr Leu Ala Val Arg Met Asp Arg His Cys
50 55 60
Asp Asn Ala Glu Lys Ile Ala Glu Phe Leu Asp Ser Arg Pro Glu Val
65 70 75 80
Ser Thr Val Leu Tyr Pro Gly Leu Lys Asn His Pro Gly His Glu Val
85 90 95
Ala Ala Lys Gln Met Lys Arg Phe Gly Gly Met Ile Ser Val Arg Phe
100 105 110
Ala Gly Gly Glu Glu Ala Ala Lys Lys Phe Cys Thr Ser Thr Lys Leu
115 120 125
Ile Cys Leu Ala Glu Ser Leu Gly Gly Val Glu Ser Leu Leu Glu His
130 135 140
Pro Ala Thr Met Thr His Gln Ser Ala Ala Gly Ser Gln Leu Glu Val
145 150 155 160
Pro Arg Asp Leu Val Arg Ile Ser Ile Gly Ile Glu Asp Ile Glu Asp
165 170 175
Leu Leu Ala Asp Val Glu Gln Ala Leu Asn Asn Leu
180 185
<210>77
<211>617
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(594)
<223>FRXA00254
<400>77
cag cca cta aaa ctc ggc gca cac gca gtc ttg cac tcc acc acc aag 48
Gln Pro Leu Lys Leu Gly Ala His Ala Val Leu His Ser Thr Thr Lys
1 5 10 15
tac atc gga gga cac tcc gac gtt gtt ggc ggc ctt gtg gtt acc aac 96
Tyr Ile Gly Gly His Ser Asp Val Val Gly Gly Leu Val Val Thr Asn
20 25 30
gac cag gaa atg gac gaa gaa ctg ctg ttc atg cag ggc ggc atc gga 144
Asp Gln Glu Met Asp Glu Glu Leu Leu Phe Met Gln Gly Gly Ile Gly
35 40 45
ccg atc cca tca gtt ttc gat gca tac ctg acc gcc cgt ggc ctc aag 192
Pro Ile Pro Ser Val Phe Asp Ala Tyr Leu Thr Ala Arg Gly Leu Lys
50 55 60
acc ctt gca gtg cgc atg gat cgc cac tgc gac aac gca gaa aag atc 240
Thr Leu Ala Val Arg Met Asp Arg His Cys Asp Asn Ala Glu Lys Ile
65 70 75 80
gcg gaa ttc ctg gac tcc cgc cca gag gtc tcc acc gtg ctc tac cca 288
Ala Glu Phe Leu Asp Ser Arg Pro Glu Val Ser Thr Val Leu Tyr Pro
85 90 95
ggt ctg aag aac cac cca ggc cac gaa gtc gca gcg aag cag atg aag 336
Gly Leu Lys Asn His Pro Gly His Glu Val Ala Ala Lys Gln Met Lys
100 105 110
cgc ttc ggc ggc atg atc tcc gtc cgt ttc gca ggc ggc gaa gaa gca 384
Arg Phe Gly Gly Met Ile Ser Val Arg Phe Ala Gly Gly Glu Glu Ala
115 120 125
gct aag aag ttc tgt acc tcc acc aaa ctg atc tgt ctg gcc gag tcc 432
Ala Lys Lys Phe Cys Thr Ser Thr Lys Leu Ile Cys Leu Ala Glu Ser
130 135 140
ctc ggt ggc gtg gaa tcc ctc ctg gag cac cca gca acc atg acc cac 480
Leu Gly Gly Val Glu Ser Leu Leu Glu His Pro Ala Thr Met Thr His
145 150 155 160
cag tca gct gcc ggc tct cag ctc gag gtt ccc cgc gac ctc gtg cgc 528
Gln Ser Ala Ala Gly Ser Gln Leu Glu Val Pro Arg Asp Leu Val Arg
165 170 175
atc tcc att ggt att gaa gac att gaa gac ctg ctc gca gat gtc gag 576
Ile Ser Ile Gly Ile Glu Asp Ile Glu Asp Leu Leu Ala Asp Val Glu
180 185 190
cag gcc ctc aat aac ctt tagaaactat ttggcggcaa gca 617
Gln Ala Leu Asn Asn Leu
195
<210>78
<211>198
<212>PRT
<213>谷氨酸棒杆菌
<400>78
Gln Pro Leu Lys Leu Gly Ala His Ala Val Leu His Ser Thr Thr Lys
1 5 10 15
Tyr Ile Gly Gly His Ser Asp Val Val Gly Gly Leu Val Val Thr Asn
20 25 30
Asp Gln Glu Met Asp Glu Glu Leu Leu Phe Met Gln Gly Gly Ile Gly
35 40 45
Pro Ile Pro Ser Val Phe Asp Ala Tyr Leu Thr Ala Arg Gly Leu Lys
50 55 60
Thr Leu Ala Val Arg Met Asp Arg His Cys Asp Asn Ala Glu Lys Ile
65 70 75 80
Ala Glu Phe Leu Asp Ser Arg Pro Glu Val Ser Thr Val Leu Tyr Pro
85 90 95
Gly Leu Lys Asn His Pro Gly His Glu Val Ala Ala Lys Gln Met Lys
100 105 110
Arg Phe Gly Gly Met Ile Ser Val Arg Phe Ala Gly Gly Glu Glu Ala
115 120 125
Ala Lys Lys Phe Cys Thr Ser Thr Lys Leu Ile Cys Leu Ala Glu Ser
130 135 140
Leu Gly Gly Val Glu Ser Leu Leu Glu His Pro Ala Thr Met Thr His
145 150 155 160
Gln Ser Ala Ala Gly Ser Gln Leu Glu Val Pro Arg Asp Leu Val Arg
165 170 175
Ile Ser Ile Gly Ile Glu Asp Ile Glu Asp Leu Leu Ala Asp Val Glu
180 185 190
Gln Ala Leu Asn Asn Leu
195
<210>79
<211>1170
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1147)
<223>RXA02532
<400>79
gatgaatttt tacccaccat ctgtacctat taaccctgcg tggcgtccac ccacagtaac 60
tgtgcaagcg ggacggccag ccagaactcc tggtgcgccg atg aac cca cct atc 115
Met Asn Pro Pro Ile
1 5
acg ttg tcc agc act tat gtt cat gat tca gaa aaa gct tat ggg cgc 163
Thr Leu Ser Ser Thr Tyr Val His Asp Ser Glu Lys Ala Tyr Gly Arg
10 15 20
gat ggc aat gat gga tgg ggt gca ttt gag gct gcc atg gga act cta 211
Asp Gly Asn Asp Gly Trp Gly Ala Phe Glu Ala Ala Met Gly Thr Leu
25 30 35
gat ggt ggg ttc gcg gta tct tat tct tca ggt ttg gca gcg gca acg 259
Asp Gly Gly Phe Ala Val Ser Tyr Ser Ser Gly Leu Ala Ala Ala Thr
40 45 50
tcg att gct gat ttg gtt cct act ggt ggc aca gtt gtt tta cct aaa 307
Ser Ile Ala Asp Leu Val Pro Thr Gly Gly Thr Val Val Leu Pro Lys
55 60 65
gct gcc tat tat ggc gtg acc aat att ttc gcc agg atg gaa gcc cgc 355
Ala Ala Tyr Tyr Gly Val Thr Asn Ile Phe Ala Arg Met Glu Ala Arg
70 75 80 85
gga agg ctg aag gtt cga act gtt gat gca gac aat acc gaa gaa gtg 403
Gly Arg Leu Lys Val Arg Thr Val Asp Ala Asp Asn Thr Glu Glu Val
90 95 100
att gct gct gct caa ggt gca gat gtg gtg tgg gtg gaa tcg atc gct 451
Ile Ala Ala Ala Gln Gly Ala Asp Val Val Trp Val Glu Ser Ile Ala
105 110 115
aat ccg acg atg gtg gta gct gat atc cct gca ata gtc gac ggt gtg 499
Asn Pro Thr Met Val Val Ala Asp Ile Pro Ala Ile Val Asp Gly Val
120 125 130
cgt ggg ctt gga gtt ttg act gtc gtt gac gcg act ttc gca acg cca 547
Arg Gly Leu Gly Val Leu Thr Val Val Asp Ala Thr Phe Ala Thr Pro
135 140 145
ctt cgt caa cgt cca ttg gaa ctt ggt gct gat att gtg ctt tac tcg 595
Leu Arg Gln Arg Pro Leu Glu Leu Gly Ala Asp Ile Val Leu Tyr Ser
150 155 160 165
gca acc aaa ctt atc ggt gga cac tct gat ctt ctt ctt gga gtc gca 643
Ala Thr Lys Leu Ile Gly Gly His Ser Asp Leu Leu Leu Gly Val Ala
170 175 180
gtg tgc aag tct gag cac cat gcg cag ttt ctt gcc act cac cgt cat 691
Val Cys Lys Ser Glu His His Ala Gln Phe Leu Ala Thr His Arg His
185 190 195
gat cat ggt tca gtg ccg gga ggt ctt gaa gcg ttt ctt gct ctc cgt 739
Asp His Gly Ser Val Pro Gly Gly Leu Glu Ala Phe Leu Ala Leu Arg
200 205 210
gga ttg tat tcc ttg gcg gtg cgt ctt gat cga gca gaa tcc aac gca 787
Gly Leu Tyr Ser Leu Ala Val Arg Leu Asp Arg Ala Glu Ser Asn Ala
215 220 225
gca gaa ctt tcg cgg cga ctt aac gcg cat cct tcg gtt acc cgc gtc 835
Ala Glu Leu Ser Arg Arg Leu Asn Ala His Pro Ser Val Thr Arg Val
230 235 240 245
aat tat cca gga ctt cct gat gat ccc caa cat gaa aaa gcc gtg cga 883
Asn Tyr Pro Gly Leu Pro Asp Asp Pro Gln His Glu Lys Ala Val Arg
250 255 260
gtc cta ccc tct gga tgt gga aac atg ttg tca ttt gag ctt gat gca 931
Val Leu Pro Ser Gly Cys Gly Asn Met Leu Ser Phe Glu Leu Asp Ala
265 270 275
aca cct gaa cga act gat gag att ctc gaa agc ctg tca ctt tta acc 979
Thr Pro Glu Arg Thr Asp Glu Ile Leu Glu Ser Leu Ser Leu Leu Thr
280 285 290
cac gcg acc agt tgg gga ggt gtg gaa aca gcc att gaa cgt cgc acc 1027
His Ala Thr Ser Trp Gly Gly Val Glu Thr Ala Ile Glu Arg Arg Thr
295 300 305
agg cgg gat gct gaa gtg gtg gca gaa gta ccg atg act ctt tgc cgc 1075
Arg Arg Asp Ala Glu Val Val Ala Glu Val Pro Met Thr Leu Cys Arg
310 315 320 325
gtt tcc gta gga att gaa gac gtt gaa gat cta tgg gaa gac ctc aac 1123
Val Ser Val Gly Ile Glu Asp Val Glu Asp Leu Trp Glu Asp Leu Asn
330 335 340
gcc tca atc gac aaa gtt ctg ggt tagaactcgt agccagtaac cag 1170
Ala Ser Ile Asp Lys Val Leu Gly
345
<210>80
<211>349
<212>PRT
<213>谷氨酸棒杆菌
<400>80
Met Asn Pro Pro Ile Thr Leu Ser Ser Thr Tyr Val His Asp Ser Glu
1 5 10 15
Lys Ala Tyr Gly Arg Asp Gly Asn Asp Gly Trp Gly Ala Phe Glu Ala
20 25 30
Ala Met Gly Thr Leu Asp Gly Gly Phe Ala Val Ser Tyr Ser Ser Gly
35 40 45
Leu Ala Ala Ala Thr Ser Ile Ala Asp Leu Val Pro Thr Gly Gly Thr
50 55 60
Val Val Leu Pro Lys Ala Ala Tyr Tyr Gly Val Thr Asn Ile Phe Ala
65 70 75 80
Arg Met Glu Ala Arg Gly Arg Leu Lys Val Arg Thr Val Asp Ala Asp
85 90 95
Asn Thr Glu Glu Val Ile Ala Ala Ala Gln Gly Ala Asp Val Val Trp
100 105 110
Val Glu Ser Ile Ala Asn Pro Thr Met Val Val Ala Asp Ile Pro Ala
115 120 125
Ile Val Asp Gly Val Arg Gly Leu Gly Val Leu Thr Val Val Asp Ala
130 135 140
Thr Phe Ala Thr Pro Leu Arg Gln Arg Pro Leu Glu Leu Gly Ala Asp
145 150 155 160
Ile Val Leu Tyr Ser Ala Thr Lys Leu Ile Gly Gly His Ser Asp Leu
165 170 175
Leu Leu Gly Val Ala Val Cys Lys Ser Glu His His Ala Gln Phe Leu
180 185 190
Ala Thr His Arg His Asp His Gly Ser Val Pro Gly Gly Leu Glu Ala
195 200 205
Phe Leu Ala Leu Arg Gly Leu Tyr Ser Leu Ala Val Arg Leu Asp Arg
210 215 220
Ala Glu Ser Asn Ala Ala Glu Leu Ser Arg Arg Leu Asn Ala His Pro
225 230 235 240
Ser Val Thr Arg Val Asn Tyr Pro Gly Leu Pro Asp Asp Pro Gln His
245 250 255
Glu Lys Ala Val Arg Val Leu Pro Ser Gly Cys Gly Asn Met Leu Ser
260 265 270
Phe Glu Leu Asp Ala Thr Pro Glu Arg Thr Asp Glu Ile Leu Glu Ser
275 280 285
Leu Ser Leu Leu Thr His Ala Thr Ser Trp Gly Gly Val Glu Thr Ala
290 295 300
Ile Glu Arg Arg Thr Arg Arg Asp Ala Glu Val Val Ala Glu Val Pro
305 310 315 320
Met Thr Leu Cys Arg Val Ser Val Gly Ile Glu Asp Val Glu Asp Leu
325 330 335
Trp Glu Asp Leu Asn Ala Ser Ile Asp Lys Val Leu Gly
340 345
<210>81
<211>861
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(838)
<223>RXS03159
<400>81
aggggctagt tttacacaaa agtggacagc ttggtctatc attgccagaa gaccggtcct 60
tttagggcca tagaattctg attacaggag ttgatctacc ttg tct ttt gac cca 115
Leu Ser Phe Asp Pro
1 5
aac acc cag ggt ttc tcc act gca tcg att cac gct ggg tat gag cca 163
Asn Thr Gln Gly Phe Ser Thr Ala Ser Ile His Ala Gly Tyr Glu Pro
10 15 20
gac gac tac tac ggt tcg att aac acc cca atc tat gcc tcc acc acc 211
Asp Asp Tyr Tyr Gly Ser Ile Asn Thr Pro Ile Tyr Ala Ser Thr Thr
25 30 35
ttc gcg cag aac gct cca aac gaa ctg cgc aaa ggc tac gag tac acc 259
Phe Ala Gln Asn Ala Pro Asn Glu Leu Arg Lys Gly Tyr Glu Tyr Thr
40 45 50
cgt gtg ggc aac ccc acc atc gtg gca tta gag cag acc gtc gca gca 307
Arg Val Gly Asn Pro Thr Ile Val Ala Leu Glu Gln Thr Val Ala Ala
55 60 65
ctc gaa ggc gca aag tat ggc cgc gca ttc tcc tcc ggc atg gct gca 355
Leu Glu Gly Ala Lys Tyr Gly Arg Ala Phe Ser Ser Gly Met Ala Ala
70 75 80 85
acc gac atc ctg ttc cgc atc atc ctc aag ccg ggc gat cac atc gtc 403
Thr Asp Ile Leu Phe Arg Ile Ile Leu Lys Pro Gly Asp His Ile Val
90 95 100
ctc ggc aac gat gct tac ggc gga acc tac cgc ctg atc gac acc gta 451
Leu Gly Asn Asp Ala Tyr Gly Gly Thr Tyr Arg Leu Ile Asp Thr Val
105 110 115
ttc acc gca tgg ggc gtc gaa tac acc gtt gtt gat acc tcc gtc gtg 499
Phe Thr Ala Trp Gly Val Glu Tyr Thr Val Val Asp Thr Ser Val Val
120 125 130
gaa gag gtc aag gca gcg atc aag gac aac acc aag ctg atc tgg gtg 547
Glu Glu Val Lys Ala Ala Ile Lys Asp Asn Thr Lys Leu Ile Trp Val
135 140 145
gaa acc cca acc aac cca gca ctt ggc atc acc gac atc gaa gca gta 595
Glu Thr Pro Thr Asn Pro Ala Leu Gly Ile Thr Asp Ile Glu Ala Val
150 155 160 165
gca aag ctc acc gaa ggc acc aac gcc aag ttg gtt gtt gac aac acc 643
Ala Lys Leu Thr Glu Gly Thr Asn Ala Lys Leu Val Val Asp Asn Thr
170 175 180
ttg gca tcc cca tac ctg cag cag cca cta aaa ctc ggc gca cac gca 691
Leu Ala Ser Pro Tyr Leu Gln Gln Pro Leu Lys Leu Gly Ala His Ala
185 190 195
agt cct tgc act cca cca cca agt aca tcg aag gac act ccg acg ttg 739
Ser Pro Cys Thr Pro Pro Pro Ser Thr Ser Lys Asp Thr Pro Thr Leu
200 205 210
ttg gcg gcc ttg tgg gta cca acg acc agg aaa tgg acg aag aac tgc 787
Leu Ala Ala Leu Trp Val Pro Thr Thr Arg Lys Trp Thr Lys Asn Cys
215 220 225
tgt tca tgc agg gcg gca tcg gac cga tcc cat cag ttt tcg atg cat 835
Cys Ser Cys Arg Ala Ala Ser Asp Arg Ser His Gln Phe Ser Met His
230 235 240 245
acc tgaccgcccg tggcctcaag acc 861
Thr
<210>82
<211>246
<212>PRT
<213>谷氨酸棒杆菌
<400>82
Leu Ser Phe Asp Pro Asn Thr Gln Gly Phe Ser Thr Ala Ser Ile His
1 5 10 15
Ala Gly Tyr Glu Pro Asp Asp Tyr Tyr Gly Ser Ile Asn Thr Pro Ile
20 25 30
Tyr Ala Ser Thr Thr Phe Ala Gln Asn Ala Pro Asn Glu Leu Arg Lys
35 40 45
Gly Tyr Glu Tyr Thr Arg Val Gly Asn Pro Thr Ile Val Ala Leu Glu
50 55 60
Gln Thr Val Ala Ala Leu Glu Gly Ala Lys Tyr Gly Arg Ala Phe Ser
65 70 75 80
Ser Gly Met Ala Ala Thr Asp Ile Leu Phe Arg Ile Ile Leu Lys Pro
85 90 95
Gly Asp His Ile Val Leu Gly Asn Asp Ala Tyr Gly Gly Thr Tyr Arg
100 105 110
Leu Ile Asp Thr Val Phe Thr Ala Trp Gly Val Glu Tyr Thr Val Val
115 120 125
Asp Thr Ser Val Val Glu Glu Val Lys Ala Ala Ile Lys Asp Asn Thr
130 135 140
Lys Leu Ile Trp Val Glu Thr Pro Thr Asn Pro Ala Leu Gl yIle Thr
145 150 155 160
Asp Ile Glu Ala Val Ala Lys Leu Thr Glu Gly Thr Asn Ala Lys Leu
165 170 175
Val Val Asp Asn Thr Leu Ala Ser Pro Tyr Leu Gln Gln Pro Leu Lys
180 185 190
Leu Gly Ala His Ala Ser Pro Cys Thr Pro Pro Pro Ser Thr Ser Lys
195 200 205
Asp Thr Pro Thr Leu Leu Ala Ala Leu Trp Val Pro Thr Thr Arg Lys
210 215 220
Trp Thr Lys Asn Cys Cys Ser Cys Arg Ala Ala Ser Asp Arg Ser His
225 230 235 240
Gln Phe Ser Met His Thr
245
<210>83
<211>703
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(703)
<223>FRXA02768
<220>
<223>所有的n=任意核苷酸
<220>
<223>所有的Xaa=任意氨基酸
<400>83
aggggctagt tttacacaaa agtggacagc ttggtctatc attgccagaa gaccggtcct 60
tttagggcca tagaattctg attacaggag ttgatctacc ttg tct ttt gac cca 115
Leu Ser Phe Asp Pro
1 5
aac acc cag ggt ttc tcc act gca tcg att cac gct ggg tat gag cca 163
Asn Thr Gln Gly Phe Ser Thr Ala Ser Ile His Ala Gly Tyr Glu Pro
10 15 20
gac gac tac tac ggt tcg att aac acc cca atc tat gcc tcc acc acc 211
Asp Asp Tyr Tyr Gly Ser Ile Asn Thr Pro Ile Tyr Ala Ser Thr Thr
25 30 35
ttc gcg cag aac gct cca aac gaa ctg cgc aaa ggc tac gag tac acc 259
Phe Ala Gln Asn Ala Pro Asn Glu Leu Arg Lys Gly Tyr Glu Tyr Thr
40 45 50
cgt gtg ggc aac ccc acc atc gtg gca tta gag cag acc gtc gca gca 307
Arg Val Gly Asn Pro Thr Ile Val Ala Leu Glu Gln Thr Val Ala Ala
55 60 65
ctc gaa ggc gca aag tat ggc cgc gca ttc tcc tcc ggc atg gct gca 355
Leu Glu Gly Ala Lys Tyr Gly Arg Ala Phe Ser Ser Gly Met Ala Ala
70 75 80 85
acc gac atc ctg ttc cgc atc atc ctc aag ccg ggc gat cac atc gtc 403
Thr Asp Ile Leu Phe Arg Ile Ile Leu Lys Pro Gly Asp His Ile Val
90 95 100
ctc ggc aac gat gct tac ggc gga acc tac cgc ctg atc gac acc gta 451
Leu Gly Asn Asp Ala Tyr Gly Gly Thr Tyr Arg Leu Ile Asp Thr Val
105 110 115
ttc acc gca tgg ggc gtc gaa tac acc gtt gtt gat acc tcc gtc gtg 499
Phe Thr Ala Trp Gly Val Glu Tyr Thr Val Val Asp Thr Ser Val Val
120 125 130
gaa gag gtc aag gca gcg atc aag gac aac acc aag gct gat ctt ggt 547
Glu Glu Val Lys Ala Ala Ile Lys Asp Asn Thr Lys Ala Asp Lau Gly
135 140 145
gga aac ccc aac caa ccc agc act ttg gca tta ccc gac atc gaa gca 595
Gly Asn Pro Asn Gln Pro Ser Thr Leu Ala Leu Pro Asp Ile Glu Ala
150 155 160 165
gtn tgc aaa act tca ccc gaa agg cac caa ccc caa gct tgt tgt ttg 643
Val Cys Lys Thr Ser Pro Glu Arg His Gln Pro Gln Ala Cys Cys Leu
170 175 180
aca aca cct tcg cat tcc cca tac ctg cag can cca ctt aaa ant tnn 691
Thr Thr Pro Ser His Ser Pro Tyr Leu Gln Xaa Pro Leu Lys Xaa Xaa
185 190 195
gng cac acg cag 703
Xaa His Thr Gln
200
<210>84
<211>201
<212>PRT
<213>谷氨酸棒杆菌
<220>
<223>All occurrences of Xaa=any amino acid
<400>84
Leu Ser Phe Asp Pro Asn Thr Gln Gly Phe Ser Thr Ala Ser Ile His
1 5 10 15
Ala Gly Tyr Glu Pro Asp Asp Tyr Tyr Gly Ser Ile Asn Thr Pro Ile
20 25 30
Tyr Ala Ser Thr Thr Phe Ala Gln Asn Ala Pro Asn Glu Leu Arg Lys
35 40 45
Gly Tyr Glu Tyr Thr Arg Val Gly Asn Pro Thr Ile Val Ala Lau Glu
50 55 60
Gln Thr Val Ala Ala Leu Glu Gly Ala Lys Tyr Gly Arg Ala Phe Ser
65 70 75 80
Ser Gly Met Ala Ala Thr Asp Ile Leu Phe Arg Ile Ile Lau Lys Pro
85 90 95
Gly Asp His Ile Val Leu Gly Asn Asp Ala Tyr Gly Gly Thr Tyr Arg
100 105 110
Leu Ile Asp Thr Val Phe Thr Ala Trp Gly Val Glu Tyr Thr Val Val
115 120 125
Asp Thr Ser Val Val Glu Glu Val Lys Ala Ala Ile Lys Asp Asn Thr
130 135 140
Lys Ala Asp Leu Gly Gly Asn Pro Asn Gln Pro Ser Thr Leu Ala Leu
145 150 155 160
Pro Asp Ile Glu Ala Val Cys Lys Thr Ser Pro Glu Arg His Gln Pro
165 170 175
Gln Ala Cys Cys Leu Thr Thr Pro Ser His Ser Pro Tyr Leu Gln Xaa
180 185 190
Pro Leu Lys Xaa Xaa Xaa His Thr Gln
195 200
<210>85
<211>1113
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1090)
<223>RXA00216
<400>85
gtgttgctcg cggccaggca gcagtgctgt acctgcctga cgcggatggt gacatcgttc 60
