CN1596267A - 来自谷氨酸棒状杆菌的编码调节蛋白的基因 - Google Patents
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Abstract
本发明涉及新的核酸分子,涉及其用于构建遗传改良的微生物的用途,还涉及借助于所述遗传改良的微生物制备精细化学品的方法,尤其是制备氨基酸的方法。
Description
发明背景
细胞中天然进行的代谢过程所产生的特定产物和副产物已用于工业的许多分支中,包括食品工业、动物饲料工业、化妆品工业及药学工业。这些分子合起来称为“精细化学品”,其包括有机酸(蛋白原的和非蛋白原的氨基酸)、核苷酸和核苷、脂类和脂肪酸、二醇、碳水化合物、芳香化合物、维生素和辅因子、以及酶。它们最好的产生是通过大规模培养已研发的在每一特定情况下可产生并分泌大量目标分子的细菌。一种对此目的尤其有用的生物体是谷氨酸棒状杆菌(Corynebacterium glutamicum),其为革兰氏阳性的非致病菌。通过菌株选择,已开发了许多突变菌株,它们产生多种目标化合物。但是,为提高特定分子产量的菌株选择是一种费时且困难的方法。
发明简述
本发明提供了新的可用于谷氨酸棒状杆菌或相关菌种的鉴定或分类的核酸分子。谷氨酸棒状杆菌为革兰氏阳性需氧细菌,其广泛用在工业上用于大规模产生多种精细化学品,也用于降解碳氢化合物(如在原油泄漏物中),以及用于氧化萜类化合物。本发明的核酸分子因此可用于鉴定能如通过发酵法用来产生精细化学品的微生物。虽然谷氨酸棒状杆菌自身为非致病的,但它与其它棒状杆菌如对人致病的重要病原菌白喉棒状杆菌(Corynebacterium diphtheriae)(白喉的病原菌)有亲缘关系。所以鉴定存在棒状杆菌的能力也具有显著的临床重要性,例如诊断应用。而且,所述核酸分子可作为对谷氨酸棒状杆菌或相关生物进行基因组作图的参考点。
这些新核酸分子编码的蛋白质文中称为代谢调节(MR)蛋白。这些MR蛋白可以例如发挥参与对细胞的正常代谢功能起关键作用的蛋白的转录、翻译或翻译后调节的功能。由于可得到用于谷氨酸棒状杆菌中的克隆载体(如在Sinskey等人,美国专利号4,649,119中所公开的)和用于遗传操作谷氨酸棒状杆菌和相关短杆菌种(如lactofermentum)的技术(Yoshihama等人,细菌学杂志(J.Bacteriol.)162:(1985)591-597;Katsumata等人,细菌学杂志(J.Bacteriol.)159:(1984)306-311;和Santamaria等人,普通微生物学杂志(J.Gen.Microbiol.)130:(1984)2237-2246),本发明的核酸分子可用于该生物体的遗传操作,使其成为一种或多种精细化学品的较好和更有效的生产者。
通过操作本发明的基因可以直接或间接地导致精细化学品产量、生产率和/或生产效率的提高。更具体地,对通常调节精细化学品代谢途径中精细化学品的产量、生产率和/或生产效率的谷氨酸棒状杆菌MR蛋白的修饰可能对所述生物体的一种或多种这些所需化合物的总生产率有直接的影响。对参与这些代谢途径的蛋白的修饰可能也对目的精细化学品的产量、生产率和/或生产效率有间接影响。代谢调节不可避免地是复杂的并且影响不同途径的调节机制在许多地方可重叠以致可以根据特定细胞事件快速调节一种以上的代谢途径。这使得可修饰一种代谢途径中的调节蛋白也可影响许多其他代谢途径,它们中的一些可能参与目的精细化学品的生物合成或降解。在这种间接方式中,对MR蛋白的作用进行调节可能影响精细化学品的生产率,所述该精细化学品产生的代谢途径不同于受所述MR蛋白直接调节的代谢途径。
可以使用本发明的核酸和蛋白分子以便直接提高谷氨酸棒状杆菌的一种或多种目的精细化学品的产量、生产率和/或生产效率。可通过本领域已知的基因重组技术操作一种或多种本发明的调节蛋白,从而使得它们的功能被调节。参与抑制编码氨基酸生物合成所需的酶的基因转录使得所述氨基酸不再能够抑制该转录的MR蛋白的突变可导致例如,所述氨基酸生产率的提高。所以,MR蛋白活性的修饰(其导致参与目的精细化学品生物合成的谷氨酸棒状杆菌蛋白的翻译增强或翻译后修饰的活化)接下来可增加所述化学品的生产率。相反的情况可同样有用:通过增强对转录或翻译的抑制或者通过对参与调节化合物降解途径的谷氨酸棒状杆菌蛋白的翻译后负修饰,可增加所述化学品的生产率。在任何一种情况下,可以增加该目的精细化学品的总产量或生产率。
同样,本发明蛋白和核苷酸分子中的所述修饰可通过间接机制提高精细化学品的产量、生产率和/或生产效率。特定化合物的代谢不可避免地与细胞中其他生物合成和降解途径相关,并且代谢途径中的必需辅因子、中间物或底物都可能被另一个代谢途径所提供或限制。所以调节本发明的一种或多种调节蛋白能够影响精细化学品的其他生物合成或降解途径的活性的效率。除此之外,对一种或多种调节蛋白的操作可增加细胞在培养物中,特别是在大规模发酵培养物(其中的生长条件可能不是最优的)中的生长和增殖的总体能力。例如通过突变通常抑制核苷生物合成的本发明MR蛋白,使得所述蛋白具有较低的抑制活性,从而可增加核苷的生物合成和细胞分裂,其中所述核苷生物合成的抑制为对营养的非最优胞外提供而产生的反应(从而阻止细胞分裂)。在那些导致培养物中细胞生长和分裂增加的MR蛋白的修饰可以导致培养物中一种或多种目标精细化学品的产量、生产率和/或生产效率增加,这至少是因为培养物中生产所述化学品的细胞数目的增加。
本发明提供了编码本文中称作代谢调节(MR)蛋白的新核酸分子并且所述蛋白例如能够进行和谷氨酸棒状杆菌中代谢途径的转录、翻译或翻译后调节相关的酶促步骤。本文中编码MR蛋白的核酸分子被称作MR核酸分子。在一个优选的实施方案中,MR蛋白参与一个或多个代谢途径的转录、翻译或翻译后调节。这些蛋白的实例是表1中列出的基因编码的蛋白。
因此,本发明的一个方面涉及这样的分离核酸分子(例如cDNA),其包含编码MR蛋白或其生物学活性部分的核苷酸序列和适合作为检测或扩增编码MR的核酸(例如DNA或mRNA)的引物或杂交探针的核酸片段。在尤其优选的实施方案中,该分离的核酸分子包含任何集合A所列出的核苷酸序列或任何这些核苷酸序列的编码区或其互补序列。在其他优选的实施方案中,该分离的核酸分子编码集合B中列出的任何氨基酸序列。本发明优选的MR蛋白同样优选地具有至少一种本文所描述的MR活性。
集合A定义了下面的表1中描述的序列表的核酸序列以及相关位置的序列修饰。
集合B定义了下面的表1中描述的序列表的多肽序列以及相关位置的序列修饰。
在另一个实施方案中,分离的核酸分子长度至少为15个核苷酸并且在严紧条件下与含有集合A的核苷酸序列的核酸分子杂交。分离的核酸分子优选与天然存在的核酸分子对应。分离的核酸更优选编码天然存在的谷氨酸棒状杆菌MR蛋白或其生物活性部分。
本发明的另一个方面涉及包含本发明核酸分子的载体(例如重组表达载体),还涉及已导入所述载体的宿主细胞。在一个实施方案中,通过使用在合适的培养基中培养的宿主细胞制备了MR蛋白。然后可从培养基或宿主细胞中分离该MR蛋白。
本发明的另一个方面涉及已导入MR基因的或其中MR基因被修饰的经遗传修饰的微生物。在一个实施方案中,通过导入至少一种作为转基因的编码突变MR序列的本发明核酸分子而修饰了所述微生物的基因组。在另一个实施方案中,通过用经修饰的MR基因进行同源重组而修饰(例如功能性破坏)了所述微生物基因组中的内源性MR基因。在一个优选的实施方案中,微生物属于棒状杆菌属或短杆菌(Brevibacterium)属,尤其优选谷氨酸棒状杆菌。在一个优选的实施方案中,该微生物也被用于制备目标化合物,如氨基酸、尤其是赖氨酸。
另一个优选的实施方案是具有一种或多种集合A中所描述核酸分子的宿主细胞。可用技术人员已知的多种方法制备这些宿主细胞。例如,可通过携带几种本发明核酸分子的载体转染这些细胞。然而,也可使用每次导入一种本发明核酸分子的载体从而同时或相继使用多种载体。从而可构建携带许多、高达数百的本发明核酸序列的宿主细胞。这种积累通常能够对宿主细胞精细化学品的生产率产生超加性效果。
本发明的另一方面涉及分离的MR蛋白或其部分,例如其生物活性部分。在一个优选的实施方案中,分离的MR蛋白或其部分在转录中、翻译中或翻译后调节谷氨酸棒状杆菌的一种或多种代谢途径。在另一个优选的实施方案中,分离的MR蛋白和其部分与集合B的氨基酸序列足够同源,因为该蛋白或其部分仍然能够转录地、翻译地或翻译后地调节谷氨酸棒状杆菌中的一种或多种代谢途径。
此外,本发明涉及分离的MR蛋白制品。在优选的实施方案中,该MR蛋白含有集合B的氨基酸序列。在另一个优选的实施方案中,本发明涉及分离的全长蛋白,其和集合B的全长氨基酸序列(其由集合A中的可读框编码)充分同源。
可以将MR多肽或其生物活性部分与非MR多肽功能性地连接以产生融合蛋白。在优选的实施方案中,该融合蛋白具有不同于单独的MR蛋白的活性,在其它优选的实施方案中,该融合蛋白转录地、翻译地或翻译后地调节谷氨酸棒状杆菌的一种或多种代谢途径。在尤其优选的实施方案中,所述融合蛋白向宿主细胞的整合调节细胞对目的化合物的生产率。
本发明的另一个方面涉及制备精细化学品的制备方法。该方法涉及含有载体的细胞的培养,这导致本发明MR核酸分子的表达而产生精细化学品。在一个优选的实施方案中,该方法还包括得到含有此类载体的细胞的步骤,所述细胞用载体转染导致MR核酸表达。在另一个优选的实施方案中,所述方法还包括从培养物中得到精细化学品的步骤。在一个优选的实施方案中,该细胞属于棒状杆菌属或短杆菌属。
本发明的另一个方面涉及调节来自微生物的分子生产率的方法。这些方法包括将细胞与调节MR蛋白活性或MR核酸表达的物质接触,使得和没有所述物质存在时相比细胞相关的活性受到调节。在一个优选的实施方案中,细胞关于谷氨酸棒状杆菌中代谢途径的一种或多种调节系统受到调节,使得该微生物的目标精细化学品的产量或生产率提高。调节MR蛋白活性的物质刺激例如MR蛋白的活性或MR核酸的表达。刺激MR蛋白活性或MR核酸表达的物质的例子包括小分子、活性MR蛋白和编码MR蛋白并已导入细胞的核酸分子。抑制MR活性或MR表达的物质的例子包括小分子和反义MR核酸分子。
本发明的另一个方面涉及调节细胞的目的化合物产量的方法,该方法包括向细胞中导入MR野生型基因或MR突变基因,该基因仍位于分立的质粒上或者整合到宿主细胞的基因组中。向基因组的整合可随机发生或者通过同源重组发生,由此天然基因被整合的拷贝所代替,导致细胞的目标化合物的生产率被调节。在一个优选的实施方案中,所述产量增加了。在另一个优选的实施方案中,化学品是精细化学品,其在一个特别优选的实施方案中是氨基酸。在一个特别优选的实施方案中,该氨基酸是L-赖氨酸。
发明详述
本发明提供了参与谷氨酸棒状杆菌代谢调节,包括参与精细化学品代谢调节的MR核酸和MR蛋白分子。本发明的这些分子可用于直接调节(例如当对赖氨酸代谢途径的调节蛋白活性的调节对该微生物的赖氨酸产量、生产率和/或生产效率具有直接影响)或间接调节微生物如谷氨酸棒状杆菌的精细化学品的生产率,间接调节仍然增加目标化合物的产量、生产率和/或生产效率(例如,当对核苷酸生物合成蛋白的调节对细菌的有机酸或脂肪酸的生产率具有影响时,这可能是由于所述化学品的生物合成或降解途径中伴随的调节修饰对核苷酸生物合成的修饰调节的一种反应)。本发明的诸方面进一步阐述如下。
I.精细化学品
术语“精细化学品”为本领域所认知,其包括已用于多种工业的由生物体产生的分子,其中所述工业例如但不限于药学工业、农业工业和化妆品工业。此类化合物包括有机酸如酒石酸、衣康酸和二氨基庚二酸、蛋白原的和非蛋白原的氨基酸、嘌呤和嘧啶碱基、核苷和核苷酸(如在Rehm等人编辑的生物技术(Biotechnology)第6卷中Kuninaka,A.(1996)核苷酸和相关化合物(Nucleotides and related compounds),561-612页,VCH:Weinheim及其中所含参考文献中所述)、脂类、饱和和不饱和脂肪酸(如花生四烯酸)、二醇(如丙二醇和丁二醇)、碳水化合物(如透明质酸和海藻糖)、芳香化合物(如芳香胺、香草醛和靛蓝)、维生素和辅因子(如在Ullmann工业化学百科全书(Ullmann’s Encyclopedia of Industrial Chemistry),A27卷,“维生素”(“Vitamins”),443-613页(1996)VCH:Weinheim及其中的参考文献;以及Ong,A.S.,Niki,E.和Packer,L.(1995)UNESCO/马来西亚科学和技术协会和亚州自由基研究协会联盟举办的“营养、脂类、健康和疾病”论文集(“Nutrition,Lipids,Health,and Disease”Proceedings ofthe UNESCO/Confederation of Scientific and Technological Associationsin Malaysia,and the Society for Free Radical Research-Asia),1994年9月1-3日在马来西亚槟榔屿举行,AOCS Press,(1995))、酶以及在Gutcho(1983)通过发酵的化学品(Chemicals by Fermentation),Noyes DataCorporation,ISBN:0818805086和其中的参考文献中所述的所有其它化学品。这些精细化学品中的某些化学品的代谢和用途进一步阐述如下。
A.氨基酸代谢和用途
氨基酸包含所有蛋白质的基本结构单元,并且为所有生物体的正常细胞功能所必需。术语“氨基酸”为本领域所认知。蛋白原的氨基酸有20种,为蛋白质的结构单元,在蛋白质中它们通过肽键连接,非蛋白原的氨基酸(已知成百种非蛋白原的氨基酸)在蛋白质中通常不存在(参见Ulmann工业化学百科全书(Ulmann工业化学百科全书(Ulmann’s Encyclopediaof Industrial Chemistry)),A2卷,57-97页,VCH:Weinheim(1985))。氨基酸可为D-或L-构型,尽管在天然存在的蛋白质中发现的唯一类型通常为L-氨基酸。已充分说明了20种蛋白原氨基酸中每一种在原核和真核细胞中的生物合成和降解途径(参见例如,Stryer,L.生物化学(Biochemistry),第三版,578-590页(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),这样称呼是因为由于它们生物合成的复杂性,必须从食物中摄取它们,它们被简单的生物合成途径转化为其余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、丝氨酸和酪氨酸)。高等动物能够合成这些氨基酸中的一部分氨基酸,但必需氨基酸必须从食物中摄取,以进行正常的蛋白质合成。
除了它们在蛋白质生物合成中的功能外,这些氨基酸本身还是令人感兴趣的化学品,并且许多已发现在食品、动物饲料、化学品、化妆品、农业和药学工业中具有多种应用。赖氨酸不仅对人的营养而且对单胃动物如禽和猪都是一种重要的氨基酸。谷氨酸盐是最常用的调味添加剂(谷氨酸单钠盐,MSG),并且广泛用于食品工业,天冬氨酸、苯丙氨酸、甘氨酸和半胱氨酸也是如此。甘氨酸、L-甲硫氨酸和色氨酸均用于药学工业。谷氨酰胺、缬氨酸、亮氨酸、异亮氨酸、组氨酸、精氨酸、脯氨酸、丝氨酸和丙氨酸用于药学工业和化妆品工业。苏氨酸、色氨酸和D/L-甲硫氨酸是广泛使用的动物饲料添加剂。(Rehm等人(编辑),生物技术(Biotechnology),第6卷,第14a章中的Leuchtenberger,W.(1996)Amino aids-technicalproduction and use,466-502页,VCH:Weinheim)。还发现这些氨基酸可用作前体,用于合成合成的氨基酸和蛋白质,如N-乙酰半胱氨酸、S-羧甲基-L-半胱氨酸、(S)-5-羟色氨以及在其他Ulmann工业化学百科全书(Ulmann’s Encyclopedia of Industrial Chemistry),A2卷,57-97页,VCH:Weinheim,1985中所述的物质。
在可产生它们的生物体如细菌中已充分描述了这些天然氨基酸的生物合成(细菌氨基酸生物合成及其调节的综述,见Umbarger,H.E.(1978)Ann.Rev.Biochem.47:533-606)。谷氨酸通过还原胺化α-酮戊二酸(柠檬酸循环中的中间体)而合成。谷氨酰胺、脯氨酸和精氨酸接下来由谷氨酸产生。丝氨酸的生物合成为三步,由3-磷酸甘油(糖酵解中间体)开始,在氧化、氨基转移和水解步骤后产生该氨基酸。半胱氨酸和甘氨酸两者均产生自丝氨酸;在丝氨酸转羟甲基酶催化的反应中前者通过高半胱氨酸与丝氨酸缩合,后者通过侧链β-碳原子转移至四氢叶酸。苯丙氨酸和酪氨酸在9步的生物合成途径中合成自糖分解和磷酸戊糖途径的前体赤藓糖4-磷酸和磷酸烯醇丙酮酸,它们的不同仅在预苯酸合成后的最后两步。色氨酸也从这两种起始分子产生,但其合成为11步的途径。酪氨酸也可在苯丙氨酸羟化酶催化的反应中由苯丙氨酸合成。丙氨酸、缬氨酸和亮氨酸均为丙酮酸盐(糖酵解的终产物)的生物合成产物。天冬氨酸形成自草酰乙酸(柠檬酸循环的中间体)。天冬酰胺、甲硫氨酸、苏氨酸和赖氨酸均通过转化天冬氨酸而形成。异亮氨酸从苏氨酸形成。组氨酸在一个复杂的9步途径中从一种活化糖--5-磷酸核糖-1-焦磷酸形成。
为细胞蛋白质合成所需的过量氨基酸不能被贮藏,并被降解以提供用于细胞主要代谢途径的中间体(综述参见Stryer,L.生物化学(Biochemistry)第三版,21章,“氨基酸降解和尿素循环”495-516页(1988))。虽然细胞可将不想要的氨基酸转化为有用的代谢中间体,从所需用于合成氨基酸的能量、前体分子和酶来说,氨基酸的产生是昂贵的。因此对于氨基酸的生物合成受到反馈抑制调节并不让人吃惊,其中特定氨基酸的存在起到减缓或完全终止其自身产生的作用(关于氨基酸生物合成途径中的反馈机制的综述参见Stryer,L.生物化学(Biochemistry)第三版,24章:”氨基酸和血红素的生物合成”575-600页(1988))。因此,任一特定氨基酸的输出受到细胞中存在的该氨基酸量的限制。
B.维生素、辅因子和营养成分的代谢和用途
维生素、辅因子和营养成分包含高等动物不能合成且必须摄取的另一类分子,虽然它们易于被其它生物体如细菌合成。这些分子自身为生物活性物质或为在多种代谢途径中作为电子载体或中间体的生物活性物质的前体。除了它们的营养价值外,这些化合物作为着色剂、抗氧化剂和催化剂或其它操作辅助剂也具有重要的工业价值(这些化合物的结构、活性和工业应用的概述参见例如,Ullman工业化学百科全书(Ullman’s Encyclopediaof Industrial Chemistry),“维生素”A27卷,443-613页,VCH:Weinheim,1996)。术语”维生素”为本领域认知,包括生物体行使正常功能所需的营养素,且该生物体自身不能合成它们。维生素组和包含辅因子和营养性化合物。术语“辅因子”包括进行正常酶促活性所需的非蛋白原化合物。此类化合物可为有机的或无机的,本发明的辅因子优选有机的。术语“营养成分”包括促进植物和动物尤其是人的健康食物添加剂。此类分子的实例为维生素、抗氧化剂和某些脂类(如多不饱和脂肪酸)。
已大量描述了这些分子在可产生它们的生物体如细菌中的生物合成(Ullman工业化学百科全书(Ullman’s Encyclopedia of IndustrialChemistry),“维生素”A27卷,443-613页,VCH:Weinheim,1996;Michal,G.(1999)生物化学途径:生物化学和分子生物学图集(BiochemicalPathways:An Atlas of Biochemistry and Molecular Biology),John Wiley&Sons;Ong,A.S.,Niki,E.和Packer,L.(1995)UNESCO/马来西亚科学和技术协会和亚州自由基研究协会联盟举办的“营养、脂类、健康和疾病”论文集(“Nutrition,Lipids,Health,and Disease”Proceedings of theUNESCO/Confederation of Scientific and Technological Associations inMalaysia,and the Society for Free Radical Research-Asia),1994年9月1-3日在马来西亚槟榔屿举行,AOCS Press:Champaign,IL X,374 S)。
硫胺素(维生素B1)由嘧啶和噻唑部分通过化学偶联产生。核黄素(维生素B2)由鸟苷-5’-三磷酸(GTP)和核糖-5’-磷酸合成。核黄素接下来用于合成黄素单核苷酸(FMN)和黄素腺嘌呤二核苷酸(FAD)。合起来称为‘维生素B6’的化合物类(例如,吡哆素、吡哆胺、5’-磷酸吡哆醛和商业上使用的维生素B6盐酸盐)均为共同结构单元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)可通过化学合成或发酵产生。泛酸生物合成的最后步骤由ATP-驱动的β-丙氨酸和泛解酸的缩合组成。负责转化为泛解酸、β-丙氨酸和泛酸缩合生物合成步骤的酶是已知的。泛酸盐的代谢活性形式为辅酶A,其生物合成由5步酶促步骤进行。泛酸盐、5’-磷酸吡哆醛、半胱氨酸和ATP为辅酶A的前体。这些酶不仅催化泛酸盐的形成,而且催化(R)-泛解酸、(R)-pantolacton、(R)-泛醇(维生素B5原)、泛酰巯基乙胺(及其衍生物)和辅酶A的产生。
已详细研究了微生物中生物素从前体分子庚二酰辅酶A的生物合成,并鉴定了几种所涉及的基因。发现许多相应的蛋白质也参与Fe-簇合成,属于nifS蛋白质类。硫辛酸衍生自辛酸,并在能量代谢中作为辅酶,此时其成为丙酮酸脱氢酶复合体和α-酮戊二酸脱氢酶复合体的一部分。叶酸盐为一组这样的物质,其均为叶酸衍生物,来自于L-谷氨酸、对氨基苯甲酸和6-甲基蝶呤。已详细研究了某些微生物中始于生物转化代谢中间体鸟苷-5’-三磷酸(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。
类可啉(如钴胺素且尤其是维生素B12)和卟啉属于一组四吡咯环体系的化学品。维生素B12的生物合成太复杂,目前尚未彻底解释清楚,但许多涉及的酶和物质已知。烟酸(烟酸盐)和烟酰胺为吡啶衍生物,也称为‘烟酸’。烟酸为重要的辅酶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)和它们的还原形式的前体。
这些化合物的大规模产生很大程度上依赖于无细胞的化学合成,虽然这些化学品中的一些如核黄素、维生素B6、泛酸盐和生物素也已通过微生物的大规模培养物产生。只有维生素B12是仅通过发酵产生的,因为其合成的复杂性。体外方法需要耗费材料和时间,通常以高的代价耗费。
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它们相应的蛋白质为治疗肿瘤和病毒感染的重要靶标。术语“嘌呤”或“嘧啶”包括构成核酸、辅酶和核苷酸的含氮碱基。术语“核苷酸”包括核酸分子的基本结构单元,其由含氮碱基、戊糖(对RNA而言,该糖为核糖,对DNA而言,该糖为D-脱氧核糖)和磷酸组成。术语“核苷”包括作为核苷酸前体的分子,但其不含核苷酸所具有的磷酸单元。通过抑制这些分子的生物合成,或抑制它们形成核酸分子的代谢,可抑制RNA和DNA合成;通过以靶向癌细胞的方式抑制这种活性,可抑制肿瘤细胞分裂和复制的能力。
此外,有不形成核酸分子但作为能量贮存体(即AMP)或作为辅酶(即FAD和NAD)的核苷酸。
一些出版物已描述了这些化学品通过影响嘌呤和/或嘧啶代谢对这些医学适应症的用途(例如Christopherson,R.I.和Lyons,S.D.(1990)“作为化学治疗剂的嘧啶和嘌呤从头生物合成的强力抑制剂”医学研究述评(Med.Res.Reviews)10:505-548)。对参与嘌呤和嘧啶代谢的酶的研究集中在研发可用作如免疫抑制剂或抗增生剂的新药上(Smith,J.L.,(1995)“核苷酸合成中的酶”结构生物学最新观点(Curr.Opin.Struct.Biol.)5:752-757;(1995)Biochem Soc.Transact.23:877-902)。但是,嘌呤和嘧啶碱基、核苷和核苷酸具有其它应用:用作一些精细化学品(例如,硫胺素、S-腺苷-甲硫氨酸、叶酸盐或核黄素)生物合成的中间体,作为细胞的能量载体(例如,ATP或GTP),和本身作为化学品,通常作为增味剂(例如,IMP或GMP)或用于一些医药用途(参见例如,Kuninaka,A.(1996)生物技术中的核苷酸和相关化合物(Nucleotides and Related Compounds inBiotechnology)第6卷,Rehm等人编辑.VCH:Weinheim,561-612页)。同样,参与嘌呤、嘧啶、核苷或核苷酸代谢的酶越来越多地用作靶,研发用于作物保护的抗这些酶的化学品包括杀真菌剂、除草剂和杀虫剂。
已阐释了这些化合物在细菌中的代谢(综述参见例如,核酸研究和分子生物学进展(Progress in Nucleic Acid Research and Molecular Biology),第42卷,Academic Press中的Zalkin,H.和Dixon,J.E.(1992)“嘌呤核苷酸的从头生物合成”,259-287页;和生物化学途径:生物化学和分子生物学图集(Biochemical Pathways:An Atlas of Biochemistry and MolecularBiology),Wiley:纽约一书中的Michal,G.(1999)“核苷酸和核苷”,第8章)。嘌呤代谢已是深入研究的主题,其对细胞的正常功能是必需的。高等动物中受损的嘌呤代谢可引起严重疾病,如痛风。嘌呤核苷酸从核糖-5-磷酸开始,经过一系列步骤,通过中间体化合物次黄苷-5’-磷酸(IMP),产生鸟苷-5’-单磷酸(GMP)或腺苷-5’-单磷酸(AMP),由此易于形成用作核苷酸的三磷酸形式。这些化合物也用作能量贮存体,这样它们的降解可提供细胞内用于许多不同生物化学过程的能量。嘧啶的生物合成由核糖-5-磷酸形成尿苷-5’-单磷酸(UMP)而进行。UMP接下来转化为胞苷-5’-三磷酸(CTP)。所有这些核苷酸的脱氧形式通过一步还原步骤从核苷酸的二磷酸核糖形式产生为核苷酸的二磷酸脱氧核糖形式。通过磷酸化后,这些分子可参与DNA合成。
D.海藻糖的代谢和用途
海藻糖由两个葡萄糖分子通过α,α-1,1连接组成。它通常作为甜味剂用于食品工业,为干的或冷冻食品和饮料中的添加剂。但它也用于药学、化妆品和生物技术工业(参见例如,Nishimoto等人(1998)美国专利号5,759,610;Singer,M.A.和Lindquist,S.生物技术趋势(Trends Biotech.)16(1998):460-467;Paiva,C.L.A.和Panek,A.D.Biotech.Ann.Rev.2(1996):293-314;和Shiosaka,M.J.Japan 172:(1997)97-102)。海藻糖通过酶从许多微生物产生,并天然释放入环境介质中,从中可用本领域公知的方法收集海藻糖。
II.代谢调节机制
所有活细胞都有复杂的分解代谢和合成代谢能力,其中许多代谢途径相互关联。为了维持该异常复杂的代谢网络的各部分之间的平衡,细胞采用了精密调节的调节系统。通过独立地或同时地调节酶合成和酶活性,细胞能够调节完全不同的代谢途径的活性以便满足细胞不断变化的需要。