ttggatcagg caccatctgc cacacggagt cttaagaaaa ttg ggc gct tat ggt 115
Leu Gly Ala Tyr Gly
1 5
tta ggt gag ctt cct gga aaa tcc gcc gcg gaa gcc gcc gac att att 163
Leu Gly Glu Leu Pro Gly Lys Ser Ala Ala Glu Ala Ala Asp Ile Ile
10 15 20
cag ggt gaa acg ggc gat ctt ctc cat att cct cag ctt ccg gcg cga 211
Gln Gly Glu Thr Gly Asp Leu Leu His Ile Pro Gln Leu Pro Ala Arg
25 30 35
ggt ttg ggt gct gat ctg atc ggt cga acc gtc ggt ctg ctg gac atg 259
Gly Leu Gly Ala Asp Leu Ile Gly Arg Thr Val Gly Leu Leu Asp Met
40 45 50
atc aac gtt gat cgc ggg gcc cga tct tgg gtg atg agc aca cgc ccc 307
Ile Asn Val Asp Arg Gly Ala Arg Ser Trp Val Met Ser Thr Arg Pro
55 60 65
agc aga ttg acg cac ctg acc ggc gat ttc ctt gac atg gat ttg gat 355
Ser Arg Leu Thr His Leu Thr Gly Asp Phe Leu Asp Met Asp Leu Asp
70 75 80 85
gcg tgc gag gaa acc tgg gga acg ggc gtc gac aag cta aaa atc caa 403
Ala Cys Glu Glu Thr Trp Gly Thr Gly Val Asp Lys Leu Lys Ile Gln
90 95 100
gtt gct ggt ccc tgg act tta ggt gcg cgc att gag ttg gcc aat ggc 451
Val Ala Gly Pro Trp Thr Leu Gly Ala Arg Ile Glu Leu Ala Asn Gly
105 110 115
cat cgc gtt ttg tct gat cgc ggt gcg atg cgt gat ctc acg cag gcg 499
His Arg Val Leu Ser Asp Arg Gly Ala Met Arg Asp Leu Thr Gln Ala
120 125 130
ctg atc gcc ggc atc gat gcg cat gca cgc aag gtt gct ggg cga ttt 547
Leu Ile Ala Gly Ile Asp Ala His Ala Arg Lys Val Ala Gly Arg Phe
135 140 145
cgc gcc gaa gtg cag gtg caa att gat gag ccg gag ctg aaa tcg ctt 595
Arg Ala Glu Val Gln Val Gln Ile Asp Glu Pro Glu Leu Lys Ser Leu
150 155 160 165
atc gac ggc tcc ctc cct ggc act tcc acc ttt gac att att cct gcg 643
Ile Asp Gly Ser Leu Pro Gly Thr Ser Thr Phe Asp Ile Ile Pro Ala
170 175 180
gtg aat gtc gct gat gcc agt gaa cgt ttg cag cag gtc ttt agc tcg 691
Val Asn Val Ala Asp Ala Ser Glu Arg Leu Gln Gln Val Phe Ser Ser
185 190 195
att gag ggg ccg aca tat ctc aac ctc acc ggc cag att cct act tgg 739
Ile Glu Gly Pro Thr Tyr Leu Asn Leu Thr Gly Gln Ile Pro Thr Trp
200 205 210
gat gtg gct cgg ggt gcg ggc gcc gat act gtg cag att tcc atg gat 787
Asp Val Ala Arg Gly Ala Gly Ala Asp Thr Val Gln Ile Ser Met Asp
215 220 225
caa gtc cgt gga aat gaa cat ttg gat ggt ttt ggt gaa acc atc acc 835
Gln Val Arg Gly Asn Glu His Leu Asp Gly Phe Gly Glu Thr Ile Thr
230 235 240 245
agt gga att cgt ctt ggt ttg ggc att acg aca gga aaa gat gtc gta 883
Ser Gly Ile Arg Leu Gly Leu Gly Ile Thr Thr Gly Lys Asp Val Val
250 255 260
gat gaa ctg ctc gag cga ccg cgg caa aag gcc gtt gag gta gca cgc 93l
Asp Glu Leu Leu Glu Arg Pro Arg Gln Lys Ala Val Glu Val Ala Arg
265 270 275
ttt ttt gat cgt tta ggt gtg ggc cga aac tat ctc gtg gat gct gtt 979
Phe Phe Asp Arg Leu Gly Val Gly Arg Asn Tyr Leu Val Asp Ala Val
280 285 290
gat att cat ccg ggt gag gat ttg gtg cag ggg acc atc acc gag gcc 1027
Asp Ile His Pro Gly Glu Asp Leu Val Gln Gly Thr Ile Thr Glu Ala
295 300 305
gcg cag gct tat cgc atg gcc cgg gtg atg tcg gag atg ttg tcg aag 1075
Ala Gln Ala Tyr Arg Met Ala Arg Val Met Ser Glu Met Leu Ser Lys
310 315 320 325
gat tca tgc gac ctt taaggcttta ccggcgctgg gtg 1113
Asp Ser Cys Asp Leu
330
<210>86
<211>330
<212>PRT
<213>谷氨酸棒杆菌
<400>86
Leu Gly Ala Tyr Gly Leu Gly Glu Leu Pro Gly Lys Ser Ala Ala Glu
1 5 10 15
Ala Ala Asp Ile Ile Gln Gly Glu Thr Gly Asp Leu Leu His Ile Pro
20 25 30
Gln Leu Pro Ala Arg Gly Leu Gly Ala Asp Leu Ile Gly Arg Thr Val
35 40 45
Gly Leu Leu Asp Met Ile Asn Val Asp Arg Gly Ala Arg Ser Trp Val
50 55 60
Met Ser Thr Arg Pro Ser Arg Leu Thr His Leu Thr Gly Asp Phe Leu
65 70 75 80
Asp Met Asp Leu Asp Ala Cys Glu Glu Thr Trp Gly Thr Gly Val Asp
85 90 95
Lys Leu Lys Ile Gln Val Ala Gly Pro Trp Thr Leu Gly Ala Arg Ile
100 105 110
Glu Leu Ala Asn Gly His Arg Val Leu Ser Asp Arg Gly Ala Met Arg
115 120 125
Asp Leu Thr Gln Ala Leu Ile Ala Gly Ile Asp Ala His Ala Arg Lys
130 135 140
Val Ala Gly Arg Phe Arg Ala Glu Val Gln Val Gln Ile Asp Glu Pro
145 150 155 160
Glu Leu Lys Ser Leu Ile Asp Gly Ser Leu Pro Gly Thr Ser Thr Phe
165 170 175
Asp Ile Ile Pro Ala Val Asn Val Ala Asp Ala Ser Glu Arg Leu Gln
180 185 190
Gln Val Phe Ser Ser Ile Glu Gly Pro Thr Tyr Leu Asn Leu Thr Gly
195 200 205
Gln Ile Pro Thr Trp Asp Val Ala Arg Gly Ala Gly Ala Asp Thr Val
210 215 220
Gln Ile Ser Met Asp Gln Val Arg Gly Asn Glu His Leu Asp Gly Phe
225 230 235 240
Gly Glu Thr Ile Thr Ser Gly Ile Arg Leu Gly Leu Gly Ile Thr Thr
245 250 255
Gly Lys Asp Val Val Asp Glu Leu Leu Glu Arg Pro Arg Gln Lys Ala
260 265 270
Val Glu Val Ala Arg Phe Phe Asp Arg Leu Gly Val Gly Arg Asn Tyr
275 280 285
Leu Val Asp Ala Val Asp Ile His Pro Gly Glu Asp Leu Val Gln Gly
290 295 300
Thr Ile Thr Glu Ala Ala Gln Ala Tyr Arg Met Ala Arg Val Met Ser
305 310 315 320
Glu Met Leu Ser Lys Asp Ser Cys Asp Leu
325 330
<210>87
<211>551
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(528)
<223>RXA02197
<400>87
gcc gaa cgc atg cgc ttt agc ttc cca cgc cag cag cgc ggc agg ttc 48
Ala Glu Arg Met Arg Phe Ser Phe Pro Arg Gln Gln Arg Gly Arg Phe
1 5 10 15
ttg tgc atc gcg gat ttc att cgc cca cgc gag caa gct gtc aag gac 96
Leu Cys Ile Ala Asp Phe Ile Arg Pro Arg Glu Gln Ala Val Lys Asp
20 25 30
ggc caa gtg gac gtc atg cca ttc cag ctg gtc acc atg ggt aat cct 144
Gly Gln Val Asp Val Met Pro Phe Gln Leu Val Thr Met Gly Asn Pro
35 40 45
att gct gat ttc gcc aac gag ttg ttc gca gcc aat gaa tac cgc gag 192
Ile Ala Asp Phe Ala Asn Glu Leu Phe Ala Ala Asn Glu Tyr Arg Glu
50 55 60
tac ttg gaa gtt cac ggc atc ggc gtg cag ctc acc gaa gca ttg gcc 240
Tyr Leu Glu Val His Gly Ile Gly Val Gln Lau Thr Glu Ala Leu Ala
65 70 75 80
gag tac tgg cac tcc cga gtg cgc agc gaa ctc aag ctg aac gac ggt 288
Glu Tyr Trp His Ser Arg Val Arg Ser Glu Leu Lys Leu Asn Asp Gly
85 90 95
gga tct gtc gct gat ttt gat cca gaa gac aag acc aag ttc ttc gac 336
Gly Ser Val Ala Asp Phe Asp Pro Glu Asp Lys Thr Lys Phe Phe Asp
100 105 110
ctg gat tac cgc ggc gcc cgc ttc tcc ttt ggt tac ggt tct tgc cct 384
Leu Asp Tyr Arg Gly Ala Arg Phe Ser Phe Gly Tyr Gly Ser Cys Pro
115 120 125
gat ctg gaa gac cgc gca aag ctg gtg gaa ttg ctc gag cca ggc cgt 432
Asp Leu Glu Asp Arg Ala Lys Leu Val Glu Leu Leu Glu Pro Gly Arg
130 135 140
atc ggc gtg gag ttg tcc gag gaa ctc cag ctg cac cca gag cag tcc 480
Ile Gly Val Glu Leu Ser Glu Glu Leu Gln Leu His Pro Glu Gln Ser
145 150 155 160
aca gac gcg ttt gtg ctc tac cac cca gag gca aag tac ttt aac gtc 528
Thr Asp Ala Phe Val Leu Tyr His Pro Glu Ala Lys Tyr Phe Asn Val
165 170 175
taacaccttt gagagggaaa act 551
<210>88
<211>176
<212>PRT
<213>谷氨酸棒杆菌
<400>88
Ala Glu Arg Met Arg Phe Ser Phe Pro Arg Gln Gln Arg Gly Arg Phe
1 5 10 15
Leu Cys Ile Ala Asp Phe Ile Arg Pro Arg Glu Gln Ala Val Lys Asp
20 25 30
Gly Gln Val Asp Val Met Pro Phe Gln Leu Val Thr Met Gly Asn Pro
35 40 45
Ile Ala Asp Phe Ala Asn Glu Leu Phe Ala Ala Asn Glu Tyr Arg Glu
50 55 60
Tyr Leu Glu Val His Gly Ile Gly Val Gln Leu Thr Glu Ala Leu Ala
65 70 75 80
Glu Tyr Trp His Ser Arg Val Arg Ser Glu Leu Lys Leu Asn Asp Gly
85 90 95
Gly Ser Val Ala Asp Phe Asp Pro Glu Asp Lys Thr Lys Phe Phe Asp
100 105 110
Leu Asp Tyr Arg Gly Ala Arg Phe Ser Phe Gly Tyr Gly Ser Cys Pro
115 120 125
Asp Leu Glu Asp Arg Ala Lys Leu Val Glu Leu Leu Glu Pro Gly Arg
130 135 140
Ile Gly Val Glu Leu Ser Glu Glu Leu Gln Leu His Pro Glu Gln Ser
145 150 155 160
Thr Asp Ala Phe Val Leu Tyr His Pro Glu Ala Lys Tyr Phe Asn Val
165 170 175
<210>89
<211>2599
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(2599)
<223>RXN02198
<400>89
agactagtgg cgctttgcct gtgttgctta ggcggcgttg aaaatgaact acgaatgaaa 60
agttcgggaa ttgtctaatc cgtactaagc tgtctacaca atg tct act tca gtt 115
Met Ser Thr Ser Val
1 5
act tca cca gcc cac aac aac gca cat tcc tcc gaa ttt ttg gat gcg 163
Thr Ser Pro Ala His Asn Asn Ala His Ser Ser Glu Phe Leu Asp Ala
10 15 20
ttg gca aac cat gtg ttg atc ggc gac ggc gcc atg ggc acc cag ctc 211
Leu Ala Asn His Val Leu Ile Gly Asp Gly Ala Met Gly Thr Gln Leu
25 30 35
caa ggc ttt gac ctg gac gtg gaa aag gat ttc ctt gat ctg gag ggg 259
Gln Gly Phe Asp Leu Asp Val Glu Lys Asp Phe Leu Asp Leu Glu Gly
40 45 50
tgt aat gag att ctc aac gac acc cgc cct gat gtg ttg agg cag att 307
Cys Asn Glu Ile Leu Asn Asp Thr Arg Pro Asp Val Leu Arg Gln Ile
55 60 65
cac cgc gcc tac ttt gag gcg gga gct gac ttg gtt gag acc aat act 355
His Arg Ala Tyr Phe Glu Ala Gly Ala Asp Leu Val Glu Thr Asn Thr
70 75 80 85
ttt ggt tgc aac ctg ccg aac ttg gcg gat tat gac atc gct gat cgt 403
Phe Gly Cys Asn Leu Pro Asn Leu Ala Asp Tyr Asp Ile Ala Asp Arg
90 95 100
tgc cgt gag ctt gcc tac aag ggc act gca gtg gct agg gaa gtg gct 451
Cys Arg Glu Leu Ala Tyr Lys Gly Thr Ala Val Ala Arg Glu Val Ala
105 110 115
gat gag atg ggg ccg ggc cga aac ggc atg cgg cgt ttc gtg gtt ggt 499
Asp Glu Met Gly Pro Gly Arg Asn Gly Met Arg Arg Phe Val Val Gly
120 125 130
tcc ctg gga cct gga acg aag ctt cca tcg ctg ggc cat gca ccg tat 547
Ser Leu Gly Pro Gly Thr Lys Leu Pro Ser Leu Gly His Ala Pro Tyr
135 140 145
gca gat ttg cgt ggg cac tac aag gaa gca gcg ctt ggc atc atc gac 595
Ala Asp Leu Arg Gly His Tyr Lys Glu Ala Ala Leu Gly Ile Ile Asp
150 155 160 165
ggt ggt ggc gat gcc ttt ttg att gag act gct cag gac ttg ctt cag 643
Gly Gly Gly Asp Ala Phe Leu Ile Glu Thr Ala Gln Asp Leu Leu Gln
170 175 180
gtc aag gct gcg gtt cac ggc gtt caa gat gcc atg gct gaa ctt gat 691
Val Lys Ala Ala Val His Gly Val Gln Asp Ala Met Ala Glu Leu Asp
185 190 195
aca ttc ttg ccc att att tgc cac gtc acc gta gag acc acc ggc acc 739
Thr Phe Leu Pro Ile Ile Cys His Val Thr Val Glu Thr Thr Gly Thr
200 205 210
atg ctc atg ggt tct gag atc ggt gcc gcg ttg aca gcg ctg cag cca 787
Met Leu Met Gly Ser Glu Ile Gly Ala Ala Leu Thr Ala Leu Gln Pro
215 220 225
ctg ggt atc gac atg att ggt ctg aac tgc gcc acc ggc cca gat gag 835
Leu Gly Ile Asp Met Ile Gly Leu Asn Cys Ala Thr Gly Pro Asp Glu
230 235 240 245
atg agc gag cac ctg cgt tac ctg tcc aag cac gcc gat att cct gtg 883
Met Ser Glu His Leu Arg Tyr Leu Ser Lys His Ala Asp Ile Pro Val
250 255 260
tcg gtg atg cct aac gca ggt ctt cct gtc ctg ggt aaa aac ggt gca 931
Ser Val Met Pro Asn Ala Gly Leu Pro Val Leu Gly Lys Asn Gly Ala
265 270 275
gaa tac cca ctt gag gct gag gat ttg gcg cag gcg ctg gct gga ttc 979
Glu Tyr Pro Leu Glu Ala Glu Asp Leu Ala Gln Ala Leu Ala Gly Phe
280 285 290
gtc tcc gaa tat ggc ctg tcc atg gtg ggt ggt tgt tgt ggc acc aca 1027
Val Ser Glu Tyr Gly Leu Ser Met Val Gly Gly Cys Cys Gly Thr Thr
295 300 305
cct gag cac atc cgt gcg gtc cgc gat gcg gtg gtt ggt gtt cca gag 1075
Pro Glu His Ile Arg Ala Val Arg Asp Ala Val Val Gly Val Pro Glu
310 315 320 325
cag gaa acc tcc aca ctg acc aag atc cct gca ggc cct gtt gag cag 1123
Gln Glu Thr Ser Thr Leu Thr Lys Ile Pro Ala Gly Pro Val Glu Gln
330 335 340
gcc tcc cgc gag gtg gag aaa gag gac tcc gtc gcg tcg ctg tac acc 1171
Ala Ser Arg Glu Val Glu Lys Glu Asp Ser Val Ala Ser Leu Tyr Thr
345 350 355
tcg gtg cca ttg tcc cag gaa acc ggc att tcc atg atc ggt gag cgc 1219
Ser Val Pro Leu Ser Gln Glu Thr Gly Ile Ser Met Ile Gly Glu Arg
360 365 370
acc aac tcc aac ggt tcc aag gca ttc cgt gag gca atg ctg tct ggc 1267
Thr Asn Ser Asn Gly Ser Lys Ala Phe Arg Glu Ala Met Leu Ser Gly
375 380 385
gat tgg gaa aag tgt gtg gat att gcc aag cag caa acc cgc gat ggt 1315
Asp Trp Glu Lys Cys Val Asp Ile Ala Lys Gln Gln Thr Arg Asp Gly
390 395 400 405
gca cac atg ctg gat ctt tgt gtg gat tac gtg gga cga gac ggc acc 1363
Ala His Met Leu Asp Leu Cys Val Asp Tyr Val Gly Arg Asp Gly Thr
410 415 420
gcc gat atg gcg acc ttg gca gca ctt ctt gct acc agc tcc act ttg 1411
Ala Asp Met Ala Thr Leu Ala Ala Leu Leu Ala Thr Ser Ser Thr Leu
425 430 435
cca atc atg att gac tcc acc gag cca gag gtt att cgc aca ggc ctt 1459
Pro Ile Met Ile Asp Ser Thr Glu Pro Glu Val Ile Arg Thr Gly Leu
440 445 450
gag cac ttg ggt gga cga agc atc gtt aac tcc gtc aac ttt gaa gac 1507
Glu His Leu Gly Gly Arg Ser Ile Val Asn Ser Val Asn Phe Glu Asp
455 460 465
ggc gat ggc cct gag tcc cgc tac cag cgc atc atg aaa ctg gta aag 1555
Gly Asp Gly Pro Glu Ser Arg Tyr Gln Arg Ile Met Lys Leu Val Lys
470 475 480 485
cag cac ggt gcg gcc gtg gtt gcg ctg acc att gat gag gaa ggc cag 1603
Gln His Gly Ala Ala Val Val Ala Leu Thr Ile Asp Glu Glu Gly Gln
490 495 500
gca cgt acc gct gag cac aag gtg cgc att gct aaa cga ctg att gac 1651
Ala Arg Thr Ala Glu His Lys Val Arg Ile Ala Lys Arg Leu Ile Asp
505 510 515
gat atc acc ggc agc tac ggc ctg gat atc aaa gac atc gtt gtg gac 1699
Asp Ile Thr Gly Ser Tyr Gly Leu Asp Ile Lys Asp Ile Val Val Asp
520 525 530
tgc ctg acc ttc ccg atc tct act ggc cag gaa gaa acc agg cga gat 1747
Cys Leu Thr Phe Pro Ile Ser Thr Gly Gln Glu Glu Thr Arg Arg Asp
535 540 545
ggc att gaa acc atc gaa gcc atc cgc gag ctg aag aag ctc tac cca 1795
Gly Ile Glu Thr Ile Glu Ala Ile Arg Glu Leu Lys Lys Leu Tyr Pro
550 555 560 565
gaa atc cac acc acc ctg ggt ctg tcc aat att tcc ttc ggc ctg aac 1843
Glu Ile His Thr Thr Leu Gly Leu Ser Asn Ile Ser Phe Gly Leu Asn
570 575 580
cct gct gca cgc cag gtt ctt aac tct gtg ttc ctc aat gag tgc att 1891
Pro Ala Ala Arg Gln Val Leu Asn Ser Val Phe Leu Asn Glu Cys Ile
585 590 595
gag gct ggt ctg gac tct gcg att gcg cac agc tcc aag att ttg ccg 1939
Glu Ala Gly Leu Asp Ser Ala Ile Ala His Ser Ser Lys Ile Leu Pro
600 605 610
atg aac cgc att gat gat cgc cag cgc gaa gtg gcg ttg gat atg gtc 1987
Met Asn Arg Ile Asp Asp Arg Gln Arg Glu Val Ala Leu Asp Met Val
615 620 625
tat gat cgc cgc acc gag gat tac gat ccg ctg cag gaa ttc atg cag 2035
Tyr Asp Arg Arg Thr Glu Asp Tyr Asp Pro Leu Gln Glu Phe Met Gln
630 635 640 645
ctg ttt gag ggc gtt tct gct gcc gat gcc aag gat gct cgc gct gaa 2083
Leu Phe Glu Gly Val Ser Ala Ala Asp Ala Lys Asp Ala Arg Ala Glu
650 655 660
cag ctg gcc gct atg cct ttg ttt gag cgt ttg gca cag cgc atc atc 2131
Gln Leu Ala Ala Met Pro Leu Phe Glu Arg Leu Ala Gln Arg Ile Ile
665 670 675
gac ggc gat aag aat ggc ctt gag gat gat ctg gaa gca ggc atg aag 2179
Asp Gly Asp Lys Asn Gly Leu Glu Asp Asp Leu Glu Ala Gly Met Lys
680 685 690
gag aag tct cct att gcg atc atc aac gag gac ctt ctc aac ggc atg 2227
Glu Lys Ser Pro Ile Ala Ile Ile Asn Glu Asp Leu Leu Asn Gly Met
695 700 705
aag acc gtg ggt gag ctg ttt ggt tcc gga cag atg cag ctg cca ttc 2275
Lys Thr Val Gly Glu Leu Phe Gly Ser Gly Gln Met Gln Leu Pro Phe
710 715 720 725
gtg ctg caa tcg gca gaa acc atg aaa act gcg gtg gcc tat ttg gaa 2323
Val Leu Gln Ser Ala Glu Thr Met Lys Thr Ala Val Ala Tyr Leu Glu
730 735 740
ccg ttc atg gaa gag gaa gca gaa gct acc gga tct gcg cag gca gag 2371
Pro Phe Met Glu Glu Glu Ala Glu Ala Thr Gly Ser Ala Gln Ala Glu
745 750 755
ggc aag ggc aaa atc gtc gtg gcc acc gtc aag ggt gac gtg cac gat 2419
Gly Lys Gly Lys Ile Val Val Ala Thr Val Lys Gly Asp Val His Asp
760 765 770
atc ggc aag aac ttg gtg gac atc att ttg tcc aac aac ggt tac gac 2467
Ile Gly Lys Asn Leu Val Asp Ile Ile Leu Ser Asn Asn Gly Tyr Asp
775 780 785
gtg gtg aac ttg ggc atc aag cag cca ctg tcc gcc atg ttg gaa gca 2515
Val Val Asn Leu Gly Ile Lys Gln Pro Leu Ser Ala Met Leu Glu Ala
790 795 800 805
gcg gaa gaa cac aaa gca gac gtc atc ggc atg tcg gga ctt ctt gtg 2563
Ala Glu Glu His Lys Ala Asp Val Ile Gly Met Ser Gly Leu Leu Val
810 815 820
aag tcc acc gtg gtg atg aag caa acc atc agc gac 2599
Lys Ser Thr Val Val Met Lys Gln Thr Ile Ser Asp
825 830
<210>90
<211>833
<212>PRT
<213>谷氨酸棒杆菌
<400>90
Met Ser Thr Ser Val Thr Ser Pro Ala His Asn Asn Ala His Ser Ser
1 5 10 15
Glu Phe Leu Asp Ala Leu Ala Asn His Val Leu Ile Gly Asp Gly Ala
20 25 30
Met Gly Thr Gln Leu Gln Gly Phe Asp Leu Asp Val Glu Lys Asp Phe
35 40 45
Leu Asp Leu Glu Gly Cys Asn Glu Ile Leu Asn Asp Thr Arg Pro Asp
50 55 60
Val Leu Arg Gln Ile His Arg Ala Tyr Phe Glu Ala Gly Ala Asp Leu
65 70 75 80
Val Glu Thr Asn Thr Phe Gly Cys Asn Leu Pro Asn Leu Ala Asp Tyr
85 90 95
Asp Ile Ala Asp Arg Cys Arg Glu Leu Ala Tyr Lys Gly Thr Ala Val
100 105 110
Ala Arg Glu Val Ala Asp Glu Met Gly Pro Gly Arg Asn Gly Met Arg
115 120 125
Arg Phe Val Val Gly Ser Leu Gly Pro Gly Thr Lys Leu Pro Ser Leu
130 135 140
Gly His Ala Pro Tyr Ala Asp Leu Arg Gly His Tyr Lys Glu Ala Ala
145 150 155 160
Leu Gly Ile Ile Asp Gly Gly Gly Asp Ala Phe Leu Ile Glu Thr Ala
165 170 175
Gln Asp Leu Leu Gln Val Lys Ala Ala Val His Gly Val Gln Asp Ala
180 185 190
Met Ala Glu Leu Asp Thr Phe Leu Pro Ile Ile Cys His Val Thr Val
195 200 205
Glu Thr Thr Gly Thr Met Leu Met Gly Ser Glu Ile Gly Ala Ala Leu
210 215 220
Thr Ala Leu Gln Pro Leu Gly Ile Asp Met Ile Gly Leu Asn Cys Ala
225 230 235 240
Thr Gly Pro Asp Glu Met Ser Glu His Leu Arg Tyr Leu Ser Lys His
245 250 255
Ala Asp Ile Pro Val Ser Val Met Pro Asn Ala Gly Leu Pro Val Leu
260 265 270
Gly Lys Asn Gly Ala Glu Tyr Pro Leu Glu Ala Glu Asp Leu Ala Gln
275 280 285
Ala Leu Ala Gly Phe Val Ser Glu Tyr Gly Leu Ser Met Val Gly Gly
290 295 300
Cys Cys Gly Thr Thr Pro Glu His Ile Arg Ala Val Arg Asp Ala Val
305 310 315 320
Val Gly Val Pro Glu Gln Glu Thr Ser Thr Leu Thr Lys Ile Pro Ala
325 330 335
Gly Pro Val Glu Gln Ala Ser Arg Glu Val Glu Lys Glu Asp Ser Val
340 345 350
Ala Ser Leu Tyr Thr Ser Val Pro Leu Ser Gln Glu Thr Gly Ile Ser
355 360 365
Met Ile Gly Glu Arg Thr Asn Ser Asn Gly Ser Lys Ala Phe Arg Glu
370 375 380
Ala Met Leu Ser Gly Asp Trp Glu Lys Cys Val Asp Ile Ala Lys Gln
3853 90 395 400
Gln Thr Arg Asp Gly Ala His Met Leu Asp Leu Cys Val Asp Tyr Val
405 410 415
Gly Arg Asp Gly Thr Ala Asp Met Ala Thr Leu Ala Ala Leu Leu Ala
420 425 430
Thr Ser Ser Thr Leu Pro Ile Met Ile Asp Ser Thr Glu Pro Glu Val
435 440 445
Ile Arg Thr Gly Leu Glu His Leu Gly Gly Arg Ser Ile Val Asn Ser
450 455 460
Val Asn Phe Glu Asp Gly Asp Gly Pro Glu Ser Arg Tyr Gln Arg Ile
465 470 475 480
Met Lys Leu Val Lys Gln His Gly Ala Ala Val Val Ala Leu Thr Ile
485 490 495
Asp Glu Glu Gly Gln Ala Arg Thr Ala Glu His Lys Val Arg Ile Ala
500 505 510
Lys Arg Leu Ile Asp Asp Ile Thr Gly Ser Tyr Gly Leu Asp Ile Lys
515 520 525
Asp Ile Val Val Asp Cys Leu Thr Phe Pro Ile Ser Thr Gly Gln Glu
530 535 540
Glu Thr Arg Arg Asp Gly Ile Glu Thr Ile Glu Ala Ile Arg Glu Leu
5455 50 555 560
Lys Lys Leu Tyr Pro Glu Ile His Thr Thr Leu Gly Leu Ser Asn Ile
565 570 575
Ser Phe Gly Leu Asn Pro Ala Ala Arg Gln Val Leu Asn Ser Val Phe
580 585 590
Leu Asn Glu Cys Ile Glu Ala Gly Leu Asp Ser Ala Ile Ala His Ser
595 600 605
Ser Lys Ile Leu Pro Met Asn Arg Ile Asp Asp Arg Gln Arg Glu Val
610 615 620
Ala Leu Asp Met Val Tyr Asp Arg Arg Thr Glu Asp Tyr Asp Pro Leu
625 630 635 640
Gln Glu Phe Met Gln Leu Phe Glu Gly Val Ser Ala Ala Asp Ala Lys
645 650 655
Asp Ala Arg Ala Glu Gln Leu Ala Ala Met Pro Leu Phe Glu Arg Leu
660 665 670
Ala Gln Arg Ile Ile Asp Gly Asp Lys Asn Gly Leu Glu Asp Asp Leu
675 680 685
Glu Ala Gly Met Lys Glu Lys Ser Pro Ile Ala Ile Ile Asn Glu Asp
690 695 700
Leu Leu Asn Gly Met Lys Thr Val Gly Glu Leu Phe Gly Ser Gly Gln
705 710 715 720
Met Gln Leu Pro Phe Val Leu Gln Ser Ala Glu Thr Met Lys Thr Ala
725 730 735
Val Ala Tyr Leu Glu Pro Phe Met Glu Glu Glu Ala Glu Ala Thr Gly
740 745 750
Ser Ala Gln Ala Glu Gly Lys Gly Lys Ile Val Val Ala Thr Val Lys
755 760 765
Gly Asp Val His Asp Ile Gly Lys Asn Leu Val Asp Ile Ile Leu Ser
770 775 780
Asn Asn Gly Tyr Asp Val Val Asn Leu Gly Ile Lys Gln Pro Leu Ser
785 790 795 800
Ala Met Leu Glu Ala Ala Glu Glu His Lys Ala Asp Val Ile Gly Met
805 810 815
Ser Gly Leu Leu Val Lys Ser Thr Val Val Met Lys Gln Thr Ile Ser
820 825 830
Asp
<210>91
<211>2578
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(2578)
<223>FRXA02198
<400>91
agactagtgg cgctttgcct gtgttgctta ggcggcgttg aaaatgaact acgaatgaaa 60
agttcgggaa ttgtctaatc cgtactaagc tgtctacaca atg tct act tca gtt 115
Met Ser Thr Ser Val
1 5
act tca cca gcc cac aac aac gca cat tcc tcc gaa ttt ttg gat gcg 163
Thr Ser Pro Ala His Asn Asn Ala His Ser Ser Glu Phe Leu Asp Ala
10 15 20
ttg gca aac cat gtg ttg atc ggc gac ggc gcc atg ggc acc cag ctc 211
Leu Ala Asn His Val Leu Ile Gly Asp Gly Ala Met Gly Thr Gln Leu
25 30 35
caa ggc ttt gac ctg gac gtg gaa aag gat ttc ctt gat ctg gag ggg 259
Gln Gly Phe Asp Leu Asp Val Glu Lys Asp Phe Leu Asp Leu Glu Gly
40 45 50
tgt aat gag att ctc aac gac acc cgc cct gat gtg ttg agg cag att 307
Cys Asn Glu Ile Leu Asn Asp Thr Arg Pro Asp Val Leu Arg Gln Ile
55 60 65
cac cgc gcc tac ttt gag gcg gga gct gac ttg gtt gag acc aat act 355
His Arg Ala Tyr Phe Glu Ala Gly Ala Asp Leu Val Glu Thr Asn Thr
70 75 80 85
ttt ggt tgc aac ctg ccg aac ttg gcg gat tat gac atc gct gat cgt 403
Phe Gly Cys Asn Leu Pro Asn Leu Ala Asp Tyr Asp Ile Ala Asp Arg
90 95 100
tgc cgt gag ctt gcc tac aag ggc act gca gtg gct agg gaa gtg gct 451
Cys Arg Glu Leu Ala Tyr Lys Gly Thr Ala Val Ala Arg Glu Val Ala
105 110 115
gat gag atg ggg ccg ggc cga aac ggc atg cgg cgt ttc gtg gtt ggt 499
Asp Glu Met Gly Pro Gly Arg Asn Gly Met Arg Arg Phe Val Val Gly
120 125 130
tcc ctg gga cct gga acg aag ctt cca tcg ctg ggc cat gca ccg tat 547
Ser Leu Gly Pro Gly Thr Lys Leu Pro Ser Leu Gly His Ala Pro Tyr
135 140 145
gca gat ttg cgt ggg cac tac aag gaa gca gcg ctt ggc atc atc gac 595
Ala Asp Leu Arg Gly His Tyr Lys Glu Ala Ala Leu Gly Ile Ile Asp
150 155 160 165
ggt ggt ggc gat gcc ttt ttg att gag act gct cag gac ttg ctt cag 643
Gly Gly Gly Asp Ala Phe Leu Ile Glu Thr Ala Gln Asp Leu Leu Gln
170 175 180
gtc aag gct gcg gtt cac ggc gtt caa gat gcc atg gct gaa ctt gat 691
Val Lys Ala Ala Val His Gly Val Gln Asp Ala Met Ala Glu Leu Asp
185 190 195
aca ttc ttg ccc att att tgc cac gtc acc gta gag acc acc ggc acc 739
Thr Phe Leu Pro Ile Ile Cys His Val Thr Val Glu Thr Thr Gly Thr
200 205 210
atg ctc atg ggt tct gag atc ggt gcc gcg ttg aca gcg ctg cag cca 787
Met Leu Met Gly Ser Glu Ile Gly Ala Ala Leu Thr Ala Leu Gln Pro
215 220 225
ctg ggt atc gac atg att ggt ctg aac tgc gcc acc ggc cca gat gag 835
Leu Gly Ile Asp Met Ile Gly Leu Asn Cys Ala Thr Gly Pro Asp Glu
230 235 240 245
atg agc gag cac ctg cgt tac ctg tcc aag cac gcc gat att cct gtg 883
Met Ser Glu His Leu Arg Tyr Leu Ser Lys His Ala Asp Ile Pro Val
250 255 260
tcg gtg atg cct aac gca ggt ctt cct gtc ctg ggt aaa aac ggt gca 931
Ser Val Met Pro Asn Ala Gly Leu Pro Val Leu Gly Lys Asn Gly Ala
265 270 275
gaa tac cca ctt gag gct gag gat ttg gcg cag gcg ctg gct gga ttc 979
Glu Tyr Pro Leu Glu Ala Glu Asp Leu Ala Gln Ala Leu Ala Gly Phe
280 285 290
gtc tcc gaa tat ggc ctg tcc atg gtg ggt ggt tgt tgt ggc acc aca 1027
Val Ser Glu Tyr Gly Leu Ser Met Val Gly Gly Cys Cys Gly Thr Thr
295 300 305
cct gag cac atc cgt gcg gtc cgc gat gcg gtg gtt ggt gtt cca gag 1075
Pro Glu His Ile Arg Ala Val Arg Asp Ala Val Val Gly Val Pro Glu
310 315 320 325
cag gaa acc tcc aca ctg acc aag atc cct gca ggc cct gtt gag cag 1123
Gln Glu Thr Ser Thr Leu Thr Lys Ile Pro Ala Gly Pro Val Glu Gln
330 335 340
gcc tcc cgc gag gtg gag aaa gag gac tcc gtc gcg tcg ctg tac acc 1171
Ala Ser Arg Glu Val Glu Lys Glu Asp Ser Val Ala Ser Leu Tyr Thr
345 350 355
tcg gtg cca ttg tcc cag gaa acc ggc att tcc atg atc ggt gag cgc 1219
Ser Val Pro Leu Ser Gln Glu Thr Gly Ile Ser Met Ile Gly Glu Arg
360 365 370
acc aac tcc aac ggt tcc aag gca ttc cgt gag gca atg ctg tct ggc 1267
Thr Asn Ser Asn Gly Ser Lys Ala Phe Arg Glu Ala Met Leu Ser Gly
375 380 385
gat tgg gaa aag tgt gtg gat att gcc aag cag caa acc cgc gat ggt 1315
Asp Trp Glu Lys Cys Val Asp Ile Ala Lys Gln Gln Thr Arg Asp Gly
390 395 400 405
gca cac atg ctg gat ctt tgt gtg gat tac gtg gga cga gac ggc acc 1363
Ala His Met Leu Asp Leu Cys Val Asp Tyr Val Gly Arg Asp Gly Thr
410 415 420
gcc gat atg gcg acc ttg gca gca ctt ctt gct acc agc tcc act ttg 1411
Ala Asp Met Ala Thr Leu Ala Ala Leu Leu Ala Thr Ser Ser Thr Leu
425 430 435
cca atc atg att gac tcc acc gag cca gag gtt att cgc aca ggc ctt 1459
Pro Ile Met Ile Asp Ser Thr Glu Pro Glu Val Ile Arg Thr Gly Leu
440 445 450
gag cac ttg ggt gga cga agc atc gtt aac tcc gtc aac ttt gaa gac 1507
Glu His Leu Gly Gly Arg Ser Ile Val Asn Ser Val Asn Phe Glu Asp
455 460 465
ggc gat ggc cct gag tcc cgc tac cag cgc atc atg aaa ctg gta aag 1555
Gly Asp Gly Pro Glu Ser Arg Tyr Gln Arg Ile Met Lys Leu Val Lys
470 475 480 485
cag cac ggt gcg gcc gtg gtt gcg ctg acc att gat gag gaa ggc cag 1603
Gln His Gly Ala Ala Val Val Ala Leu Thr Ile Asp Glu Glu Gly Gln
490 495 500
gca cgt acc gct gag cac aag gtg cgc att gct aaa cga ctg att gac 1651
Ala Arg Thr Ala Glu His Lys Val Arg Ile Ala Lys Arg Leu Ile Asp
505 510 515
gat atc acc ggc agc tac ggc ctg gat atc aaa gac atc gtt gtg gac 1699
Asp Ile Thr Gly Ser Tyr Gly Leu Asp Ile Lys Asp Ile Val Val Asp
520 525 530
tgc ctg acc ttc ccg atc tct act ggc cag gaa gaa acc agg cga gat 1747
Cys Leu Thr Phe Pro Ile Ser Thr Gly Gln Glu Glu Thr Arg Arg Asp
535 540 545
ggc att gaa acc atc gaa gcc atc cgc gag ctg aag aag ctc tac cca 1795
Gly Ile Glu Thr Ile Glu Ala Ile Arg Glu Leu Lys Lys Leu Tyr Pro
550 555 560 565
gaa atc cac acc acc ctg ggt ctg tcc aat att tcc ttc ggc ctg aac 1843
Glu Ile His Thr Thr Leu Gly Leu Ser Asn Ile Ser Phe Gly Leu Asn
570 575 580
cct gct gca cgc cag gtt ctt aac tct gtg ttc ctc aat gag tgc att 1891
Pro Ala Ala Arg Gln Val Leu Asn Ser Val Phe Leu Asn Glu Cys Ile
585 590 595
gag gct ggt ctg gac tct gcg att gcg cac agc tcc aag att ttg ccg 1939
Glu Ala Gly Leu Asp Ser Ala Ile Ala His Ser Ser Lys Ile Leu Pro
600 605 610
atg aac cgc att gat gat cgc cag cgc gaa gtg gcg ttg gat atg gtc 1987
Met Asn Arg Ile Asp Asp Arg Gln Arg Glu Val Ala Leu Asp Met Val
615 620 625
tat gat cgc cgc acc gag gat tac gat ccg ctg cag gaa ttc atg cag 2035
Tyr Asp Arg Arg Thr Glu Asp Tyr Asp Pro Leu Gln Glu Phe Met Gln
630 635 640 645
ctg ttt gag ggc gtt tct gct gcc gat gcc aag gat gct cgc gct gaa 2083
Leu Phe Glu Gly Val Ser Ala Ala Asp Ala Lys Asp Ala Arg Ala Glu
650 655 660
cag ctg gcc gct atg cct ttg ttt gag cgt ttg gca cag cgc atc atc 2131
Gln Leu Ala Ala Met Pro Leu Phe Glu Arg Leu Ala Gln Arg Ile Ile
665 670 675
gac ggc gat aag aat ggc ctt gag gat gat ctg gaa gca ggc atg aag 2179
Asp Gly Asp Lys Asn Gly Leu Glu Asp Asp Leu Glu Ala Gly Met Lys
680 685 690
gag aag tct cct att gcg atc atc aac gag gac ctt ctc aac ggc atg 2227
Glu Lys Ser Pro Ile Ala Ile Ile Asn Glu Asp Leu Leu Asn Gly Met
695 700 705
aag acc gtg ggt gag ctg ttt ggt tcc gga cag atg cag ctg cca ttc 2275
Lys Thr Val Gly Glu Leu Phe Gly Ser Gly Gln Met Gln Leu Pro Phe
710 715 720 725
gtg ctg caa tcg gca gaa acc atg aaa act gcg gtg gcc tat ttg gaa 2323
Val Leu Gln Ser Ala Glu Thr Met Lys Thr Ala Val Ala Tyr Leu Glu
730 735 740
ccg ttc atg gaa gag gaa gca gaa gct acc gga tct gcg cag gca gag 2371
Pro Phe Met Glu Glu Glu Ala Glu Ala Thr Gly Ser Ala Gln Ala Glu
745 750 755
ggc aag ggc aaa atc gtc gtg gcc acc gtc aag ggt gac gtg cac gat 2419
Gly Lys Gly Lys Ile Val Val Ala Thr Val Lys Gly Asp Val His Asp
760 765 770
atc ggc aag aac ttg gtg gac atc att ttg tcc aac aac ggt tac gac 2467
Ile Gly Lys Asn Leu Val Asp Ile Ile Leu Ser Asn Asn Gly Tyr Asp
775 780 785
gtg gtg aac ttg ggc atc aag cag cca ctg tcc gcc atg ttg gaa gca 2515
Val Val Asn Leu Gly Ile Lys Gln Pro Leu Ser Ala Met Leu Glu Ala
790 795 800 805
gcg gaa gaa cac aaa gca gac gtc atc ggc atg tcg gga ctt ctt gtg 2563
Ala Glu Glu His Lys Ala Asp Val Ile Gly Met Ser Gly Leu Leu Val
810 815 820
aag tcc acc gtg gtg 2578
Lys Ser Thr Val Val
825
<210>92
<211>826
<212>PRT
<213>谷氨酸棒杆菌
<400>92
Met Ser Thr Ser Val Thr Ser Pro Ala His Asn Asn Ala His Ser Ser
1 5 10 15
Glu Phe Leu Asp Ala Leu Ala Asn His Val Leu Ile Gly Asp Gly Ala
20 25 30
Met Gly Thr Gln Leu Gln Gly Phe Asp Leu Asp Val Glu Lys Asp Phe
35 40 45
Leu Asp Leu Glu Gly Cys Asn Glu Ile Leu Asn Asp Thr Arg Pro Asp
50 55 60
Val Leu Arg Gln Ile His Arg Ala Tyr Phe Glu Ala Gly Ala Asp Leu
65 70 75 80
Val Glu Thr Asn Thr Phe Gly Cys Asn Leu Pro Asn Leu Ala Asp Tyr
85 90 95
Asp Ile Ala Asp Arg Cys Arg Glu Leu Ala Tyr Lys Gly Thr Ala Val
100 105 110
Ala Arg Glu Val Ala Asp Glu Met Gly Pro Gly Arg Asn Gly Met Arg
115 120 125
Arg Phe Val Val Gly Ser Leu Gly Pro Gly Thr Lys Leu Pro Ser Leu
130 135 140
Gly His Ala Pro Tyr Ala Asp Leu Arg Gly His Tyr Lys Glu Ala Ala
145 150 155 160
Leu Gly Ile Ile Asp Gly Gly Gly Asp Ala Phe Leu Ile Glu Thr Ala
165 170 175
Gln Asp Leu Leu Gln Val Lys Ala Ala Val His Gly Val Gln Asp Ala
180 185 190
Met Ala Glu Leu Asp Thr Phe Leu Pro Ile Ile Cys His Val Thr Val
195 200 205
Glu Thr Thr Gly Thr Met Leu Met Gly Ser Glu Ile Gly Ala Ala Leu
210 215 220
Thr Ala Leu Gln Pro Leu Gly Ile Asp Met Ile Gly Leu Asn Cys Ala
225 230 235 240
Thr Gly Pro Asp Glu Met Ser Glu His Leu Arg Tyr Leu Ser Lys His
245 250 255
Ala Asp Ile Pro Val Ser Val Met Pro Asn Ala Gly Leu Pro Val Leu
260 265 270
Gly Lys Asn Gly Ala Glu Tyr Pro Leu Glu Ala Glu Asp Leu Ala Gln
275 280 285
Ala Leu Ala Gly Phe Val Ser Glu Tyr Gly Leu Ser Met Val Gly Gly
290 295 300
Cys Cys Gly Thr Thr Pro Glu His Ile Arg Ala Val Arg Asp Ala Val
305 310 315 320
Val Gly Val Pro Glu Gln Glu Thr Ser Thr Leu Thr Lys Ile Pro Ala
325 330 335
Gly Pro Val Glu Gln Ala Ser Arg Glu Val Glu Lys Glu Asp Ser Val
340 345 350
Ala Ser Leu Tyr Thr Ser Val Pro Leu Ser Gln Glu Thr Gly Ile Ser
355 360 365
Met Ile Gly Glu Arg Thr Asn Ser Asn Gly Ser Lys Ala Phe Arg Glu
370 375 380
Ala Met Leu Ser Gly Asp Trp Glu Lys Cys Val Asp Ile Ala Lys Gln
385 390 395 400
Gln Thr Arg Asp Gly Ala His Met Leu Asp Leu Cys Val Asp Tyr Val
405 410 415
Gly Arg Asp Gly Thr Ala Asp Met Ala Thr Leu Ala Ala Leu Leu Ala
420 425 430
Thr Ser Ser Thr Leu Pro Ile Met Ile Asp Ser Thr Glu Pro Glu Val
435 440 445
Ile Arg Thr Gly Leu Glu His Leu Gly Gly Arg Ser Ile Val Asn Ser
450 455 460
Val Asn Phe Glu Asp Gly Asp Gly Pro Glu Ser Arg Tyr Gln Arg Ile
465 470 475 480
Met Lys Leu Val Lys Gln His Gly Ala Ala Val Val Ala Leu Thr Ile
485 490 495
Asp Glu Glu Gly Gln Ala Arg Thr Ala Glu His Lys Val Arg Ile Ala
500 505 510
Lys Arg Leu Ile Asp Asp Ile Thr Gly Ser Tyr Gly Leu Asp Ile Lys
515 520 525
Asp Ile Val Val Asp Cys Leu Thr Phe Pro Ile Ser Thr Gly Gln Glu
530 535 540
Glu Thr Arg Arg Asp Gly Ile Glu Thr Ile Glu Ala Ile Arg Glu Leu
545 550 555 560
Lys Lys Leu Tyr Pro Glu Ile His Thr Thr Leu Gly Leu Ser Asn Ile
565 570 575
Ser Phe Gly Leu Asn Pro Ala Ala Arg Gln Val Leu Asn Ser Val Phe
580 585 590
Leu Asn Glu Cys Ile Glu Ala Gly Leu Asp Ser Ala Ile Ala His Ser
595 600 605
Ser Lys Ile Leu Pro Met Asn Arg Ile Asp Asp Arg Gln Arg Glu Val
610 615 620
Ala Leu Asp Met Val Tyr Asp Arg Arg Thr Glu Asp Tyr Asp Pro Leu
625 630 635 640
Gln Glu Phe Met Gln Leu Phe Glu Gly Val Ser Ala Ala Asp Ala Lys
645 650 655
Asp Ala Arg Ala Glu Gln Leu Ala Ala Met Pro Leu Phe Glu Arg Leu
660 665 670
Ala Gln Arg Ile Ile Asp Gly Asp Lys Asn Gly Leu Glu Asp Asp Leu
675 680 685
Glu Ala Gly Met Lys Glu Lys Ser Pro Ile Ala Ile Ile Asn Glu Asp
690 695 700
Leu Leu Asn Gly Met Lys Thr Val Gly Glu Leu Phe Gly Ser Gly Gln
705 710 715 720
Met Gln Leu Pro Phe Val Leu Gln Ser Ala Glu Thr Met Lys Thr Ala
725 730 735
Val Ala Tyr Leu Glu Pro Phe Met Glu Glu Glu Ala Glu Ala Thr Gly
740 745 750
Ser Ala Gln Ala Glu Gly Lys Gly Lys Ile Val Val Ala Thr Val Lys
755 760 765
Gly Asp Val His Asp Ile Gly Lys Asn Leu Val Asp Ile Ile Leu Ser
770 775 780
Asn Asn Gly Tyr Asp Val Val Asn Leu Gly Ile Lys Gln Pro Leu Ser
785 790 795 800
Ala Met Leu Glu Ala Ala Glu Glu His Lys Ala Asp Val Ile Gly Met
805 810 815
Ser Gly Leu Leu Val Lys Ser Thr Val Val
820 825
<210>93
<211>621
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(598)
<223>RXN03074
<400>93
tttgtgggca atctggtttt ttcgtaattg tgtgggatga atctcttaaa aattcacatt 60
tagcaggaca agcatactgt tttagttcta tgctgtgggc atg act caa agt gct 115
Met Thr Gln Ser Ala
1 5
cca gaa ttc att gcc acc gca gac ctc gta gac atc atc ggc gac aac 163