可以在转录水平或翻译水平或者这两种水平上诱导或者抑制酶的合成(综述见Lewin,B.(1990)基因IV(Genes IV),第3部分:“通过转录调控原核细胞基因(Controlling prokaryotic genes by transcription)”,牛津大学出版社,牛津,213-301页,及其参考文献,和Michal,G.(1999),生物化学途径:生物化学和分子生物学图集(Biochemical Pathways:An Atlas ofBiochemistry and Molecular Biology),John Wiley&Sons)。所有这些已知的调节过程都被应答多种外部影响(例如温度、营养供给或光)的其他基因介导。参与这种类型调节的蛋白因子的例子包括转录因子。它们是结合DNA从而导致基因表达或者增加(正调节,如大肠杆菌ara操纵子)或者降低(负调节,如大肠杆菌lac操纵子)的蛋白质。这些调节表达的转录因子本身可受到调节。例如,它们的活性可受到结合DNA结合蛋白的低分子量化合物的调节,从而所述蛋白对DNA上合适的结合位点的结合被刺激(如ara操纵子的阿拉伯糖)或抑制(如lac操纵子的乳糖)(见,例如,Helmann,J.D.和Chamberlin,M.J.(1988),“细菌σ因子的结构和功能(Structure and function of bacterial sigma factors)”Ann.Rev.Biochem.57:839-872;Adhya,S.(1995)“今日lac和gal操纵子(The lac and galoperons today)”以及Boos,W.等人,“麦芽糖体系(The maltose system)”,这两篇文章在大肠杆菌中基因表达的调节(Regulation of Gene Expressionin Escherichia coli)一书中,由Lin,E.C.C.和Lynch,A.S.编辑,Chapman&Hall:New York,181-200页和201-229页;和Moran,C.P.(1993)“RNA聚合酶和转录因子(RNA polymerase and transcription factors)”,在:枯草芽孢杆菌和其他革兰氏阳性菌(Bacillus subtilis and other Gram-positivebacteria)一书中,由Sonenshein,A.L.编辑ASM:Washington D.C.653-667页)。
蛋白质的合成不仅在转录水平而且在翻译水平受到调节。可以通过许多机制进行调节,包括对核糖体结合一种或多种mRNA能力的修饰、核糖体与mRNA的结合、保持或去除mRNA二级结构、对特定基因使用常见或不常见密码子、一种或多种tRNA的丰度和特定调节机制如弱化作用进行调节(见Vellanoweth,R.I.(1993)枯草芽孢杆菌和其他革兰氏阳性菌中的翻译及其调节(Translation and its regulation in Bacillus subtilis andother Gram-positive bacteria),Sonenshein,A.L.等人编辑ASM:Washington,D.C.,699-711页及其参考文献)。
可以对各种代谢途径中的单个蛋白(相继调节)或同时对多种蛋白(协同调节)进行转录和翻译调节。表达受到协同方式调节的基因通常位于基因组中操纵子或调节子接近处。这种基因转录和基因翻译的上调或下调受到多种因素如底物(在一种或多种代谢途径中使用的前体和中间体)、分解代谢物(与从复杂有机分子如糖的降解的能量产生相关的生物化学途径产生的分子)和终产物(在代谢途径末得到的分子)的细胞或细胞外的量控制。编码特定代谢途径的活性所需的酶的基因表达被所述代谢途径的大量底物分子所诱导。相应地,该基因表达被胞内大量该途径的终产物所抑制(Snyder,L.和Champness,W.(1977),细菌的分子生物学(TheMolecular Biology of Bacteria)ASM:Washington)。基因表达同样可被其他外部或内部因素如环境条件(例如热、氧化性应急或饥饿)所调节。这种全局环境变化导致专门的调节基因表达的变化,这些调节基因通过结合DNA而直接或间接(通过其他基因或蛋白)引发基因表达,从而诱导或抑制转录(见,例如,Lin,E.C.C.和Lynch,A.S.编辑(1995),大肠杆菌中基因表达的调节(Regulation of Gene Expression in Escherichia coli),Chapman&Hall:New York)。
调节细胞代谢的另一个机制发生在蛋白质水平。该调节通过其他酶的活性或通过结合阻止或赋予该蛋白正常功能的低分子量组分进行。通过结合低分子量化合物进行蛋白调节的例子包括GTP或NAD的结合。低分子量化学物质的结合通常是可逆的,例如GTP结合蛋白。这些蛋白存在两种状态(结合GTP或GDP),一种状态是蛋白质的活性形式,另一中是无活性形式。
蛋白活性被其他酶的作用调节,其中其他酶通常通过对蛋白的共价修饰(即氨基酸残基如组氨酸或天冬氨酸的磷酸化或甲基化)调节。这种共价修饰通常是可逆的并且受到具有相反活性的酶影响。一个例子是激酶和磷酸酶在蛋白质磷酸化中的相反活性:蛋白激酶在靶蛋白上(例如丝氨酸或苏氨酸)磷酸化特定残基,而蛋白磷酸酶从所述蛋白除去磷酸基团。调节其他蛋白活性的酶本身通常受到外界刺激的调节。这些刺激被作为传感器的蛋白所介导。被这些传感器蛋白介导外界信号的一个熟知的机制是二聚体化,但也知道其他机制(见,例如Msadek,T.等(1993)“双组分调节体系(Two-component Regulatory Systems)”在:枯草芽孢杆菌和其他革兰氏阳性菌(Bacillus subtilis and Other Gram-Positive Bacteria)一书中,Sonenshein,A.L.等人编辑,ASM:Washington,729-745页及其参考文献)。
详细了解调控微生物中细胞代谢的调节网络对通过发酵高产量地生产化学品是关键的。可以去除或降低下调代谢途径的调控系统以便提高目标化学品的合成,相应地,可以组成性地活化或优化上调目标产物的代谢途径的调控系统的活性(如Hirose,Y.和Okada,H.(1979)“氨基酸的微生物生产(Microbial Production of Amino Acids)”,在:Peppler,H.J.和Perlman,D.(编辑)微生物技术(Microbial Technology)第二版,第1卷,第7章,Academic Press,New York)。
III.本发明的组成要件和方法
本发明至少部分是基于本文中称作MR核酸和MR蛋白分子的新分子的检测,这些分子通过转录、翻译或翻译后的措施调节谷氨酸棒状杆菌中的一种或多种代谢途径。在一个实施方案中,MR分子转录地、翻译地或翻译后地调节谷氨酸棒状杆菌中的代谢途径。在一个优选的实施方案中,调节谷氨酸棒状杆菌中的一种或多种代谢途径的本发明MR分子的活性影响所述生物体的目标精细化学品的生产率。在一个尤其优选的实施方案中,本发明的MR分子具有经调节的活性,使得被本发明MR蛋白调节的谷氨酸棒状杆菌代谢途径的效率或产量受到调节,并且这直接或间接地调节谷氨酸棒状杆菌的目标精细化学品的产量、生产率和/或生产效率。
术语“MR蛋白”或“MR多肽”包括转录中、翻译中或者翻译后调节谷氨酸棒状杆菌中代谢途径的蛋白。MR蛋白的例子包括表1和集合A中所列出的MR基因编码的那些蛋白。术语“MR基因”和“MR核酸序列”包括编码MR蛋白的核酸序列,其含有一个编码区和相应的非翻译5’和3’序列区。MR基因的例子在表1中列出。术语“生产率”为本领域已知并且包括发酵产物的浓度(例如在预定的时间段和预定的发酵体积产生的目标精细化学品(例如kg产物/h/l))的浓度。术语“生产效率”包括得到特定产物量所需时间(例如,细胞达到精细化学品的特定生产率所需时间)。术语“产量”或“产物/碳产量”为本领域已知并且包括将碳源转化成产物(即精细化学品)的效率。这例如通常表达为kg产物/kg碳源。增加化合物的产量或生产率为在预定期间内及特定的培养体积中增加了所得分子的量或该化合物适宜得到的分子的量。术语“生物合成”和“生物合成途径”为本领域已知并且包括细胞在例如多步过程或高度调节的过程中从中间物合成化合物,优选有机化合物。术语“降解”和“降解途径”为本领域已知并且包括细胞在例如多步过程或高度调节的过程中将化合物,优选有机化合物裂解成降解产物(用更通俗的术语:更小或更简单的分子)。术语“代谢”为本领域已知并且包括生物体中发生的全部生物化学反应。特定化合物的代谢(例如氨基酸如甘氨酸的代谢)因此包括细胞中该化合物的所有生物合成、修饰和降解途径。术语“调节”为本领域已知并且包括调控另一蛋白质活性的蛋白质的活性。术语“转录调节”为本领域已知并且包括抑制或活化编码靶蛋白的DNA转化成mRNA的蛋白质的活性。术语“翻译调节”为本领域已知并且包括抑制或活化编码靶蛋白的mRNA转化成蛋白分子的蛋白质的活性。术语“翻译后调节”为本领域已知并且包括通过共价修饰靶蛋白(例如通过甲基化、糖基化或磷酸化)抑制或提高靶蛋白活性的蛋白质的活性。
在另一个实施方案中,微生物如谷氨酸棒状杆菌中本发明的MR分子能够调节目标分子例如精细化学品的生产率。借助于基因重组技术,可操作一种或多种本发明的代谢途径调节蛋白使得它们的功能被调节。可以例如提高生物合成酶的效率或者破坏其变构调控区以便阻止该化合物产生的反馈抑制。所以,可以通过替换、缺失或添加来删除或修饰降解酶使得其对目标化合物的降解活性降低而不损害细胞的存活。在任何情况下,可以增加任何所述目标精细化学品的总产量或生产率。
还可能的是本发明的蛋白和核苷酸分子的这些修饰能够间接提高精细化学品的生产率。细胞中代谢途径的调节机制不可避免地相关联,一种代谢途径的激活能够伴随着抑制或激活另一种代谢途径。调节本发明的一种或多种蛋白的活性能影响其他精细化学品生物合成或降解途径活性的生产率或效率。降低一种MR蛋白抑制编码氨基酸生物合成中特定蛋白的基因转录使得可能同时抑制其他氨基酸生物合成途径,因为这些代谢途径相互关联。通过调节本发明MR蛋白可能使细胞生长和细胞分裂与细胞胞外环境以一定程度解偶联;当用于生长和细胞分裂的胞外条件不是最佳时某种MR蛋白通常抑制核苷酸的生物合成,通过影响该MR蛋白使其失去抑制核苷酸合成的功能,可能使得即使胞外条件较差时仍能生长。这对于大规模发酵培养尤其重要,大规模发酵的温度、营养供给或通气通常不是最佳的,但是当所述因子的细胞调节系统被去除后仍然能够促进生长和细胞分裂。
制备本发明核酸序列的一个适宜的起点是谷氨酸棒状杆菌菌株的基因组,可以从美国典型培养物保藏中心得到该菌株(名称是ATCC 13032)。
用常规方法,通过表1所示的修饰可以从这些核酸序列制备本发明的核酸序列。
本发明的MR蛋白或其生物学活性部分或片段能够在转录中、翻译中或翻译后调节谷氨酸棒状杆菌的代谢途径或可以具有表1列出的一种或多种活性。
下面的分段详细描述本发明的各方面:
A.分离的核酸分子
本发明的一个方面涉及编码MR多肽或其生物活性部分的分离核酸分子及核酸分子片段,其中所述核酸分子片段足以用作杂交探针或引物,用来鉴定或扩增编码MR的核酸(例如MR DNA)。如此处所用,术语“核酸分子”是指包括DNA分子(例如cDNA或基因组DNA)和RNA分子(例如mRNA)和用核苷酸类似物产生的DNA或RNA类似物。该术语也包括位于基因编码区3’和5’端的非翻译区:基因编码区5’端上游的至少约100个核苷酸序列和基因编码区3’端下游的至少约20个核苷酸序列。核酸分子可为单链或双链,但优选地为双链DNA。“分离的”核酸分子为从其它核酸分子分离的核酸分子,其存在于核酸的天然源中。优选地,“分离的”核酸不具有该核酸所来自的生物体基因组DNA中天然存在于该核酸侧翼的序列(即位于该核酸5’和3’端的序列)。例如,在不同实施方案中,该分离的MR核酸分子可包含少于约5kb,4kb,3kb,2kb,1kb,0.5kb或0.1kb的该核酸所来自的细胞(如谷氨酸棒状杆菌细胞)基因组DNA中天然存在于该核酸分子侧翼的核苷酸序列。而且,“分离的”核酸分子如cDNA分子当通过重组技术产生时可基本上不含有其它细胞物质或培养基,当通过化学合成产生时,可基本上不含有化学前体或其它化学品。
本发明的核酸分子,如具有的核苷酸序列为集合A的核酸分子或其部分可用标准的分子生物学技术和本文提供的序列信息制备。例如,使用集合A全部或部分序列作为杂交探针和标准的杂交技术(例如,如在Sambrook,J.,Fritsh,E.F.和Maniatis,T.分子克隆:实验室手册(Molecular Cloning:A Laboratory Manual).第二版,冷泉港实验室,冷泉港实验室出版社,冷泉港,纽约,1989中所述),可自谷氨酸棒状杆菌文库分离谷氨酸棒状杆菌MR DNA。而且,包含集合A的全部序列或其部分的核酸分子可使用根据该序列设计的寡核苷酸引物,应用聚合酶链反应分离(例如,包含集合A的全部或部分的核酸分子可使用根据该同样序列设计的寡核苷酸引物,应用聚合酶链反应分离)。例如,可从正常的内皮细胞分离mRNA(例如,通过Chirgwin等人(1979)生物化学(Biochemistry)18:5294-5299的硫氰酸胍提取法)以及用反转录酶(例如,莫洛尼鼠类白血病毒反转录酶,可从Gibco/BRL,Bethesda,MD获得;或AMV反转录酶,可从Seikagaku America,Inc.,St.Petersburg,FL获得)可制备cDNA。用于聚合酶链反应扩增的合成寡核苷酸引物可根据集合A所示的核苷酸序列之一设计。本发明的核酸可用cDNA或另外用基因组DNA作为模板、使用合适的寡核苷酸引物按照标准的PCR扩增技术来扩增。这样扩增的核酸可克隆入合适的载体并通过DNA序列分析鉴定。对应于MR核苷酸序列的寡核苷酸可通过标准的合成技术制备,例如用DNA自动合成仪制备。
在一个优选的实施方案中,本发明的分离核酸分子包含集合A中所示的任一核苷酸序列。
在另一个优选的实施方案中,本发明的分离的核酸分子含有与集合A中所示核酸序列之一互补的核酸分子,所述核酸分子与集合A中所示核酸序列之一足够互补使其与集合A中所示序列之一杂交而得到稳定双链体。
在一个实施方案中,本发明的核酸分子编码这样的蛋白质或其部分,其包括的氨基酸序列充分同源于集合B的一个氨基酸序列,以至于该蛋白质或其部分仍然能够在转录中、翻译中或翻译后调节谷氨酸棒状杆菌的代谢途径。如此处所用,术语“充分同源”是指蛋白质或其部分的氨基酸序列具有最小数的与集合B中氨基酸序列相同或等同的氨基酸残基(例如,氨基酸残基具有与集合B中序列之一的氨基酸残基相似的侧链),以至于该蛋白质或其部分能够在转录中、翻译中或翻译后调节谷氨酸棒状杆菌的代谢途径。此类代谢途径中的蛋白质成员如此处所述,可调节一种或多种精细化学品的生物合成或降解。此类活性的实例也在此进行了描述。因此,“MR蛋白的功能”涉及一种或多种精细化学品代谢途径的总体调节。MR蛋白活性的实例在表1中给出。
本发明MR核酸分子编码的蛋白部分优选地是任何MR蛋白的具生物学活性的部分。文中所用术语“MR蛋白的生物学活性部分”包含MR蛋白的一部分,例如结构域或基序,其能够在转录中、翻译中或翻译后调节谷氨酸棒状杆菌的代谢途径或者具有表1中所示活性。为了确定MR蛋白或者其生物学活性部分能否在转录中、翻译中或翻译后调节谷氨酸棒状杆菌的代谢途径,可以进行酶活性分析。这些分析方法在实施例部分的实施例8中详细描述,技术人员熟知这些方法。
除了群体中存在的MR序列的天然变体外,技术人员还理解可通过突变向集合A的核苷酸序列中导入变化,由此导致所编码的MR蛋白的氨基酸序列改变,但MR蛋白的功能不变。例如,可在集合A的核苷酸序列中进行在“非必需”氨基酸残基处导致氨基酸替换的核苷酸替换。可以修饰任一MR蛋白(集合B)的野生型序列中的“非必需”氨基酸残基而不改变所述MR蛋白的活性,但“必需”氨基酸残基为MR蛋白活性所需。然而,其它氨基酸残基(例如,在具有MR活性的结构域中不保守或仅为半保守的那些氨基酸残基)对活性可为非必需的,并因此可被改变,而此类氨基酸残基的改变不改变MR活性。
编码同源于集合B的蛋白质序列的MR蛋白的分离核酸分子可通过将一个或多个核苷酸替换、添加或删除导入集合A的核苷酸序列而产生,这样将一个或多个氨基酸的替换、添加或删除导入所编码的蛋白质。可通过标准技术将突变导入集合A的核苷酸序列之一,如定点诱变和PCR-介导的诱变。优选地,在一处或多处预测的非必需氨基酸残基处进行保守氨基酸替换。“保守氨基酸替换”为将氨基酸残基用具有相似侧链的氨基酸残基替换。具有相似侧链的氨基酸残基类已在本领域定义过。这些分类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)、具有不带电荷的极性侧链的氨基酸(例如,甘氨酸、天冬酰胺、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-分支侧链的氨基酸(例如,苏氨酸、缬氨酸、异亮氨酸)和具有芳香侧链的氨基酸(例如,酪氨酸、苯丙氨酸、色氨酸、组氨酸)。因此,在MR蛋白中预测的非必需氨基酸残基优选地用来自同一侧链类的另一氨基酸残基替换。在另一实施方案中,可如通过饱和诱变沿着全部或部分MR编码序列随机导入突变,并对所得突变体筛选文中所述的MR活性,以鉴定保留MR活性的突变体。对任一集合A的序列诱变后,可重组表达编码蛋白,并用例如文中所述的试验(参见实施例部分的实施例8)确定所述蛋白质的活性。
B.重组表达载体和宿主细胞
本发明的另一方面涉及包含编码MR蛋白(或其部分)的核酸或包含基因组合且其中至少一种基因编码MR蛋白的载体,优选表达载体。如此处所用,术语“载体”是指这样的核酸分子,其可转运与其连接的另一核酸。一种类型的载体为“质粒”,是指其中可连接额外DNA片段的环状双链DNA环。另一种类型的载体为病毒载体,其中可将额外DNA片段连接入病毒基因组。某些载体可在它们所导入的宿主细胞中自主复制(例如,具有细菌复制起点的细菌载体和游离型哺乳动物载体)。其它载体(例如,非游离型哺乳动物载体)在导入宿主细胞后整合入宿主细胞的基因组,并因此随着宿主基因组而复制。而且,某些载体可指导与它们操作性连接的基因的表达。此类载体此处称为“表达载体”。一般,用于重组DNA技术中的表达载体通常为质粒形式。在本说明书中,“质粒”和“载体”可互换使用,因为质粒是最经常使用的载体形式。但是,本发明旨在包括其它形式的表达载体,如病毒载体(例如,复制缺陷型反转录病毒、腺病毒和腺相关病毒),它们起到类似的作用。
本发明的重组表达载体以适于在宿主细胞中表达核酸的形式包含本发明的核酸,这就是说重组表达载体包括一个或多个调节序列,其选择是基于用作表达的宿主细胞进行的,它们可操作地与待表达的核酸序列连接。在重组表达载体中,术语“功能性地连接”是指目的核苷酸序列与调节序列以允许该核苷酸序列表达(例如,在体外转录/翻译系统中,或当载体导入宿主细胞时在宿主细胞中表达)的方式连接。术语“调节序列”是指包括启动子、增强子和其它表达调控元件(例如,多聚腺苷酸化信号)。此类调节序列如在Goeddel;基因表达技术:酶学方法(GeneExpressionTechnology:Methods in Enzymology)185,Academic Press,SanDiego,CA(1990)中描述。调节序列包括在许多类型宿主细胞中控制核苷酸序列组成型表达的调节序列和仅在某些宿主细胞中控制核苷酸序列直接表达的调节序列。本领域普通技术人员了解到表达载体的设计可取决于如对欲转化宿主细胞的选择、所需的蛋白质表达程度等因素。本发明的表达载体可导入宿主细胞,以产生由此处所述核酸所编码的蛋白质或肽,包括融合蛋白或融合肽(例如,MR蛋白、MR蛋白的突变形式、融合蛋白等)。
本发明的重组表达载体可设计用于在原核或真核细胞中表达MR蛋白。例如,MR基因可在细菌细胞如谷氨酸棒状杆菌、昆虫细胞(使用杆状病毒表达载体)、酵母细胞和其它真菌细胞(参见Romanos,M.A.等人(1992)“在酵母中外源基因的表达:综述”(“Foreign gene Expression in yeast:areview”),酵母(Yeast)8:423-488;J.W.Bennet&L.L.Lasure编辑的更多的用于真菌的基因操作(More Gene Manipulations in Fungi),AcademicPress:San Diego一书中的van den Hondel,C.A.M.J.J.等人(1991)“丝状真菌中的异源基因表达”(“Heterologous gene Expression in filamentousfungi”),396-428页;以及Peberdy,J.F.等人编辑的真菌的应用分子遗传学(Applied Molecular Genetics of Fungi),Cambridge University Press:Cambridge一书中的van den Hondel,C.A.M.J.J.和Punt,P.J.(1991)“丝状真菌的基因转移体系和载体开发”(Gene transfer systems and vectordevelopment for filamentous fungi),1-28页)、藻类和单细胞植物细胞(参见Schmidt,R.和Willmitzer,L.(1988),根瘤农杆菌介导的在鼠耳芥叶和子叶外植体中的高效转化,植物细胞报道(Plant Cell Rep.):583-586)、或哺乳动物细胞中表达。合适的宿主细胞在Goeddel,基因表达技术:酶学方法(Gene Expression Technology:Methods in Enzymology)185,AcademicPress,San Diego,CA(1990)中进一步进行了讨论。另外,可体外转录并翻译重组表达载体,如使用T7启动子调节序列和T7聚合酶。
在原核生物中蛋白质的表达主要用含有组成型或诱导型启动子的载体进行,控制融合或非融合蛋白的表达。融合载体将一些氨基酸添加至其中编码的蛋白质上,通常添加至重组蛋白的氨基末端。此类融合载体一般具有三个任务:1)用于增强重组蛋白的表达;2)用于增加重组蛋白的溶解性;和3)在亲和层析中作为配体而利于重组蛋白的纯化。通常,于融合部分和重组蛋白的连接处导入蛋白酶剪切位点到融合表达载体中,这使得在该融合蛋白纯化后,可将重组蛋白从融合单元分离。这些酶和它们的对应识别序列包括Xa因子、凝血酶和肠激酶。
通常的融合表达载体包括pGEX(Pharmacia Biotech Inc;Smith,D.B.和Johnson,K.S.(1988)基因(Gene)67:31-40)、pMAL(New EnglandBiolabs,Beverly,MA)和pRIT5(Pharmacia,Piscataway,NJ),它们分别将谷胱甘肽S-转移酶(GST)、麦芽糖E结合蛋白和蛋白A结合至重组靶蛋白。在一个实施方案中,将MR蛋白的编码序列克隆入pGEX表达载体,以产生编码融合蛋白的载体,其从N-末端至C-末端包含GST-凝血酶切割位点-X蛋白。该融合蛋白使用谷胱甘肽-琼脂糖树脂通过亲和层析纯化。通过将该融合蛋白用凝血酶切割可得到不与GST融合的重组MR蛋白。
合适的诱导型非融合大肠杆菌表达载体的实例包括pTrc(Amann等人(1988)基因(Gene)69:301-315)和pET 11d(Studier等人,基因表达技术:酶学方法(Gene Expression Technology:Methods in Enzymology)185,Academic Press,San Diego,California(1990)60-89)。pTrc载体中目标基因的表达依赖于宿主RNA聚合酶从杂合的trp-lac融合启动子的转录。pET 11d载体中目标基因的表达依赖于共表达的病毒RNA聚合酶(T7 gn1)介导的从T7-gn10-lac融合启动子的转录。该病毒聚合酶由宿主菌株BL21(DE3)或HMS174(DE3)中居住的、具有位于lacUV 5启动子转录调控下的T7 gn1基因的λ前噬菌体提供。
最大化重组蛋白表达的一个策略是将该蛋白质在具有对所述重组蛋白进行蛋白酶切割的能力缺损的宿主细菌中表达(Gottesman,S.,基因表达技术:酶学方法(Gene Expression Technology:Methods in Enzymology)185,Academic Press,San Diego,California(1990)119-128)。另一策略是改变欲插入表达载体的核酸的核酸序列,以使用于每一氨基酸的单个密码子为选择用于表达的细菌如谷氨酸棒状杆菌所优选使用的密码子(Wada等人(1992)核酸研究(Nucleic Acids Res.)20:2111-2118)。本发明核酸序列的此改变可应用标准的DNA合成技术实施。
在另一实施方案中,MR蛋白表达载体为酵母表达载体。用于在酿酒酵母中表达的载体包括pYepSec1(Baldari,等人(1987)Embo J.6:229-234),pMFa(Kurjan和Herskowitz,(1982)细胞(Cell)30:933-943),pJRY88(Schultz等人(1987)基因(Gene)54:113-123),和pYES2(InvitrogenCorporation,San Diego,CA)。适用于其它真菌如丝状真菌的载体和用于构建载体的方法包括在:J.F.Peberdy等人编辑的真菌的应用分子遗传学(Applied Molecular Genetics of Fungi),Cambridge University Press:Cambridge一书中的van den Hondel,C.A.M.J.J.和Punt,P.J.(1991)“丝状真菌的基因转移体系和载体开发”(Gene transfer systems and vectordevelopment for filamentous fungi),1-28页中详述的那些。
另外,本发明的MR蛋白可使用杆状病毒表达载体在昆虫细胞中表达。可用于在培养的昆虫细胞(例如Sf 9细胞)中表达蛋白质的杆状病毒载体包括pAc系列(Smith等人(1983)Mol.Cell Biol.3:2156-2165)和pVL系列(Lucklow和Summers(1989)病毒学(Virology)170:31-39)。
在另一实施方案中,本发明的MR蛋白可在单细胞植物细胞(如藻类)或在来自高等植物(如种子植物,诸如作物)的植物细胞中表达。植物表达载体的实例包括在:Becker,D.,KeMRer,E.,Schell,J.和Masterson,R.(1992)“具有靠近左边界的选择性标记的新植物双元载体(“New plantbinary vectors with selectable markers located proximal to the leftborder”),植物分子生物学(Plant Mol.Biol.)20:1195-1197;和Bevan,M.W.(1984)“用于植物转化的双元农杆菌载体”(“Binary Agrobacteriumvectors for plant transformation”),核酸研究(Nucl.Acid.Res.)12:8711-8721中详述的那些。
在另一实施方案中,本发明的核酸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)自然(Nature)329:840)和pMT2PC(Kaufman等人(1987)EMBO J.6:187-195)。当用于哺乳动物细胞时,表达载体的调控功能通常由病毒调控元件提供。例如,常用的启动子来自多瘤、腺病毒2、巨细胞病毒和猿猴病毒40。用于原核和真核细胞的其它合适表达系统参见Sambrook,J.,Fritsh,E.F.和Maniatis,T.分子克隆:实验室手册(Molecular Cloning:ALaboratory Manual),第二版,Cold Spring Harbor Laboratory,ColdSpring Harbor Laboratory Press,Cold Spring Harbor,NY,1989中的第16和17章。