Pro Glu Phe Ile Ala Thr Ala Asp Leu Val Asp Ile Ile Gly Asp Asn
10 15 20
gcg caa tca tgc gac act cag ttt caa aac ctt gga ggt gcc aca gaa 211
Ala Gln Ser Cys Asp Thr Gln Phe Gln Asn Leu Gly Gly Ala Thr Glu
25 30 35
ttc cac gga ata ata acc acc gtg aaa tgc ttc caa gac aac gcc ctc 259
Phe His Gly Ile Ile Thr Thr Val Lys Cys Phe Gln Asp Asn Ala Leu
40 45 50
ctg aaa tcc atc ctg agc gag gat aat cct ggg gga gtg ctg gtt atc 307
Leu Lys Ser Ile Leu Ser Glu Asp Asn Pro Gly Gly Val Leu Val Ile
55 60 65
gat ggc gac gca tcc gtg cac acc gcg cta gtt ggc gac atc att gca 355
Asp Gly Asp Ala Ser Val His Thr Ala Leu Val Gly Asp Ile Ile Ala
70 75 80 85
gga ctt gga aaa gat cat ggt tgg tcc gga gta att gtc aac gga gca 403
Gly Leu Gly Lys Asp His Gly Trp Ser Gly Val Ile Val Asn Gly Ala
90 95 100
att cga gac tcc gca gtc atc ggc acc atg acc ttt ggt tgt aaa gcc 451
Ile Arg Asp Ser Ala Val Ile Gly Thr Met Thr Phe Gly Cys Lys Ala
105 110 115
ctt gga acc aac ccg cgg aaa tcc act aaa act ggt tcc ggc gaa cga 499
Leu Gly Thr Asn Pro Arg Lys Ser Thr Lys Thr Gly Ser Gly Glu Arg
120 125 130
gac gta gtg gta tcg att ggt ggc att gac ttc att cct ggt cat tac 547
Asp Val Val Val Ser Ile Gly Gly Ile Asp Phe Ile Pro Gly His Tyr
135 140 145
gtc tac gcg gac tct gac gga att atc gtc acc gag gcg cca att aag 595
Val Tyr Ala Asp Ser Asp Gly Ile Ile Val Thr Glu Ala Pro Ile Lys
150 155 160 165
cag taatttgttt tgacgacgca gta 621
Gln
<210>94
<211>166
<212>PRT
<213>谷氨酸棒杆菌
<400>94
Met Thr Gln Ser Ala Pro Glu Phe Ile Ala Thr Ala Asp Leu Val Asp
1 5 10 15
Ile Ile Gly Asp Asn Ala Gln Ser Cys Asp Thr Gln Phe Gln Asn Leu
20 25 30
Gly Gly Ala Thr Glu Phe His Gly Ile Ile Thr Thr Val Lys Cys Phe
35 40 45
Gln Asp Asn Ala Leu Leu Lys Ser Ile Leu Ser Glu Asp Asn Pro Gly
50 55 60
Gly Val Leu Val Ile Asp Gly Asp Ala Ser Val His Thr Ala Leu Val
65 70 75 80
Gly Asp Ile Ile Ala Gly Leu Gly Lys Asp His Gly Trp Ser Gly Val
85 90 95
Ile Val Asn Gly Ala Ile Arg Asp Ser Ala Val Ile Gly Thr Met Thr
100 105 110
Phe Gly Cys Lys Ala Leu Gly Thr Asn Pro Arg Lys Ser Thr Lys Thr
115 120 125
Gly Ser Gly Glu Arg Asp Val Val Val Ser Ile Gly Gly Ile Asp Phe
130 135 140
Ile Pro Gly His Tyr Val Tyr Ala Asp Ser Asp Gly Ile Ile Val Thr
145 150 155 160
Glu Ala Pro Ile Lys Gln
165
<210>95
<211>621
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(598)
<223>FRXA02906
<400>95
tttgtgggca atctggtttt ttcgtaattg tgtgggatga atctcttaaa aattcacatt 60
tagcaggaca agcatactgt tttagttcta tgctgtgggc atg act caa agt gct 115
Met Thr Gln Ser Ala
1 5
cca gaa ttc att gcc acc gca gac ctc gta gac atc atc ggc gac aac 163
Pro Glu Phe Ile Ala Thr Ala Asp Leu Val Asp Ile Ile Gly Asp Asn
10 15 20
gcg caa tca tgc gac act cag ttt caa aac ctt gga ggt gcc aca gaa 211
Ala Gln Ser Cys Asp Thr Gln Phe Gln Asn Leu Gly Gly Ala Thr Glu
25 30 35
ttc cac gga ata ata acc acc gtg aaa tgc ttc caa gac aac gcc ctc 259
Phe His Gly Ile Ile Thr Thr Val Lys Cys Phe Gln Asp Asn Ala Leu
40 45 50
ctg aaa tcc atc ctg agc gag gat aat cct ggg gga gtg ctg gtt atc 307
Leu Lys Ser Ile Leu Ser Glu Asp Asn Pro Gly Gly Val Leu Val Ile
55 60 65
gat ggc gac gca tcc gtg cac acc gcg cta gtt ggc gac atc att gca 355
Asp Gly Asp Ala Ser Val His Thr Ala Leu Val Gly Asp Ile Ile Ala
70 75 80 85
gga ctt gga aaa gat cat ggt tgg tcc gga gta att gtc aac gga gca 403
Gly Leu Gly Lys Asp His Gly Trp Ser Gly Val Ile Val Asn Gly Ala
90 95 100
att cga gac tcc gca gtc atc ggc acc atg acc ttt ggt tgt aaa gcc 451
Ile Arg Asp Ser Ala Val Ile Gly Thr Met Thr Phe Gly Cys Lys Ala
105 110 115
ctt gga acc aac ccg cgg aaa tcc act aaa act ggt tcc ggc gaa cga 499
Leu Gly Thr Asn Pro Arg Lys Ser Thr Lys Thr Gly Ser Gly Glu Arg
120 125 130
gac gta gtg gta tcg att ggt ggc att gac ttc att cct ggt cat tac 547
Asp Val Val Val Ser Ile Gly Gly Ile Asp Phe Ile Pro Gly His Tyr
135 140 145
gtc tac gcg gac tct gac gga att atc gtc acc gag gcg cca att aag 595
Val Tyr Ala Asp Ser Asp Gly Ile Ile Val Thr Glu Ala Pro Ile Lys
150 155 160 165
cag taatttgttt tgacgacgca gta 621
Gln
<210>96
<211>166
<212>PRT
<213>谷氨酸棒杆菌
<400>96
Met Thr Gln Ser Ala Pro Glu Phe Ile Ala Thr Ala Asp Leu Val Asp
1 5 10 15
Ile Ile Gly Asp Asn Ala Gln Ser Cys Asp Thr Gln Phe Gln Asn Leu
20 25 30
Gly Gly Ala Thr Glu Phe His Gly Ile Ile Thr Thr Val Lys Cys Phe
35 40 45
Gln Asp Asn Ala Leu Leu Lys Ser Ile Leu Ser Glu Asp Asn Pro Gly
50 55 60
Gly Val Leu Val Ile Asp Gly Asp Ala Ser Val His Thr Ala Leu Val
65 70 75 80
Gly Asp Ile Ile Ala Gly Leu Gly Lys Asp His Gly Trp Ser Gly Val
85 90 95
Ile Val Asn Gly Ala Ile Arg Asp Ser Ala Val Ile Gly Thr Met Thr
100 105 110
Phe Gly Cys Lys Ala Leu Gly Thr Asn Pro Arg Lys Ser Thr Lys Thr
115 120 125
Gly Ser Gly Glu Arg Asp Val Val Val Ser Ile Gly Gly Ile Asp Phe
130 135 140
Ile Pro Gly His Tyr Val Tyr Ala Asp Ser Asp Gly Ile Ile Val Thr
145 150 155 160
Glu Ala Pro Ile Lys Gln
165
<210>97
<211>1557
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1534)
<223>RXN00132
<400>97
aacagcttca atcaattcgg tgtccactcc aacatgtaga gtggtgcgcg ttaaaaaagt 60
tttcctaatt ttcattttct taaaaggagc tcgccaggac atg gca cag gtt atg 115
Met Ala Gln Val Met
1 5
gac ttc aag gtt gcc gat ctt tca cta gca gag gca gga cgt cac cag 163
Asp Phe Lys Val Ala Asp Leu Ser Leu Ala Glu Ala Gly Arg His Gln
10 15 20
att cgt ctt gca gag tat gag atg cca ggt ctc atg cag ttg cgc aag 211
Ile Arg Leu Ala Glu Tyr Glu Met Pro Gly Leu Met Gln Leu Arg Lys
25 30 35
gaa ttc gca gac gag cag cct ttg aag ggc gcc cga att gct ggt tct 259
Glu Phe Ala Asp Glu Gln Pro Leu Lys Gly Ala Arg Ile Ala Gly Ser
40 45 50
atc cac atg acg gtc cag acc gcc gtg ctt att gag acc ctc act gct 307
Ile His Met Thr Val Gln Thr Ala Val Leu Ile Glu Thr Leu Thr Ala
55 60 65
ttg ggc gct gag gtt cgt tgg gct tcc tgc aac att ttc tcc acc cag 355
Leu Gly Ala Glu Val Arg Trp Ala Ser Cys Asn Ile Phe Ser Thr Gln
70 75 80 85
gat gag gct gca gcg gct atc gtt gtc ggc tcc ggc acc gtc gaa gag 403
Asp Glu Ala Ala Ala Ala Ile Val Val Gly Ser Gly Thr Val Glu Glu
90 95 100
cca gct ggt gtt cca gta ttc gcg tgg aag ggt gag tca ctg gag gag 451
Pro Ala Gly Val Pro Val Phe Ala Trp Lys Gly Glu Ser Leu Glu Glu
105 110 115
tac tgg tgg tgc atc aac cag atc ttc agc tgg ggc gat gag ctg cca 499
Tyr Trp Trp Cys Ile Asn Gln Ile Phe Ser Trp Gly Asp Glu Leu Pro
120 125 130
aac atg atc ctc gac gac ggc ggt gac gcc acc atg gct gtt att cgc 547
Asn Met Ile Leu Asp Asp Gly Gly Asp Ala Thr Met Ala Val Ile Arg
135 140 145
ggt cgc gaa tac gag cag gct ggt ctg gtt cca cca gca gag gcc aac 595
Gly Arg Glu Tyr Glu Gln Ala Gly Leu Val Pro Pro Ala Glu Ala Asn
150 155 160 165
gat tcc gat gag tac atc gca ttc ttg ggc atg ctg cgt gag gtt ctt 643
Asp Ser Asp Glu Tyr Ile Ala Phe Leu Gly Met Leu Arg Glu Val Leu
170 175 180
gct gca gag cct ggc aag tgg ggc aag atc gct gag gcc gtt aag ggt 691
Ala Ala Glu Pro Gly Lys Trp Gly Lys Ile Ala Glu Ala Val Lys Gly
185 190 195
gtc acc gag gaa acc acc acc ggt gtg cac cgc ctg tac cac ttc gct 739
Val Thr Glu Glu Thr Thr Thr Gly Val His Arg Leu Tyr His Phe Ala
200 205 210
gaa gaa ggc gtg ctg cct ttc cca gcg atg aac gtc aac gac gct gtc 787
Glu Glu Gly Val Leu Pro Phe Pro Ala Met Asn Val Asn Asp Ala Val
215 220 225
acc aag tcc aag ttt gat aac aag tac ggc acc cgc cac tcc ctg atc 835
Thr Lys Ser Lys Phe Asp Asn Lys Tyr Gly Thr Arg His Ser Leu Ile
230 235 240 245
gac ggc atc aac cgc gcc act gac atg ctc atg ggc ggc aag aac gtg 883
Asp Gly Ile Asn Arg Ala Thr Asp Met Leu Met Gly Gly Lys Asn Val
250 255 260
ctt gtc tgc ggt tac ggc gat gtc ggc aag ggc tgc gct gag gct ttc 931
Leu Val Cys Gly Tyr Gly Asp Val Gly Lys Gly Cys Ala Glu Ala Phe
265 270 275
gac ggc cag ggc gct cgc gtc aag gtc acc gaa gct gac cca atc aac 979
Asp Gly Gln Gly Ala Arg Val Lys Val Thr Glu Ala Asp Pro Ile Asn
280 285 290
gct ctt cag gct ctg atg gat ggc tac tct gtg gtc acc gtt gat gag 1027
Ala Leu Gln Ala Leu Met Asp Gly Tyr Ser Val Val Thr Val Asp Glu
295 300 305
gcc atc gag gac gcc gac atc gtg atc acc gcg acc ggc aac aag gac 1075
Ala Ile Glu Asp Ala Asp Ile Val Ile Thr Ala Thr Gly Asn Lys Asp
310 315 320 325
atc att tcc ttc gag cag atg ctc aag atg aag gat cac gct ctg ctg 1123
Ile Ile Ser Phe Glu Gln Met Leu Lys Met Lys Asp His Ala Leu Leu
330 335 340
ggc aac atc ggt cac ttt gat aat gag atc gat atg cat tcc ctg ttg 1171
Gly Asn Ile Gly His Phe Asp Asn Glu Ile Asp Met His Ser Leu Leu
345 350 355
cac cgc gac gac gtc acc cgc acc acg atc aag cca cag gtc gac gag 1219
His Arg Asp Asp Val Thr Arg Thr Thr Ile Lys Pro Gln Val Asp Glu
360 365 370
ttc acc ttc tcc acc ggt cgc tcc atc atc gtc ctg tcc gaa ggt cgc 1267
Phe Thr Phe Ser Thr Gly Arg Ser Ile Ile Val Leu Ser Glu Gly Arg
375 380 385
ctg ttg aac ctt ggc aac gcc acc gga cac cca tca ttt gtc atg tcc 1315
Leu Leu Asn Leu Gly Asn Ala Thr Gly His Pro Ser Phe Val Met Ser
390 395 400 405
aac tct ttc gcc gat cag acc att gcg cag atc gaa ctg ttc caa aac 1363
Asn Ser Phe Ala Asp Gln Thr Ile Ala Gln Ile Glu Leu Phe Gln Asn
410 415 420
gaa gga cag tac gag aac gag gtc tac cgt ctg cct aag gtt ctc gac 1411
Glu Gly Gln Tyr Glu Asn Glu Val Tyr Arg Leu Pro Lys Val Leu Asp
425 430 435
gaa aag gtg gca cgc atc cac gtt gag gct ctc ggc ggt cag ctc acc 1459
Glu Lys Val Ala Arg Ile His Val Glu Ala Leu Gly Gly Gln Leu Thr
440 445 450
gaa ctg acc aag gag cag gct gag tac atc ggc gtt gac gtt gca ggc 1507
Glu Leu Thr Lys Glu Gln Ala Glu Tyr Ile Gly Val Asp Val Ala Gly
455 460 465
cca ttc aag ccg gag cac tac cgc tac taatgattgt cagcattgag gga 1557
Pro Phe Lys Pro Glu His Tyr Arg Tyr
470 475
<210>98
<211>478
<212>PRT
<213>谷氨酸棒杆菌
<400>98
Met Ala Gln Val Met Asp Phe Lys Val Ala Asp Leu Ser Leu Ala Glu
1 5 10 15
Ala Gly Arg His Gln Ile Arg Leu Ala Glu Tyr Glu Met Pro Gly Leu
20 25 30
Met Gln Leu Arg Lys Glu Phe Ala Asp Glu Gln Pro Leu Lys Gly Ala
35 40 45
Arg Ile Ala Gly Ser Ile His Met Thr Val Gln Thr Ala Val Leu Ile
50 55 60
Glu Thr Leu Thr Ala Leu Gly Ala Glu Val Arg Trp Ala Ser Cys Asn
65 70 75 80
Ile Phe Ser Thr Gln Asp Glu Ala Ala Ala Ala Ile Val Val Gly Ser
85 90 95
Gly Thr Val Glu Glu Pro Ala Gly Val Pro Val Phe Ala Trp Lys Gly
100 105 110
Glu Ser Leu Glu Glu Tyr Trp Trp Cys Ile Asn Gln Ile Phe Ser Trp
115 120 125
Gly Asp Glu Leu Pro Asn Met Ile Leu Asp Asp Gly Gly Asp Ala Thr
130 135 140
Met Ala Val Ile Arg Gly Arg Glu Tyr Glu Gln Ala Gly Leu Val Pro
145 150 155 160
Pro Ala Glu Ala Asn Asp Ser Asp Glu Tyr Ile Ala Phe Leu Gly Met
165 170 175
Leu Arg Glu Val Leu Ala Ala Glu Pro Gly Lys Trp Gly Lys Ile Ala
180 185 190
Glu Ala Val Lys Gly Val Thr Glu Glu Thr Thr Thr Gly Val His Arg
195 200 205
Leu Tyr His Phe Ala Glu Glu Gly Val Leu Pro Phe Pro Ala Met Asn
210 215 220
Val Asn Asp Ala Val Thr Lys Ser Lys Phe Asp Asn Lys Tyr Gly Thr
225 230 235 240
Arg His Ser Leu Ile Asp Gly Ile Asn Arg Ala Thr Asp Met Leu Met
245 250 255
Gly Gly Lys Asn Val Leu Val Cys Gly Tyr Gly Asp Val Gly Lys Gly
260 265 270
Cys Ala Glu Ala Phe Asp Gly Gln Gly Ala Arg Val Lys Val Thr Glu
275 280 285
Ala Asp Pro Ile Asn Ala Leu Gln Ala Leu Met Asp Gly Tyr Ser Val
290 295 300
Val Thr Val Asp Glu Ala Ile Glu Asp Ala Asp Ile Val Ile Thr Ala
305 310 315 320
Thr Gly Asn Lys Asp Ile Ile Ser Phe Glu Gln Met Leu Lys Met Lys
325 330 335
Asp His Ala Leu Leu Gly Asn Ile Gly His Phe Asp Asn Glu Ile Asp
340 345 350
Met His Ser Leu Leu His Arg Asp Asp Val Thr Arg Thr Thr Ile Lys
355 360 365
Pro Gln Val Asp Glu Phe Thr Phe Ser Thr Gly Arg Ser Ile Ile Val
370 375 380
Leu Ser Glu Gly Arg Leu Leu Asn Leu Gly Asn Ala Thr Gly His Pro
385 390 395 400
Ser Phe Val Met Ser Asn Ser Phe Ala Asp Gln Thr Ile Ala Gln Ile
405 410 415
Glu Leu Phe Gln Asn Glu Gly Gln Tyr Glu Asn Glu Val Tyr Arg Leu
420 425 430
Pro Lys Val Leu Asp Glu Lys Val Ala Arg Ile His Val Glu Ala Leu
435 440 445
Gly Gly Gln Leu Thr Glu Leu Thr Lys Glu Gln Ala Glu Tyr Ile Gly
450 455 460
Val Asp Val Ala Gly Pro Phe Lys Pro Glu His Tyr Arg Tyr
465 470 475
<210>99
<211>128
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(105)
<223>FRXA00132
<400>99
cac gtt gag gct ctc ggc ggt cag ctc acc gaa ctg acc aag gag cag 48
His Val Glu Ala Leu Gly Gly Gln Leu Thr Glu Leu Thr Lys Glu Gln
1 5 10 15
gct gag tac atc ggc gtt gac gtt gca ggc cca ttc aag ccg gag cac 96
Ala Glu Tyr Ile Gly Val Asp Val Ala Gly Pro Phe Lys Pro Glu His
20 25 30
tac cgc tac taatgattgt cagcattgag gga 128
Tyr Arg Tyr
35
<210>100
<211>35
<212>PRT
<213>谷氨酸棒杆菌
<400>100
His Val Glu Ala Leu Gly Gly Gln Leu Thr Glu Leu Thr Lys Glu Gln
1 5 10 15
Ala Glu Tyr Ile Gly Val Asp Val Ala Gly Pro Phe Lys Pro Glu His
20 25 30
Tyr Arg Tyr
35
<210>101
<211>1396
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1396)
<223>FRXA01371
<400>101
aacagcttca atcaattcgg tgtccactcc aacatgtaga gtggtgcgcg ttaaaaaagt 60
tttcctaatt ttcattttct taaaaggagc tcgccaggac atg gca cag gtt atg 115
Met Ala Gln Val Met
1 5
gac ttc aag gtt gcc gat ctt tca cta gca gag gca gga cgt cac cag 163
Asp Phe Lys Val Ala Asp Leu Ser Leu Ala Glu Ala Gly Arg His Gln
10 15 20
att cgt ctt gca gag tat gag atg cca ggt ctc atg cag ttg cgc aag 211
Ile Arg Leu Ala Glu Tyr Glu Met Pro Gly Leu Met Gln Leu Arg Lys
25 30 35
gaa ttc gca gac gag cag cct ttg aag ggc gcc cga att gct ggt tct 259
Glu Phe Ala Asp Glu Gln Pro Leu Lys Gly Ala Arg Ile Ala Gly Ser
40 45 50
atc cac atg acg gtc cag acc gcc gtg ctt att gag acc ctc act gct 307
Ile His Met Thr Val Gln Thr Ala Val Leu Ile Glu Thr Leu Thr Ala
55 60 65
ttg ggc gct gag gtt cgt tgg gct tcc tgc aac att ttc tcc acc cag 355
Leu Gly Ala Glu Val Arg Trp Ala Ser Cys Asn Ile Phe Ser Thr Gln
70 75 80 85
gat gag gct gca gcg gct atc gtt gtc ggc tcc ggc acc gtc gaa gag 403
Asp Glu Ala Ala Ala Ala Ile Val Val Gly Ser Gly Thr Val Glu Glu
90 95 100
cca gct ggt gtt cca gta ttc gcg tgg aag ggt gag tca ctg gag gag 451
Pro Ala Gly Val Pro Val Phe Ala Trp Lys Gly Glu Ser Leu Glu Glu
105 110 115
tac tgg tgg tgc atc aac cag atc ttc agc tgg ggc gat gag ctg cca 499
Tyr Trp Trp Cys Ile Asn Gln Ile Phe Ser Trp Gly Asp Glu Leu Pro
120 125 130
aac atg atc ctc gac gac ggc ggt gac gcc acc atg gct gtt att cgc 547
Asn Met Ile Leu Asp Asp Gly Gly Asp Ala Thr Met Ala Val Ile Arg
135 140 145
ggt cgc gaa tac gag cag gct ggt ctg gtt cca cca gca gag gcc aac 595
Gly Arg Glu Tyr Glu Gln Ala Gly Leu Val Pro Pro Ala Glu Ala Asn
150 155 160 165
gat tcc gat gag tac atc gca ttc ttg ggc atg ctg cgt gag gtt ctt 643
Asp Ser Asp Glu Tyr Ile Ala Phe Leu Gly Met Leu Arg Glu Val Leu
170 175 180
gct gca gag cct ggc aag tgg ggc aag atc gct gag gcc gtt aag ggt 691
Ala Ala Glu Pro Gly Lys Trp Gly Lys Ile Ala Glu Ala Val Lys Gly
185 190 195
gtc acc gag gaa acc acc acc ggt gtg cac cgc ctg tac cac ttc gct 739
Val Thr Glu Glu Thr Thr Thr Gly Val His Arg Leu Tyr His Phe Ala
200 205 210
gaa gaa ggc gtg ctg cct ttc cca gcg atg aac gtc aac gac gct gtc 787
Glu Glu Gly Val Leu Pro Phe Pro Ala Met Asn Val Asn Asp Ala Val
215 220 225
acc aag tcc aag ttt gat aac aag tac ggc acc cgc cac tcc ctg atc 835
Thr Lys Ser Lys Phe Asp Asn Lys Tyr Gly Thr Arg His Ser Leu Ile
230 235 240 245
gac ggc atc aac cgc gcc act gac atg ctc atg ggc ggc aag aac gtg 883
Asp Gly Ile Asn Arg Ala Thr Asp Met Leu Met Gly Gly Lys Asn Val
250 255 260
ctt gtc tgc ggt tac ggc gat gtc ggc aag ggc tgc gct gag gct ttc 931
Leu Val Cys Gly Tyr Gly Asp Val Gly Lys Gly Cys Ala Glu Ala Phe
265 270 275
gac ggc cag ggc gct cgc gtc aag gtc acc gaa gct gac cca atc aac 979
Asp Gly Gln Gly Ala Arg Val Lys Val Thr Glu Ala Asp Pro Ile Asn
280 285 290
gct ctt cag gct ctg atg gat ggc tac tct gtg gtc acc gtt gat gag 1027
Ala Leu Gln Ala Leu Met Asp Gly Tyr Ser Val Val Thr Val Asp Glu
295 300 305
gcc atc gag gac gcc gac atc gtg atc acc gcg acc ggc aac aag gac 1075
Ala Ile Glu Asp Ala Asp Ile Val Ile Thr Ala Thr Gly Asn Lys Asp
310 315 320 325
atc att tcc ttc gag cag atg ctc aag atg aag gat cac gct ctg ctg 1123
Ile Ile Ser Phe Glu Gln Met Leu Lys Met Lys Asp His Ala Leu Leu
330 335 340
ggc aac atc ggt cac ttt gat aat gag atc gat atg cat tcc ctg ttg 1171
Gly Asn Ile Gly His Phe Asp Asn Glu Ile Asp Met His Ser Leu Leu
345 350 355
cac cgc gac gac gtc acc cgc acc acg atc aag cca cag gtc gac gag 1219
His Arg Asp Asp Val Thr Arg Thr Thr Ile Lys Pro Gln Val Asp Glu
360 365 370
ttc acc ttc tcc acc ggt cgc tcc atc atc gtc ctg tcc gaa ggt cgc 1267
Phe Thr Phe Ser Thr Gly Arg Ser Ile Ile Val Leu Ser Glu Gly Arg
375 380 385
ctg ttg aac ctt ggc aac gcc acc gga cac cca tca ttt gtc atg tcc 1315
Leu Leu Asn Leu Gly Asn Ala Thr Gly His Pro Ser Phe Val Met Ser
390 395 400 405
aac tct ttc gcc gat cag acc att gcg cag atc gaa ctg ttc caa aac 1363
Asn Ser Phe Ala Asp Gln Thr Ile Ala Gln Ile Glu Leu Phe Gln Asn
410 415 420
gaa gga cag tac gag aac gag gtc tac cgt ctg 1396
Glu Gly Gln Tyr Glu Asn Glu Val Tyr Arg Leu
425 430
<210>102
<211>432
<212>PRT
<213>谷氨酸棒杆菌
<400>102
Met Ala Gln Val Met Asp Phe Lys Val Ala Asp Leu Ser Leu Ala Glu
1 5 10 15
Ala Gly Arg His Gln Ile Arg Leu Ala Glu Tyr Glu Met Pro Gly Leu
20 25 30
Met Gln Leu Arg Lys Glu Phe Ala Asp Glu Gln Pro Leu Lys Gly Ala
35 40 45
Arg Ile Ala Gly Ser Ile His Met Thr Val Gln Thr Ala Val Leu Ile
50 55 60
Glu Thr Leu Thr Ala Leu Gly Ala Glu Val Arg Trp Ala Ser Cys Asn
65 70 75 80
Ile Phe Ser Thr Gln Asp Glu Ala Ala Ala Ala Ile Val Val Gly Ser
85 90 95
Gly Thr Val Glu Glu Pro Ala Gly Val Pro Val Phe Ala Trp Lys Gly
100 105 110
Glu Ser Leu Glu Glu Tyr Trp Trp Cys Ile Asn Gln Ile Phe Ser Trp
115 120 125
Gly Asp Glu Leu Pro Asn Met Ile Leu Asp Asp Gly Gly Asp Ala Thr
130 135 140
Met Ala Val Ile Arg Gly Arg Glu Tyr Glu Gln Ala Gly Leu Val Pro
145 150 155 160
Pro Ala Glu Ala Asn Asp Ser Asp Glu Tyr Ile Ala Phe Leu Gly Met
165 170 175
Leu Arg Glu Val Leu Ala Ala Glu Pro Gly Lys Trp Gly Lys Ile Ala
180 185 190
Glu Ala Val Lys Gly Val Thr Glu Glu Thr Thr Thr Gly Val His Arg
195 200 205
Leu Tyr His Phe Ala Glu Glu Gly Val Leu Pro Phe Pro Ala Met Asn
210 215 220
Val Asn Asp Ala Val Thr Lys Ser Lys Phe Asp Asn Lys Tyr Gly Thr
225 230 235 240
Arg His Ser Leu Ile Asp Gly Ile Asn Arg Ala Thr Asp Met Leu Met
245 250 255
Gly Gly Lys Asn Val Leu Val Cys Gly Tyr Gly Asp Val Gly Lys Gly
260 265 270
Cys Ala Glu Ala Phe Asp Gly Gln Gly Ala Arg Val Lys Val Thr Glu
275 280 285
Ala Asp Pro Ile Asn Ala Leu Gln Ala Leu Met Asp Gly Tyr Ser Val
290 295 300
Val Thr Val Asp Glu Ala Ile Glu Asp Ala Asp Ile Val Ile Thr Ala
305 310 315 320
Thr Gly Asn Lys Asp Ile Ile Ser Phe Glu Gln Met Leu Lys Met Lys
325 330 335
Asp His Ala Leu Leu Gly Asn Ile Gly His Phe Asp Asn Glu Ile Asp
340 345 350
Met His Ser Leu Leu His Arg Asp Asp Val Thr Arg Thr Thr Ile Lys
355 360 365
Pro Gln Val Asp Glu Phe Thr Phe Ser Thr Gly Arg Ser Ile Ile Val
370 375 380
Leu Ser Glu Gly Arg Leu Leu Asn Leu Gly Asn Ala Thr Gly His Pro
385 390 395 400
Ser Phe Val Met Ser Asn Ser Phe Ala Asp Gln Thr Ile Ala Gln Ile
405 410 415
Glu Leu Phe Gln Asn Glu Gly Gln Tyr Glu Asn Glu Val Tyr Arg Leu
420 425 430
<210>103
<211>2358
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(2335)
<223>RXN02085
<400>103
cacccggtga tttcgcgaac cttgaaacat cgtcagaaga ttgccgtgcg tcctagccgg 60
gatccgcacg ttcggctcaa gcagaaagtc tttaactcac atg act tcc aac ttt 115
Met Thr Ser Asn Phe
1 5
tct tcc act gtc gct ggt ctt cct cgc atc gga gcg aag cgt gaa ctg 163
Ser Ser Thr Val Ala Gly Leu Pro Arg Ile Gly Ala Lys Arg Glu Leu
10 15 20
aag ttc gcg ctc gaa ggc tac tgg aat gga tca att gaa ggt cgc gaa 211
Lys Phe Ala Leu Glu Gly Tyr Trp Asn Gly Ser Ile Glu Gly Arg Glu
25 30 35
ctt gcg cag acc gcc cgc caa ttg gtc aac act gca tcg gat tct ttg 259
Leu Ala Gln Thr Ala Arg Gln Leu Val Asn Thr Ala Ser Asp Ser Leu
40 45 50
tct gga ttg gat tcc gtt ccg ttt gca gga cgt tcc tac tac gac gca 307
Ser Gly Leu Asp Ser Val Pro Phe Ala Gly Arg Ser Tyr Tyr Asp Ala
55 60 65
atg ctc gat acc gcc gct att ttg ggt gtg ctg ccg gag cgt ttt gat 355
Met Leu Asp Thr Ala Ala Ile Leu Gly Val Leu Pro Glu Arg Phe Asp
70 75 80 85
gac atc gct gat cat gaa aac gat ggt ctc cca ctg tgg att gac cgc 403
Asp Ile Ala Asp His Glu Asn Asp Gly Leu Pro Leu Trp Ile Asp Arg
90 95 100
tac ttt ggc gct gct cgc ggt act gag acc ctg cct gca cag gca atg 451
Tyr Phe Gly Ala Ala Arg Gly Thr Glu Thr Leu Pro Ala Gln Ala Met
105 110 115
acc aag tgg ttt gat acc aac tac cac tac ctc gtg ccg gag ttg tct 499
Thr Lys Trp Phe Asp Thr Asn Tyr His Tyr Leu Val Pro Glu Leu Ser
120 125 130
gcg gat aca cgt ttc gtt ttg gat gcg tcc gcg ctg att gag gat ctc 547
Ala Asp Thr Arg Phe Val Leu Asp Ala Ser Ala Leu Ile Glu Asp Leu
135 140 145
cgt tgc cag cag gtt cgt ggc gtt aat gcc cgc cct gtt ctg gtt ggt 595
Arg Cys Gln Gln Val Arg Gly Val Asn Ala Arg Pro Val Leu Val Gly
150 155 160 165
cca ctg act ttc ctt tcc ctt gct cgc acc act gat ggt tcc aat cct 643
Pro Leu Thr Phe Leu Ser Leu Ala Arg Thr Thr Asp Gly Ser Asn Pro
170 175 180
ttg gat cac ctg cct gca ctg ttt gag gtc tac gag cgc ctc atc aag 691
Leu Asp His Leu Pro Ala Leu Phe Glu Val Tyr Glu Arg Leu Ile Lys
185 190 195
tct ttc gat act gag tgg gtt cag atc gat gag cct gcg ttg gtc acc 739
Ser Phe Asp Thr Glu Trp Val Gln Ile Asp Glu Pro Ala Leu Val Thr
200 205 210
gat gtt gct cct gag gtt ttg gag cag gtc cgc gct ggt tac acc act 787
Asp Val Ala Pro Glu Val Leu Glu Gln Val Arg Ala Gly Tyr Thr Thr
215220225
ttg gct aag cgc gat ggc gtg ttt gtc aat act tac ttc ggc tct ggc 835
Leu Ala Lys Arg Asp Gly Val Phe Val Asn Thr Tyr Phe Gly Ser Gly
230 235 240 245
gat cag gcg ctg aac act ctt gcg ggc atc ggc ctt ggc gcg att ggc 883
Asp Gln Ala Leu Asn Thr Leu Ala Gly Ile Gly Leu Gly Ala Ile Gly
250 255 260
gtt gac ttg gtc acc cat ggc gtc act gag ctt gct gcg tgg aag ggt 931
Val Asp Leu Val Thr His Gly Val Thr Glu Leu Ala Ala Trp Lys Gly
265 270 275
gag gag ctg ctg gtt gcg ggc atc gtt gat ggt cgt aac att tgg cgc 979
Glu Glu Leu Leu Val Ala Gly Ile Val Asp Gly Arg Asn Ile Trp Arg
280 285 290
acc gac ctg tgt gct gct ctt gct tcc ctg aag cgc ctg gca gct cgc 1027
Thr Asp Leu Cys Ala Ala Leu Ala Ser Leu Lys Arg Leu Ala Ala Arg
295 300 305
ggc cca atc gca gtg tct acc tct tgt tca ctg ctg cac gtt cct tac 1075
Gly Pro Ile Ala Val Ser Thr Ser Cys Ser Leu Leu His Val Pro Tyr
310 315 320 325
acc ctc gag gct gag aac att gag cct gag gtc cgc gac tgg ctt gcc 1123
Thr Leu Glu Ala Glu Asn Ile Glu Pro Glu Val Arg Asp Trp Leu Ala
330 335 340
ttc ggc tcg gag aag atc acc gag gtc aag ctg ctt gcc gac gcc cta 1171
Phe Gly Ser Glu Lys Ile Thr Glu Val Lys Leu Leu Ala Asp Ala Leu
345 350 355
gcc ggc aac atc gac gcg gct gcg ttc gat gcg gcg tcc gca gca att 1219
Ala Gly Asn Ile Asp Ala Ala Ala Phe Asp Ala Ala Ser Ala Ala Ile
360 365 370
gct tct cga cgc acc tcc cca cgc acc gca cca atc acg cag gaa ctc 1267
Ala Ser Arg Arg Thr Ser Pro Arg Thr Ala Pro Ile Thr Gln Glu Leu
375 380 385
cct ggc cgt agc cgt gga tcc ttc gac act cgt gtt acg ctg cag gag 1315
Pro Gly Arg Ser Arg Gly Ser Phe Asp Thr Arg Val Thr Leu Gln Glu
390 395 400 405
aag tca ctg gag ctt cca gct ctg cca acc acc acc att ggt tct ttc 1363
Lys Ser Leu Glu Leu Pro Ala Leu Pro Thr Thr Thr Ile Gly Ser Phe
410 415 420
cca cag acc cca tcc att cgt tct gct cgc gct cgt ctg cgc aag gaa 1411
Pro Gln Thr Pro Ser Ile Arg Ser Ala Arg Ala Arg Leu Arg Lys Glu
425 430 435
tcc atc act ttg gag cag tac gaa gag gca atg cgc gaa gaa atc gat 1459
Ser Ile Thr Leu Glu Gln Tyr Glu Glu Ala Met Arg Glu Glu Ile Asp
440 445 450
ctg gtc atc gcc aag cag gaa gaa ctt ggt ctt gat gtg ttg gtt cac 1507
Leu Val Ile Ala Lys Gln Glu Glu Leu Gly Leu Asp Val Leu Val His
455 460 465
ggt gag cca gag cgc aac gac atg gtt cag tac ttc tct gaa ctt ctc 1555
Gly Glu Pro Glu Arg Asn Asp Met Val Gln Tyr Phe Ser Glu Leu Leu
470 475 480 485
gac ggt ttc ctc tca acc gcc aac ggc tgg gtc caa agc tac ggc tcc 1603
Asp Gly Phe Leu Ser Thr Ala Asn Gly Trp Val Gln Ser Tyr Gly Ser
490 495 500
cgc tgt gtt cgt cct cca gtg ttg ttc gga aac gtt tcc cgc cca gcg 1651
Arg Cys Val Arg Pro Pro Val Leu Phe Gly Asn Val Ser Arg Pro Ala
505 510 515
cca atg act gtc aag tgg ttc cag tac gca cag agc ctg acc cag aag 1699
Pro Met Thr Val Lys Trp Phe Gln Tyr Ala Gln Ser Leu Thr Gln Lys
520 525 530
cat gtc aag gga atg ctc acc ggt cca gtc acc atc ctt gca tgg tcc 1747
His Val Lys Gly Met Leu Thr Gly Pro Val Thr Ile Leu Ala Trp Ser
535 540 545
ttc gtt cgc gat gat cag ccg ctg gct acc act gct gac cag gtt gca 1795
Phe Val Arg Asp Asp Gln Pro Leu Ala Thr Thr Ala Asp Gln Val Ala
550 555 560 565
ctg gca ctg cgc gat gaa att aac gat ctc atc gag gct ggc gcg aag 1843
Leu Ala Leu Arg Asp Glu Ile Asn Asp Leu Ile Glu Ala Gly Ala Lys
570 575 580
atc atc cag gtg gat gag cct gcg att cgt gaa ctg ttg ccg cta cga 1891
Ile Ile Gln Val Asp Glu Pro Ala Ile Arg Glu Leu Leu Pro Leu Arg
585 590 595
gac gtc gat aag cct gcc tac ctg cag tgg tcc gtg gac tcc ttc cgc 1939
Asp Val Asp Lys Pro Ala Tyr Leu Gln Trp Ser Val Asp Ser Phe Arg
600 605 610
ctg gcg act gcc ggc gca ccc gac gac gtc caa atc cac acc cac atg 1987
Leu Ala Thr Ala Gly Ala Pro Asp Asp Val Gln Ile His Thr His Met
615 620 625
tgc tac tcc gag ttc aac gaa gtg atc tcc tcg gtc atc gcg ttg gat 2035
Cys Tyr Ser Glu Phe Asn Glu Val Ile Ser Ser Val Ile Ala Leu Asp
630 635 640 645
gcc gat gtc acc acc atc gaa gca gca cgt tcc gac atg cag gtc ctc 2083
Ala Asp Val Thr Thr Ile Glu Ala Ala Arg Ser Asp Met Gln Val Leu
650 655 660
gct gct ctg aaa tct tcc ggc ttc gag ctc ggc gtc gga cct ggt gtg 2131
Ala Ala Leu Lys Ser Ser Gly Phe Glu Leu Gly Val Gly Pro Gly Val
665 670 675
tgg gat atc cac tcc ccg cgc gtt cct tcc gcg cag aaa gtg gac ggt 2179
Trp Asp Ile His Ser Pro Arg Val Pro Ser Ala Gln Lys Val Asp Gly
680 685 690
ctc ctc gag gct gca ctg cag tcc gtg gat cct cgc cag ctg tgg gtc 2227
Leu Leu Glu Ala Ala Leu Gln Ser Val Asp Pro Arg Gln Leu Trp Val
695 700 705
aac cca gac tgt ggt ctg aag acc cgt gga tgg cca gaa gtg gaa gct 2275
Asn Pro Asp Cys Gly Leu Lys Thr Arg Gly Trp Pro Glu Val Glu Ala
710 715 720 725
tcc cta aag gtt ctc gtt gag tcc gct aag cag gct cgt gag aaa atc 2323
Ser Leu Lys Val Leu Val Glu Ser Ala Lys Gln Ala Arg Glu Lys Ile
730 735 740
gga gca act atc taaattgggt taccgctagg aac 2358
Gly Ala Thr Ile
745
<210>104
<211>745
<212>PRT
<213>谷氨酸棒杆菌
<400>104
Met Thr Ser Asn Phe Ser Ser Thr Val Ala Gly Leu Pro Arg Ile Gly
1 5 10 15
Ala Lys Arg Glu Leu Lys Phe Ala Leu Glu Gly Tyr Trp Asn Gly Ser
20 25 30
Ile Glu Gly Arg Glu Leu Ala Gln Thr Ala Arg Gln Leu Val Asn Thr
35 40 45
Ala Ser Asp Ser Leu Ser Gly Leu Asp Ser Val Pro Phe Ala Gly Arg
50 55 60
Ser Tyr Tyr Asp Ala Met Leu Asp Thr Ala Ala Ile Leu Gly Val Leu
65 70 75 80
Pro Glu Arg Phe Asp Asp Ile Ala Asp His Glu Asn Asp Gly Leu Pro
85 90 95
Leu Trp Ile Asp Arg Tyr Phe Gly Ala Ala Arg Gly Thr Glu Thr Leu
100 105 110
Pro Ala Gln Ala Met Thr Lys Trp Phe Asp Thr Asn Tyr His Tyr Leu
115 120 125
Val Pro Glu Leu Ser Ala Asp Thr Arg Phe Val Leu Asp Ala Ser Ala
130 135 140
Leu Ile Glu Asp Leu Arg Cys Gln Gln Val Arg Gly Val Asn Ala Arg
145 150 155 160
Pro Val Leu Val Gly Pro Leu Thr Phe Leu Ser Leu Ala Arg Thr Thr
165 170 175
Asp Gly Ser Asn Pro Leu Asp His Lau Pro Ala Leu Phe Glu Val Tyr
180 185 190
Glu Arg Leu Ile Lys Ser Phe Asp Thr Glu Trp Val Gln Ile Asp Glu
195 200 205
Pro Ala Leu Val Thr Asp Val Ala Pro Glu Val Leu Glu Gln Val Arg
210 215 220
Ala Gly Tyr Thr Thr Leu Ala Lys Arg Asp Gly Val Phe Val Asn Thr
225 230 235 240
Tyr Phe Gly Ser Gly Asp Gln Ala Leu Asn Thr Leu Ala Gly Ile Gly
245 250 255
Leu Gly Ala Ile Gly Val Asp Leu Val Thr His Gly Val Thr Glu Leu
260 265 270
Ala Ala Trp Lys Gly Glu Glu Leu Leu Val Ala Gly Ile Val Asp Gly
275 280 285
Arg Asn Ile Trp Arg Thr Asp Leu Cys Ala Ala Leu Ala Ser Leu Lys
290 295 300
Arg Leu Ala Ala Arg Gly Pro Ile Ala Val Ser Thr Ser Cys Ser Leu
305 310 315 320
Leu His Val Pro Tyr Thr Leu Glu Ala Glu Asn Ile Glu Pro Glu Val
325 330 335
Arg Asp Trp Leu Ala Phe Gly Ser Glu Lys Ile Thr Glu Val Lys Leu
340 345 350
Leu Ala Asp Ala Leu Ala Gly Asn Ile Asp Ala Ala Ala Phe Asp Ala
355 360 365
Ala Ser Ala Ala Ile Ala Ser Arg Arg Thr Ser Pro Arg Thr Ala Pro
370 375 380
Ile Thr Gln Glu Leu Pro Gly Arg Ser Arg Gly Ser Phe Asp Thr Arg
385 390 395 400
Val Thr Leu Gln Glu Lys Ser Leu Glu Leu Pro Ala Leu Pro Thr Thr
405 410 415
Thr Ile Gly Ser Phe Pro Gln Thr Pro Ser Ile Arg Ser Ala Arg Ala
420 425 430
Arg Leu Arg Lys Glu Ser Ile Thr Leu Glu Gln Tyr Glu Glu Ala Met
435 440 445
Arg Glu Glu Ile Asp Leu Val Ile Ala Lys Gln Glu Glu Leu Gly Leu
450 455 460
Asp Val Leu Val His Gly Glu Pro Glu Arg Asn Asp Met Val Gln Tyr
465 470 475 480
Phe Ser Glu Leu Leu Asp Gly Phe Leu Ser Thr Ala Asn Gly Trp Val
485 490 495
Gln Ser Tyr Gly Ser Arg Cys Val Arg Pro Pro Val Leu Phe Gly Asn
500 505 510
Val Ser Arg Pro Ala Pro Met Thr Val Lys Trp Phe Gln Tyr Ala Gln
515 520 525
Ser Leu Thr Gln Lys His Val Lys Gly Met Leu Thr Gly Pro Val Thr
530 535 540
Ile Leu Ala Trp Ser Phe Val Arg Asp Asp Gln Pro Leu Ala Thr Thr
545 550 555 560
Ala Asp Gln Val Ala Leu Ala Leu Arg Asp Glu Ile Asn Asp Leu Ile
565 570 575
Glu Ala Gly Ala Lys Ile Ile Gln Val Asp Glu Pro Ala Ile Arg Glu
580 585 590
Leu Leu Pro Leu Arg Asp Val Asp Lys Pro Ala Tyr Leu Gln Trp Ser
595 600 605
Val Asp Ser Phe Arg Leu Ala Thr Ala Gly Ala Pro Asp Asp Val Gln
610 615 620
Ile His Thr His Met Cys Tyr Ser Glu Phe Asn Glu Val Ile Ser Ser
625 630 635 640
Val Ile Ala Leu Asp Ala Asp Val Thr Thr Ile Glu Ala Ala Arg Ser
645 650 655
Asp Met Gln Val Leu Ala Ala Leu Lys Ser Ser Gly Phe Glu Leu Gly
660 665 670
Val Gly Pro Gly Val Trp Asp Ile His Ser Pro Arg Val Pro Ser Ala
675 680 685
Gln Lys Val Asp Gly Leu Leu Glu Ala Ala Leu Gln Ser Val Asp Pro
690 695 700
Arg Gln Leu Trp Val Asn Pro Asp Cys Gly Leu Lys Thr Arg Gly Trp
705 710 715 720
Pro Glu Val Glu Ala Ser Leu Lys Val Leu Val Glu Ser Ala Lys Gln
725 730 735
Ala Arg Glu Lys Ile Gly Ala Thr Ile
740 745
<210>105
<211>1923
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1900)
<223>FRXA02085
<400>105
cacccggtga tttcgcgaac cttgaaacat cgtcagaaga ttgccgtgcg tcctagccgg 60
gatccgGaGg ttcggctcaa gcagaaagtc tttaactcac atg act tcc aac ttt 115
Met Thr Ser Asn Phe
1 5
tct tcc act gtc gct ggt ctt cct cgc atc gga gcg aag cgt gaa ctg 163
Ser Ser Thr Val Ala Gly Leu Pro Arg Ile Gly Ala Lys Arg Glu Leu
10 15 20
aag ttc gcg ctc gaa ggc tac tgg aat gga tca att gaa ggt cgc gaa 211
Lys Phe Ala Leu Glu Gly Tyr Trp Asn Gly Ser Ile Glu Gly Arg Glu
25 30 35
ctt gcg cag acc gcc cgc caa ttg gtc aac act gca tcg gat tct ttg 259
Leu Ala Gln Thr Ala Arg Gln Leu Val Asn Thr Ala Ser Asp Ser Leu
40 45 50
tct gga ttg gat tcc gtt ccg ttt gca gga cgt tcc tac tac gac gca 307
Ser Gly Leu Asp Ser Val Pro Phe Ala Gly Arg Ser Tyr Tyr Asp Ala
55 60 65
atg ctc gat acc gcc gct att ttg ggt gtg ctg ccg gag cgt ttt gat 355
Met Leu Asp Thr Ala Ala Ile Leu Gly Val Leu Pro Glu Arg Phe Asp
70 75 80 85
gac atc gct gat cat gaa aac gat ggt ctc cca ctg tgg att gac cgc 403
Asp Ile Ala Asp His Glu Asn Asp Gly Leu Pro Leu Trp Ile Asp Arg
90 95 100
tac ttt ggc gct gct cgc ggt act gag acc ctg cct gca cag gca atg 451
Tyr Phe Gly Ala Ala Arg Gly Thr Glu Thr Leu Pro Ala Gln Ala Met
105 110 115
acc aag tgg ttt gat acc aac tac cac tac ctc gtg ccg gag ttg tct 499
Thr Lys Trp Phe Asp Thr Asn Tyr His Tyr Leu Val Pro Glu Leu Ser
120 125 130
gcg gat aca cgt ttc gtt ttg gat gcg tcc gcg ctg att gag gat ctc 547
Ala Asp Thr Arg Phe Val Leu Asp Ala Ser Ala Leu Ile Glu Asp Leu
135 140 145
cgt tgc cag cag gtt cgt ggc gtt aat gcc cgc cct gtt ctg gtt ggt 595
Arg Cys Gln Gln Val Arg Gly Val Asn Ala Arg Pro Val Leu Val Gly
150 155 160 165
cca ctg act ttc ctt tcc ctt gct cgc acc act gat ggt tcc aat cct 643
Pro Leu Thr Phe Leu Ser Leu Ala Arg Thr Thr Asp Gly Ser Asn Pro
170 175 180
ttg gat cac ctg cct gca ctg ttt gag gtc tac gag cgc ctc atc aag 691