在另一实施方案中,重组哺乳动物表达载体可优选地在特定细胞类型中指导核酸的表达(例如,使用组织-特异性调控元件来表达核酸)。组织-特异性调控元件为本领域公知。合适的组织-特异性启动子的非限制性实例包括白蛋白启动子(肝-特异性;Pinkert等人(1987)基因进展(Genes Dev.)1:268-277)、淋巴-特异性启动子(Calame and Eaton(1988)Adv.Immunol.43:235-275),特别是T细胞受体启动子(Winoto和Baltimore(1989)EMBOJ.8:729-733)和免疫球蛋白启动子(Banerji等人(1983)细胞(Cell)33:729-740;Queen和Baltimore(1983)细胞(Cell)33:741-748)、神经元-特异性启动子(例如,神经丝启动子;Byrne和Ruddle(1989)PNAS86:5473-5477)、胰-特异性启动子(Edlund等人(1985)科学(Science)230:912-916)、以及乳腺-特异性启动子(例如,奶乳清启动子;美国专利号4,873,316和欧洲申请公开号264,166)。也同样包括调节发育的启动子,如鼠hox启动子(Kessel和Gruss(1990)科学(Science)249:374-379)和甲胎蛋白启动子(Campes和Tilghman(1989)基因进展(Genes Dev.)3:537-546)。
本发明还提供了含本发明的DNA分子的重组表达载体,其中该DNA分子以反义方向克隆在表达载体中。也就是,DNA分子功能性地连接到调控序列上,该连接方式使得可以表达产生与MR mRNA反义的RNA分子(通过转录该DNA分子)。可以对功能性地连接反义方向克隆的核酸的调控序列进行选择以控制反义RNA分子在多种细胞类型中持续表达,例如,可选择病毒启动子和/或增强子或调控序列以控制反义RNA实现组成型组织特异性或细胞类型特异性表达。反义表达载体的形式可为重组质粒、噬菌粒或减毒病毒,其中在高效调控区域的控制下产生反义核酸,其活性由载体所导入细胞的类型决定。反义基因的基因表达调控的讨论见Weintraub,H.等人,反义RNA作为遗传分析的分子工具,Reviews-Trendsin Genetics,卷1(1)1986。
本发明的另一方面涉及已导入本发明重组表达载体的宿主细胞。术语“宿主细胞”和“重组宿主细胞”在此处可互换使用。当然,该术语不仅指特定的受试细胞,也指该细胞的后代或潜在后代。由于突变或环境因素在传代过程中可发生某些改变,这样,后代实际上可能并非与亲代细胞相同,但其仍包括在此处所用术语的范围之内。
宿主细胞可为任意的原核或真核细胞。例如,MR蛋白可在细菌细胞如谷氨酸棒状杆菌、昆虫细胞、酵母或哺乳动物细胞(如中国仓鼠卵巢(CHO)细胞或COS细胞)中表达。其他合适的宿主细胞为技术人员公知。与谷氨酸棒状杆菌有关的、可常规用作本发明核酸和蛋白质分子的宿主细胞的微生物如表3所示。
载体DNA可通过常规转化或转染技术导入原核或真核细胞中。如此处所用,术语“转化”和“转染”意指本领域内公知的各种将外源核酸(例如DNA)导入宿主细胞的技术,包括磷酸钙或氯化钙共沉淀、DEAE-葡聚糖-介导的转染、脂转染、电穿孔。转化或转染宿主细胞的合适方法可在Sambrook等人(分子克隆:实验室手册(Molecular Cloning:A LaboratoryManual).第二版,Cold Spring Harbor Laboratory,Cold Spring HarborLaboratory Press,Cold Spring Harbor,NY,1989)和其它的实验室手册中找到。
对于哺乳动物细胞的稳定转染,已知根据所用的表达载体和所用的转染技术,仅一小部分的细胞可将外源DNA整合到基因组中。一般将编码选择标记的基因(如抗生素抗性基因)与目的基因一同导入宿主细胞以鉴定和筛选整合体。优选的选择标记包括那些可赋予药物抗性的基因,所述药物如G418、潮霉素和氨甲蝶呤。编码选择标记的核酸可与编码MR蛋白质的基因位于同一载体上被导入宿主细胞或可以位于不同的载体上导入。经导入核酸稳定转染的细胞可通过药物筛选进行鉴定(例如,掺入了选择标志基因的细胞将存活,而其他细胞死亡)。
通过制备出含至少部分MR基因的载体,所述部分MR基因中已经导入缺失、添加或替代因而可以改变MR基因(如功能性破坏)来生成同源重组微生物。优选地该MR基因为谷氨酸棒状杆菌MR基因,但也可为来自相关细菌或甚至来自哺乳动物、酵母或昆虫的同系物。在一个优选的实施方案中,载体设计为在同源重组后可功能性破坏内源MR基因(即,不再编码功能蛋白质;也称为“敲除”载体)。另外,载体可设计为在同源重组后内源MR基因发生突变或改变但仍编码功能蛋白质(例如,可改变上游调控区域由此改变内源MR蛋白的表达)。在此同源重组载体中,改变的MR基因部分在其5′和3′端侧翼为MR基因的其它核酸,从而允许在载体携带的外源MR基因和微生物中的内源MR基因之间发生同源重组。此其它的两翼MR核酸的长度应足以使与内源基因的重组成功进行。通常,载体内包括几千碱基的侧翼DNA(5′及3′端)(参见例如,Thomas,K.R.,和Capecchi,M.R.(1987)细胞(Cell)51:503中对同源重组载体的描述)。使用本领域公知的方法,将该载体导入微生物(例如,通过电穿孔)和细胞,筛选其中导入的MR基因已与内源MR基因同源重组的微生物和细胞。
在另一实施方案中,产生的重组微生物可含有可调控导入基因表达的所选系统。例如,将在载体中包含的MR基因插入乳糖操纵子的控制下,使得MR基因仅在存在IPTG时表达。此类调控系统为本领域熟知。
本发明的宿主细胞如培养的原核或真核宿主细胞可用于产生(即表达)MR蛋白。因此,本发明还提供了使用本发明的宿主细胞来产生MR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(其中已导入了编码MR蛋白的重组表达载体,或其中基因组已导入了编码野生型或改变的MR蛋白的基因),直至产生MR蛋白。在另一实施方案中,该方法还包括从培养基或宿主细胞分离MR蛋白。
C.本发明的用途和方法
文中所述的核酸分子、蛋白质、蛋白质同系物、融合蛋白、引物、载体和宿主细胞可用于一种或多种下述方法中:鉴定谷氨酸棒状杆菌及相关生物体;绘制与谷氨酸棒状杆菌相关的生物体的基因组图谱;鉴定并定位目标谷氨酸棒状杆菌序列;进化研究;确定为MR蛋白所需的功能区域;调节MR蛋白活性;调节MR途径的活性;以及调节目标化合物如精细化学品的细胞生产率。本发明的MR核酸分子具有多种用途。首先,它们可用于鉴定生物体为谷氨酸棒状杆菌或其近亲。它们也可用于在混合的微生物群体中鉴定谷氨酸棒状杆菌或其相关菌的存在。本发明提供了许多谷氨酸棒状杆菌基因的核酸序列。通过将微生物单一群体或混合群体的培养物提取的基因组DNA在严格条件下与包含对谷氨酸棒状杆菌独特的谷氨酸棒状杆菌基因区域的探针杂交,可确定该生物体是否存在。虽然谷氨酸棒状杆菌自身为非致病的,但它与致病种如白喉棒状杆菌有亲缘关系。这种生物体的检测具有显著的临床重要性。
本发明的核酸和蛋白质分子也可用作基因组特异性区域的标记。它不仅可用于基因组图谱,也可用于谷氨酸棒状杆菌蛋白质的功能研究。例如,可切割谷氨酸棒状杆菌基因组,并将片段与DNA-结合蛋白孵育来确定与特定的谷氨酸棒状杆菌DNA-结合蛋白结合的基因组区域。另外,那些结合蛋白的片段可用本发明的核酸分子探测,优选地用具有易于检测的标记物的核酸分子探测;此核酸分子与基因组片段的结合可将片段定位于谷氨酸棒状杆菌的基因组图谱中,并且当用不同的酶进行多次时,利于快速确定蛋白质结合的核酸序列。而且,本发明的核酸分子可充分同源于亲缘种的序列,这样这些核酸分子可用作在亲缘菌如乳糖发酵短杆菌中构建基因组图谱的标记。
本发明的MR核酸分子也用于进化研究和蛋白质结构研究。大量的原核和真核细胞应用本发明分子参与的代谢过程;通过将本发明核酸分子的序列与那些来自其它生物体的编码类似酶的序列比较,可评估所述生物体的进化关系。所以,此比较使得可评估哪些区域保守和哪些区域不保守,这有利于确定那些对酶功能所必需的蛋白质区域。这种确定对蛋白质工程研究有价值,并可指出该蛋白质可耐受何种诱变且不丢失其功能。
本发明MR核酸分子的操作可导致产生与野生型MR蛋白功能不同的MR蛋白。这些蛋白质可在功效或活性上提高,在细胞内可以较通常多的量存在,或可在功效或活性上降低。
这些活性的改变可以是使得谷氨酸棒状杆菌的一种或多种精细化学品的产量、生产率和/或生产效率提高的改变。通过优化活化编码目标精细化学品的生物合成蛋白的基因的转录或翻译或者通过影响或删除抑制这种基因转录或翻译的MR蛋白的活性,可能增加该生物合成途径的活性或活性率,归功于例如一种限制酶的量增加。相应地,可以通过改变MR蛋白的活性,使得其组成性地转录后失活参与目标精细化学品的降解途径的蛋白或者通过改变MR蛋白的活性,使得其组成性地抑制这种基因的转录或翻译以增加细胞的所述精细化学品的产量和/或生产率,这归功于该化合物的降解减少。
调节一种或多种MR蛋白的活性使得可能间接刺激细胞的一种或多种精细化学品的生产或者提高生产率,因为各种代谢途径相关联。可例如通过活化赖氨酸生物合成中一种或多种酶的表达来增加产量、生产率和/或生产效率以同时增加其他化合物的表达,例如其他氨基酸,当需要更大量的赖氨酸时细胞通常需要更大量的这些化合物。还可以改变整个细胞中的代谢调节,使得发酵培养物(其中营养和氧气的供给可能较差并且该环境中还可能大量存在毒性废物)的环境条件中的细胞的生长和复制改善。从而可例如改善培养物中细胞的生长和增殖,即使生长条件不是最佳,通过诱变某种MR蛋白(其针对胞外培养基中的高水平废产物抑制细胞膜生长所需的分子的合成以便阻碍非最佳生长条件中的细胞生长和细胞分裂)使得所述蛋白不再能够抑制所述合成。这种生长增加或这种存活增加将同样增加发酵培养物中目标精细化学品的产量和/或生产率,因为在给培养物中相对更多数目的细胞生产该化合物。
上述用于增加谷氨酸棒状杆菌目标化合物产量的MR蛋白的诱变策略不用于限制;对这些诱变策略的变化对技术人员是显而易见的。本文公开的这些策略和机制使得可以使用本发明的核酸和蛋白质分子以产生表达突变的MR核酸和蛋白质分子的谷氨酸棒状杆菌或相关的细菌菌株,从而提高目标化合物的产量、生产率和/或产生效率。该目标化合物可为谷氨酸棒状杆菌的任一天然产物,其包括生物合成途径的终产物和天然代谢途径的中间体,该目标化合物也可为在谷氨酸棒状杆菌代谢中非天然存在的分子,但该分子为本发明的谷氨酸棒状杆菌菌株产生。
不应将下面的实施例理解为对限制对本发明的进一步阐述。在该申请中引用的所有参考文献、专利申请、专利、公开的专利申请、表的全部内容此处引用作为参考。
实施例
实施例1:谷氨酸棒状杆菌ATCC 13032的总基因组DNA的制备
谷氨酸棒状杆菌(ATCC 13032)的培养物在30℃于BHI培养基(Difco)中剧烈振摇过夜。离心收获细胞,弃上清并将细胞重悬于5ml缓冲液-I(培养物原初体积的5%—所有陈述的体积按100ml培养物体积计算)。缓冲液-I的组成:140.34g/l蔗糖,2.46g/l MgSO4·7H2O,10ml/l KH2PO4溶液(100g/l,用KOH调节至pH6.7),50ml/l M12浓缩物(10g/l(NH4)2SO4,1g/lNaCl,2g/l MgSO4·7H2O,0.2g/l CaCl2,0.5g/l酵母提取物(Difco),10ml/l痕量元素混合物(200mg/l FeSO4·H2O,10mg/l ZnSO4·7H2O,3mg/lMnCl2·4H2O,30mg/l H3BO3,20mmg/l CoCl2·6H2O,1mg/l NiCl2·6H2O,3mg/l Na2MoO4·2H2O,500mg/l络合剂(EDTA或柠檬酸),100ml/l维生素-混合物(0.2mg/l生物素,0.2mg/l叶酸,20mg/l对氨基苯甲酸,20mg/l核黄素,40mg/l Ca-泛酸盐,140mg/l烟酸,40mg/l维生素B6盐酸盐,200mg/l肌醇)。向悬液中加入溶菌酶至终浓度2.5mg/ml。在37℃孵育约4小时后,细胞壁降解,离心收获所得的原生质体。沉淀用5ml缓冲液-I洗一次并用5ml TE-缓冲液(10mM Tris-HCl,1mM EDTA,pH8)洗一次。将沉淀重悬于4ml TE-缓冲液,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后,将悬液在37℃孵育约18小时。使用标准方法用酚、酚-氯仿-异戊醇和氯仿-异戊醇提取而纯化DNA。然后,加入1/50体积的3M醋酸钠和2体积的乙醇,于-20℃孵育30分钟,并在高速离心机上使用SS34转子(Sorvall)于12,000转/分离心30分钟,沉淀DNA。将该DNA溶于含20μg/ml RNaseA的1ml TE-缓冲液中,并用1000ml TE-缓冲液于4℃透析至少3小时。在此期间,更换缓冲液3次。向分装的0.4ml透析过的DNA溶液中加入0.4ml 2M LiCl和0.8ml乙醇。于-20℃孵育30分钟后,离心(13,000转/分,Biofuge Fresco,Heraeus,Hanau,德国)收集DNA。将该DNA沉淀溶于TE-缓冲液。用此方法制备的DNA可用于所有目的,包括Southern印迹或构建基因组文库。
实施例2:在大肠杆菌中构建谷氨酸棒状杆菌(ATCC13032)的基因组文库
从如实施例1所述制备的DNA开始,按照公知的且已建立的方法(参见例如,Sambrook,J.等人(1989)“分子克隆:实验室手册”(“MolecularCloning:A Laboratory Manual”),Cold Spring Harbor Laboratory Press,或Ausubel,F.M.等人(1994)“分子生物学最新方法”(“Current Protocolsin Molecular Biology”),John Wiley&Sons.)构建粘粒和质粒文库。
可使用任一质粒或粘粒。特别使用的是质粒pBR322(Sutcliffe,J.G.(1979)美国国家科学院院报(Proc.Natl.Acad.Sci.USA),75:3737-3741);pACYC177(Change&Cohen(1978)细菌学杂志(J.Bacteriol)134:1141-1156)、pBS系列的质粒(pBSSK+,pBSSK-及其它;Stratagene,LaJolla,USA)、或粘粒如SuperCos1(Stratagene,LaJolla,USA)或Lorist6(Gibson,T.J.,Rosenthal A.和Waterson,R.H.(1987)基因(Gene)53:283-286)。
实施例3:DNA序列测定和功能的计算分析
如实施例2所述的基因组文库按照标准方法用于DNA序列测定,尤其是使用ABI377序列测定仪通过链终止法测定(参见例如,Fleischman,R.D.等人(1995),流感嗜血菌(Haemophilus Influenzae)Rd的全基因组随机序列测定和组装,科学(Science),269:496-512)。使用具有以下核苷酸序列的测序引物:5’-GGAAACAGTATGACCATG-3’或5’-GTAAAACGACGGCCAGT-3’。
实施例4:体内诱变
谷氨酸棒状杆菌的体内诱变可通过将质粒(或其它载体)DNA在不能维持其遗传信息完整性的大肠杆菌或其它微生物(例如Bacillus spp.或酵母如酿酒酵母)中传代而实施。常见的突变基因菌株在用于DNA修复系统的基因中有突变(例如mutHLS,mutD,mutT等;参考文献见大肠杆菌和沙门菌(Escherichia coli and Salmonella),ASM:Washington一书中的Rupp,W.D.(1996)DNA修复机制,2277-2294页)。此类菌株为技术人员熟知。此类菌株的用途在例如Greener,A.和Callahan,M.(1994)Strategies 7:32-34中进行了阐述。
实施例5:在大肠杆菌和谷氨酸棒状杆菌之间的DNA转移
几种棒状杆菌和短杆菌种类含有自主复制的内源质粒(例如pHM1519或pBL1)(综述参见例如,Martin,J.F.等人(1987)生物技术(Biotechnology),5:137-146)。使用标准的大肠杆菌载体易于构建大肠杆菌和谷氨酸棒状杆菌的穿梭载体(Sambrook,J.等人(1989),“分子克隆:实验室手册”(“Molecular Cloning:A Laboratory Manual”),Cold Spring HarborLaboratory Press,或Ausubel,F.M.等人(1994)“最新分子生物学方法”(“Current Protocols in Molecular Biology”),John Wiley&Sons),其中加入复制原点和来自谷氨酸棒状杆菌的适当标记。此类复制原点优选地取自从棒状杆菌和短杆菌分离的内源质粒。这些种类中作为转化标记尤其有用的是卡那霉素抗性基因(如衍生自Tn5或Tn903转座子的基因)或氯霉素抗性基因(Winnacker,E.L.(1987),From Genes to Clones-Introductionto Gene Technology,VCH,Weinheim)。在文献中有许多在大肠杆菌和谷氨酸棒状杆菌中均复制的多种穿梭载体的例子,且其可用于多个目的,包括基因过表达(参考文献参见例如,Yoshihama,M.等人(1985)细菌学杂志(J.Bacteriol.)162:591-597,Martin J.F.等人(1987)生物技术(Biotechnology),5:137-146和Eikmanns,B.J.等人(1992)基因(Gene),102:93-98)。
使用标准技术,可将目的基因克隆入上述的穿梭载体之一并可将此杂合载体导入谷氨酸棒状杆菌菌株。谷氨酸棒状杆菌的转化可通过原生质体转化(Kastsumata,R.等人(1984)细菌学杂志(J.Bacteriol.)159,306-311)、电穿孔(Liebl,E.等人(1989)FEMS Microbiol.Letters,53:399-303)以及当使用特别的载体时,也通过接合(如在Sch_fer,A等人(1990)细菌学杂志(J.Bacteriol.)172:1663-1666中所述)来完成。也可转移谷氨酸棒状杆菌的穿梭载体至大肠杆菌,其为通过制备来自谷氨酸棒状杆菌的质粒DNA(使用本领域公知的标准方法)并将其转化至大肠杆菌。该转化步骤可使用标准方法进行,但有利的是使用Mcr-缺陷的大肠杆菌菌株,如NM522(Gough&Murray(1983)分子生物学杂志(J.Mol.Biol.)166:1-19)。
实施例6:突变蛋白质的表达的确定
在转化的宿主细胞中对突变蛋白质活性的观察依赖于突变蛋白质以与野生型蛋白质相似的方式和相似的量表达的事实。确定突变基因转录水平(指示可用于翻译为基因产物的mRNA量)的有用方法是进行Northern印迹(参考文献见例如,Ausubel等人(1988)分子生物学最新方法(CurrentProtocols in Molecular Biology),Wiley:New York),其中设计为与目的基因结合的引物用可检测标志(通常为放射性的或化学发光的)标记,这样当提取生物体培养物的总RNA、在凝胶上运行、转移至稳定的基质并与该探针孵育时,探针的结合和结合量指示了该基因的存在和其mRNA量。该信息是突变基因转录程度的证据。通过几种方法可从谷氨酸棒状杆菌中分离总细胞RNA,这些方法均为本领域熟知,如在Bormann,E.R.等人(1992)分子微生物学(Mol.Microbiol.)6:317-326中所描述。
为了确定从该mRNA翻译的蛋白质的存在或相对量,可使用标准技术如Western印迹(参见例如,Ausubel等人(1988)分子生物学最新方法(Current Protocols in Molecular Biology),Wiley:New York)。在该方法中,提取细胞总蛋白质、通过凝胶电泳分离、转移至基质如硝化纤维并与探针孵育,其中所述探针如与目标蛋白质特异性结合的抗体。该探针通常用易于检测到的化学发光或显色标记物标记。所观察到的标记物的存在和量指示了在细胞中目标突变蛋白质的存在和量。
实施例7:遗传改变的谷氨酸棒状杆菌的生长—培养基和培养条件
遗传修饰的棒状杆菌培养在合成或天然的生长培养基中。用于棒状杆菌的许多种不同生长培养基为人们熟知并易于获得(Lieb等人(1989)Appl.Microbiol.Biotechnol.,32:205-210;Von der Osten等人(1998)Biotechnology Letters,11:11-16;Patent DE 4,120,867;在原核生物(TheProcaryotes),Volume II,Balows,A.等人编辑.Springer-Verlag一书中的Liebl(1992)“棒状杆菌属”(“The Genus Corynebacteria”))。这些培养基由一种或多种碳源、氮源、无机盐、维生素和痕量元素组成。优选的碳源是糖类,如单糖、二糖或或多糖。优良碳源的实例是葡萄糖、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、淀粉和纤维素。也可通过复合化合物如糖蜜或其它来自糖加工的副产品提供糖至培养基。提供不同碳源的混合物也可是有利的。其它可能的碳源为醇类和有机酸,如甲醇、乙醇、醋酸或乳酸。氮源通常为有机或无机氮化合物,或为包含这些化合物的物质。示例性氮源包括氨气或胺盐如NH4Cl或(NH4)2SO4、NH4OH、硝酸盐、尿素、氨基酸和复合氮源如玉米浆、大豆粉、大豆蛋白质、酵母提取物、肉羹等。
可包括在培养基中的无机盐化合物包括钙、镁、钠、钴、钼、钾、锰、锌、铜和铁的氯化物、亚磷酸盐或硫酸盐。可向培养基中加入螯合物以维持溶液中的金属离子。尤其有用的螯合物包括二羟基酚,如儿茶酚或原儿荼酚盐,和有机酸如柠檬酸。培养基一般也包括其它生长因子如维生素或生长促进剂,其实例包括生物素、核黄素、硫胺素、叶酸、烟酸、泛酸盐和维生素B6。生长因子和盐通常来自复合的培养基组成,如酵母提取物、糖蜜、玉米浆等。培养基化合物的精确组成强烈取决于直接实验并在每一具体情况下单独确定。有关培养基优化的信息在课本“Applied Microbiol.Physiology,A Practical Approach”(P.M.Rhodes,P.F.Stanbury编辑,IRL Press(1997)53-73页,ISBN 0 19 9635773)中可找到。也可从供应商处得到生长培养基,如Standard 1(Merck)或BHI(brain heart infusion,DIFCO)和其它。
通过加热(1.5巴,121℃加热20分钟)或过滤除菌对所有的培养基组成成份进行灭菌。组成成份可一起灭菌,或需要的话分开灭菌。所有的培养基组成成份可在培养开始时存在,或它们可任选地连续加入或分批加入。
对每一实验分别定义培养条件。温度应在15℃和45℃的范围之间。在实验中温度可保持恒定或可改变。培养基的pH应在5至8.5的范围之间,优选地在7.0附近,可通过向培养基中添加缓冲液而维持。用于此目的的示例性缓冲液为磷酸钾缓冲液。可备选地或同时使用合成缓冲液如MOPS、HEPES、ACES和其它。在生长过程中通过加入NaOH或NH4OH也可维持恒定的培养pH。如果使用复合的培养基组成如酵母提取物,可减轻对额外的缓冲液的需要,因为许多复合化合物具有高缓冲能力。如果使用发酵罐培养微生物,用气态氨也可控制pH。
培养时间通常在几小时到几天之间。选择培养时间以允许肉汤中累积最大量的产物。可在多种容器如不同规格的微滴定板、玻璃管、玻璃烧瓶或玻璃或金属发酵罐中实施公开的生长实验。对于筛选大量的克隆,应在微滴定板、玻璃管或在有或没有挡板的摇瓶中培养微生物。优选地使用100ml摇瓶,其中装有10%(体积)的所需生长培养基。该烧瓶应在旋转振荡器(振幅25mm)上以100-300转/分摇动。可通过维持潮湿环境减少蒸发损失;备选地,对蒸发损失应进行数学上的校正。
如果试验遗传修饰的克隆,也应试验未修饰的对照克隆或含基本质粒的无任何插入的对照克。使用已于30℃孵育的在琼脂板上生长的细胞接种培养基至OD600 0.5-1.5,其中所述琼脂板如CM板(10g/l葡萄糖,2.5g/lNaCl,2g/l尿素,10g/l聚胨,5g/l酵母抽取物,5g/l肉羹,22g/l琼脂,用2M NaOH调节至pH6.8)。培养基的接种通过导入来自CM板的谷氨酸棒状杆菌细胞的盐水悬液或添加该细菌的原液体培养物而完成。
实施例8:突变蛋白质功能的体外分析
本领域已建立了对酶活性和动力学的确定。确定特定的改变酶的活性的实验应加以修改,使适应野生型酶特异活性的确定,这在本领域普通技术人员的能力范围内。可在下列参考文献中找到对酶从总体上的概观,以及有关结构、动力学、原理、方法、应用和对许多酶活性进行确定的例子的具体说明,例如Dixon,M.和Webb,E.C.,(1979)酶(Enzymes).Longmans:伦敦;Fersht,(1985)酶结构和机制(Enzyme Structure andMechanism).Freeman:纽约;Walsh,(1979)酶促反应机制(EnzymaticReaction Mechanisms).Freeman:San Francisco;Price,N.C.,Stevens,L.(1982)酶学基础(Fundamentals of Enzymology).Oxford Univ.Press:Oxford;Boyer,P.D.编辑(1983)酶(The Enzymes),第三版,Academic Press:纽约;Bisswanger,H.,(1994)酶动力学(Enzymkinetik),第二版.VCH:Weinheim(ISBN 3527300325);Bergmeyer,H.U.,Bergmeyer,J.,Graβl,M.编辑(1983-1986)酶促分析方法(Methods of Enzymatic Analysis),第三版,I-XII卷,Verlag Chemie:Weinheim;以及Ullmann工业化学百科全书(Ullmann’s Encyclopedia of Industrial Chemistry)(1987)A9卷,“酶”.VCH:Weinheim,352-363页。
可通过几种已建立的方法测定结合DNA的蛋白质的活性,如DNA带移动试验(也称为凝胶阻滞试验)。此类蛋白质对其它分子表达的影响可使用报告基因试验测定(如在Kolmar,H.等人(1995)EMBO J.14:3895-3904及其中所引用的参考文献中所述)。报告基因测试系统为人们所熟知,并被建立以应用于原核和真核细胞,使用的酶如β-半乳糖苷酶、绿色荧光蛋白和几种其它的酶。
可根据如在生物膜,分子结构和功能(Biomembranes.MolecularStructure and Function),Springer:Heidelberg一书中的Gennis,R.B.(1989)“Pores,Channels and Transporters”,85-137;199-234和270-322页中所述的技术进行膜转运蛋白活性的测定。
实施例9:分析突变蛋白质对目标产物产生的影响
谷氨酸棒状杆菌中的遗传修饰对目标化合物(如氨基酸)产生的影响可通过将经修饰的微生物在合适的条件(如上文中所描述的那些条件)下生长并分析用于增加目标产物(即氨基酸)产生的培养基和/或细胞组成来确定。此类分析技术为技术人员熟知,包括光谱学、薄层色谱法、多种染色法、酶促和微生物学方法、以及分析色谱法如高效液相色谱法(参见例如,Ullman工业化学百科全书(Ullman,Encyclopedia of IndustrialChemistry),A2卷,89-90和443-613页,VCH:Weinheim(1985);Fallon,A.等人(1987)生物化学和分子生物学实验室技术(Laboratory Techniques inBiochemistry and Molecular Biology)中的“HPLC在生物化学中的应用”(“Applications of HPLC in Biochemistry”),17卷;Rehm等人(1993)生物技术(Biotechnology),3卷,III章“产物回收和纯化”(“Productrecovery and purification”),469-714页,VCH:Weinheim;Belter,P.A.等人(1988)生物分离:生物技术中的下游加工(Bioseparations:downstreamprocessing for biotechnology),John Wiley and Sons;Kennedy,J.F.和Cabral,J.M.S.(1992)生物物质的回收方法(Recovery processes forbiological materials)John Wiley and Sons;在Ulmann工业化学百科全书(Ulmann’s Encyclopedia of Industrial Chemistry)一书中的Shaeiwitz,J.A.和Henry,J.D.(1988)生物化学分离(Biochemical separations),B3卷,11章,1-27页,VCH:Weinheim;以及Dechow,F.J.(1989)生物技术中的分离和纯化技术(Separation and purification techniques inbiotech nology),Noyes Publications.)。
除了测定发酵终产物外,还可分析用于产生目标化合物的代谢途径的其它组成如中间体和副产物来确定化合物的总体产生力、化合物的产率和/或生产效率。分析方法包括测定培养基中的营养素水平(例如,糖类、碳水化合物、氮源、磷酸盐和其它离子)、测定生物量组成和生长、分析生物合成途径中共同代谢物的产生及测定发酵过程中产生的气体。这些测定的标准方法在应用微生物生理学,实用方法(Applied Microbial Physiology,APractical Approach),P.M.Rhodes和P.F.Stanbury编辑,IRL Press,103-129;131-163和165-192页(ISBN:0199635773)及其引用的参考文献中列出。
实施例10:从谷氨酸棒状杆菌培养物中纯化目标产物
可通过多种本领域熟知的方法实施从谷氨酸棒状杆菌细胞或上述培养物上清回收目标产物。如果目标产物不从细胞中分泌,可低速离心从培养物中收获细胞、用标准技术如机械力或超声裂解细胞。离心去除细胞碎片,保留含有可溶蛋白质的上清级分用于进一步纯化目标化合物。如果化合物从谷氨酸棒状杆菌细胞分泌,则低速离心从培养物中去除细胞,并保留上清级分用于进一步纯化。
将来自两种中任一纯化方法的上清级分用合适的树脂进行层析,其中目标分子保留在层析树脂上而样本中的许多杂质不保留在层析树脂上,或者杂质保留在层析树脂上而样本不保留。此层析步骤必需时可重复,其中使用相同或不同的层析树脂。技术人员在选择合适的层析树脂以及将它们最有效地用于特定的待纯化分子方面是很精通的。可通过过滤或超滤浓缩纯化产物,并贮存在使产物稳定性最高的温度下。
有大量的纯化方法为本领域公知,上述纯化方法不用于限制。此类纯化技术例如在Bailey,J.E.和Ollis,D.F.生物化学工程基础(BiochemicalEngineering Fundamentals),McGraw-Hill:纽约(1986)中描述。
分离化合物的特性和纯度可通过本领域的标准技术确定。这些技术包括高效液相色谱(HPLC)、光谱法、染色法、薄层色谱法、NIRS、酶促试验或微生物学方法。此类分析方法在Patek等人(1994)Appl.Environ.Microbiol.60:133-140;Malakhova等人(1996)生物技术(Biotekhnologiya)11:27-32;和Schmidt等人(1998)生物加工工程(Bioprocess Engineer)19:67-70.Ulmann工业化学百科全书(Ulmann’s Encyclopedia of Industrial Chemistry)(1996)A27卷,VCH:Weinheim,89-90页,521-540页,540-547页,559-566页,575-581页和581-587页;Michal,G.(1999)生物化学途径:生物化学和分子生物学图集(Biochemical Pathways:An Atlas of Biochemistry and MolecularBiology),John Wiley and Sons;生物化学和分子生物学中的实验室技术(Laboratory Techniques in Biochemistry and Molecular Biology),17卷中的Fallon,A.等人(1987)HPLC在生物化学中的应用(Applications ofHPLC in Biochemistry)。
等同物
技术人员知道或通过简单的常规方法可确定许多于文中所描述本发明具体实施方案的等同物。此类等同物旨在包括在下面的专利权利要求书中。
表1中的信息理解如下:
在第1列中,“DNA ID”,每种情况下相关数字指所公开序列表的SEQ ID NO。所以,“DNA ID”列中的“5”指SEQ ID NO:5。
在第2列中,“AA ID”,每种情况下相关数字指所公开序列的SEQ IDNO。“AA ID”列中的“6”指SEQ ID NO:6。
在第3列中,列出了“标识”——每个序列的明确内部名称。
在第4列中,“AA pos”,每种情况下相关数字指相同行中多肽序列“AA ID”的氨基酸位置。所以,“AA pos”列中“26”是相应所示的多肽序列的氨基酸位置26。位置计算从N末端的+1开始。
在第5列中,“AA野生型”,每种情况下相关数字指相应野生型菌株在第4列中所示位置的氨基酸(用单字母代码)。
在第6列中,“AA突变株”,每种情况下相关数字指相应突变菌株在第4列中所示位置的氨基酸(用单字母代码)。
在第7列中,列出了“功能”——相应多肽序列的生理学功能。
蛋白原氨基酸的单字母代码:
A 丙氨酸
C 半胱氨酸
D 天冬氨酸
E 谷氨酸
F 苯丙氨酸
G 甘氨酸
H 组氨酸
I 异亮氨酸
K 赖氨酸
L 亮氨酸
M 甲硫氨酸
N 天冬酰胺
P 脯氨酸
Q 谷氨酰胺
R 精氨酸
S 丝氨酸
T 苏氨酸
V 缬氨酸
W 色氨酸
Y 酪氨酸
表1
编码调节蛋白的基因
DNA ID: AA ID: 标识 AA位置: AA野生型 AA突变体功能:
1 2 RXA00205 220 G D 核糖操纵子阻抑蛋白
3 4 RXA00253 177 T I 四环素阻抑蛋白E类
5 6 RXA00291 219 E K 传感蛋白激酶DPIB(EC 2.7.3.-)
7 8 RXA00319 63 A V 低分子量磷酸酪氨酸蛋白磷酸酶(EC 3.1.3.48)
9 10 RXA00486 80 R H 转录调节蛋白,LYSR家族
11 12 RXA00593 18 A V 细胞分裂转录因子,WHMD
13 14 RXA00609 107 A T 转录调节蛋白
15 16 RXA00651 374 G D 感觉转导蛋白激酶(EC 2.7.3.-)
17 18 RXA00719 306 D N GTP-结合蛋白
468 A T GTP-结合蛋白
19 20 RXA00813 340 G D 分泌性蛋白激酶
21 22 RXA01081 102 R C 转录调节蛋白
23 24 RXA01315 20 A T 转录调节子,TETR家族
101 G D 转录调节子,TETR家族
25 26 RXA01573 474 P S 2’,3’-环核苷酸2’-磷酸二酯酶(EC 3.1.4.16)
27 28 RXA01607 142 R W 转录调节蛋白
179 A T 转录调节蛋白
29 30 RXA01759 63 S F 转录调节蛋白GLTC
191 P S 转录调节蛋白GLTC
31 32 RXA01826 603 P S 丝氨酸/苏氨酸蛋白激酶(EC 2.7.1.37)
33 34 RXA02097 28 A V 转录调节子
427 G R 转录调节子
35 36 RXA02210 4 A V 转录调节子,TETR家族
37 38 RXA02362 525 M I 转录调节子
39 40 RXA02376 150 H R GTP-结合蛋白
41 42 RXA02627 338 E K DTXR/铁调节的脂蛋白前体
43 44 RXA02667 193 G D 转录调节子
45 46 RXA02758 110 G E 磷丝氨酸磷酸酶(EC 3.1.3.3)
47 48 RXA02910 186 D N 转录调节蛋白,LYSR家族
49 50 RXA03100 170 R C 酰胺脲结合蛋白前体
176 S F 酰胺脲结合蛋白前体
51 52 RXA03127 142 E K 转录调节蛋白
53 54 RXA03136 205 P S 转录活化子,LUXR家族
385 D N 转录活化子,LUXR家族
55 56 RXA03201 206 D N UDP-葡萄糖4-差向异构酶(EC 5.1.3.2)
57 58 RXA03407 70 A V 转录调节子
59 60 RXA03629 36 S N 转录调节子
61 62 RXA03928 170 E R 转录调节子
171 A S 转录调节子
172 M H 转录调节子
63 64 RXA04129 54 A V 转录调节子,ARSR家族
65 66 RXA04350 150 R H 转录调节蛋白
67 68 RXA04363 81 S F 转录阻抑蛋白
69 70 RXA04620 260 R H 转录调节蛋白
71 72 RXA06017 33 A V 可能的过氧化氢诱导的基因活化子
73 74 RXA07002 158 P S 传感蛋白BAES(EC 2.7.3.-)
75 76 RXA07003 126 L F 转录调节蛋白
195 R K 转录调节蛋白
序列表
<110>巴斯福股份公司
<120>来自谷氨酸棒状杆菌的编码调节蛋白的基因
<130>O.Z.0050/52970
<160>76
<210>1
<211>1294
<212>DNA
<213>谷氨酸棒状杆菌(Corynebacterium glutamicum)
<220>
<221>CDS
<222>(101)..(1264)
<223>RXA00205
<400>1
gaaatcaagt ggcctagatc cattgacact tagactgtga cctaggcttg actttcgtgg 60
gggagtgggg ataagttcat cttaaacaca atgcaatcga ttg cat tta cgt tcc 115
Leu His Leu Arg Ser
1 5
tta tcc cac aat agg ggt acc ttc cag aaa gtt ggt gag gag atg gct 163
Leu Ser His Asn Arg Gly Thr Phe Gln Lys Val Gly Glu Glu Met Ala
10 15 20
tcc gaa acc tcc agc ccg aag aag cgg gcc acc acg ctc aaa gac atc 211
Ser Glu Thr Ser Ser Pro Lys Lys Arg Ala Thr Thr Leu Lys Asp Ile
25 30 35
gcg caa gca aca cag ctt tca gtc agc acg gtg tcc cgg gca ttg gcc 259
Ala Gln Ala Thr Gln Leu Ser Val Ser Thr Val Ser Arg Ala Leu Ala
40 45 50
aac aac gcg agc att ccg gaa tcc aca cgc atc cga gtg gtt gaa gcc 307
Asn Asn Ala Ser Ile Pro Glu Ser Thr Arg Ile Arg Val Val Glu Ala
55 60 65
gct caa aag ctg aac tac cgt ccc aat gcc caa gct cgt gca ttg cgg 355
Ala Gln Lys Leu Asn Tyr Arg Pro Asn Ala Gln Ala Arg Ala Leu Arg
70 75 80 85
aag tcg agg aca gac acc atc ggt gtc atc att cca aac att gag aac 403
Lys Ser Arg Thr Asp Thr Ile Gly Val Ile Ile Pro Asn Ile Glu Asn
90 95 100
cca tat ttc tcc tca cta gca gca tcg att caa aaa gct gct cgt gaa 451
Pro Tyr Phe Ser Ser Leu Ala Ala Ser Ile Gln Lys Ala Ala Arg Glu
105 110 115
gct ggg gtg tcc acc att ttg tcc aac tct gaa gaa aac cca gag ctg 499
Ala Gly Val Ser Thr Ile Leu Ser Asn Ser Glu Glu Asn Pro Glu Leu
120 125 130
ctt ggt cag act ttg gcg atc atg gat gac caa cgc ctc gat gga atc 547
Leu Gly Gln Thr Leu Ala Ile Met Asp Asp Gln Arg Leu Asp Gly Ile
135 140 145
atc gtg gtg cca cac att cag tca gag gaa caa gtc act gac ttg gtt 595
Ile Val Val Pro His Ile Gln Ser Glu Glu Gln Val Thr Asp Leu Val
150 155 160 165
aac agg gga gtg cca gta gtg ctg gca gac cgt agt ttt gtt aac tcg 643
Asn Arg Gly Val Pro Val Val Leu Ala Asp Arg Ser Phe Val Asn Ser
170 175 180
tct att cct tcg gtt acc tca gat cca gtt ccg ggc atg act gaa gct 691
Ser Ile Pro Ser Val Thr Ser Asp Pro Val Pro Gly Met Thr Glu Ala
185 190 195
gtg gac tta ctc ctg gca gct gac gtg caa ttg ggc tac ctt gcc ggc 739
Val Asp Leu Leu Leu Ala Ala Asp Val Gln Leu Gly Tyr Leu Ala Gly
200 205 210
ccg cag gat act tcc act ggt cag ctg cgt ctt aac act ttt gaa aga 787
Pro Gln Asp Thr Ser Thr Gly Gln Leu Arg Leu Asn Thr Phe Glu Arg
215 220 225
cta tgc gtg gac cgc ggc atc gtc gga gca tct gtc tat tac ggt ggc 835
Leu Cys Val Asp Arg Gly Ile Val Gly Ala Ser Val Tyr Tyr Gly Gly
230 235 240 245
tac cgc caa gaa tct gga tat gac ggc atc aag gtg ctg atc aag cag 883
Tyr Arg Gln Glu Ser Gly Tyr Asp Gly Ile Lys Val Leu Ile Lys Gln
250 255 260
gga gcc aat gcg att atc gct ggt gac tcc atg atg acc atc ggt gcg 931
Gly Ala Asn Ala Ile Ile Ala Gly Asp Ser Met Met Thr Ile Gly Ala
265 270 275
ttg ttg gct ctt cat gag atg aat ttg aag atc ggt gag gat gtg cag 979
Leu Leu Ala Leu His Glu Met Asn Leu Lys Ile Gly Glu Asp Val Gln
280 285 290
ctc att ggg ttt gat aac aac cca att ttc cgg ctg cag aat cca ccg 1027
Leu Ile Gly Phe Asp Asn Asn Pro Ile Phe Arg Leu Gln Asn Pro Pro
295 300 305
ctg agc atc att gac cag cac gta caa gag atc ggt aag cgt gcg ttt 1075
Leu Ser Ile Ile Asp Gln His Val Gln Glu Ile Gly Lys Arg Ala Phe
310 315 320 325
gag att ctg cag aag ctg atc aat ggg gac acc gcg caa aaa tct gtg 1123
Glu Ile Leu Gln Lys Leu Ile Asn Gly Asp Thr Ala Gln Lys Ser Val
330 335 340
gtg att cca acg cag ctc agc atc aat gga tca acg gcg gtt tcc caa 1171
Val Ile Pro Thr Gln Leu Ser Ile Asn Gly Ser Thr Ala Val Ser Gln
345 350 355
aag gcg gcc gca aag gca gca aaa gca gcc caa aaa gca gcc gcg aaa 1219
Lys Ala Ala Ala Lys Ala Ala Lys Ala Ala Gln Lys Ala Ala Ala Lys
360 365 370
gcc gca cag aac acg caa cac gag gtg agc cta gat ggt gaa ctc 1264
Ala Ala Gln Asn Thr Gln His Glu Val Ser Leu Asp Gly Glu Leu
375 380 385
tgaacaagcg cttcatcagc atgatcctgc 1294
<210>2
<211>388
<212>PRT
<213>谷氨酸棒状杆菌
<400>2
Leu His Leu Arg Ser Leu Ser His Asn Arg Gly Thr Phe Gln Lys Val
1 5 10 15
Gly Glu Glu Met Ala Ser Glu Thr Ser Ser Pro Lys Lys Arg Ala Thr
20 25 30
Thr Leu Lys Asp Ile Ala Gln Ala Thr Gln Leu Ser Val Ser Thr Val
35 40 45
Ser Arg Ala Leu Ala Asn Asn Ala Ser Ile Pro Glu Ser Thr Arg Ile
50 55 60
Arg Val Val Glu Ala Ala Gln Lys Leu Asn Tyr Arg Pro Asn Ala Gln
65 70 75 80
Ala Arg Ala Leu Arg Lys Ser Arg Thr Asp Thr Ile Gly Val Ile Ile
85 90 95
Pro Asn Ile Glu Asn Pro Tyr Phe Ser Ser Leu Ala Ala Ser Ile Gln
100 105 110
Lys Ala Ala Arg Glu Ala Gly Val Ser Thr Ile Leu Ser Asn Ser Glu
115 120 125
Glu Asn Pro Glu Leu Leu Gly Gln Thr Leu Ala Ile Met Asp Asp Gln
130 135 140
Arg Leu Asp Gly Ile Ile Val Val Pro His Ile Gln Ser Glu Glu Gln
145 150 155 160
Val Thr Asp Leu Val Asn Arg Gly Val Pro Val Val Leu Ala Asp Arg
165 170 175
Ser Phe Val Asn Ser Ser Ile Pro Ser Val Thr Ser Asp Pro Val Pro
180 185 190
Gly Met Thr Glu Ala Val Asp Leu Leu Leu Ala Ala Asp Val Gln Leu
195 200 205
Gly Tyr Leu Ala Gly Pro Gln Asp Thr Ser Thr Gly Gln Leu Arg Leu
210 215 220
Asn Thr Phe Glu Arg Leu Cys Val Asp Arg Gly Ile Val Gly Ala Ser
225 230 235 240
Val Tyr Tyr Gly Gly Tyr Arg Gln Glu Ser Gly Tyr Asp Gly Ile Lys
245 250 255
Val Leu Ile Lys Gln Gly Ala Asn Ala Ile Ile Ala Gly Asp Ser Met
260 265 270
Met Thr Ile Gly Ala Leu Leu Ala Leu His Glu Met Asn Leu Lys Ile
275 280 285
Gly Glu Asp Val Gln Leu Ile Gly Phe Asp Asn Asn Pro Ile Phe Arg
290 295 300
Leu Gln Asn Pro Pro Leu Ser Ile Ile Asp Gln His Val Gln Glu Ile
305 310 315 320
Gly Lys Arg Ala Phe Glu Ile Leu Gln Lys Leu Ile Asn Gly Asp Thr
325 330 335
Ala Gln Lys Ser Val Val Ile Pro Thr Gln Leu Ser Ile Asn Gly Ser
340 345 350
Thr Ala Val Ser Gln Lys Ala Ala Ala Lys Ala Ala Lys Ala Ala Gln
355 360 365
Lys Ala Ala Ala Lys Ala Ala Gln Asn Thr Gln His Glu Val Ser Leu
370 375 380
Asp Gly Glu Leu
385
<210>3
<211>868
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(838)
<223>RXA00253
<400>3
acccatgtta gatgttttat tcagggattt tagttgatat gtccagtatc tcgctgaaaa 60
cgctggttgt cttgtagaaa aaggcgtaac gtcatataac atg cct agc gaa act 115
Met Pro Ser Glu Thr
1 5
atg aaa cca gcc gta gcg tca act ctg gcg gcc act tcc acg gga cgt 163
Met Lys Pro Ala Val Ala Ser Thr Leu Ala Ala Thr Ser Thr Gly Arg
10 15 20
cgt cct gga cgc ccc acc caa cgt atc ctt tcc gtc gaa tcc ata gtg 211
Arg Pro Gly Arg Pro Thr Gln Arg Ile Leu Ser Val Glu Ser Ile Val
25 30 35
gag cgc act tta aac att gcc ggc cgc gaa gga ttc gct gcc gtg acc 259
Glu Arg Thr Leu Asn Ile Ala Gly Arg Glu Gly Phe Ala Ala Val Thr
40 45 50
atg aac cgc ctc gcc cga gac atg ggt gtc acc cct cgc gca ctg tat 307
Met Asn Arg Leu Ala Arg Asp Met Gly Val Thr Pro Arg Ala Leu Tyr
55 60 65
aac cat gtg cta aat cgt caa gaa atc att gat cgc gtc tgg gtg cgc 355
Asn His Val Leu Asn Arg Gln Glu Ile Ile Asp Arg Val Trp Val Arg
70 75 80 85
atc atc gat gat atc aag gtg ccc gat ctt gat ccg gac aat tgg cgg 403
Ile Ile Asp Asp Ile Lys Val Pro Asp Leu Asp Pro Asp Asn Trp Arg
90 95 100
caa tct att cat acg ctg tgg agc tca ttg cgc gac caa ttc cgt gag 451
Gln Ser Ile His Thr Leu Trp Ser Ser Leu Arg Asp Gln Phe Arg Glu
105 110 115
act cca cgt gtt ctt ctg gtc gcg ctg gat gaa cag atc tct act cag 499
Thr Pro Arg Val Leu Leu Val Ala Leu Asp Glu Gln Ile Ser Thr Gln
120 125 130
ggc act tcc cca ctg cga atc gcg ggt gcg gag gag tcc ttg aag ttc 547
Gly Thr Ser Pro Leu Arg Ile Ala Gly Ala Glu Glu Ser Leu Lys Phe
135 140 145
ttg act gat atc ggg ctg tcc ctc aag gaa gca acc atc atc cgg gag 595
Leu Thr Asp Ile Gly Leu Ser Leu Lys Glu Ala Thr Ile Ile Arg Glu
150 155 160 165
atg atg atg gct gat gtc ttc agc ttc acc ctg act tct gac tac acc 643
Met Met Met Ala Asp Val Phe Ser Phe Thr Leu Thr Ser Asp Tyr Thr
170 175 180
ttt gac aat cgt cca gag ggc gaa aag ccg gat gtg ttt gct ccg gtt 691
Phe Asp Asn Arg Pro Glu Gly Glu Lys Pro Asp Val Phe Ala Pro Val
185 190 195