Leu Asp His Leu Pro Ala Leu Phe Glu Val Tyr Glu Arg Leu Ile Lys
185 190 195
tct ttc gat act gag tgg gtt cag atc gat gag cct gcg ttg gtc acc 739
Ser Phe Asp Thr Glu Trp Val Gln Ile Asp Glu Pro Ala Leu Val Thr
200 205 210
gat gtt gct cct gag gtt ttg gag cag gtc cgc gct ggt tac acc act 787
Asp Val Ala Pro Glu Val Leu Glu Gln Val Arg Ala Gly Tyr Thr Thr
215 220 225
ttg gct aag cgc gat ggc gtg ttt gtc aat act tac ttc ggc tct ggc 835
Leu Ala Lys Arg Asp Gly Val Phe Val Asn Thr Tyr Phe Gly Ser Gly
230 235 240 245
gat cag gcg ctg aac act ctt gcg ggc atc ggc ctt ggc gcg att ggc 883
Asp Gln Ala Leu Asn Thr Leu Ala Gly Ile Gly Leu Gly Ala Ile Gly
250 255 260
gtt gac ttg gtc acc cat ggc gtc act gag ctt gct gcg tgg aag ggt 931
Val Asp Leu Val Thr His Gly Val Thr Glu Leu Ala Ala Trp Lys Gly
265 270 275
gag gag ctg ctg gtt gcg ggc atc gtt gat ggt cgt aac att tgg cgc 979
Glu Glu Leu Leu Val Ala Gly Ile Val Asp Gly Arg Asn Ile Trp Arg
280 285 290
acc gac ctg tgt gct gct ctt gct tcc ctg aag cgc ctg gca gct cgc 1027
Thr Asp Leu Cys Ala Ala Leu Ala Ser Leu Lys Arg Leu Ala Ala Arg
295 300 305
ggc cca atc gca gtg tct acc tct tgt tca ctg ctg cac gtt cct tac 1075
Gly Pro Ile Ala Val Ser Thr Ser Cys Ser Leu Leu His Val Pro Tyr
310 315 320 325
acc ctc gag gct gag aac att gag cct gag gtc cgc gac tgg ctt gcc 1123
Thr Leu Glu Ala Glu Asn Ile Glu Pro Glu Val Arg Asp Trp Leu Ala
330 335 340
ttc ggc tcg gag aag atc acc gag gtc aag ctg ctt gcc gac gcc cta 1171
Phe Gly Ser Glu Lys Ile Thr Glu Val Lys Leu Leu Ala Asp Ala Leu
345 350 355
gcc ggc aac atc gac gcg gct gcg ttc gat gcg gcg tcc gca gca att 1219
Ala Gly Asn Ile Asp Ala Ala Ala Phe Asp Ala Ala Ser Ala Ala Ile
360 365 370
gct tct cga cgc acc tcc cca cgc acc gca cca atc acg cag gaa ctc 1267
Ala Ser Arg Arg Thr Ser Pro Arg Thr Ala Pro Ile Thr Gln Glu Leu
375 380 385
cct ggc cgt agc cgt gga tcc ttc gac act cgt gtt acg ctg cag gag 1315
Pro Gly Arg Ser Arg Gly Ser Phe Asp Thr Arg Val Thr Leu Gln Glu
390 395 400 405
aag tca ctg gag ctt cca gct ctg cca acc acc acc att ggt tct ttc 1363
Lys Ser Leu Glu Leu Pro Ala Leu Pro Thr Thr Thr Ile Gly Ser Phe
410 415 420
cca cag acc cca tcc att cgt tct gct cgc gct cgt ctg cgc aag gaa 1411
Pro Gln Thr Pro Ser Ile Arg Ser Ala Arg Ala Arg Leu Arg Lys Glu
425 430 435
tcc atc act ttg gag cag tac gaa gag gca atg cgc gaa gaa atc gat 1459
Ser Ile Thr Leu Glu Gln Tyr Glu Glu Ala Met Arg Glu Glu Ile Asp
440 445 450
ctg gtc atc gcc aag cag gaa gaa ctt ggt ctt gat gtg ttg gtt cac 1507
Leu Val Ile Ala Lys Gln Glu Glu Leu Gly Leu Asp Val Leu Val His
455 460 465
ggt gag cca gag cgc aac gac atg gtt cag tac ttc tct gaa ctt ctc 1555
Gly Glu Pro Glu Arg Asn Asp Met Val Gln Tyr Phe Ser Glu Leu Leu
470 475 480 485
gac ggt ttc ctc tca acc gcc aac ggc tgg gtc caa agc tac ggc tcc 1603
Asp Gly Phe Leu Ser Thr Ala Asn Gly Trp Val Gln Ser Tyr Gly Ser
490 495 500
cgc tgt gtt cgt cct cca gtg ttg ttc gga aac gtt tcc cgc cca gcg 1651
Arg Cys Val Arg Pro Pro Val Leu Phe Gly Asn Val Ser Arg Pro Ala
505 510 515
cca atg act gtc aag tgg ttc cag tac gca cag agc ctg acc cag aag 1699
Pro Met Thr Val Lys Trp Phe Gln Tyr Ala Gln Ser Leu Thr Gln Lys
520 525 530
cat gtc aag gga atg ctc acc ggt cca gtc acc atc ctt gca tgg tcc 1747
His Val Lys Gly Met Leu Thr Gly Pro Val Thr Ile Leu Ala Trp Ser
535 540 545
ttc gtt cgc gat gat cag ccg ctg gct acc act gct gac cag gtt gca 1795
Phe Val Arg Asp Asp Gln Pro Leu Ala Thr Thr Ala Asp Gln Val Ala
550 555 560 565
ctg gca ctg cgc gat gaa att aac gat ctc atc gag gct ggc gcg aag 1843
Leu Ala Leu Arg Asp Glu Ile Asn Asp Leu Ile Glu Ala Gly Ala Lys
570 575 580
atc atc cag gtg gat gag cct gcg att cgt gaa ctg ttg ccc gct acg 1891
Ile Ile Gln Val Asp Glu Pro Ala Ile Arg Glu Leu Leu Pro Ala Thr
585 590 595
aga cgt cga taagcctgcc tacctgcagt ggt 1923
Arg Arg Arg
600
<210>106
<211>600
<212>PRT
<213>谷氨酸棒杆菌
<400>106
Met Thr Ser Asn Phe Ser Ser Thr Val Ala Gly Leu Pro Arg Ile Gly
1 5 10 15
Ala Lys Arg Glu Leu Lys Phe Ala Leu Glu Gly Tyr Trp Asn Gly Ser
20 25 30
Ile Glu Gly Arg Glu Leu Ala Gln Thr Ala Arg Gln Leu Val Asn Thr
35 40 45
Ala Ser Asp Ser Leu Ser Gly Leu Asp Ser Val Pro Phe Ala Gly Arg
50 55 60
Ser Tyr Tyr Asp Ala Met Leu Asp Thr Ala Ala Ile Leu Gly Val Leu
65 70 75 80
Pro Glu Arg Phe Asp Asp Ile Ala Asp His Glu Asn Asp Gly Leu Pro
85 90 95
Leu Trp Ile Asp Arg Tyr Phe Gly Ala Ala Arg Gly Thr Glu Thr Leu
100 105 110
Pro Ala Gln Ala Met Thr Lys Trp Phe Asp Thr Asn Tyr His Tyr Leu
115 120 125
Val Pro Glu Leu Ser Ala Asp Thr Arg Phe Val Leu Asp Ala Ser Ala
130 135 140
Leu Ile Glu Asp Leu Arg Cys Gln Gln Val Arg Gly Val Asn Ala Arg
145 150 155 160
Pro Val Leu Val Gly Pro Leu Thr Phe Leu Ser Leu Ala Arg Thr Thr
165 170 175
Asp Gly Ser Asn Pro Leu Asp His Leu Pro Ala Leu Phe Glu Val Tyr
180 185 190
Glu Arg Leu Ile Lys Ser Phe Asp Thr Glu Trp Val Gln Ile Asp Glu
195 200 205
Pro Ala Leu Val Thr Asp Val Ala Pro Glu Val Leu Glu Gln Val Arg
210 215 220
Ala Gly Tyr Thr Thr Leu Ala Lys Arg Asp Gly Val Phe Val Asn Thr
225 230 235 240
Tyr Phe Gly Ser Gly Asp Gln Ala Leu Asn Thr Leu Ala Gly Ile Gly
245 250 255
Leu Gly Ala Ile Gly Val Asp Leu Val Thr His Gly Val Thr Glu Leu
260 265 270
Ala Ala Trp Lys Gly Glu Glu Leu Leu Val Ala Gly Ile Val Asp Gly
275 280 285
Arg Asn Ile Trp Arg Thr Asp Leu Cys Ala Ala Leu Ala Ser Leu Lys
290 295 300
Arg Leu Ala Ala Arg Gly Pro Ile Ala Val Ser Thr Ser Cys Ser Leu
305 310 315 320
Leu His Val Pro Tyr Thr Leu Glu Ala Glu Asn Ile Glu Pro Glu Val
325 330 335
Arg Asp Trp Leu Ala Phe Gly Ser Glu Lys Ile Thr Glu Val Lys Leu
340 345 350
Leu Ala Asp Ala Leu Ala Gly Asn Ile Asp Ala Ala Ala Phe Asp Ala
355 360 365
Ala Ser Ala Ala Ile Ala Ser Arg Arg Thr Ser Pro Arg Thr Ala Pro
370 375 380
Ile Thr Gln Glu Leu Pro Gly Arg Ser Arg Gly Ser Phe Asp Thr Arg
385 390 395 400
Val Thr Leu Gln Glu Lys Ser Leu Glu Leu Pro Ala Leu Pro Thr Thr
405 410 415
Thr Ile Gly Ser Phe Pro Gln Thr Pro Ser Ile Arg Ser Ala Arg Ala
420 425 430
Arg Leu Arg Lys Glu Ser Ile Thr Leu Glu Gln Tyr Glu Glu Ala Met
435 440 445
Arg Glu Glu Ile Asp Leu Val Ile Ala Lys Gln Glu Glu Leu Gly Leu
450 455 460
Asp Val Leu Val His Gly Glu Pro Glu Arg Asn Asp Met Val Gln Tyr
465 470 475 480
Phe Ser Glu Leu Leu Asp Gly Phe Leu Ser Thr Ala Asn Gly Trp Val
485 490 495
Gln Ser Tyr Gly Ser Arg Cys Val Arg Pro Pro Val Leu Phe Gly Asn
500 505 510
Val Ser Arg Pro Ala Pro Met Thr Val Lys Trp Phe Gln Tyr Ala Gln
515 520 525
Ser Leu Thr Gln Lys His Val Lys Gly Met Leu Thr Gly Pro Val Thr
530 535 540
Ile Leu Ala Trp Ser Phe Val Arg Asp Asp Gln Pro Leu Ala Thr Thr
545 550 555 560
Ala Asp Gln Val Ala Leu Ala Leu Arg Asp Glu Ile Asn Asp Leu Ile
565 570 575
Glu Ala Gly Ala Lys Ile Ile Gln Val Asp Glu Pro Ala Ile Arg Glu
580 585 590
Leu Leu Pro Ala Thr Arg Arg Arg
595 600
<210>107
<211>603
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(580)
<223>FRXA02086
<400>107
gatgatcagc cgctggctac cactgctgac caggttgcac tggcactgcg cgatgaaatt 60
aacgatctca tcgaggctgg cgcgaagatc atccaggtgg atg agc ctg cga ttc 115
Met Ser Leu Arg Phe
1 5
gtg aac tgt tgc ccg cta cga gac gtc gat aag cct gcc tac ctg cag 163
Val Asn Cys Cys Pro Leu Arg Asp Val Asp Lys Pro Ala Tyr Leu Gln
10 15 20
tgg tcc gtg gac tcc ttc cgc ctg gcg act gcc ggc gca ccc gac gac 211
Trp Ser Val Asp Ser Phe Arg Leu Ala Thr Ala Gly Ala Pro Asp Asp
25 30 35
gtc caa atc cac acc cac atg tgc tac tcc gag ttc aac gaa gtg atc 259
Val Gln Ile His Thr His Met Cys Tyr Ser Glu Phe Asn Glu Val Ile
40 45 50
tcc tcg gtc atc gcg ttg gat gcc gat gtc acc acc atc gaa gca gca 307
Ser Ser Val Ile Ala Leu Asp Ala Asp Val Thr Thr Ile Glu Ala Ala
55 60 65
cgt tcc gac atg cag gtc ctc gct gct ctg aaa tct tcc ggc ttc gag 355
Arg Ser Asp Met Gln Val Leu Ala Ala Leu Lys Ser Ser Gly Phe Glu
70 75 80 85
ctc ggc gtc gga cct ggt gtg tgg gat atc cac tcc ccg cgc gtt cct 403
Leu Gly Val Gly Pro Gly Val Trp Asp Ile His Ser Pro Arg Val Pro
90 95 100
tcc gcg cag aaa gtg gac ggt ctc ctc gag gct gca ctg cag tcc gtg 451
Ser Ala Gln Lys Val Asp Gly Leu Leu Glu Ala Ala Leu Gln Ser Val
105 110 115
gat cct cgc cag ctg tgg gtc aac cca gac tgt ggt ctg aag acc cgt 499
Asp Pro Arg Gln Leu Trp Val Asn Pro Asp Cys Gly Leu Lys Thr Arg
120 125 130
gga tgg cca gaa gtg gaa gct tcc cta aag gtt ctc gtt gag tcc gct 547
Gly Trp Pro Glu Val Glu Ala Ser Leu Lys Val Leu Val Glu Ser Ala
135 140 145
aag cag gct cgt gag aaa atc gga gca act atc taaattgggt taccgctagg 600
Lys Gln Ala Arg Glu Lys Ile Gly Ala Thr Ile
150 155 160
aac 603
<210>108
<211>160
<212>PRT
<213>谷氨酸棒杆菌
<400>108
Met Ser Leu Arg Phe Val Asn Cys Cys Pro Leu Arg Asp Val Asp Lys
1 5 10 15
Pro Ala Tyr Leu Gln Trp Ser Val Asp Ser Phe Arg Leu Ala Thr Ala
20 25 30
Gly Ala Pro Asp Asp Val Gln Ile His Thr His Met Cys Tyr Ser Glu
35 40 45
Phe Asn Glu Val Ile Ser Ser Val Ile Ala Leu Asp Ala Asp Val Thr
50 55 60
Thr Ile Glu Ala Ala Arg Ser Asp Met Gln Val Leu Ala Ala Leu Lys
65 70 75 80
Ser Ser Gly Phe Glu Leu Gly Val Gly Pro Gly Val Trp Asp Ile His
85 90 95
Ser Pro Arg Val Pro Ser Ala Gln Lys Val Asp Gly Leu Leu Glu Ala
100 105 110
Ala Leu Gln Ser Val Asp Pro Arg Gln Leu Trp Val Asn Pro Asp Cys
115 120 125
Gly Leu Lys Thr Arg Gly Trp Pro Glu Val Glu Ala Ser Leu Lys Val
130 135 140
Leu Val Glu Ser Ala Lys Gln Ala Arg Glu Lys Ile Gly Ala Thr Ile
145 150 155 160
<210>109
<211>1326
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1303)
<223>RXN02648
<400>109
atgaataaaa ttccgggtgc agtgaccgta ggtgaggtaa acgcggttag agtcgaatga 60
gagtttgata ctttctttcg acttttagat tggattttca atg agc cag aac cgc 115
Met Ser Gln Asn Arg
1 5
atc agg acc act cac gtt ggt tcc ttg ccc cgt acc cca gag cta ctt 163
Ile Arg Thr Thr His Val Gly Ser Leu Pro Arg Thr Pro Glu Leu Leu
10 15 20
gat gca aac atc aag cgt tct aac ggt gag att ggg gag gag gaa ttc 211
Asp Ala Asn Ile Lys Arg Ser Asn Gly Glu Ile Gly Glu Glu Glu Phe
25 30 35
ttc cag att ctg cag tct tct gta gat gac gtg atc aag cgc cag gtt 259
Phe Gln Ile Leu Gln Ser Ser Val Asp Asp Val Ile Lys Arg Gln Val
40 45 50
gac ctg ggt atc gac atc ctt aac gag ggc gaa tac ggc cac gtc acc 307
Asp Leu Gly Ile Asp Ile Leu Asn Glu Gly Glu Tyr Gly His Val Thr
55 60 65
tcc ggt gca gtt gac ttc ggt gca tgg tgg aac tac tcc ttc acc cgc 355
Ser Gly Ala Val Asp Phe Gly Ala Trp Trp Asn Tyr Ser Phe Thr Arg
70 75 80 85
ctg ggc gga ctg acc atg acc gat acc gac cgt tgg gca agc cag gaa 403
Leu Gly Gly Leu Thr Met Thr Asp Thr Asp Arg Trp Ala Ser Gln Glu
90 95 100
gca gtg cgt tcc acc cct ggc aac atc gag ctg acc agc ttc tct gat 451
Ala Val Arg Ser Thr Pro Gly Asn Ile Glu Leu Thr Ser Phe Ser Asp
105 110 115
cgt cgc gac cgc gca ttg ttc agc gaa gca tac gag gat cca gta tct 499
Arg Arg Asp Arg Ala Leu Phe Ser Glu Ala Tyr Glu Asp Pro Val Ser
120 125 130
ggc atc ttc acc ggt cgc gct tct gtg ggc aac cca gag ttc acc gga 547
Gly Ile Phe Thr Gly Arg Ala Ser Val Gly Asn Pro Glu Phe Thr Gly
135 140 145
cct att acc tac att ggc cag gaa gaa act cag acg gat gtt gat ctg 595
Pro Ile Thr Tyr Ile Gly Gln Glu Glu Thr Gln Thr Asp Val Asp Leu
150 155 160 165
ctg aag aag ggc atg aac gca gcg gga gct acc gac ggc ttc gtt gca 643
Leu Lys Lys Gly Met Asn Ala Ala Gly Ala Thr Asp Gly Phe Val Ala
170 175 180
gca cta tcc cca gga tct gca gct cga ttg acc aac aag ttc tac gac 691
Ala Leu Ser Pro Gly Ser Ala Ala Arg Leu Thr Asn Lys Phe Tyr Asp
185 190 195
act gat gaa gaa gtc gtc gca gca tgt gct gat gcg ctt tcc cag gaa 739
Thr Asp Glu Glu Val Val Ala Ala Cys Ala Asp Ala Leu Ser Gln Glu
200 205 210
tac aag atc atc acc gat gca ggt ctg acc gtt cag ctc gac gca ccg 787
Tyr Lys Ile Ile Thr Asp Ala Gly Leu Thr Val Gln Leu Asp Ala Pro
215 220 225
gac ttg gca gaa gca tgg gat cag atc aac cca gag cca agc gtg aag 835
Asp Leu Ala Glu Ala Trp Asp Gln Ile Asn Pro Glu Pro Ser Val Lys
230 235 240 245
gat tac ttg gac tgg atc ggt aca cgc atc gat gcc atc aac agt gca 883
Asp Tyr Leu Asp Trp Ile Gly Thr Arg Ile Asp Ala Ile Asn Ser Ala
250 255 260
gtg aag ggc ctt cca aag gaa cag acc cgc ctg cac atc tgc tgg ggc 93l
Val Lys Gly Leu Pro Lys Glu Gln Thr Arg Leu His Ile Cys Trp Gly
265 270 275
tct tgg cac gga cca cac gtc act gac atc cca ttc ggt gac atc att 979
Ser Trp His Gly Pro His Val Thr Asp Ile Pro Phe Gly Asp Ile Ile
280 285 290
ggt gag atc ctg cgc gca gag gtc ggt ggc ttc tcc ttc gaa ggc gca 1027
Gly Glu Ile Leu Arg Ala Glu Val Gly Gly Phe Ser Phe Glu Gly Ala
295 300 305
tct cct cgt cac gca cac gag tgg cgt gta tgg gaa gaa aac aag ctt 1075
Ser Pro Arg His Ala His Glu Trp Arg Val Trp Glu Glu Asn Lys Leu
310 315 320 325
cct gaa ggc tct gtt atc tac cct ggt gtt gtg tct cac tcc atc aac 1123
Pro Glu Gly Ser Val Ile Tyr Pro Gly Val Val Ser His Ser Ile Asn
330 335 340
gct gtg gag cac cca cgc ctg gtt gct gat cgt atc gtt cag ttc gcc 1171
Ala Val Glu His Pro Arg Leu Val Ala Asp Arg Ile Val Gln Phe Ala
345 350 355
aag ctt gtt ggc cct gag aac gtc att gcg tcc act gac tgt ggt ctg 1219
Lys Leu Val Gly Pro Glu Asn Val Ile Ala Ser Thr Asp Cys Gly Leu
360 365 370
ggc gga cgt ctg cat tcc cag atc gca tgg gca aag ctg gag tcc cta 1267
Gly Gly Arg Leu His Ser Gln Ile Ala Trp Ala Lys Leu Glu Ser Leu
375 380 385
gta gag ggc gct cgc att gca tca aag gaa ctg ttc taagctagac 1313
Val Glu Gly Ala Arg Ile Ala Ser Lys Glu Leu Phe
390 395 400
aacgagggtt gct 1326
<210>110
<211>401
<212>PRT
<213>谷氨酸棒杆菌
<400>110
Met Ser Gln Asn Arg Ile Arg Thr Thr His Val Gly Ser Leu Pro Arg
1 5 10 15
Thr Pro Glu Leu Leu Asp Ala Asn Ile Lys Arg Ser Asn Gly Glu Ile
20 25 30
Gly Glu Glu Glu Phe Phe Gln Ile Leu Gln Ser Ser Val Asp Asp Val
35 40 45
Ile Lys Arg Gln Val Asp Leu Gly Ile Asp Ile Leu Asn Glu Gly Glu
50 55 60
Tyr Gly His Val Thr Ser Gly Ala Val Asp Phe Gly Ala Trp Trp Asn
65 70 75 80
Tyr Ser Phe Thr Arg Leu Gly Gly Leu Thr Met Thr Asp Thr Asp Arg
85 90 95
Trp Ala Ser Gln Glu Ala Val Arg Ser Thr Pro Gly Asn Ile Glu Leu
100 105 110
Thr Ser Phe Ser Asp Arg Arg Asp Arg Ala Leu Phe Ser Glu Ala Tyr
115 120 125
Glu Asp Pro Val Ser Gly Ile Phe Thr Gly Arg Ala Ser Val Gly Asn
130 135 140
Pro Glu Phe Thr Gly Pro Ile Thr Tyr Ile Gly Gln Glu Glu Thr Gln
145 150 155 160
Thr Asp Val Asp Leu Leu Lys Lys Gly Met Asn Ala Ala Gly Ala Thr
165 170 175
Asp Gly Phe Val Ala Ala Leu Ser Pro Gly Ser Ala Ala Arg Leu Thr
180 185 190
Asn Lys Phe Tyr Asp Thr Asp Glu Glu Val Val Ala Ala Cys Ala Asp
195 200 205
Ala Leu Ser Gln Glu Tyr Lys Ile Ile Thr Asp Ala Gly Leu Thr Val
210 215 220
Gln Leu Asp Ala Pro Asp Leu Ala Glu Ala Trp Asp Gln Ile Asn Pro
225 230 235 240
Glu Pro Ser Val Lys Asp Tyr Leu Asp Trp Ile Gly Thr Arg Ile Asp
245 250 255
Ala Ile Asn Ser Ala Val Lys Gly Leu Pro Lys Glu Gln Thr Arg Leu
260 265 270
His Ile Cys Trp Gly Ser Trp His Gly Pro His Val Thr Asp Ile Pro
275 280 285
Phe Gly Asp Ile Ile Gly Glu Ile Leu Arg Ala Glu Val Gly Gly Phe
290 295 300
Ser Phe Glu Gly Ala Ser Pro Arg His Ala His Glu Trp Arg Val Trp
305 310 315 320
Glu Glu Asn Lys Leu Pro Glu Gly Ser Val Ile Tyr Pro Gly Val Val
325 330 335
Ser His Ser Ile Asn Ala Val Glu His Pro Arg Leu Val Ala Asp Arg
340 345 350
Ile Val Gln Phe Ala Lys Leu Val Gly Pro Glu Asn Val Ile Ala Ser
355 360 365
Thr Asp Cys Gly Leu Gly Gly Arg Leu His Ser Gln Ile Ala Trp Ala
370 375 380
Lys Leu Glu Ser Leu Val Glu Gly Ala Arg Ile Ala Ser Lys Glu Leu
385 390 395 400
Phe
<210>111
<211>548
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(1)..(525)
<223>FRXA02648
<400>111
gac gca ccg gac ttg gca gaa gca tgg gat cag atc aac cca gag cca 48
Asp Ala Pro Asp Leu Ala Glu Ala Trp Asp Gln Ile Asn Pro Glu Pro
1 5 10 15
agc gtg aag gat tac ttg gac tgg atc ggt aca cgc atc gat gcc atc 96
Ser Val Lys Asp Tyr Leu Asp Trp Ile Gly Thr Arg Ile Asp Ala Ile
20 25 30
aac agt gca gtg aag ggc ctt cca aag gaa cag acc cgc ctg cac atc 144
Asn Ser Ala Val Lys Gly Leu Pro Lys Glu Gln Thr Arg Leu His Ile
35 40 45