cct aag cca tgg ctt gat gag aac cca gat gtg gaa gcg cca ctg acc 739
Pro Lys Pro Trp Leu Asp Glu Asn Pro Asp Val Glu Ala Pro Leu Thr
200 205 210
cgt aaa gca gtc gaa gag tcc gtc tca act tct gac gaa ctc ttc ggc 787
Arg Lys Ala Val Glu Glu Ser Val Ser Thr Ser Asp Glu Leu Phe Gly
215 220 225
tac atg gtg gag gct cgc att gct tat att gaa aag ctg ctt gcc gcc 835
Tyr Met Val Glu Ala Arg Ile Ala Tyr Ile Glu Lys Leu Leu Ala Ala
230 235 240 245
aaa tagtttctaa aggttattga gggcctgctc 868
Lys
<210>4
<211>246
<212>PRT
<213>谷氨酸棒状杆菌
<400>4
Met Pro Ser Glu Thr Met Lys Pro Ala Val Ala Ser Thr Leu Ala Ala
1 5 10 15
Thr Ser Thr Gly Arg Arg Pro Gly Arg Pro Thr Gln Arg Ile Leu Ser
20 25 30
Val Glu Ser Ile Val Glu Arg Thr Leu Asn Ile Ala Gly Arg Glu Gly
35 40 45
Phe Ala Ala Val Thr Met Asn Arg Leu Ala Arg Asp Met Gly Val Thr
50 55 60
Pro Arg Ala Leu Tyr Asn His Val Leu Asn Arg Gln Glu Ile Ile Asp
65 70 75 80
Arg Val Trp Val Arg Ile Ile Asp Asp Ile Lys Val Pro Asp Leu Asp
85 90 95
Pro Asp Asn Trp Arg Gln Ser Ile His Thr Leu Trp Ser Ser Leu Arg
100 105 110
Asp Gln Phe Arg Glu Thr Pro Arg Val Leu Leu Val Ala Leu Asp Glu
115 120 125
Gln Ile Ser Thr Gln Gly Thr Ser Pro Leu Arg Ile Ala Gly Ala Glu
130 135 140
Glu Ser Leu Lys Phe Leu Thr Asp Ile Gly Leu Ser Leu Lys Glu Ala
145 150 155 160
Thr Ile Ile Arg Glu Met Met Met Ala Asp Val Phe Ser Phe Thr Leu
165 170 175
Thr Ser Asp Tyr Thr Phe Asp Asn Arg Pro Glu Gly Glu Lys Pro Asp
180 185 190
Val Phe Ala Pro Val Pro Lys Pro Trp Leu Asp Glu Asn Pro Asp Val
195 200 205
Glu Ala Pro Leu Thr Arg Lys Ala Val Glu Glu Ser Val Ser Thr Ser
210 215 220
Asp Glu Leu Phe Gly Tyr Met Val Glu Ala Arg Ile Ala Tyr Ile Glu
225 230 235 240
Lys Leu Leu Ala Ala Lys
245
<210>5
<211>1783
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1753)
<223>RXA00291
<400>5
tgaggtagtg cgcaaaataa gacttttgtg cattatgatc agaattgttg gcctgggact 60
tcgcttcacg ctctgctgat aatcgccccc gggggtagac atg tct gtt ggt gga 115
Met Ser Val Gly Gly
1 5
tcc gac tgg aaa aac ttc aag gag gtg gac atc att cgc ttt gct acc 163
Ser Asp Trp Lys Asn Phe Lys Glu Val Asp Ile Ile Arg Phe Ala Thr
10 15 20
cga ata ctg gtg att caa gtg gct acc gtc gcg ttg gtg gta gct att 211
Arg Ile Leu Val Ile Gln Val Ala Thr Val Ala Leu Val Val Ala Ile
25 30 35
tgc acc gga att ttc gca gtt ttg atg atg gat cag atg aaa act gag 259
Cys Thr Gly Ile Phe Ala Val Leu Met Met Asp Gln Met Lys Thr Glu
40 45 50
gcc gag cac aca gcg ctg tcc atc gga cgt tcg gtg gca tcc aac ccg 307
Ala Glu His Thr Ala Leu Ser Ile Gly Arg Ser Val Ala Ser Asn Pro
55 60 65
cag atc cgc gag gaa gta gcg ctt gat act caa aca gga gca aac cca 355
Gln Ile Arg Glu Glu Val Ala Leu Asp Thr Gln Thr Gly Ala Asn Pro
70 75 80 85
tcg gcc gaa gaa tta gcc gat gga gat atc caa gcg gtt gca cag gcg 403
Ser Ala Glu Glu Leu Ala Asp Gly Asp Ile Gln Ala Val Ala Gln Ala
90 95 100
gcc aat gaa cgc act gga gct ttg ttt gtc gtt atc act gac ggt tta 451
Ala Asn Glu Arg Thr Gly Ala Leu Phe Val Val Ile Thr Asp Gly Leu
105 110 115
ggt atc cgc ctg tcc cac cca gat gag gaa cgt ctg ggg gag cag gtg 499
Gly Ile Arg Leu Ser His Pro Asp Glu Glu Arg Leu Gly Glu Gln Val
120 125 130
agc act agc ttt gag gct gcc atg cgg ggt gaa gaa acc atg gcg tgg 547
Ser Thr Ser Phe Glu Ala Ala Met Arg Gly Glu Glu Thr Met Ala Trp
135 140 145
gag act ggg acc ctc ggt gcg tcc gcg cga gca aaa gtg cct atc ttt 595
Glu Thr Gly Thr Leu Gly Ala Ser Ala Arg Ala Lys Val Pro Ile Phe
150 155 160 165
gcg ccg gat tct agt gtt cca gtc ggt gag gtc agt gtt ggg ttt gag 643
Ala Pro Asp Ser Ser Val Pro Val Gly Glu Val Ser Val Gly Phe Glu
170 175 180
cga gac agt gtg tat tcc cgc ctg ccc atg ttc ctc gcc gcc ctt gct 691
Arg Asp Ser Val Tyr Ser Arg Leu Pro Met Phe Leu Ala Ala Leu Ala
185 190 195
ctt att tct gtg ttg gga atc ctt atc ggc gtg ggt gta gcc atg ggc 739
Leu Ile Ser Val Leu Gly Ile Leu Ile Gly Val Gly Val Ala Met Gly
200 205 210
atg cga cgc cgt tgg gaa cgc gtg acc ttg ggt ttg cag ccg gag gag 787
Met Arg Arg Arg Trp Glu Arg Val Thr Leu Gly Leu Gln Pro Glu Glu
215 220 225
cta gtg acc ctt gtg caa aat cag act gca gtc atc gat ggc att gat 835
Leu Val Thr Leu Val Gln Asn Gln Thr Ala Val Ile Asp Gly Ile Asp
230 235 240 245
gag ggc gtg ctg gcg ctg agc cca aac gga aca att ggg gtg cat aat 883
Glu Gly Val Leu Ala Leu Ser Pro Asn Gly Thr Ile Gly Val His Asn
250 255 260
gag cag gcg caa tcc atg att ggt gca ggt cct atg agt ggc agg acg 931
Glu Gln Ala Gln Ser Met Ile Gly Ala Gly Pro Met Ser Gly Arg Thr
265 270 275
ttg aaa gaa cta ggg ctt gac ctg ggt ctt gat ggc gtt gta ttg cat 979
Leu Lys Glu Leu Gly Leu Asp Leu Gly Leu Asp Gly Val Val Leu His
280 285 290
ggt cag cat ccg gaa acc gtt gcc cat aac ggc agg atc ctc tat ctg 1027
Gly Gln His Pro Glu Thr Val Ala His Asn Gly Arg Ile Leu Tyr Leu
295 300 305
gat ttc cac ccc gtg cgc cgt ggg gat caa gat tta ggc tac gtg gta 1075
Asp Phe His Pro Val Arg Arg Gly Asp Gln Asp Leu Gly Tyr Val Val
310 315 320 325
acc atc cgc gat cgt acc gac atc att gaa ctc agt gaa cgc ctc gac 1123
Thr Ile Arg Asp Arg Thr Asp Ile Ile Glu Leu Ser Glu Arg Leu Asp
330 335 340
tct gtg cgc acc atg acc cac gca ctc cgc gcc cag cgc cac gag ttt 1171
Ser Val Arg Thr Met Thr His Ala Leu Arg Ala Gln Arg His Glu Phe
345 350 355
gcc aac cgc atc cac acc gca aca ggg ctt atc gac gcc ggc cgc gtc 1219
Ala Asn Arg Ile His Thr Ala Thr Gly Leu Ile Asp Ala Gly Arg Val
360 365 370
cac gac gcg gca gag ttt cta ggc gat ata tcc cgc aac ggg gga cag 1267
His Asp Ala Ala Glu Phe Leu Gly Asp Ile Ser Arg Asn Gly Gly Gln
375 380 385
tca cat cca ttg atc gga tca gcg cac ctc aat gaa gca ttt ttg agc 1315
Ser His Pro Leu Ile GIy Ser Ala His Leu Asn Glu Ala Phe Leu Ser
390 395 400 405
tca ttt tta agt act gct tct att tcg gca tct gaa aag ggc gtt agt 1363
Ser Phe Leu Ser Thr Ala Ser Ile Ser Ala Ser Glu Lys Gly Val Ser
410 415 420
ctg cgc atc aac tct gac acg ctc atc ctt ggc act gtt aaa gat cca 1411
Leu Arg Ile Asn Ser Asp Thr Leu Ile Leu Gly Thr Val Lys Asp Pro
425 430 435
gaa gat gta gca acc att ttg ggt aat tta atc aac aat gcc atc gac 1459
Glu Asp Val Ala Thr Ile Leu Gly Asn Leu Ile Asn Asn Ala Ile Asp
440 445 450
gcc gcg gtg gca ggt gaa gcc cca cgg tgg att gag ctt acg ttg atg 1507
Ala Ala Val Ala Gly Glu Ala Pro Arg Trp Ile Glu Leu Thr Leu Met
455 460 465
gat gat gcc gat acg ctg gtc att tct gtt gca gat tct ggt cct gga 1555
Asp Asp Ala Asp Thr Leu Val Ile Ser Val Ala Asp Ser Gly Pro Gly
470 475 480 485
atc cca gag ggc gtg gat gta ttt gcc aca gcc acc cag ata gga gac 1603
Ile Pro Glu Gly Val Asp Val Phe Ala Thr Ala Thr Gln Ile Gly Asp
490 495 500
tct gaa gat aat gaa cgc acc cac ggg cat ggc att ggt cta aaa ctg 1651
Ser Glu Asp Asn Glu Arg Thr His Gly His Gly Ile Gly Leu Lys Leu
505 510 515
tgc cgg gct ttg gct aga tca cat ggt ggc gat gtc tgg gtg att gat 1699
Cys Arg Ala Leu Ala Arg Ser His Gly Gly Asp Val Trp Val Ile Asp
520 525 530
aga gga acc gaa gat ggc gct gta ttt gga gtg aaa cta ccg gga gta 1747
Arg Gly Thr Glu Asp Gly Ala Val Phe Gly Val Lys Leu Pro Gly Val
535 540 545
atg gag taatggatca aacacttaaa gttttagtaa 1783
Met Glu
550
<210>6
<211>551
<212>PRT
<213>谷氨酸棒状杆菌
<400>6
Met Ser Val Gly Gly Ser Asp Trp Lys Asn Phe Lys Glu Val Asp Ile
I 5 10 15
Ile Arg Phe Ala Thr Arg Ile Leu Val Ile Gln Val Ala Thr Val Ala
20 25 30
Leu Val Val Ala Ile Cys Thr Gly Ile Phe Ala Val Leu Met Met Asp
35 40 45
Gln Met Lys Thr Glu Ala Glu His Thr Ala Leu Ser Ile Gly Arg Ser
50 55 60
Val Ala Ser Asn Pro Gln Ile Arg Glu Glu Val Ala Leu Asp Thr Gln
65 70 75 80
Thr Gly Ala Asn Pro Ser Ala Glu Glu Leu Ala Asp Gly Asp Ile Gln
85 90 95
Ala Val Ala Gln Ala Ala Asn Glu Arg Thr Gly Ala Leu Phe Val Val
100 105 110
Ile Thr Asp Gly Leu Gly Ile Arg Leu Ser His Pro Asp Glu Glu Arg
115 120 125
Leu Gly Glu Gln Val Ser Thr Ser Phe Glu Ala Ala Met Arg Gly Glu
130 135 140
Glu Thr Met Ala Trp Glu Thr Gly Thr Leu Gly Ala Ser Ala Arg Ala
145 150 155 160
Lys Val Pro Ile Phe Ala Pro Asp Ser Ser Val Pro Val Gly Glu Val
165 170 175
Ser Val Gly Phe Glu Arg Asp Ser Val Tyr Ser Arg Leu Pro Met Phe
180 185 190
Leu Ala Ala Leu Ala Leu Ile Ser Val Leu Gly Ile Leu Ile Gly Val
195 200 205
Gly Val Ala Met Gly Met Arg Arg Arg Trp Glu Arg Val Thr Leu Gly
210 215 220
Leu Gln Pro Glu Glu Leu Val Thr Leu Val Gln Asn Gln Thr Ala Val
225 230 235 240
Ile Asp Gly Ile Asp Glu Gly Val Leu Ala Leu Ser Pro Asn Gly Thr
245 250 255
Ile Gly Val His Asn Glu Gln Ala Gln Ser Met Ile Gly Ala Gly Pro
260 265 270
Met Ser Gly Arg Thr Leu Lys Glu Leu Gly Leu Asp Leu Gly Leu Asp
275 280 285
Gly Val Val Leu His Gly Gln His Pro Glu Thr Val Ala His Asn Gly
290 295 300
Arg Ile Leu Tyr Leu Asp Phe His Pro Val Arg Arg Gly Asp Gln Asp
305 310 315 320
Leu Gly Tyr Val Val Thr Ile Arg Asp Arg Thr Asp Ile Ile Glu Leu
325 330 335
Ser Glu Arg Leu Asp Ser Val Arg Thr Met Thr His Ala Leu Arg Ala
340 345 350
Gln Arg His Glu Phe Ala Asn Arg Ile His Thr Ala Thr Gly Leu Ile
355 360 365
Asp Ala Gly Arg Val His Asp Ala Ala Glu Phe Leu Gly Asp Ile Ser
370 375 380
Arg Asn Gly Gly Gln Ser His Pro Leu Ile Gly Ser Ala His Leu Asn
385 390 395 400
Glu Ala Phe Leu Ser Ser Phe Leu Ser Thr Ala Ser Ile Ser Ala Ser
405 410 415
Glu Lys Gly Val Ser Leu Arg Ile Asn Ser Asp Thr Leu Ile Leu Gly
420 425 430
Thr Val Lys Asp Pro Glu Asp Val Ala Thr Ile Leu Gly Asn Leu Ile
435 440 445
Asn Asn Ala Ile Asp Ala Ala Val Ala Gly Glu Ala Pro Arg Trp Ile
450 455 460
Glu Leu Thr Leu Met Asp Asp Ala Asp Thr Leu Val Ile Ser Val Ala
465 470 475 480
Asp Ser Gly Pro Gly Ile Pro Glu Gly Val Asp Val Phe Ala Thr Ala
485 490 495
Thr Gln Ile Gly Asp Ser Glu Asp Asn Glu Arg Thr His Gly His Gly
500 505 510
Ile Gly Leu Lys Leu Cys Arg Ala Leu Ala Arg Ser His Gly Gly Asp
515 520 525
Val Trp Val Ile Asp Arg Gly Thr Glu Asp Gly Ala Val Phe Gly Val
530 535 540
Lys Leu Pro Gly Val Met Glu
545 550
<210>7
<211>628
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(598)
<223>RXA00319
<400>7
atcgattgtg ttgccgtaac ctggggctac ggcagcaaaa ctgaatggga cgctgcccgc 60
tacaccgtga gcaccgcaga agaattagaa aggatcatcc atg act ggg cct aaa 115
Met Thr Gly Pro Lys
1 5
act tcg cta cct gtg gaa att gtt ttc gta tgc acc gga aac att tgc 163
Thr Ser Leu Pro Val Glu Ile Val Phe Val Cys Thr Gly Asn Ile Cys
10 15 20
cga tcc ccc atg tcg gaa gtc atc gcg aag gca aaa gcg gaa gaa gct 211
Arg Ser Pro Met Ser Glu Val Ile Ala Lys Ala Lys Ala Glu Glu Ala
25 30 35
ggc ttg gaa gac aac gtc att ttc tcc tcc tgt ggc atg ggc aat tgg 259
Gly Leu Glu Asp Asn Val Ile Phe Ser Ser Cys Gly Met Gly Asn Trp
40 45 50
cac gtt ggc caa cct gct gac aag cga gct ctc gcg gaa ctg aaa tca 307
His Val Gly Gln Pro Ala Asp Lys Arg Ala Leu Ala Glu Leu Lys Ser
55 60 65
gcc ggt tac aac ggc gac acc cac cgc gca gca caa ctt ggt ccc gag 355
Ala Gly Tyr Asn Gly Asp Thr His Arg Ala Ala Gln Leu Gly Pro Glu
70 75 80 85
cac atg cgc gca gat ctc ttc gtc gcg cta gat tcc ggc cac gcc ggt 403
His Met Arg Ala Asp Leu Phe Val Ala Leu Asp Ser Gly His Ala Gly
90 95 100
gag ctc gcc gca acg ggt gtt ccc aac gac aaa atc cgc ctc atg cgt 451
Glu Leu Ala Ala Thr Gly Val Pro Asn Asp Lys Ile Arg Leu Met Arg
105 110 115
tcc ttc gac cca gag tcc aac ccc acc gac gat gtc gca gac cct tac 499
Ser Phe Asp Pro Glu Ser Asn Pro Thr Asp Asp Val Ala Asp Pro Tyr
120 125 130
tac ggc aca tcc cag gat ttc gtg ctc acc cgt gaa aac atc gaa gat 547
Tyr Gly Thr Ser Gln Asp Phe Val Leu Thr Arg Glu Asn Ile Glu Asp
135 140 145
gct atg ccg ggc ctt ttg gag tgg gtc aga gat cac atc cgc act gat 595
Ala Met Pro Gly Leu Leu Glu Trp Val Arg Asp His Ile Arg Thr Asp
150 155 160 165
tct taggtctttg agctaaaaag tcctaaggta 628
Ser
210>8
<211>166
<212>PRT
<213>谷氨酸棒状杆菌
<400>8
Met Thr Gly Pro Lys Thr Ser Leu Pro Val Glu Ile Val Phe Val Cys
1 5 10 15
Thr Gly Asn Ile Cys Arg Ser Pro Met Ser Glu Val Ile Ala Lys Ala
20 25 30
Lys Ala Glu Glu Ala Gly Leu Glu Asp Asn Val Ile Phe Ser Ser Cys
35 40 45
Gly Met Gly Asn Trp His Val Gly Gln Pro Ala Asp Lys Arg Ala Leu
50 55 60
Ala Glu Leu Lys Ser Ala Gly Tyr Asn Gly Asp Thr His Arg Ala Ala
65 70 75 80
Gln Leu Gly Pro Glu His Met Arg Ala Asp Leu Phe Val Ala Leu Asp
85 90 95
Ser Gly His Ala Gly Glu Leu Ala Ala Thr Gly Val Pro Asn Asp Lys
100 105 110
Ile Arg Leu Met Arg Ser Phe Asp Pro Glu Ser Asn Pro Thr Asp Asp
115 120 125
Val Ala Asp Pro Tyr Tyr Gly Thr Ser Gln Asp Phe Val Leu Thr Arg
130 135 140
Glu Asn Ile Glu Asp Ala Met Pro Gly Leu Leu Glu Trp Val Arg Asp
145 150 155 160
His Ile Arg Thr Asp Ser
165
<210>9
<211>1039
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1009)
<223>RXA00486
<400>9
gtttattcat gcccttgatt attgccaaag aaacctttaa ggactagatc gaaaaacagc 60
caactatagt taagtaatac tgaacaattt tggaggtgtc gtg ctc aat ctc aac 115
Val Leu Asn Leu Asn
1 5
cgc tta cac atc ctg cag gaa ttc cac cgc ctg gga acg att aca gca 163
Arg Leu His Ile Leu Gln Glu Phe His Arg Leu Gly Thr Ile Thr Ala
10 15 20
gtg gcg gaa tcc atg aac tac agc cgc tct gcc atc tcc caa caa atg 211
Val Ala Glu Ser Met Asn Tyr Ser Arg Ser Ala Ile Ser Gln Gln Met
25 30 35
gcg ctg ctg gaa aaa gaa att ggt gtg aaa ctc ttt gaa aaa agc ggc 259
Ala Leu Leu Glu Lys Glu Ile Gly Val Lys Leu Phe Glu Lys Ser Gly
40 45 50
cga aac ctc tac ttc aca gaa caa ggc gaa gtg ttg gcc tca gaa aca 307
Arg Asn Leu Tyr Phe Thr Glu Gln Gly Glu Val Leu Ala Ser Glu Thr
55 60 65
cat gcg atc atg gca gca gtc gac cat gcc cgc gca gcc gtt cta gat 355
His Ala Ile Met Ala Ala Val Asp His Ala Arg Ala Ala Val Leu Asp
70 75 80 85
tcg ctg tct gaa gtg tcc gga acg ctg aaa gtc acc tcc ttc caa tcc 403
Ser Leu Ser Glu Val Ser Gly Thr Leu Lys Val Thr Ser Phe Gln Ser
90 95 100
ctg ctg ttc acc ctt gcc ccg aaa gcc atc gcg cgc ctg acc gag aaa 451
Leu Leu Phe Thr Leu Ala Pro Lys Ala Ile Ala Arg Leu Thr Glu Lys
105 110 115
tac cca cac ctg caa gta gaa atc tcc caa cta gaa gtc acc gca gcg 499
Tyr Pro His Leu Gln Val Glu Ile Ser Gln Leu Glu Val Thr Ala Ala
120 125 130
ctc gaa gaa ctc cgc gcc cgc cgc gtc gac gtc gca ctc ggc gag gaa 547
Leu Glu Glu Leu Arg Ala Arg Arg Val Asp Val Ala Leu Gly Glu Glu
135 140 145
tac ccc gtg gaa gtc ccc ctt gtt gag gcc agc att cac cgc gaa gtc 595
Tyr Pro Val Glu Val Pro Leu Val Glu Ala Ser Ile His Arg Glu Val
150 155 160 165
ctc ttc gaa gac ccc atg ctg ctc gtc acc cca gca agc ggc cca tac 643
Leu Phe Glu Asp Pro Met Leu Leu Val Thr Pro Ala Ser Gly Pro Tyr
170 175 180
tct ggc ctc acc ctg cca gaa ctc cgc gac atc ccc atc gcc atc gat 691
Ser Gly Leu Thr Leu Pro Glu Leu Arg Asp Ile Pro Ile Ala Ile Asp
185 190 195
cca ccc gac ctt ccc gcg ggc gaa tgg gtc cat agg ctc tgc cgg cgc 739
Pro Pro Asp Leu Pro Ala Gly Glu Trp Val His Arg Leu Cys Arg Arg
200 205 210
gcc ggg ttt gag ccc cgc gtg acc ttt gaa acc agc gat ccc atg ctc 787
Ala Gly Phe Glu Pro Arg Val Thr Phe Glu Thr Ser Asp Pro Met Leu
215 220 225
caa gca cac ctc gtg cgt agc ggc ttg gcc gtg aca ttt tcc ccc aca 835
Gln Ala His Leu Val Arg Ser Gly Leu Ala Val Thr Phe Ser Pro Thr
230 235 240 245
ctg ctc acc ccg atg ctg gaa agc gtg cac atc cag ccg ctg ccc ggc 883
Leu Leu Thr Pro Met Leu Glu Ser Val His Ile Gln Pro Leu Pro Gly
250 255 260
aac ccc acg cgc acg ctc tac acc gcg gtc agg gaa ggg cgc cag ggg 931
Asn Pro Thr Arg Thr Leu Tyr Thr Ala Val Arg Glu Gly Arg Gln Gly
265 270 275
cat cca gcc att aaa gct ttt cga cga gcc ctc gcc cat gtg gcc aaa979
His Pro Ala Ile Lys Ala Phe Arg Arg Ala Leu Ala His Val Ala Lys
280 285 290
gaa tct tat ttg gag gct cgt cta gta gag tgagttcttg tgagccttca 1029
Glu Ser Tyr Leu Glu Ala Arg Leu Val Glu
295 300
gacaaatcat 1039
<210>10
<211>303
<212>PRT
<213>谷氨酸棒状杆菌
<400>10
Val Leu Asn Leu Asn Arg Leu His Ile Leu Gln Glu Phe His Arg Leu
1 5 10 15
Gly Thr Ile Thr Ala Val Ala Glu Ser Met Asn Tyr Ser Arg Ser Ala
20 25 30
Ile Ser Gln Gln Met Ala Leu Leu Glu Lys Glu Ile Gly Val Lys Leu
35 40 45
Phe Glu Lys Ser Gly Arg Asn Leu Tyr Phe Thr Glu Gln Gly Glu Val
50 55 60
Leu Ala Ser Glu Thr His Ala Ile Met Ala Ala Val Asp His Ala Arg
65 70 75 80
Ala Ala Val Leu Asp Ser Leu Ser Glu Val Ser Gly Thr Leu Lys Val
85 90 95
Thr Ser Phe Gln Ser Leu Leu Phe Thr Leu Ala Pro Lys Ala Ile Ala
100 105 110
Arg Leu Thr Glu Lys Tyr Pro His Leu Gln Val Glu Ile Ser Gln Leu
115 120 125
Glu Val Thr Ala Ala Leu Glu Glu Leu Arg Ala Arg Arg Val Asp Val
130 135 140
Ala Leu Gly Glu Glu Tyr Pro Val Glu Val Pro Leu Val Glu Ala Ser
145 150 155 160
Ile His Arg Glu Val Leu Phe Glu Asp Pro Met Leu Leu Val Thr Pro
165 170 175
Ala Ser Gly Pro Tyr Ser Gly Leu Thr Leu Pro Glu Leu Arg Asp Ile
180 185 190
Pro Ile Ala Ile Asp Pro Pro Asp Leu Pro Ala Gly Glu Trp Val His
195 200 205
Arg Leu Cys Arg Arg Ala Gly Phe Glu Pro Arg Val Thr Phe Glu Thr
210 215 220
Ser Asp Pro Met Leu Gln Ala His Leu Val Arg Ser Gly Leu Ala Val
225 230 235 240
Thr Phe Ser Pro Thr Leu Leu Thr Pro Met Leu Glu Ser Val His Ile
245 250 255
Gln Pro Leu Pro Gly Asn Pro Thr Arg Thr Leu Tyr Thr Ala Val Arg
260 265 270
Glu Gly Arg Gln Gly His Pro Ala Ile Lys Ala Phe Arg Arg Ala Leu
275 280 285
Ala His Val Ala Lys Glu Ser Tyr Leu Glu Ala Arg Leu Val Glu
290 295 300
<210>11
<211>478
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(448)
<223>RXA00593
<400>11
ttggtgtgat gcattcgaca gcaaattggc tgtgtgacta cacttgcgag tgtattaagt 60
attaggccgt gcatatgtag cgcattttaa ggagattgtc atg acg tct gtg att 115
Met Thr Ser Val Ile
1 5
cca gag cag cgc aac aac ccc ttt tat agg gac agc gcc aca att gct 163
Pro Glu Gln Arg Asn Asn Pro Phe Tyr Arg Asp Ser Ala Thr Ile Ala
10 15 20
tcc tcg gac cac aca gag cgt ggt gag tgg gtc act cag gca aag tgt 211
Ser Ser Asp His Thr Glu Arg Gly Glu Trp Val Thr Gln Ala Lys Cys
25 30 35
cga aat ggc gac cca gat gca ttg ttt gtt cgt ggt gca gcg caa cgc 259
Arg Asn Gly Asp Pro Asp Ala Leu Phe Val Arg Gly Ala Ala Gln Arg
40 45 50
cga gca gca gca att tgc cgc cac tgc cct gta gcc atg cag tgc tgc 307
Arg Ala Ala Ala Ile Cys Arg His Cys Pro Val Ala Met Gln Cys Cys
55 60 65
gcc gat gcc tta gat aac aag gtg gaa ttc gga gtc tgg gga ggc ctg 355
Ala Asp Ala Leu Asp Asn Lys Val Glu Phe Gly Val Trp Gly Gly Leu
70 75 80 85
acc gag cgc cag cgc cgt gca ttg ctt cga aag aag ccg cac att act 403
Thr Glu Arg Gln Arg Arg Ala Leu Leu Arg Lys Lys Pro His Ile Thr
90 95 100
aac tgg gct gaa tat ttg gct cag ggg ggc gag atc gcc ggg gtt 448
Asn Trp Ala Glu Tyr Leu Ala Gln Gly Gly Glu Ile Ala Gly Val
105 110 115
taattaattt caagggctgg ccattaacgt 478
<210>12
<211>116
<212>PRT
<213>谷氨酸棒状杆菌
<400>12
Met Thr Ser Val Ile Pro Glu Gln Arg Asn Asn Pro Phe Tyr Arg Asp
1 5 10 15
Ser Ala Thr Ile Ala Ser Ser Asp His Thr Glu Arg Gly Glu Trp Val
20 25 30
Thr Gln Ala Lys Cys Arg Asn Gly Asp Pro Asp Ala Leu Phe Val Arg
35 40 45
Gly Ala Ala Gln Arg Arg Ala Ala Ala Ile Cys Arg His Cys Pro Val
50 55 60
Ala Met Gln Cys Cys Ala Asp Ala Leu Asp Asn Lys Val Glu Phe Gly
65 70 75 80
Val Trp Gly Gly Leu Thr Glu Arg Gln Arg Arg Ala Leu Leu Arg Lys
85 90 95
Lys Pro His Ile Thr Asn Trp Ala Glu Tyr Leu Ala Gln Gly Gly Glu
100 105 110
Ile Ala Gly Val
115
<210>13
<211>796
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101..(766)
<223>RXA00609
<400>13
gaagcccacg gcggccgagc tttcgtcaat tccacaccag gtctaggatc cattttcggc 60
ctggaaatcc ccgcaccaga acaatcaaag gaatacaccc atg agc aag atc ctg l15
Met Ser Lys Ile Leu
1 5
ctc gct gaa gat gac gcc ggc atc gca gat ttc atc gtt cgt ggc ctc 163
Leu Ala Glu Asp Asp Ala Gly Ile Ala Asp Phe Ile Val Arg Gly Leu
10 15 20
atc cgc gaa ggc ttc gaa tgc gag gtc acc gaa tcc ggc gcc gaa gct 211
Ile Arg Glu Gly Phe Glu Cys Glu Val Thr Glu Ser Gly Ala Glu Ala
25 30 35
ttc gcc cgc gca cat tcc ggc gat ttc gat ctc atg gtt tta gac ctc 259
Phe Ala Arg Ala His Ser Gly Asp Phe Asp Leu Met Val Leu Asp Leu
40 45 50
ggc ctc ccc cac atg gac ggc acg gat gtc cta gag caa tta aga aat 307
Gly Leu Pro His Met Asp Gly Thr Asp Val Leu Glu Gln Leu Arg Asn
55 60 65
ctg cag gtc acg cta cct atc att gtg ctc acg gca cgc acc aac atc 355
Leu Gln Val Thr Leu Pro Ile Ile Val Leu Thr Ala Arg Thr Asn Ile
70 75 80 85
gag gac cgc ctc cgc acc ctc gag ggc ggc gcc gac gat tac atg ccc 403
Glu Asp Arg Leu Arg Thr Leu Glu Gly Gly Ala Asp Asp Tyr Met Pro
90 95 100
aaa cca ttc caa ttc gca gaa tta ctg gcc cgc atc aaa ctc cgc ctc 451
Lys Pro Phe Gln Phe Ala Glu Leu Leu Ala Arg Ile Lys Leu Arg Leu
105 110 115
gcc aaa cac act cct cag gaa acg ccg acc gat gcg cgc gtg cta cga 499
Ala Lys His Thr Pro Gln Glu Thr Pro Thr Asp Ala Arg Val Leu Arg
120 125 130
aac ggc gat ttg gag ctc gat ctt cgt acc cag cgt gtg ctc atc gac 547
Asn Gly Asp Leu Glu Leu Asp Leu Arg Thr Gln Arg Val Leu Ile Asp
135 140 145
ggc tcc tgg cac gac ctt tcc cgc cgc gaa gtc gat ctg ctc gaa acc 595
Gly Ser Trp His Asp Leu Ser Arg Arg Glu Val Asp Leu Leu Glu Thr
150 155 160 165
ctc atg cga cac cca ggg caa atc ctc tcc cga gtc caa ctc ctc cga 643
Leu Met Arg His Pro Gly Gln Ile Leu Ser Arg Val Gln Leu Leu Arg
170 175 180
ctg gtg tgg gac atg gat tgg gac ccc ggc tca aac gtg gtg gac gta 691
Leu Val Trp Asp Met Asp Trp Asp Pro Gly Ser Asn Val Val Asp Val
185 190 195
tat atc cgc gcg ttg agg aag aaa atc ggt gcc cat cgg gtc gaa acc 739
Tyr Ile Arg Ala Leu Arg Lys Lys Ile Gly Ala His Arg Val Glu Thr
200 205 210
atc cga gga tct ggc tac cgg ctg cgc taactgcaga acgagaccaa 786
Ile Arg Gly Ser Gly Tyr Arg Leu Arg
215 220
aaacctaaaa 796
<210>14
<211>222
<212>PRT
<213>谷氨酸棒状杆菌
<400>14
Met Ser Lys Ile Leu Leu Ala Glu Asp Asp Ala Gly Ile Ala Asp Phe
1 5 10 15
Ile Val Arg Gly Leu Ile Arg Glu Gly Phe Glu Cys Glu Val Thr Glu
20 25 30
Ser Gly Ala Glu Ala Phe Ala Arg Ala His Ser Gly Asp Phe Asp Leu
35 40 45
Met Val Leu Asp Leu Gly Leu Pro His Met Asp Gly Thr Asp Val Leu
50 55 60
Glu Gln Leu Arg Asn Leu Gln Val Thr Leu Pro Ile Ile Val Leu Thr
65 70 75 80
Ala Arg Thr Asn Ile Glu Asp Arg Leu Arg Thr Leu Glu Gly Gly Ala
85 90 95
Asp Asp Tyr Met Pro Lys Pro Phe Gln Phe Ala Glu Leu Leu Ala Arg
. 100 105 110
Ile Lys Leu Arg Leu Ala Lys His Thr Pro Gln Glu Thr Pro Thr Asp
115 120 125
Ala Arg Val Leu Arg Asn Gly Asp Leu Glu Leu Asp Leu Arg Thr Gln
130 135 140
Arg Val Leu Ile Asp Gly Ser Trp His Asp Leu Ser Arg Arg Glu Val
145 150 155 160
Asp Leu Leu Glu Thr Leu Met Arg His Pro Gly Gln Ile Leu Ser Arg
165 170 175
Val Gln Leu Leu Arg Leu Val Trp Asp Met Asp Trp Asp Pro Gly Ser
180 185 190
Asn Val Val Asp Val Tyr Ile Arg Ala Leu Arg Lys Lys Ile Gly Ala
195 200 205
His Arg Val Glu Thr Ile Arg Gly Ser Gly Tyr Arg Leu Arg
210 215 220
<210>15
<211>1498
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1468)
<223>RXA00651
<400>15
cgcgtcgatg ggaaggccgt cagtaattac ttccggggct gcctcggtgg tggtctctgg 60
ggttgcttca ggttccgccg gggtacaagc ggtgagcatg atg gaa gca gcg agg 115
Met Glu Ala Ala Arg
1 5
ata gta ggt aat gta cga cgc atg cag tca agc cta gat cgt gtg tcg 163
Ile Val Gly Asn Val Arg Arg Met Gln Ser Ser Leu Asp Arg Val Ser
10 15 20
gaa acc gga cgc aat gag ctc gat gtt gaa acc ctt gtg aag aag ggg 211
Glu Thr Gly Arg Asn Glu Leu Asp Val Glu Thr Leu Val Lys Lys Gly
25 30 35
aat caa ccg ggc gcg atg agc tat cgc aac agt atc cac att ttg aca 259
Asn Gln Pro Gly Ala Met Ser Tyr Arg Asn Ser Ile His Ile Leu Thr
40 45 50
gcc tcg ctg ctg gtc gtg ggg ttg gga gct tcc gcc cgc ctg acg ctg 307
Ala Ser Leu Leu Val Val Gly Leu Gly Ala Ser Ala Arg Leu Thr Leu
55 60 65
ccg atg ttt gcg ctg tcg tgc gtg ctg ttg ttt gtg tgg ggt ttt ctg 355
Pro Met Phe Ala Leu Ser Cys Val Leu Leu Phe Val Trp Gly Phe Leu
70 75 80 85
tac ttc tat gga tca acc aaa cgc gta gat ttg agc cac ggc atg cag 403
Tyr Phe Tyr Gly Ser Thr Lys Arg Val Asp Leu Ser His Gly Met Gln
90 95 100
ctg ggc tgg ctg ttt gtg ctg acg ctg gtg tgg att ttt atg gtg ccg 451
Leu Gly Trp Leu Phe Val Leu Thr Leu Val Trp Ile Phe Met Val Pro
105 110 115
atc gtg ccc gtg tcc att tat ctg ctg ttc ccg ctg ttt ttc ctc tat 499
Ile Val Pro Val Ser Ile Tyr Leu Leu Phe Pro Leu Phe Phe Leu Tyr
120 125 130
cta cag gtg atg cct gac gtg aga ggc att att gcg att ttg ggt gcg 547
Leu Gln Val Met Pro Asp Val Arg Gly Ile Ile Ala Ile Leu Gly Ala
135 140 145
aca gcg att gcg att gcc agc cag tat tcc gtg ggg ttg acc ttt ggt 595
Thr Ala Ile Ala Ile Ala Ser Gln Tyr Ser Val Gly Leu Thr Phe Gly
150 155 160 165
ggt gtg atg ggt ccg gtg gtc tct gcg atc gtg acc gtg gct att gat 643
Gly Val Met Gly Pro Val Val Ser Ala Ile Val Thr Val Ala Ile Asp
170 175 180
tac gcg ttc cgc acg ttg tgg cgg gtg aat aat gaa aag cag gaa ttg 691
Tyr Ala Phe Arg Thr Leu Trp Arg Val Asn Asn Glu Lys Gln Glu Leu
185 190 195
att gat cag ttg att gaa act cgc tcc cag ctg gcg gtg acg gaa cga 739
Ile Asp Gln Leu Ile Glu Thr Arg Ser Gln Leu Ala Val Thr Glu Arg
200 205 210
aat gcg ggt att gct gcg gaa cgt caa cgt att gcg cat gaa att cat 787