tgc tgg ggc tct tgg cac gga cca cac gtc act gac atc cca ttc ggt 192
Cys Trp Gly Ser Trp His Gly Pro His Val Thr Asp Ile Pro Phe Gly
50 55 60
gac atc att ggt gag atc ctg cgc gca gag gtc ggt ggc ttc tcc ttc 240
Asp Ile Ile Gly Glu Ile Leu Arg Ala Glu Val Gly Gly Phe Ser Phe
65 70 75 80
gaa ggc gca tct cct cgt cac gca cac gag tgg cgt gta tgg gaa gaa 288
Glu Gly Ala Ser Pro Arg His Ala His Glu Trp Arg Val Trp Glu Glu
85 90 95
aac aag ctt cct gaa ggc tct gtt atc tac cct ggt gtt gtg tct cac 336
Asn Lys Leu Pro Glu Gly Ser Val Ile Tyr Pro Gly Val Val Ser His
100 105 110
tcc atc aac gct gtg gag cac cca cgc ctg gtt gct gat cgt atc gtt 384
Ser Ile Asn Ala Val Glu His Pro Arg Leu Val Ala Asp Arg Ile Val
115 120 125
cag ttc gcc aag ctt gtt ggc cct gag aac gtc att gcg tcc act gac 432
Gln Phe Ala Lys Leu Val Gly Pro Glu Asn Val Ile Ala Ser Thr Asp
130 135 140
tgt ggt ctg ggc gga cgt ctg cat tcc cag atc gca tgg gca aag ctg 480
Cys Gly Leu Gly Gly Arg Leu His Ser Gln Ile Ala Trp Ala Lys Leu
145 150 155 160
gag tcc cta gta gag ggc gct cgc att gca tca aag gaa ctg ttc 525
Glu Ser Leu Val Glu Gly Ala Arg Ile Ala Ser Lys Glu Leu Phe
165 170 175
taagctagac aacgagggtt gct 548
<210>112
<211>175
<212>PRT
<213>谷氨酸棒杆菌
<400>112
Asp Ala Pro Asp Leu Ala Glu Ala Trp Asp Gln Ile Asn Pro Glu Pro
1 5 10 15
Ser Val Lys Asp Tyr Leu Asp Trp Ile Gly Thr Arg Ile Asp Ala Ile
20 25 30
Asn Ser Ala Val Lys Gly Leu Pro Lys Glu Gln Thr Arg Leu His Ile
35 40 45
Cys Trp Gly Ser Trp His Gly Pro His Val Thr Asp Ile Pro Phe Gly
50 55 60
Asp Ile Ile Gly Glu Ile Leu Arg Ala Glu Val Gly Gly Phe Ser Phe
65 70 75 80
Glu Gly Ala Ser Pro Arg His Ala His Glu Trp Arg Val Trp Glu Glu
85 90 95
Asn Lys Leu Pro Glu Gly Ser Val Ile Tyr Pro Gly Val Val Ser His
100 105 110
Ser Ile Asn Ala Val Glu His Pro Arg Leu Val Ala Asp Arg Ile Val
115 120 125
Gln Phe Ala Lys Leu Val Gly Pro Glu Asn Val Ile Ala Ser Thr Asp
130 135 140
Cys Gly Leu Gly Gly Arg Leu His Ser Gln Ile Ala Trp Ala Lys Leu
145 150 155 160
Glu Ser Leu Val Glu Gly Ala Arg Ile Ala Ser Lys Glu Leu Phe
165 170 175
<210>113
<211>784
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(784)
<223>FRXA02658
<400>113
atgaataaaa ttccgggtgc agtgaccgta ggtgaggtaa acgcggttag agtcgaatga 60
gagtttgata ctttctttcg acttttagat tggattttca atg agc cag aac cgc 115
Met Ser Gln Asn Arg
1 5
atc agg acc act cac gtt ggt tcc ttg ccc cgt acc cca gag cta ctt 163
Ile Arg Thr Thr His Val Gly Ser Leu Pro Arg Thr Pro Glu Leu Leu
10 15 20
gat gca aac atc aag cgt tct aac ggt gag att ggg gag gag gaa ttc 211
Asp Ala Asn Ile Lys Arg Ser Asn Gly Glu Ile Gly Glu Glu Glu Phe
25 30 35
ttc cag att ctg cag tct tct gta gat gac gtg atc aag cgc cag gtt 259
Phe Gln Ile Leu Gln Ser Ser Val Asp Asp Val Ile Lys Arg Gln Val
40 45 50
gac ctg ggt atc gac atc ctt aac gag ggc gaa tac ggc cac gtc acc 307
Asp Leu Gly Ile Asp Ile Leu Asn Glu Gly Glu Tyr Gly His Val Thr
55 60 65
tcc ggt gca gtt gac ttc ggt gca tgg tgg aac tac tcc ttc acc cgc 355
Ser Gly Ala Val Asp Phe Gly Ala Trp Trp Asn Tyr Ser Phe Thr Arg
70 75 80 85
ctg ggc gga ctg acc atg acc gat acc gac cgt tgg gca agc cag gaa 403
Leu Gly Gly Leu Thr Met Thr Asp Thr Asp Arg Trp Ala Ser Gln Glu
90 95 100
gca gtg cgt tcc acc cct ggc aac atc gag ctg acc agc ttc tct gat 451
Ala Val Arg Ser Thr Pro Gly Asn Ile Glu Leu Thr Ser Phe Ser Asp
105 110 115
cgt cgc gac cgc gca ttg ttc agc gaa gca tac gag gat cca gta tct 499
Arg Arg Asp Arg Ala Leu Phe Ser Glu Ala Tyr Glu Asp Pro Val Ser
120 125 130
ggc atc ttc acc ggt cgc gct tct gtg ggc aac cca gag ttc acc gga 547
Gly Ile Phe Thr Gly Arg Ala Ser Val Gly Asn Pro Glu Phe Thr Gly
135 140 145
cct att acc tac att ggc cag gaa gaa act cag acg gat gtt gat ctg 595
Pro Ile Thr Tyr Ile Gly Gln Glu Glu Thr Gln Thr Asp Val Asp Leu
150 155 160 165
ctg aag aag ggc atg aac gca gcg gga gct acc gac ggc ttc gtt gca 643
Leu Lys Lys Gly Met Asn Ala Ala Gly Ala Thr Asp Gly Phe Val Ala
170 175 180
gca cta tcc cca gga tct gca gct cga ttg acc aac aag ttc tac gac 691
Ala Leu Ser Pro Gly Ser Ala Ala Arg Leu Thr Asn Lys Phe Tyr Asp
185 190 195
act gat gaa gaa gtc gtc gca gca tgt gct gat gcg ctt tcc cag gaa 739
Thr Asp Glu Glu Val Val Ala Ala Cys Ala Asp Ala Leu Ser Gln Glu
200 205 210
tac aag atc atc acc gat gca ggt ctg acc gtt cag ctc gac gca 784
Tyr Lys Ile Ile Thr Asp Ala Gly Leu Thr Val Gln Leu Asp Ala
215 220 225
<210>114
<211>228
<212>PRT
<213>谷氨酸棒杆菌
<400>114
Met Ser Gln Asn Arg Ile Arg Thr Thr His Val Gly Ser Leu Pro Arg
1 5 10 15
Thr Pro Glu Leu Leu Asp Ala Asn Ile Lys Arg Ser Asn Gly Glu Ile
20 25 30
Gly Glu Glu Glu Phe Phe Gln Ile Leu Gln Ser Ser Val Asp Asp Val
35 40 45
Ile Lys Arg Gln Val Asp Leu Gly Ile Asp Ile Leu Asn Glu Gly Glu
50 55 60
Tyr Gly His Val Thr Ser Gly Ala Val Asp Phe Gly Ala Trp Trp Asn
65 70 75 80
Tyr Ser Phe Thr Arg Leu Gly Gly Leu Thr Met Thr Asp Thr Asp Arg
85 90 95
Trp Ala Ser Gln Glu Ala Val Arg Ser Thr Pro Gly Asn Ile Glu Leu
100 105 110
Thr Ser Phe Ser Asp Arg Arg Asp Arg Ala Leu Phe Ser Glu Ala Tyr
115 120 125
Glu Asp Pro Val Ser Gly Ile Phe Thr Gly Arg Ala Ser Val Gly Asn
130 135 140
Pro Glu Phe Thr Gly Pro Ile Thr Tyr Ile Gly Gln Glu Glu Thr Gln
145 150 155 160
Thr Asp Val Asp Leu Leu Lys Lys Gly Met Asn Ala Ala Gly Ala Thr
165 170 175
Asp Gly Phe Val Ala Ala Leu Ser Pro Gly Ser Ala Ala Arg Leu Thr
180 185 190
Asn Lys Phe Tyr Asp Thr Asp Glu Glu Val Val Ala Ala Cys Ala Asp
195 200 205
Ala Leu Ser Gln Glu Tyr Lys Ile Ile Thr Asp Ala Gly Leu Thr Val
210 215 220
Gln Leu Asp Ala
225
<210>115
<211>408
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(385)
<223>RXC02238
<400>115
ggcgcttagc caaaacatag agcggtaggg tatgcttatc cgattgagca acctttcccg 60
ctcttaacac tactgtccat atacttttga aaaggtgtca gtg acc aac gtg agc 115
Val Thr Asn Val Ser
1 5
aac gag acc aac gcc acc aag gcc gtc ttc gat ccg cca gtg ggc att 163
Asn Glu Thr Asn Ala Thr Lys Ala Val Phe Asp Pro Pro Val Gly Ile
10 15 20
acc gct cct ccg atc gat gaa ctg ctg gat aag gtc act tcc aag tac 211
Thr Ala Pro Pro Ile Asp Glu Leu Leu Asp Lys Val Thr Ser Lys Tyr
25 30 35
gcc ctc gtg atc ttc gca gcc aag cgt gcg cgc cag atc aac agc ttc 259
Ala Leu Val Ile Phe Ala Ala Lys Arg Ala Arg Gln Ile Asn Ser Phe
40 45 50
tac cat cag gca gat gag gga gta ttc gag ttc atc gga cca ttg gtt 307
Tyr His Gln Ala Asp Glu Gly Val Phe Glu Phe Ile Gly Pro Leu Val
55 60 65
act ccg cag cca ggc gaa aag cca ctt tct att gct ctg cgt gag atc 355
Thr Pro Gln Pro Gly Glu Lys Pro Leu Ser Ile Ala Leu Arg Glu Ile
70 75 80 85
aat gca ggt ctg ttg gac cac gag gaa ggt taaaagacct tataacttca 405
Asn Ala Gly Leu Leu Asp His Glu Glu Gly
90 95
cac 408
<210>116
<211>95
<212>PRT
<213>谷氨酸棒杆菌
<400>116
Val Thr Asn Val Ser Asn Glu Thr Asn Ala Thr Lys Ala Val Phe Asp
1 5 10 15
Pro Pro Val Gly Ile Thr Ala Pro Pro Ile Asp Glu Leu Leu Asp Lys
20 25 30
Val Thr Ser Lys Tyr Ala Leu Val Ile Phe Ala Ala Lys Arg Ala Arg
35 40 45
Gln Ile Asn Ser Phe Tyr His Gln Ala Asp Glu Gly Val Phe Glu Phe
50 55 60
Ile Gly Pro Leu Val Thr Pro Gln Pro Gly Glu Lys Pro Leu Ser Ile
65 70 75 80
Ala Leu Arg Glu Ile Asn Ala Gly Leu Leu Asp His Glu Glu Gly
85 90 95
<210>117
<211>1827
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1804)
<223>RXC00128
<400>117
ccattttccg tttggtcttg cctaaagaac cgcatggaaa ttatcgtgaa gcaccgatcc 60
cgttgatcgc tccagagaca ccgtgggaag gggagcagca gtg agt aaa att tcg 115
Val Ser Lys Ile Ser
1 5
acg aaa ctg aag gcc ctc acc gcg gtg ctg tct gtg acc act ctg gtg 163
Thr Lys Leu Lys Ala Leu Thr Ala Val Leu Ser Val Thr Thr Leu Val
10 15 20
gct ggg tgt tcc acg ctt ccg cag aac acg gat ccg caa gtg ctg cgc 211
Ala Gly Cys Ser Thr Leu Pro Gln Asn Thr Asp Pro Gln Val Leu Arg
25 30 35
tca ttt tcc ggg tcc caa agc aca caa gag ata gca ggg ccg acc ccg 259
Ser Phe Ser Gly Ser Gln Ser Thr Gln Glu Ile Ala Gly Pro Thr Pro
40 45 50
aat caa gat ccg gat ttg ttg atc cgc ggc ttc ttc agc gca ggt gcg 307
Asn Gln Asp Pro Asp Leu Leu Ile Arg Gly Phe Phe Ser Ala Gly Ala
55 60 65
tat ccg act cag cag tat gaa gcg gcg aag gcg tat ctg acg gaa ggg 355
Tyr Pro Thr Gln Gln Tyr Glu Ala Ala Lys Ala Tyr Leu Thr Glu Gly
70 75 80 85
acg cgc agc acg tgg aat ccg gct gcg tcg act cgt att ttg gat cgc 403
Thr Arg Ser Thr Trp Asn Pro Ala Ala Ser Thr Arg Ile Leu Asp Arg
90 95 100
att gat ctg aac act ctg cca ggt tcg acg aat gcg gaa cga acg att 451
Ile Asp Leu Asn Thr Leu Pro Gly Ser Thr Asn Ala Glu Arg Thr Ile
105 110 115
gcg atc cgt gga acg cag gtc gga acg ttg ctc agc ggt ggc gtg tat 499
Ala Ile Arg Gly Thr Gln Val Gly Thr Leu Leu Ser Gly Gly Val Tyr
120 125 130
cag ccg gag aat gcg gag ttt gaa gct gag atc acg atg cgt cgg gaa 547
Gln Pro Glu Asn Ala Glu Phe Glu Ala Glu Ile Thr Met Arg Arg Glu
135 140 145
gat ggg gag tgg cgt atc gat gct ttg ccg gac ggg att tta tta gag 595
Asp Gly Glu Trp Arg Ile Asp Ala Leu Pro Asp Gly Ile Leu Leu Glu
150 155 160 165
aga aac gat ctg cgg aac cat tac act ccg cac gat gtg tat ttc ttt 643
Arg Asn Asp Leu Arg Asn His Tyr Thr Pro His Asp Val Tyr Phe Phe
170 175 180
gat cct tct ggc cag gtg ttg gtg ggg gat cgg cgt tgg ttg ttc aat 691
Asp Pro Ser Gly Gln Val Leu Val Gly Asp Arg Arg Trp Leu Phe Asn
185 190 195
gag tcg cag tcg atg tcc acg gtg ctg atg gcc ctt ctg gtt aat ggt 739
Glu Ser Gln Ser Met Ser Thr Val Leu Met Ala Leu Leu Val Asn Gly
200 205 210
cct tcg ccg gca att tct cct ggt gtg gtc aat cag ctg tcc acg gat 787
Pro Ser Pro Ala Ile Ser Pro Gly Val Val Asn Gln Leu Ser Thr Asp
215 220 225
gcg tcg ttc gtg ggg ttc aat gat ggg gag tat cag ttc act ggt ttg 835
Ala Ser Phe Val Gly Phe Asn Asp Gly Glu Tyr Gln Phe Thr Gly Leu
230 235 240 245
gga aat ttg gat gat gat gcg cgt ttg cgt ttc gcc gcc cag gcc gtg 883
Gly Asn Leu Asp Asp Asp Ala Arg Leu Arg Phe Ala Ala Gln Ala Val
250 255 260
tgg acg ttg gcg cat gct gat gtc gca ggc ccc tac act ttg gtc gct 931
Trp Thr Leu Ala His Ala Asp Val Ala Gly Pro Tyr Thr Leu Val Ala
265 270 275
gac ggc gcg ccg ttg ctg tcg gag ttc cca acg ctc acc acc gat gac 979
Asp Gly Ala Pro Leu Leu Ser Glu Phe Pro Thr Leu Thr Thr Asp Asp
280 285 290
ctc gcc gaa tac aac cca gag gct tac acc aac acg gtg tcc acg ttg 1027
Leu Ala Glu Tyr Asn Pro Glu Ala Tyr Thr Asn Thr Val Ser Thr Leu
295 300 305
ttt gcg ttg cag gat gga tcg ttg tcg agg gtc agt tcc ggc aat gtg 1075
Phe Ala Leu Gln Asp Gly Ser Leu Ser Arg Val Ser Ser Gly Asn Val
310 315 320 325
agt cca cta cag ggc att tgg agc ggt gga gat atc gat tct gca gcg 1123
Ser Pro Leu Gln Gly Ile Trp Ser Gly Gly Asp Ile Asp Ser Ala Ala
330 335 340
att tcc tcc tcc gcc aat gtg gtg gca gcg gta cgc cac gaa aac aac 1171
Ile Ser Ser Ser Ala Asn Val Val Ala Ala Val Arg His Glu Asn Asn
345 350 355
gag gca gtg ctt act gtt ggc tcc atg gaa ggc gtg act tca gat gcg 1219
Glu Ala Val Leu Thr Val Gly Ser Met Glu Gly Val Thr Ser Asp Ala
360 365 370
ttg agg agt gaa acg atc act cgt ccc acc ttt gaa tac gcg tcg agt 1267
Leu Arg Ser Glu Thr Ile Thr Arg Pro Thr Phe Glu Tyr Ala Ser Ser
375 380 385
ggg ttg tgg gct gtg gtg gat ggg gag acg cct gtc cga gtc gca cga 1315
Gly Leu Trp Ala Val Val Asp Gly Glu Thr Pro Val Arg Val Ala Arg
390 395 400 405
tcg gca aca acc ggt gag ctc gtc cag acg gag gcg gag att gtg ctg 1363
Ser Ala Thr Thr Gly Glu Leu Val Gln Thr Glu Ala Glu Ile Val Leu
410 415 420
cca agg gat gtg acg ggt ccg atc tct gaa ttc caa ctg tca cga act 1411
Pro Arg Asp Val Thr Gly Pro Ile Ser Glu Phe Gln Leu Ser Arg Thr
425 430 435
ggg gtc cgg gcc gcc atg atc att gaa ggc aag gtg tac gtg ggc gtc 1459
Gly Val Arg Ala Ala Met Ile Ile Glu Gly Lys Val Tyr Val Gly Val
440 445 450
gta acg cgt cct ggt ccg ggc gag cgg cgc gtg aca aat atc acg gag 1507
Val Thr Arg Pro Gly Pro Gly Glu Arg Arg Val Thr Asn Ile Thr Glu
455 460 465
gtg gcg ccg agc ttg ggc gag gcg gcg ctg tcg atc aac tgg cgc cca 1555
Val Ala Pro Ser Leu Gly Glu Ala Ala Leu Ser Ile Asn Trp Arg Pro
470 475 480 485
gac ggc att ttg ctt gtg ggc acg tca att cca gag acg ccg ctg tgg 1603
Asp Gly Ile Leu Leu Val Gly Thr Ser Ile Pro Glu Thr Pro Leu Trp
490 495 500
cgc gtc gag cag gac gga tcg gcg att tcg tcg atg ccg agc ggg aat 1651
Arg Val Glu Gln Asp Gly Ser Ala Ile Ser Ser Met Pro Ser Gly Asn
505 510 515
ctc agc gcg ccg gtg gtg gcg gtg gca agt tcc gcg acg acg gtc tac 1699
Leu Ser Ala Pro Val Val Ala Val Ala Ser Ser Ala Thr Thr Val Tyr
520 525 530
gtc act gat tcg cat gcg atg ctt cag ctg ccg act gcc gat aat gat 1747
Val Thr Asp Ser His Ala Met Leu Gln Leu Pro Thr Ala Asp Asn Asp
535 540 545
att tgg cgc gag gtg ccc ggt ttg ctg ggc acg cgt gcg gcg ccg gtg 1795
Ile Trp Arg Glu Val Pro Gly Leu Leu Gly Thr Arg Ala Ala Pro Val
550 555 560 565
gtt gcg tac tgatggagct gttcttcccg cgc 1827
Val Ala Tyr
<210>118
<211>568
<212>PRT
<213>谷氨酸棒杆菌
<400>118
Val Ser Lys Ile Ser Thr Lys Leu Lys Ala Leu Thr Ala Val Leu Ser
1 5 10 15
Val Thr Thr Leu Val Ala Gly Cys Ser Thr Leu Pro Gln Asn Thr Asp
20 25 30
Pro Gln Val Leu Arg Ser Phe Ser Gly Ser Gln Ser Thr Gln Glu Ile
35 40 45
Ala Gly Pro Thr Pro Asn Gln Asp Pro Asp Leu Leu Ile Arg Gly Phe
50 55 60
Phe Ser Ala Gly Ala Tyr Pro Thr Gln Gln Tyr Glu Ala Ala Lys Ala
65 70 75 80
Tyr Leu Thr Glu Gly Thr Arg Ser Thr Trp Asn Pro Ala Ala Ser Thr
85 90 95
Arg Ile Leu Asp Arg Ile Asp Leu Asn Thr Leu Pro Gly Ser Thr Asn
100 105 110
Ala Glu Arg Thr Ile Ala Ile Arg Gly Thr Gln Val Gly Thr Leu Leu
115 120 125
Ser Gly Gly Val Tyr Gln Pro Glu Asn Ala Glu Phe Glu Ala Glu Ile
130 135 140
Thr Met Arg Arg Glu Asp Gly Glu Trp Arg Ile Asp Ala Leu Pro Asp
145 150 155 160
Gly Ile Leu Leu Glu Arg Asn Asp Leu Arg Asn His Tyr Thr Pro His
165 170 175
Asp Val Tyr Phe Phe Asp Pro Ser Gly Gln Val Leu Val Gly Asp Arg
180 185 190
Arg Trp Leu Phe Asn Glu Ser Gln Ser Met Ser Thr Val Leu Met Ala
195 200 205
Leu Leu Val Asn Gly Pro Ser Pro Ala Ile Ser Pro Gly Val Val Asn
210 215 220
Gln Leu Ser Thr Asp Ala Ser Phe Val Gly Phe Asn Asp Gly Glu Tyr
225 230 235 240
Gln Phe Thr Gly Leu Gly Asn Leu Asp Asp Asp Ala Arg Leu Arg Phe
245 250 255
Ala Ala Gln Ala Val Trp Thr Leu Ala His Ala Asp Val Ala Gly Pro
260 265 270
Tyr Thr Leu Val Ala Asp Gly Ala Pro Leu Leu Ser Glu Phe Pro Thr
275 280 285
Leu Thr Thr Asp Asp Leu Ala Glu Tyr Asn Pro Glu Ala Tyr Thr Asn
290 295 300
Thr Val Ser Thr Leu Phe Ala Leu Gln Asp Gly Ser Leu Ser Arg Val
305 310 315 320
Ser Ser Gly Asn Val Ser Pro Leu Gln Gly Ile Trp Ser Gly Gly Asp
325 330 335
Ile Asp Ser Ala Ala Ile Ser Ser Ser Ala Asn Val Val Ala Ala Val
340 345 350
Arg His Glu Asn Asn Glu Ala Val Leu Thr Val Gly Ser Met Glu Gly
355 360 365
Val Thr Ser Asp Ala Leu Arg Ser Glu Thr Ile Thr Arg Pro Thr Phe
370 375 380
Glu Tyr Ala Ser Ser Gly Leu Trp Ala Val Val Asp Gly Glu Thr Pro
385 390 395 400
Val Arg Val Ala Arg Ser Ala Thr Thr Gly Glu Leu Val Gln Thr Glu
405 410 415
Ala Glu Ile Val Leu Pro Arg Asp Val Thr Gly Pro Ile Ser Glu Phe
420 425 430
Gln Leu Ser Arg Thr Gly Val Arg Ala Ala Met Ile Ile Glu Gly Lys
435 440 445
Val Tyr Val Gly Val Val Thr Arg Pro Gly Pro Gly Glu Arg Arg Val
450 455 460
Thr Asn Ile Thr Glu Val Ala Pro Ser Leu Gly Glu Ala Ala Leu Ser
465 470 475 480
Ile Asn Trp Arg Pro Asp Gly Ile Leu Leu Val Gly Thr Ser Ile Pro
485 490 495
Glu Thr Pro Leu Trp Arg Val Glu Gln Asp Gly Ser Ala Ile Ser Ser
500 505 510
Met Pro Ser Gly Asn Leu Ser Ala Pro Val Val Ala Val Ala Ser Ser
515 520 525
Ala Thr Thr Val Tyr Val Thr Asp Ser His Ala Met Leu Gln Leu Pro
530 535 540
Thr Ala Asp Asn Asp Ile Trp Arg Glu Val Pro Gly Leu Leu Gly Thr
545 550 555 560
Arg Ala Ala Pro Val Val Ala Tyr
565
<210>119
<211>1344
<212>DNA
<213>谷氨酸棒杆菌
<220>
<221>CDS
<222>(101)..(1321)
<223>RXA02240
<400>119
cagctagacc actgacattg cagttttaga cagcttggtc tatattggtt ttttgtattt 60
aagactattt attctcaact tcttcgaaag aagggtattt gtg gct cag cca acc 115
Val Ala Gln Pro Thr
1 5
gcc gtc cgt ttg ttc acc agt gaa tct gta act gag gga cat cca gac 163
Ala Val Arg Leu Phe Thr Ser Glu Ser Val Thr Glu Gly His Pro Asp