Asn Ala Gly Ile Ala Ala Glu Arg Gln Arg Ile Ala His Glu Ile His
215 220 225
gac acg gtc gcc cag gga ctc tcc tcc att caa atg ctg ctg cat gtc 835
Asp Thr Val Ala Gln Gly Leu Ser Ser Ile Gln Met Leu Leu His Val
230 235 240 245
tct gaa cag gag att ctc gtt gct gag atg gaa gag aag cca aag gag 883
Ser Glu Gln Glu Ile Leu Val Ala Glu Met Glu Glu Lys Pro Lys Glu
250 255 260
gcg atc gtg aag aag atg cgc ctt gcc cga caa aca gcc tcc gac aat 931
Ala Ile Val Lys Lys Met Arg Leu Ala Arg Gln Thr Ala Ser Asp Asn
265 270 275
ctc agt gag gct cgc gcg atg att gcg gcg ttg caa ccg gca gcg ctg 979
Leu Ser Glu Ala Arg Ala Met Ile Ala Ala Leu Gln Pro Ala Ala Leu
280 285 290
tct aaa acc tcc ttg gaa gca gca ctt cac cgc gtc aca gaa ccg ttg 1027
Ser Lys Thr Ser Leu Glu Ala Ala Leu His Arg Val Thr Glu Pro Leu
295 300 305
ttg ggt att aat ttt gtg att tct gtc gac ggt gat gtt cgc caa ctg 1075
Leu Gly Ile Asn Phe Val Ile Ser Val Asp Gly Asp Val Arg Gln Leu
310 315 320 325
ccc atg aaa act gaa gcc acc ctt ctg cga att gct caa ggt gcg atc 1123
Pro Met Lys Thr Glu Ala Thr Leu Leu Arg Ile Ala Gln Gly Ala Ile
330 335 340
gga aat gtg gcg aaa cat tca gag gcg aaa aac tgc cac gtg aca cta 1171
Gly Asn Val Ala Lys His Ser Glu Ala Lys Asn Cys His Val Thr Leu
345 350 355
acc tac gaa gac aca gaa gta cgc ctt gat gtg gtt gat gac ggt gtg 1219
Thr Tyr Glu Asp Thr Glu Val Arg Leu Asp Val Val Asp Asp Gly Val
360 365 370
ggt ttt gag cct tcg gaa gtg tcc agt acc ccc gct ggc ctt ggc cat 1267
Gly Phe Glu Pro Ser Glu Val Ser Ser Thr Pro Ala Gly Leu Gly His
375 380 385
atc ggc tta acc gca ttg cag cag cgt gcg atg gaa ttg cac ggc gaa 1315
Ile Gly Leu Thr Ala Leu Gln Gln Arg Ala Met Glu Leu His Gly Glu
390 395 400 405
gtt ata gtg gaa tct gca tat ggg cag ggt act gcg gta tct gca gca 1363
Val Ile Val Glu Ser Ala Tyr Gly Gln Gly Thr Ala Val Ser Ala Ala
410 415 420
ttg ccg gtg gag cca cca gag ggg ttt gtc ggg gcg ccg gtt ttg gca 1411
Leu Pro Val Glu Pro Pro Glu Gly Phe Val Gly Ala Pro Val Leu Ala
425 430 435
gat tcg gac tca agt gct aca ggc gag gtt gaa cta agt tct cca act 1459
Asp Ser Asp Ser Ser Ala Thr Gly Glu Val Glu Leu Ser Ser Pro Thr
440 445 450
gac gat gag taaggctaga ctaaagtacg attcatctgc 1498
Asp Asp Glu
455
<210>16
<211>456
<212>PRT
<213>谷氨酸棒状杆菌
<400>16
Met Glu Ala Ala Arg Ile Val Gly Asn Val Arg Arg Met Gln Ser Ser
1 5 10 15
Leu Asp Arg Val Ser Glu Thr Gly Arg Asn Glu Leu Asp Val Glu Thr
20 25 30
Leu Val Lys Lys Gly Asn Gln Pro Gly Ala Met Ser Tyr Arg Asn Ser
35 40 45
Ile His Ile Leu Thr Ala Ser Leu Leu Val Val Gly Leu Gly Ala Ser
50 55 60
Ala Arg Leu Thr Leu Pro Met Phe Ala Leu Ser Cys Val Leu Leu Phe
65 70 75 80
Val Trp Gly Phe Leu Tyr Phe Tyr Gly Ser Thr Lys Arg Val Asp Leu
85 90 95
Ser His Gly Met Gln Leu Gly Trp Leu Phe Val Leu Thr Leu Val Trp
100 105 110
Ile Phe Met Val Pro Ile Val Pro Val Ser Ile Tyr Leu Leu Phe Pro
115 120 125
Leu Phe Phe Leu Tyr Leu Gln Val Met Pro Asp Val Arg Gly Ile Ile
130 135 140
Ala Ile Leu Gly Ala Thr Ala Ile Ala Ile Ala Ser Gln Tyr Ser Val
145 150 155 160
Gly Leu Thr Phe Gly Gly Val Met Gly Pro Val Val Ser Ala Ile Val
165 170 175
Thr Val Ala Ile Asp Tyr Ala Phe Arg Thr Leu Trp Arg Val Asn Asn
180 185 190
Glu Lys Gln Glu Leu Ile Asp Gln Leu Ile Glu Thr Arg Ser Gln Leu
195 200 205
Ala Val Thr Glu Arg Asn Ala Gly Ile Ala Ala Glu Arg Gln Arg Ile
210 215 220
Ala His Glu Ile His Asp Thr Val Ala Gln Gly Leu Ser Ser Ile Gln
225 230 235 240
Met Leu Leu His Val Ser Glu Gln Glu Ile Leu Val Ala Glu Met Glu
245 250 255
Glu Lys Pro Lys Glu Ala Ile Val Lys Lys Met Arg Leu Ala Arg Gln
260 265 270
Thr Ala Ser Asp Asn Leu Ser Glu Ala Arg Ala Met Ile Ala Ala Leu
275 280 285
Gln Pro Ala Ala Leu Ser Lys Thr Ser Leu Glu Ala Ala Leu His Arg
290 295 300
Val Thr Glu Pro Leu Leu Gly Ile Asn Phe Val Ile Ser Val Asp Gly
305 310 315 320
Asp Val Arg Gln Leu Pro Met Lys Thr Glu Ala Thr Leu Leu Arg Ile
325 330 335
Ala Gln Gly Ala Ile Gly Asn Val Ala Lys His Ser Glu Ala Lys Asn
340 345 350
Cys His Val Thr Leu Thr Tyr Glu Asp Thr Glu Val Arg Leu Asp Val
355 360 365
Val Asp Asp Gly Val Gly Phe Glu Pro Ser Glu Val Ser Ser Thr Pro
370 375 380
Ala Gly Leu Gly His Ile Gly Leu Thr Ala Leu Gln Gln Arg Ala Met
385 390 395 400
Glu Leu His Gly Glu Val Ile Val Glu Ser Ala Tyr Gly Gln Gly Thr
405 410 415
Ala Val Ser Ala Ala Leu Pro Val Glu Pro Pro Glu Gly Phe Val Gly
420 425 430
Ala Pro Val Leu Ala Asp Ser Asp Ser Ser Ala Thr Gly Glu Val Glu
435 440 445
Leu Ser Ser Pro Thr Asp Asp Glu
450 455
<210>17
<211>1759
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1729)
<223>RXA00719
<400>17
cagatgatgc acacatcgtg gacacctctg atatgaccat ggatcaagta cttgatcacc 60
tcatccacct agtggaagcc tccgctgaaa ggagcaacca gtg act gat aaa cac 115
Val Thr Asp Lys His
l 5
acc atg cct ggt gaa gag gac gac acc gta ttc gtc tac cac acc cac 163
Thr Met Pro Gly Glu Glu Asp Asp Thr Val Phe Val Tyr His Thr His
10 15 20
aaa ggc gaa atg gac gtc gaa ggt gcg ttt gct gac gaa gaa gaa cta 211
Lys Gly Glu Met Asp Val Glu Gly Ala Phe Ala Asp Glu Glu Glu Leu
25 30 35
gca cca cac ggc ggt tgg gct tcc gca gat ttc gac cca gca gaa ttc 259
Ala Pro His Gly Gly Trp Ala Ser Ala Asp Phe Asp Pro Ala Glu Phe
40 45 50
ggc tac gaa gac tct gac gat gac ttc gat gca gag gac ttt gac gaa 307
Gly Tyr Glu Asp Ser Asp Asp Asp Phe Asp Ala Glu Asp Phe Asp Glu
55 60 65
aca gag ttc tcc aac cct gat ttc ggc gaa gac tac tct gat gaa gac 355
Thr Glu Phe Ser Asn Pro Asp Phe Gly Glu Asp Tyr Ser Asp Glu Asp
70 75 80 85
tgg gaa gaa atc gag acc gca ttc gga ttc gac cca agc cac ctt gaa 403
Trp Glu Glu Ile Glu Thr Ala Phe Gly Phe Asp Pro Ser His Leu Glu
90 95 100
gaa gct ctc tgc acg gtc gct atc gtc gga cgc cca aat gtt ggt aaa 451
Glu Ala Leu Cys Thr Val Ala Ile Val Gly Arg Pro Asn Val Gly Lys
105 110 115
tca acc ttg gtg aac cgc ttt att gga cgt cga gaa gca gtc gtg gaa 499
Ser Thr Leu Val Asn Arg Phe Ile Gly Arg Arg Glu Ala Val Val Glu
120 125 130
gat ttc ccc ggc gta acc cgt gac cgc atc tcc tac atc tct gac tgg 547
Asp Phe Pro Gly Val Thr Arg Asp Arg Ile Ser Tyr Ile Ser Asp Trp
135 140 145
ggt gga cac cgt ttc tgg gtt cag gac aca ggc gga tgg gat cct aac 595
Gly Gly His Arg Phe Trp Val Gln Asp Thr Gly Gly Trp Asp Pro Asn
150 155 160 165
gtc aag ggc atc cac gca tcg atc gca cag caa gca gaa gtt gct atg 643
Val Lys Gly Ile His Ala Ser Ile Ala Gln Gln Ala Glu Val Ala Met
170 175 180
agc act gcc gat gtc atc gta ttc gtc gtg gac acc aag gtg ggc atc 691
Ser Thr Ala Asp Val Ile Val Phe Val Val Asp Thr Lys Val Gly Ile
185 190 195
acc gaa act gac tca gtg atg gca gca aaa ctg ttg cgc tcg gaa gtg 739
Thr Glu Thr Asp Ser Val Met Ala Ala Lys Leu Leu Arg Ser Glu Val
200 205 210
cca gtg atc ttg gtt gcg aac aaa ttc gac tcc gac agc cag tgg gct 787
Pro Val Ile Leu Val Ala Asn Lys Phe Asp Ser Asp Ser Gln Trp Ala
215 220 225
gac atg gct gag ttc tac agc ctc ggc ctt ggc gat cca tac cca gtt 835
Asp Met Ala Glu Phe Tyr Ser Leu Gly Leu Gly Asp Pro Tyr Pro Val
230 235 240 245
tca gcc cag cat gga cgt ggt ggc gct gac gtt ttg gac aaa gtc ctt 883
Ser Ala Gln His Gly Arg Gly Gly Ala Asp Val Leu Asp Lys Val Leu
250 255 260
gaa ctc ttc cca gaa gag cct cgc tcc aag tcc atc gtg gaa ggc cct 931
Glu Leu Phe Pro Glu Glu Pro Arg Ser Lys Ser Ile Val Glu Gly Pro
265 270 275
cgt cgt gtc gcc ctt gtg ggt aag cca aac gtg ggt aag tct tca ctg 979
Arg Arg Val Ala Leu Val Gly Lys Pro Asn Val Gly Lys Ser Ser Leu
280 285 290
ctc aac aag ttt gct ggc gag acc cgc tct gtc gtg gac aat gtt gca 1027
Leu Asn Lys Phe Ala Gly Glu Thr Arg Ser Val Val Asp Asn Val Ala
295 300 305
gga acc acc gtt gac ccc gtt gac tcc ctg att cag ctg gat caa aaa 1075
Gly Thr Thr Val Asp Pro Val Asp Ser Leu Ile Gln Leu Asp GIn Lys
310 315 320 325
ctg tgg aaa ttc gtg gat act gct ggt ctt cgc aaa aag gtc aag act 1123
Leu Trp Lys Phe Val Asp Thr Ala Gly Leu Arg Lys Lys Val Lys Thr
330 335 340
gca tct ggc cac gag tac tac gca tca ctg cgt acc cac ggt gcc atc 1171
Ala Ser Gly His Glu Tyr Tyr Ala Ser Leu Arg Thr His Gly Ala Ile
345 350 355
gat gca gct gag ctg tgt gtt ttg ctt atc gat tcc tcc gaa ccc atc 1219
Asp Ala Ala Glu Leu Cys Val Leu Leu Ile Asp Ser Ser Glu Pro Ile
360 365 370
acc gag cag gat cag cgc gtg ctc gca atg atc acc gat gcc ggt aag 1267
Thr Glu Gln Asp Gln Arg Val Leu Ala Met Ile Thr Asp Ala Gly Lys
375 380 385
gca ctg gtt att gcg ttc aac aag tgg gat ctc atg gat gaa gat cgc 1315
Ala Leu Val Ile Ala Phe Asn Lys Trp Asp Leu Met Asp Glu Asp Arg
390 395 400 405
cgc atc gat ttg gat cgc gaa ctt gat ctc cag ttg gca cac gtg cct 1363
Arg Ile Asp Leu Asp Arg Glu Leu Asp Leu Gln Leu Ala His Val Pro
410 415 420
tgg gca aag cgc atc aac atc tcc gcc aaa acc ggt cgt gca ctg cag 1411
Trp Ala Lys Arg Ile Asn Ile Ser Ala Lys Thr Gly Arg Ala Leu Gln
425 430 435
cgc ctc gag cca gca atg ttg gaa gcg ctc gac aac tgg gat cgc cgt 1459
Arg Leu Glu Pro Ala Met Leu Glu Ala Leu Asp Asn Trp Asp Arg Arg
440 445 450
atc tcc act ggt cag ctg aac acc tgg ctg cgt gaa gca att gct gcg 1507
Ile Ser Thr Gly Gln Leu Asn Thr Trp Leu Arg Glu Ala Ile Ala Ala
455 460 465
aac cca cca cca atg cgt ggc gga cgt ttg cct cga gtg ctg ttt gcc 1555
Asn Pro Pro Pro Met Arg Gly Gly Arg Leu Pro Arg Val Leu Phe Ala
470 475 480 485
acc cag gca tct act cag cca cca gtg atc gta ctg ttc acc acc ggc 1603
Thr Gln Ala Ser Thr Gln Pro Pro Val Ile Val Leu Phe Thr Thr Gly
490 495 500
ttc ctc gaa gca ggt tac cga cga tac ctg gag cgc aag ttc cgt gaa 1651
Phe Leu Glu Ala Gly Tyr Arg Arg Tyr Leu Glu Arg Lys Phe Arg Glu
505 510 515
cgt ttc ggc ttt gaa ggc act cca gtg cga atc gct gtg cgt gtt cgc 1699
Arg Phe Gly Phe Glu Gly Thr Pro Val Arg Ile Ala Val Arg Val Arg
520 525 530
gag cgc cgc ggc aag ggc gga aac aag cag taaagcttga ttttccctaa 1749
Glu Arg Arg Gly Lys Gly Gly Asn Lys Gln
535 540
aagcacttat 1759
<210>18
<211>543
<212>PRT
<213>谷氨酸棒状杆菌
<400>18
Val Thr Asp Lys His Thr Met Pro Gly Glu Glu Asp Asp Thr Val Phe
1 5 10 15
Val Tyr His Thr His Lys Gly Glu Met Asp Val Glu Gly Ala Phe Ala
20 25 30
Asp Glu Glu Glu Leu Ala Pro His Gly Gly Trp Ala Ser Ala Asp Phe
35 40 45
Asp Pro Ala Glu Phe Gly Tyr Glu Asp Ser Asp Asp Asp Phe Asp Ala
50 55 60
Glu Asp Phe Asp Glu Thr Glu Phe Ser Asn Pro Asp Phe Gly Glu Asp
65 70 75 80
Tyr Ser Asp Glu Asp Trp Glu Glu Ile Glu Thr Ala Phe Gly Phe Asp
85 90 95
Pro Ser His Leu Glu Glu Ala Leu Cys Thr Val Ala Ile Val Gly Arg
100 105 110
Pro Asn Val Gly Lys Ser Thr Leu Val Asn Arg Phe Ile Gly Arg Arg
115 120 125
Glu Ala Val Val Glu Asp Phe Pro Gly Val Thr Arg Asp Arg Ile Ser
130 135 140
Tyr Ile Ser Asp Trp Gly Gly His Arg Phe Trp Val Gln Asp Thr Gly
145 150 155 160
Gly Trp Asp Pro Asn Val Lys Gly Ile His Ala Ser Ile Ala Gln Gln
165 170 175
Ala Glu Val Ala Met Ser Thr Ala Asp Val Ile Val Phe Val Val Asp
180 185 190
Thr Lys Val Gly Ile Thr Glu Thr Asp Ser Val Met Ala Ala Lys Leu
195 200 205
Leu Arg Ser Glu Val Pro Val Ile Leu Val Ala Asn Lys Phe Asp Ser
210 215 220
Asp Ser Gln Trp Ala Asp Met Ala Glu Phe Tyr Ser Leu Gly Leu Gly
225 230 235 240
Asp Pro Tyr Pro Val Ser Ala Gln His Gly Arg Gly Gly Ala Asp Val
245 250 255
Leu Asp Lys Val Leu Glu Leu Phe Pro Glu Glu Pro Arg Ser Lys Ser
260 265 270
Ile Val Glu Gly Pro Arg Arg Val Ala Leu Val Gly Lys Pro Asn Val
275 280 285
Gly Lys Ser Ser Leu Leu Asn Lys Phe Ala Gly Glu Thr Arg Ser Val
290 295 300
Val Asp Asn Val Ala Gly Thr Thr Val Asp Pro Val Asp Ser Leu Ile
305 310 315 320
Gln Leu Asp Gln Lys Leu Trp Lys Phe Val Asp Thr Ala Gly Leu Arg
325 330 335
Lys Lys Val Lys Thr Ala Ser Gly His Glu Tyr Tyr Ala Ser Leu Arg
340 345 350
Thr His Gly Ala Ile Asp Ala Ala Glu Leu Cys Val Leu Leu Ile Asp
355 360 365
Ser Ser Glu Pro Ile Thr Glu Gln Asp Gln Arg Val Leu Ala Met Ile
370 375 380
Thr Asp Ala Gly Lys Ala Leu Val Ile Ala Phe Asn Lys Trp Asp Leu
385 390 395 400
Met Asp Glu Asp Arg Arg Ile Asp Leu Asp Arg Glu Leu Asp Leu Gln
405 410 415
Leu Ala His Val Pro Trp Ala Lys Arg Ile Asn Ile Ser Ala Lys Thr
420 425 430
Gly Arg Ala Leu Gln Arg Leu Glu Pro Ala Met Leu Glu Ala Leu Asp
435 440 445
Asn Trp Asp Arg Arg Ile Ser Thr Gly Gln Leu Asn Thr Trp Leu Arg
450 455 460
Glu Ala Ile Ala Ala Asn Pro Pro Pro Met Arg Gly Gly Arg Leu Pro
465 470 475 480
Arg Val Leu Phe Ala Thr Gln Ala Ser Thr Gln Pro Pro Val Ile Val
485 490 495