10 15 20
aaa ata tgt gat gct att tcc gat acc att ttg gac gcg ctg ctc gaa 211
Lys Ile Cys Asp Ala Ile Ser Asp Thr Ile Leu Asp Ala Leu Leu Glu
25 30 35
aaa gat ccg cag tcg cgc gtc gca gtg gaa act gtg gtc acc acc gga 259
Lys Asp Pro Gln Ser Arg Val Ala Val Glu Thr Val Val Thr Thr Gly
40 45 50
atc gtc cat gtt gtt ggc gag gtc cgt acc agc gct tac gta gag atc 307
Ile Val His Val Val Gly Glu Val Arg Thr Ser Ala Tyr Val Glu Ile
55 60 65
cct caa tta gtc cgc aac aag ctc atc gaa atc gga ttc aac tcc tct 355
Pro Gln Leu Val Arg Asn Lys Leu Ile Glu Ile Gly Phe Asn Ser Ser
70 75 80 85
gag gtt gga ttc gac gga cgc acc tgt ggc gtc tca gta tcc atc ggt 403
Glu Val Gly Phe Asp Gly Arg Thr Cys Gly Val Ser Val Ser Ile Gly
90 95 100
gag cag tcc cag gaa atc gct gac ggc gtg gat aac tcc gac gaa gcc 451
Glu Gln Ser Gln Glu Ile Ala Asp Gly Val Asp Asn Ser Asp Glu Ala
105 110 115
cgc acc aac ggc gac gtt gaa gaa gac gac cgc gca ggt gct ggc gac 499
Arg Thr Asn Gly Asp Val Glu Glu Asp Asp Arg Ala Gly Ala Gly Asp
120 125 130
cag ggc ctg atg ttc ggc tac gcc acc aac gaa acc gaa gag tac atg 547
Gln Gly Leu Met Phe Gly Tyr Ala Thr Asn Glu Thr Glu Glu Tyr Met
135 140 145
cct ctt cct atc gcg ttg gcg cac cga ctg tca cgt cgt ctg acc cag 595
Pro Leu Pro Ile Ala Leu Ala His Arg Leu Ser Arg Arg Leu Thr Gln
150 155 160 165
gtt cgt aaa gag ggc atc gtt cct cac ctg cgt cca gac gga aaa acc 643
Val Arg Lys Glu Gly Ile Val Pro His Leu Arg Pro Asp Gly Lys Thr
170 175 180
cag gtc acc ttc gca tac gat gcg caa gac cgc cct agc cac ctg gat 691
Gln Val Thr Phe Ala Tyr Asp Ala Gln Asp Arg Pro Ser His Leu Asp
185 190 195
acc gtt gtc atc tcc acc cag cac gac cca gaa gtt gac cgt gca tgg 739
Thr Val Val Ile Ser Thr Gln His Asp Pro Glu Val Asp Arg Ala Trp
200 205 210
ttg gaa acc caa ctg cgc gaa cac gtc att gat tgg gta atc aaa gac 787
Leu Glu Thr Gln Leu Arg Glu His Val Ile Asp Trp Val Ile Lys Asp
215 220 225
gca ggc att gag gat ctg gca acc ggt gag atc acc gtg ttg atc aac 835
Ala Gly Ile Glu Asp Leu Ala Thr Gly Glu Ile Thr Val Leu Ile Asn
230 235 240 245
cct tca ggt tcc ttc att ctg ggt ggc ccc atg ggt gat gcg ggt ctg 883
Pro Ser Gly Ser Phe Ile Leu Gly Gly Pro Met Gly Asp Ala Gly Leu
250 255 260
acc ggc cgc aag atc atc gtg gat acc tac ggt ggc atg gct cgc cat 931
Thr Gly Arg Lys Ile Ile Val Asp Thr Tyr Gly Gly Met Ala Arg His
265 270 275
ggt ggt gga gca ttc tcc ggt aag gat cca agc aag gtg gac cgc tct 979
Gly Gly Gly Ala Phe Ser Gly Lys Asp Pro Ser Lys Val Asp Arg Ser
280 285 290
gct gca tac gcc atg cgt tgg gta gca aag aac atc gtg gca gca ggc 1027
Ala Ala Tyr Ala Met Arg Trp Val Ala Lys Asn Ile Val Ala Ala Gly
295 300 305
ctt gct gat cgc gct gaa gtt cag gtt gca tac gcc att gga cgc gca 1075
Leu Ala Asp Arg Ala Glu Val Gln Val Ala Tyr Ala Ile Gly Arg Ala
310 315 320 325
aag cca gtc gga ctt tac gtt gaa acc ttt gac acc aac aag gaa ggc 1123
Lys Pro Val Gly Leu Tyr Val Glu Thr Phe Asp Thr Asn Lys Glu Gly
330 335 340
ctg agc gac gag cag att cag gct gcc gtg ttg gag gtc ttt gac ctg 1171
Leu Ser Asp Glu Gln Ile Gln Ala Ala Val Leu Glu Val Phe Asp Leu
345 350 355
cgt cca gca gca att atc cgt gag ctt gat ctg ctt cgt ccg atc tac 1219
Arg Pro Ala Ala Ile Ile Arg Glu Leu Asp Leu Leu Arg Pro Ile Tyr
360 365 370
gct gac act gct gcc tac ggc cac ttt ggt cgc act gat ttg gac ctt 1267
Ala Asp Thr Ala Ala Tyr Gly His Phe Gly Arg Thr Asp Leu Asp Leu
375 380 385
cct tgg gag gct atc gac cgc gtt gat gaa ctt cgc gca gcc ctc aag 1315
Pro Trp Glu Ala Ile Asp Arg Val Asp Glu Leu Arg Ala Ala Leu Lys
390 395 400 405
ttg gcc taaaaatctg atgtagtatcttc 1344
Leu Ala
<210>120
<211>407
<212>PRT
<213>谷氨酸棒杆菌
<400>120
Val Ala Gln Pro Thr Ala Val Arg Leu Phe Thr Ser Glu Ser Val Thr
1 5 10 15
Glu Gly His Pro Asp Lys Ile Cys Asp Ala Ile Ser Asp Thr Ile Leu
20 25 30
Asp Ala Leu Leu Glu Lys Asp Pro Gln Ser Arg Val Ala Val Glu Thr
35 40 45
Val Val Thr Thr Gly Ile Val His Val Val Gly Glu Val Arg Thr Ser
50 55 60
Ala Tyr Val Glu Ile Pro Gln Leu Val Arg Asn Lys Leu Ile Glu Ile
65 70 75 80
Gly Phe Asn Ser Ser Glu Val Gly Phe Asp Gly Arg Thr Cys Gly Val
85 90 95
Ser Val Ser Ile Gly Glu Gln Ser Gln Glu Ile Ala Asp Gly Val Asp
100 105 110
Asn Ser Asp Glu Ala Arg Thr Asn Gly Asp Val Glu Glu Asp Asp Arg
115 120 125
Ala Gly Ala Gly Asp Gln Gly Leu Met Phe Gly Tyr Ala Thr Asn Glu
130 135 140
Thr Glu Glu Tyr Met Pro Leu Pro Ile Ala Leu Ala His Arg Leu Ser
145 150 155 160
Arg Arg Leu Thr Gln Val Arg Lys Glu Gly Ile Val Pro His Leu Arg
165 170 175
Pro Asp Gly Lys Thr Gln Val Thr Phe Ala Tyr Asp Ala Gln Asp Arg
180 185 190
Pro Ser His Leu Asp Thr Val Val Ile Ser Thr Gln His Asp Pro Glu
195 200 205
Val Asp Arg Ala Trp Leu Glu Thr Gln Leu Arg Glu His Val Ile Asp
210 215 220
Trp Val Ile Lys Asp Ala Gly Ile Glu Asp Leu Ala Thr Gly Glu Ile
225 230 235 240
Thr Val Leu Ile Asn Pro Ser Gly Ser Phe Ile Leu Gly Gly Pro Met
245 250 255
Gly Asp Ala Gly Leu Thr Gly Arg Lys Ile Ile Val Asp Thr Tyr Gly
260 265 270
Gly Met Ala Arg His Gly Gly Gly Ala Phe Ser Gly Lys Asp Pro Ser
275 280 285
Lys Val Asp Arg Ser Ala Ala Tyr Ala Met Arg Trp Val Ala Lys Asn
290 295 300
Ile Val Ala Ala Gly Leu Ala Asp Arg Ala Glu Val Gln Val Ala Tyr
305 310 315 320
Ala Ile Gly Arg Ala Lys Pro Val Gly Leu Tyr Val Glu Thr Phe Asp
325 330 335
Thr Asn Lys Glu Gly Leu Ser Asp Glu Gln Ile Gln Ala Ala Val Leu
340 345 350
Glu Val Phe Asp Leu Arg Pro Ala Ala Ile Ile Arg Glu Leu Asp Leu
355 360 365
Leu Arg Pro Ile Tyr Ala Asp Thr Ala Ala Tyr Gly His Phe Gly Arg
370 375 380
Thr Asp Leu Asp Leu Pro Trp Glu Ala Ile Asp Arg Val Asp Glu Leu
385 390 395 400
Arg Ala Ala Leu Lys Leu Ala
405
<210>121
<211>23
<212>DNA
<213>人工序列
<220>
<223>人工序列说明:引物
<400>121
tcgggtatcc gcgctacactt aga 23
<210>122
<211>23
<212>DNA
<213>人工序列
<220>
<223>人工序列说明:引物
<400>122
GGAAACCGGG GCATCGAAAC TTA 23
<210>123
<211>18
<212>DNA
<213>人工序列
<220>
<223>人工序列说明:引物
<400>123
ggaaacagta tgaccatg 18
<210>124
<211>17
<212>DNA
<213>人工序列
<220>
<223>人工序列说明:引物
<400>124
gtaaaacgac ggccagt 18
<210>125
<211>4334
<212>DNA
<213>谷氨酸棒杆菌
<400>125
aaatcgcttg accattgcag gttggtttat gactgttgag ggagagactg gctcgtggcc 60
gacaatcaat gaagctatgt ctgaatttag cgtgtcacgt cagaccgtga atagagcact 120
taagtctgcg ggcattgaac ttccacgagg acgccgtaaa gcttcccagt aaatgtgcca 180
tctcgtaggc agaaaacggt tccccccgta ggggtctctc tcttggcctc ctttctaggt 240
cgggctgatt gctcttgaag ctctctaggg gggctcacac cataggcaga taacggttcc 300
ccaccggctc acctcgtaag cgcacaagga ctgctcccaa agatcttcaa agccactgcc 360
gcgactccgc ttcgcgaagc cttgccccgc ggaaatttcc tccaccgagt tcgtgcacac 420
ccctatgcca agcttctttc accctaaatt cgagagattg gattcttacc gtggaaattc 480
ttcgcaaaaa tcgtcccctg atcgcccttg cgacgttgct cgcggcggtg ccgctggttg 540
cgcttggctt gaccgacttg atcagcttgc atgcctgcag gtcgacggat ccccgggtgg 600
gaaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca 660
tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat 720
attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga tgctgattta 780
tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg 840
tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat 900
gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc 960
atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg 1020
aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg 1080
ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc 1140
gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg 1200
agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat 1260
aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac 1320
cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca 1380
gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta 1440
cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt 1500
catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag 1560
agcattacgc tgacttgacg ggacggcggc tttgttgaat aaatcgaact tttgctgagt 1620
tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt 1680
tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt 1740
tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag 1800
gcagacctca gcgcccccga attgatcagt actgcggcgt cgctgatcgc cctcgcgacg 1860
ttgtgcgggt ggcttgtccc tgagggcgct gcgacagata gctaaaaatc tgcgtcagga 1920
tcgccgtaga gcgcgcgtcg cgtcgattgg aggcttcccc tttggttgac ggtcttcaat 1980
cgctctacgg cgatcctgac gcttttttgt tgcgtaccgt cgatcgtttt atttctgtcg 2040
atcccgaaaa agtttttgcc ttttgtaaaa aacttctcgg tcgccccgca aattttcgat 2100
tccagatttt ttaaaaacca agccagaaat acgacacacc gtttgcagat aatctgtctt 2160
tcggaaaaat caagtgcgat acaaaatttt tagcacccct gagctgcgca aagtcccgct 2220
tcgtgaaaat tttcgtgccg cgtgattttc cgccaaaaac tttaacgaac gttcgttata 2280
atggtgtcat gaccttcacg acgaagtacc aaaattggcc cgaatcatca gctatggatc 2340
tctctgatgt cgcgctggag tccgacgcgc tcgatgctgc cgtcgattta aaaacggtga 2400
tcggattttt ccgagctctc gatacgacgg acgcgccagc atcacgagac tgggccagtg 2460
ccgcgagcga cctagaaact ctcgtggcgg atcttgagga gctggctgac gagctgcgtg 2520
ctcggcagcg ccaggaggac gcacagtagt ggaggatcga atcagttgcg cctactgcgg 2580
tggcctgatt cctccccggc ctgacccgcg aggacggcgc gcaaaatatt gctcagatgc 2640
gtgtcgtgcc gcagccagcc gcgagcgcgc caacaaacgc cacgccgagg agctggaggc 2700
ggctaggtcg caaatggcgc tggaagtgcg tcccccgagc gaaattttgg ccatggtcgt 2760
cacagagctg gaagcggcag cgagaattat ccgcgatcgt ggcgcggtgc ccgcaggcat 2820
gacaaacatc gtaaatgccg cgtttcgtgt ggccgtggcc gcccaggacg tgtcagcgcc 2880
gccaccacct gcaccgaatc ggcagcagcg tcgcgcgtcg aaaaagcgca caggcggcaa 2940
gaagcgataa gctgcacgaa tacctgaaaa atgttgaacg ccccgtgagc ggtaactcac 3000
agggcgtcgg ctaaccccca gtccaaacca gggagaaagc gctcaaaaat gactctagcg 3060
gattcacgag acattgacac accggcctgg aaattttccg ctgatctgtt cgacacccat 3120
cccgagctcg cgctgcgatc acgtggctgg acgagcgaag accgccgcga attcctcgct 3180
cacctgggca gagaaaattt ccagggcagc aagacccgcg acttcgccag cgcttggatc 3240
aaagacccgg acacgggaga aacacagccg aagttatacc gagttggttc aaaatcgctt 3300
gcccggtgcc agtatgttgc tctgacgcac gcgcagcacg cagccgtgct tgtcctggac 3360
attgatgtgc cgagccacca ggccggcggg aaaatcgagc acgtaaaccc cgaggtctac 3420
gcgattttgg agcgctgggc acgcctggaa aaagcgccag cttggatcgg cgtgaatcca 3480
ctgagcggga aatgccagct catctggctc attgatccgg tgtatgccgc agcaggcatg 3540
agcagcccga atatgcgcct gctggctgca acgaccgagg aaatgacccg cgttttcggc 3600
gctgaccagg ctttttcaca taggctgagc cggtggccac tgcacgtctc cgacgatccc 3660
accgcgtacc gctggcatgc ccagcacaat cgcgtggatc gcctagctga tcttatggag 3720
gttgctcgca tgatctcagg cacagaaaaa cctaaaaaac gctatgagca ggagttttct 3780
agcggacggg cacgtatcga agcggcaaga aaagccactg cggaagcaaa agcacttgcc 3840
acgcttgaag caagcctgcc gagcgccgct gaagcgtctg gagagctgat cgacggcgtc 3900
cgtgtcctct ggactgctcc agggcgtgcc gcccgtgatg agacggcttt tcgccacgct 3960
ttgactgtgg gataccagtt aaaagcggct ggtgagcgcc taaaagacac caagatcatc 4020
gacgcctacg agcgtgccta caccgtcgct caggcggtcg gagcagacgg ccgtgagcct 4080
gatctgccgc cgatgcgtga ccgccagacg atggcgcgac gtgtgcgcgg ctacgtcgct 4140
aaaggccagc cagtcgtccc tgctcgtcag acagagacgc agagcagccg agggcgaaaa 4200
gctctggcca ctatgggaag acgtggcggt aaaaaggccg cagaacgctg gaaagaccca 4260
aacagtgagt acgcccgagc acagcgagaa aaactagcta agtccagtca acgacaagct 4320
aggaaagcta aagg 4334
Claims (52)
1.经分离的核酸分子或该经分离的核酸分子的互补序列,所述的核酸分子包含如SEQ ID NO:1、SEQ ID NO:3或SEQ ID NO:5的核苷酸序列的核酸分子。
2.权利要求1的分离的核酸分子,其中所述的核酸分子编码氨基酸代谢途径中所涉及的metz蛋白。
3.分离的核酸分子或该经分离的核酸分子的互补序列,所述的核酸分子编码包含SEQ ID NO:2、SEQ ID NO:4或SEQ ID NO:6所示氨基酸序列的多肽的天然存在等位变体。
4.经分离的核酸分子或该经分离的核酸分子的互补序列,所述的核酸分子含有与SEQ ID NO:1、SEQ ID NO:3或SEQ ID NO:5所示的完整核苷酸序列具有至少50%同一性的核苷酸序列。
5.经分离的核酸分子或该经分离的核酸分子的互补序列,所述的核酸分子含有选自SEQ ID NO:1、SEQ ID NO:3或SEQ ID NO:5所示核苷酸序列中的至少15个连续的核苷酸的片段,其中,所述的片段具有metz活性。
6.分离的核酸分子,该核酸分子在严格条件下可以与权利要求1-5中任一项的核酸分子杂交。
7.分离的核酸分子,该核酸分子含有权利要求1-6,22或47中任一项所述的核酸分子和编码异源多肽的核苷酸序列。
8.包含权利要求1-6,22或47中任一项所述的核酸分子的载体。
9.权利要求8的载体,还包括至少一种另外的代谢途径的核酸分子。
10.权利要求8或9的载体,其为表达载体。
11.以权利要求8-10中任一项的表达载体转染的宿主细胞。
12.权利要求9的载体,其中所述的至少一种另外的代谢途径的核酸分子包含选自表1中列出的奇数号序列的核苷酸序列的核酸分子,但不是任何F标明的核酸分子。
13.权利要求11的宿主细胞,其中所述细胞是微生物。
14.权利要求11的宿主细胞,其中该细胞属于棒杆菌属(Corynebacterium)或者短杆菌属(Brevibacterium)。
15.权利要求11的宿主细胞,其中所述的核酸分子的表达导致该细胞精细化学物质生产的调节。
16.权利要求15的宿主细胞,其中所述的精细化学物质是氨基酸。
17.权利要求16的宿主细胞,其中所述氨基酸是甲硫氨酸或赖氨酸。
18.生产多肽的方法,包括在合适的培养基中培养权利要求11的宿主细胞,从而生产多肽。
19.经分离的多肽,其包含SEQ ID NO:2、SEQ ID NO:4或SEQID NO:6的氨基酸序列。
20.权利要求19的经分离的多肽,其中所述的多肽是参与氨基酸代谢的metz多肽。
21.权利要求20的经分离的多肽,其中所述氨基酸是甲硫氨酸或赖氨酸。
22.经分离的核酸分子或该经分离的核酸分子的互补序列,所述的核酸分子编码含有SEQ ID NO:2、SEQ ID NO:4或SEQ ID NO:6的氨基酸序列的多肽。
23.经分离的多肽,其包含下述多肽的天然存在等位变体,上述被包含的多肽含有选自SEQ ID NO:2、SEQ ID NO:4或SEQ ID NO:6所示氨基酸序列。
24.权利要求19-23,25或46的经分离的多肽,还包括异源氨基酸序列。
25.经分离的多肽或其互补序列,所述的经分离的多肽由含有与SEQ ID NO:1、SEQ ID NO:3或SEQ ID NO:5的核苷酸序列至少50%同源的核苷酸序列的核酸分子所编码。
26.生产精细化学物质的方法,包括培养权利要求11的细胞,从而产生精细化学物质。
27.权利要求26的方法,其中所述细胞在硫源存在下培养。
28.权利要求26的方法,其中所述的方法还包括从所述的培养物中回收精细化学物质的步骤。
29.权利要求26的方法,其中所述精细化学物质是氨基酸。
30.权利要求29的方法,其中所述氨基酸是甲硫氨酸或赖氨酸。
31.权利要求26的方法,其中所述的方法还包括用权利要求8-10中任一项的载体转染所说细胞的步骤,这导致产生含有该载体的细胞。
32.权利要求26的方法,其中所述的细胞属于棒杆菌属或者短杆菌属。
33.权利要求26的方法,其中所述的细胞选自以下菌株:谷氨酸棒杆菌(Corynebacterium glutamicum)、力士棒杆菌(Corynebacteriumherculis)、百合花棒杆菌(Corynebacterium lilium)、嗜乙酰乙酸棒杆菌(Corynebacterium acetoacidophilum)、醋谷棒杆菌(Corynebacterium acetoglutamicum)、嗜乙酰棒杆菌(Corynebacteriumacetophilum)、产氨棒杆菌(Corynebacterium ammoniagenes)、Corynebacteriumfujiokense、Corynebacterium nitrilophilus、产氨短杆菌(Brevibacterium ammoniagenes)、Brevibacterium butanicum、分歧短杆菌(Brevibacterium divaricatum)、黄色短杆菌(Brevibacterium flavum)、希氏短杆菌(Brevibacterium healit)、酮戊二酸短杆菌(Brevibacteriumketoglutamicum)、Brevibacterium ketosoreductum、乳发酵短杆菌(Brevibacterium lactofermentum)、扩展短杆菌(Brevibacterium linens)、解石蜡短杆菌(Brevibacterium paraffinilyticum)和表3所列菌株。
34.生产精细化学物质的方法,包括培养其基因组DNA已经通过引入权利要求1-6,22或47任一项的核酸分子被改变的细胞。
35.权利要求34所述的方法,其中所述的基因组DNA通过包含至少一种另外的代谢途径核酸分子而改变。
36.权利要求35所述的方法,其中所述的另一种代谢途径的核酸分子含有选自表1奇数号序列的核苷酸序列的核酸分子,但不是任何F标明的核酸分子。
37.权利要求36的方法,其中所述至少一种另外的代谢途径的核酸分子选自metZ,metC,metB,metA,metE,metH,hom,asd,lysC,lysC/ask,rxa00657,dapA,dapB,dapC,dapD/argD,dapE,dapF,lysA,ddh,lysE,lysG,lysR,hsk,ppc,pycA,accD,accA,accB,accC,编码葡萄糖-6-磷酸脱氢酶的gpdh基因,opcA,pgdh,ta,tk,pgl,rlpe,rpe或任何上述基因的组合。
38.权利要求36的方法,其中所述代谢途径是甲硫氨酸或赖氨酸代谢。
39.调节一种细胞精细化学物质生产的方法,包括向细胞中导入一种或多种代谢途径基因,其中所述至少一种代谢途径的基因选自权利要求1-6,22或47中任一项所示的核酸分子,从而调节精细化学物质的生产。
40.权利要求39的方法,其中所述代谢途径基因整合到细胞的基因组中。
41.权利要求39的方法,其中所述代谢途径基因在质粒上。
42.权利要求39的方法,其中所述精细化学物质是氨基酸。
43.权利要求42的方法,其中所述氨基酸是甲硫氨酸或赖氨酸。
44.权利要求39的方法,其中所述代谢途径基因选自权利要求1-6任一项的核酸分子。
45.权利要求39的方法,其中所述代谢途径基因的核苷酸序列被突变以增加精细化学物质的生产。
46.经分离的多肽,其由包含SEQ ID NO:1、SEQ ID NO:3或SEQ ID NO:5所示的核苷酸序列的核酸分子编码。
47.编码下述多肽的经分离的核酸分子或其互补序列,所述的多肽包含与SEQ ID NO:2、SEQ ID NO:4或SEQ ID NO:6的全部氨基酸序列具有至少60%同一性的氨基酸序列。
48.一种诊断受试者中白喉棒状菌(Corynebacterium diphtheriae)的存在或活性的方法,该方法包含检测权利要求1-6,22或47的至少一种核酸分子,或权利要求19-23,25或46中所述的至少一种多肽的存在,由此诊断受试者中白喉棒状菌的存在或活性。
49.包含下述核酸分子的宿主细胞,所述核酸分子包含SEQ IDNO:1,SEQ ID NO:3,或SEQ ID NO:5所示的核苷酸序列,其中所述的核酸分子被破坏。
50.包含下述核酸分子的宿主细胞,所述的核酸分子包含SEQ IDNO:1,SEQ ID NO:3,或SEQ ID NO:5所示的核苷酸序列,其中所述的核酸分子与SEQ ID NO:1所示序列相比包含一个或多个核酸修饰。
51.包含下述核酸分子的宿主细胞,所述的核酸分子包含SEQ IDNO:1,SEQ ID NO:3,或SEQ ID NO:5所示的核苷酸序列,其中所述的核酸分子的调节区域相对于所述分子的野生型调节区域而言是经过修饰的。
52.权利要求1-6,22或47的至少一种核酸分子,或权利要求19-23,25或46中所述的至少一种多肽在诊断受试者中白喉棒状菌的存在或活性中的应用。
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JP4786107B2 (ja) | 2011-10-05 |
KR101012231B1 (ko) | 2011-02-09 |
BR0017148A (pt) | 2003-03-11 |
CA2402186A1 (en) | 2001-09-13 |
WO2001066573A2 (en) | 2001-09-13 |
EP1261718B1 (en) | 2010-11-03 |
WO2001066573A3 (en) | 2002-05-10 |
CN101255431B (zh) | 2012-05-16 |
DE60045191D1 (de) | 2010-12-16 |
JP2003525623A (ja) | 2003-09-02 |
KR20020086616A (ko) | 2002-11-18 |
ZA200208060B (en) | 2003-11-10 |
AU2390301A (en) | 2001-09-17 |
CA2402186C (en) | 2011-04-05 |
ATE486941T1 (de) | 2010-11-15 |
DK1261718T3 (da) | 2011-02-14 |
BRPI0017148B1 (pt) | 2016-03-01 |
ES2354867T3 (es) | 2011-03-18 |
EP1261718A2 (en) | 2002-12-04 |
AU2001223903B2 (en) | 2006-11-02 |
CN1452659A (zh) | 2003-10-29 |
MXPA02008710A (es) | 2003-02-24 |
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