Leu Phe Thr Thr Gly Phe Leu Glu Ala Gly Tyr Arg Arg Tyr Leu Glu
500 505 510
Arg Lys Phe Arg Glu Arg Phe Gly Phe Glu Gly Thr Pro Val Arg Ile
515 520 525
Ala Val Arg Val Arg Glu Arg Arg Gly Lys Gly Gly Asn Lys Gln
530 535 540
<210>19
<211>1261
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1231)
<223>RXA00813
<400>19
tccagcggct ggctaagtcc gtggagatgc atgggctgac cggttctttg ccgagggttt 60
taagctcagc atgcgacgcg gtcctcgggg aggtggcggc atg act gac att gat 115
Met Thr Asp Ile Asp
1 5
ctg gtg gtg gaa aac gtc caa agg att atc gcc acc aaa gag aca ccg 163
Leu Val Val Glu Asn Val Gln Arg Ile Ile Ala Thr Lys Glu Thr Pro
10 15 20
ccg acc tct gcg gaa ata gcg agc ctg att cgg gaa caa gca ggc gtg 211
Pro Thr Ser Ala Glu Ile Ala Ser Leu Ile Arg Glu Gln Ala Gly Val
25 30 35
atc agt aac gag gac atc gtg atg gtg ttg cgt cga ctg cgc agt gat 259
Ile Ser Asn Glu Asp Ile Val Met Val Leu Arg Arg Leu Arg Ser Asp
40 45 50
tct gtg ggc gtg gga ccg ttg gaa tct ctg ctt gcg ctt cct ggc gtg 307
Ser Val Gly Val Gly Pro Leu Glu Ser Leu Leu Ala Leu Pro Gly Val
55 60 65
acg gat gtg ttg gtt aat gcc cat gac agc gtg tgg att gat cgc ggt 355
Thr Asp Val Leu Val Asn Ala His Asp Ser Val Trp Ile Asp Arg Gly
70 75 80 85
cag ggc gtg gag aaa gtc gac atg gat ctg ggc tca gag gag gcg gtg 403
Gln Gly Val Glu Lys Val Asp Met Asp Leu Gly Ser Glu Glu Ala Val
90 95 100
cgt cgc ctt gcc acc cgg ttg gcg ttg acc tgt ggc aga cgc tta gat 451
Arg Arg Leu Ala Thr Arg Leu Ala Leu Thr Cys Gly Arg Arg Leu Asp
105 110 115
gat gcg cag cct ttc gct gat ggc cga atc acc agg gac gac ggc agc 499
Asp Ala Gln Pro Phe Ala Asp Gly Arg Ile Thr Arg Asp Asp Gly Ser
120 125 130
gtg ttg cgc att cac gcg gtg ttg gca ccc ttg gcg gaa tcc ggc acg 547
Val Leu Arg Ile His Ala Val Leu Ala Pro Leu Ala Glu Ser Gly Thr
135 140 145
tgc atc agt gtg cga gta ctg cgt caa gca cgg ctg agc ctt gat gat 595
Cys Ile Ser Val Arg Val Leu Arg Gln Ala Arg Leu Ser Leu Asp Asp
150 155 160 165
ctt atc caa agc ggc acg gtg cct gag gac atc gcg cct gcg ctc cgg 643
Leu Ile Gln Ser Gly Thr Val Pro Glu Asp Ile Ala Pro Ala Leu Arg
170 175 180
aac atc atc aat caa cgg cgc tcg ttc ctt gtt gtc ggt ggc acc ggc 691
Asn Ile Ile Asn Gln Arg Arg Ser Phe Leu Val Val Gly Gly Thr Gly
185 190 195
aca ggg aaa acc aca ttg ctg tcc gcg atg ctc acc gaa gtt ccc gct 739
Thr Gly Lys Thr Thr Leu Leu Ser Ala Met Leu Thr Glu Val Pro Ala
200 205 210
gat caa cga atc atc tgc atc gag gac acc gca gag ctt cat ccc ggc 787
Asp Gln Arg Ile Ile Cys Ile Glu Asp Thr Ala Glu Leu His Pro Gly
215 220 225
cat cca agc acc atc aac ttg gtg tct cgc caa gca aac gtc gag ggc 835
His Pro Ser Thr Ile Asn Leu Val Ser Arg Gln Ala Asn Val Glu Gly
230 235 240 245
gcc ggc gcc gtg agc atg gcg gat ttg ttg aaa caa tcg ctg cgc atg 883
Ala Gly Ala Val Ser Met Ala Asp Leu Leu Lys Gln Ser Leu Arg Met
250 255 260
agg cct gac cgg att gtc gtc gga gag att cgc ggt gcg gaa gtc gtg 931
Arg Pro Asp Arg Ile Val Val Gly Glu Ile Arg Gly Ala Glu Val Val
265 270 275
gat ctt ttg gct gcg atg aat acc gga cac gac ggc ggt gct ggc acc 979
Asp Leu Leu Ala Ala Met Asn Thr Gly His Asp Gly Gly Ala Gly Thr
280 285 290
att cac gcg aac tcc atc tct gaa gtt ccc gcg cgc atg gaa gct ctt 1027
Ile His Ala Asn Ser Ile Ser Glu Val Pro Ala Arg Met Glu Ala Leu
295 300 305
gcg gcg acc ggc gga ttg gac cgc atg gca ttg cat tct caa ctc gcg 1075
Ala Ala Thr Gly Gly Leu Asp Arg Met Ala Leu His Ser Gln Leu Ala
310 315 320 325
gcc gca gtg gac att gtg ctg gtc atg aaa cac acc cct ttt ggc cgc 1123
Ala Ala Val Asp Ile Val Leu Val Met Lys His Thr Pro Phe Gly Arg
330 335 340
agg cta gct caa ctc ggg gtg ctc cgc gga aat cct gtg acc acg cag 1171
Arg Leu Ala Gln Leu Gly Val Leu Arg Gly Asn Pro Val Thr Thr Gln
345 350 355
gtg gtg tgg gat ttg gac cac ggc atg cac gaa ggg agc gaa gag gca 1219
Val Val Trp Asp Leu Asp His Gly Met His Glu Gly Ser Glu Glu Ala
360 365 370
tgg ttt atg ccc taggccttct tagcgtggcg gtgttgatct 1261
Trp Phe Met Pro
375
<210>20
<211>377
<212>PRT
<213>谷氨酸棒状杆菌
<400>20
Met Thr Asp Ile Asp Leu Val Val Glu Asn Val Gln Arg Ile Ile Ala
1 5 10 15
Thr Lys Glu Thr Pro Pro Thr Ser Ala Glu Ile Ala Ser Leu Ile Arg
20 25 30
Glu Gln Ala Gly Val Ile Ser Asn Glu Asp Ile Val Met Val Leu Arg
35 40 45
Arg Leu Arg Ser Asp Ser Val Gly Val Gly Pro Leu Glu Ser Leu Leu
50 55 60
Ala Leu Pro Gly Val Thr Asp Val Leu Val Asn Ala His Asp Ser Val
65 70 75 80
Trp Ile Asp Arg Gly Gln Gly Val Glu Lys Val Asp Met Asp Leu Gly
85 90 95
Ser Glu Glu Ala Val Arg Arg Leu Ala Thr Arg Leu Ala Leu Thr Cys
100 105 110
Gly Arg Arg Leu Asp Asp Ala Gln Pro Phe Ala Asp Gly Arg Ile Thr
115 120 125
Arg Asp Asp Gly Ser Val Leu Arg Ile His Ala Val Leu Ala Pro Leu
130 135 140
Ala Glu Ser Gly Thr Cys Ile Ser Val Arg Val Leu Arg Gln Ala Arg
145 150 155 160
Leu Ser Leu Asp Asp Leu Ile Gln Ser Gly Thr Val Pro Glu Asp Ile
165 170 175
Ala Pro Ala Leu Arg Asn Ile Ile Asn Gln Arg Arg Ser Phe Leu Val
180 185 190
Val Gly Gly Thr Gly Thr Gly Lys Thr Thr Leu Leu Ser Ala Met Leu
195 200 205
Thr Glu Val Pro Ala Asp Gln Arg Ile Ile Cys Ile Glu Asp Thr Ala
210 215 220
Glu Leu His Pro Gly His Pro Ser Thr Ile Asn Leu Val Ser Arg Gln
225 230 235 240
Ala Asn Val Glu Gly Ala Gly Ala Val Ser Met Ala Asp Leu Leu Lys
245 250 255
Gln Ser Leu Arg Met Arg Pro Asp Arg Ile Val Val Gly Glu Ile Arg
260 265 270
Gly Ala Glu Val Val Asp Leu Leu Ala Ala Met Asn Thr Gly His Asp
275 280 285
Gly Gly Ala Gly Thr Ile His Ala Asn Ser Ile Ser Glu Val Pro Ala
290 295 300
Arg Met Glu Ala Leu Ala Ala Thr Gly Gly Leu Asp Arg Met Ala Leu
305 310 315 320
His Ser Gln Leu Ala Ala Ala Val Asp Ile Val Leu Val Met Lys His
325 330 335
Thr Pro Phe Gly Arg Arg Leu Ala Gln Leu Gly Val Leu Arg Gly Asn
340 345 350
Pro Val Thr Thr Gln Val Val Trp Asp Leu Asp His Gly Met His Glu
355 360 365
Gly Ser Glu Glu Ala Trp Phe Met Pro
370 375
<210>21
<211>880
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(850)
<223>RXA01081
<400>21
cattggtatt tgctcgaaag cacaacttaa aacagtaata atggtaatca cattcttttc 60
tattaagtaa gcaagtacac cggcccatta aagaggcacc atg acc cca gca aac 115
Met Thr Pro Ala Asn
1 5
gaa agt cct atg act aat cca tta ggt tct gcc ccc acc cca gcc aag 163
Glu Ser Pro Met Thr Asn Pro Leu Gly Ser Ala Pro Thr Pro Ala Lys
10 15 20
cca ctt ctt gac agt gtt ctt gat gag ctc ggt caa gat atc atc agt 211
Pro Leu Leu Asp Ser Val Leu Asp Glu Leu Gly Gln Asp Ile Ile Ser
25 30 35
ggc aag gtt gct gtc gga gat acc ttc aag ctg atg gac atc ggc gag 259
Gly Lys Val Ala Val Gly Asp Thr Phe Lys Leu Met Asp Ile Gly Glu
40 45 50
cgt ttt ggc att tcc cgc aca gtg gca cgc gaa gcg atg cgc gct ttg 307
Arg Phe Gly Ile Ser Arg Thr Val Ala Arg Glu Ala Met Arg Ala Leu
55 60 65
gag cag ctc ggt ctt gtc gct tct tca cgt cgc att ggc att act gtt 355
Glu Gln Leu Gly Leu Val Ala Ser Ser Arg Arg Ile Gly Ile Thr Val
70 75 80 85
ttg cca cag gaa gag tgg gct gtt ttt gat aag tcc atc att cgc tgg 403
Leu Pro Gln Glu Glu Trp Ala Val Phe Asp Lys Ser Ile Ile Arg Trp
90 95 100
cgt ctc aat gac gaa ggt cag cgt gaa ggc cag ctt cag tct ctt acc 451
Arg Leu Asn Asp Glu Gly Gln Arg Glu Gly Gln Leu Gln Ser Leu Thr
105 110 115
gag ctt cgt att gct att gaa ccg att gcc gcg cgc agc gtt gct ctt 499
Glu Leu Arg Ile Ala Ile Glu Pro Ile Ala Ala Arg Ser Val Ala Leu
120 125 130
cac gcg tca acc gcc gag ctc gag aaa atc cgc gcg ctc gca aca gag 547
His Ala Ser Thr Ala Glu Leu Glu Lys Ile Arg Ala Leu Ala Thr Glu
135 140 145
atg cgt cag ttg ggt gaa tct ggt cag ggt gcg tcc cag cgc ttc ctc 595
Met Arg Gln Leu Gly Glu Ser Gly Gln Gly Ala Ser Gln Arg Phe Leu
150 155 160 165
gaa gcg gac gtc act ttc cac gag ctc atc ttg cgt tat tgc cac aat 643
Glu Ala Asp Val Thr Phe His Glu Leu Ile Leu Arg Tyr Cys His Asn
170 175 180
gag atg ttc gct gca ctg att ccg tcg att agc gcg gtt ctt gtc ggc 691
Glu Met Phe Ala Ala Leu Ile Pro Ser Ile Ser Ala Val Leu Val Gly
185 190 195
cgc acc gag ctc ggc ctg cag cct gat ctg ccg gcg cac gag gcg cta 739
Arg Thr Glu Leu Gly Leu Gln Pro Asp Leu Pro Ala His Glu Ala Leu
200 205 210
gac aac cac gat aag ctt gcc gac gcc ctc ctt aac cgc gac gcc gac 787
Asp Asn His Asp Lys Leu Ala Asp Ala Leu Leu Asn Arg Asp Ala Asp
215 220 225
gcc gca gaa act gcg tcc cga aac atc ctc aat gag gtg cgc agc gcg 835
Ala Ala Glu Thr Ala Ser Arg Asn Ile Leu Asn Glu Val Arg Ser Ala
230 235 240 245
ctg ggc acg ctg aac taacgtgata cgcgcactgc gtttgccact 880
Leu Gly Thr Leu Asn
250
<210>22
<211>250
<212>PRT
<213>谷氨酸棒状杆菌
<400>22
Met Thr Pro Ala Asn Glu Ser Pro Met Thr Asn Pro Leu Gly Ser Ala
1 5 10 15
Pro Thr Pro Ala Lys Pro Leu Leu Asp Ser Val Leu Asp Glu Leu Gly
20 25 30
Gln Asp Ile Ile Ser Gly Lys Val Ala Val Gly Asp Thr Phe Lys Leu
35 40 45
Met Asp Ile Gly Glu Arg Phe Gly Ile Ser Arg Thr Val Ala Arg Glu
50 55 60
Ala Met Arg Ala Leu Glu Gln Leu Gly Leu Val Ala Ser Ser Arg Arg
65 70 75 80
Ile Gly Ile Thr Val Leu Pro Gln Glu Glu Trp Ala Val Phe Asp Lys
85 90 95
Ser Ile Ile Arg Trp Arg Leu Asn Asp Glu Gly Gln Arg Glu Gly Gln
100 105 110
Leu Gln Ser Leu Thr Glu Leu Arg Ile Ala Ile Glu Pro Ile Ala Ala
115 120 125
Arg Ser Val Ala Leu His Ala Ser Thr Ala Glu Leu Glu Lys Ile Arg
130 135 140
Ala Leu Ala Thr Glu Met Arg Gln Leu Gly Glu Ser Gly Gln Gly Ala
145 150 155 160
Ser Gln Arg Phe Leu Glu Ala Asp Val Thr Phe His Glu Leu Ile Leu
165 170 175
Arg Tyr Cys His Asn Glu Met Phe Ala Ala Leu Ile Pro Ser Ile Ser
180 185 190
Ala Val Leu Val Gly Arg Thr Glu Leu Gly Leu Gln Pro Asp Leu Pro
195 200 205
Ala His Glu Ala Leu Asp Asn His Asp Lys Leu Ala Asp Ala Leu Leu
210 215 220
Asn Arg Asp Ala Asp Ala Ala Glu Thr Ala Ser Arg Asn Ile Leu Asn
225 230 235 240
Glu Val Arg Ser Ala Leu Gly Thr Leu Asn
245 250
<210>23
<211>781
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(751)
<223>RXA01315
<400>23
catgtgcggt tgtgtcctaa gtcattgatg tcacctaacg cccctaagtt cactcggtgc 60
aatattgcac tgagtgcaag tttacactag gtttacttca gtg gat att gaa gag 115
Val Asp Ile Glu Glu
1 5
cag ccc tcg tta aga gaa atc aag cgc caa atg acc ctg gaa gcg ata 163
Gln Pro Ser Leu Arg Glu Ile Lys Arg Gln Met Thr Leu Glu Ala Ile
10 15 20
gaa gat aac gca acc agg ctc att ctg gag cgt ggc ttc gac aat gtc 211
Glu Asp Asn Ala Thr Arg Leu Ile Leu Glu Arg Gly Phe Asp Asn Val
25 30 35
aca atc gaa gac atc tgc gca gag gca ggg ata tcc aag cgc aca ttc 259
Thr Ile Glu Asp Ile Cys Ala Glu Ala Gly Ile Ser Lys Arg Thr Phe
40 45 50
ttt aac tac gtg gag tcc aaa gag tct gtg gcc atc ggg cac aca gcc 307
Phe Asn Tyr Val Glu Ser Lys Glu Ser Val Ala Ile Gly His Thr Ala
55 60 65
aag ctc cca acg gat gaa gaa cgt gaa gca ttc ctg gct acg cgt cat 355
Lys Leu Pro Thr Asp Glu Glu Arg Glu Ala Phe Leu Ala Thr Arg His
70 75 80 85
gaa aat att atc gat act gta ttt gac ctg gta atc aac ctc ttt ggc 403
Glu Asn Ile Ile Asp Thr Val Phe Asp Leu Val Ile Asn Leu Phe Gly
90 95 100
aac cac gac aac tcc aag tct gga gtt gca ggc gac att atg cgt cga 451
Asn His Asp Asn Ser Lys Ser Gly Val Ala Gly Asp Ile Met Arg Arg
105 110 115
cgc aaa gag atc cgg gtg aag cat cca gaa ctg gca gtg caa cat ttc 499
Arg Lys Glu Ile Arg Val Lys His Pro Glu Leu Ala Val Gln His Phe
120 125 130
gcc agg ttc cac caa gca cgc gaa ggg cta gaa cac cta att gtt gag 547
Ala Arg Phe His Gln Ala Arg Glu Gly Leu Glu His Leu I1e Val Glu
135 140 145
tac ttc gaa aaa tgg cca ggc tcc caa cat cta gat gag cct gca gat 595
Tyr Phe Glu Lys Trp Pro Gly Ser Gln His Leu Asp Glu Pro Ala Asp
150 155 160 165
cga gaa gca atc gcc ata gtt ggc ctg ctg atc tcg gtc atg ctt caa 643
Arg Glu Ala I1e Ala Ile Val Gly Leu Leu Ile Ser Val Met Leu Gln
170 175 180
ggt tct cgt gaa tgg cac gac atg cca caa ggc acg caa gct gat ttc 691
Gly Ser Arg Glu Trp His Asp Met Pro Gln Gly Thr Gln Ala Asp Phe
185 190 195
caa gcc tgc tgt cgc aaa gca att aaa aat act ttt ctt ctt aga ggt 739
Gln Ala Cys Cys Arg Lys Ala Ile Lys Asn Thr Phe Leu Leu Arg Gly
200 205 210
gga ttt tca gaa tgacatcaca ggtcaagccg gacgacgaac 781
Gly Phe Ser Glu
215
<210>24
<211>217
<212>PRT
<213>谷氨酸棒状杆菌
<400>24
Val Asp Ile Glu Glu Gln Pro Ser Leu Arg Glu Ile Lys Arg Gln Met
1 5 10 15
Thr Leu Glu Ala Ile Glu Asp Asn Ala Thr Arg Leu Ile Leu Glu Arg
20 25 30
Gly Phe Asp Asn Val Thr Ile Glu Asp Ile Cys Ala Glu Ala Gly Ile
35 40 45
Ser Lys Arg Thr Phe Phe Asn Tyr Val Glu Ser Lys Glu Ser Val Ala
50 55 60
Ile Gly His Thr Ala Lys Leu Pro Thr Asp Glu Glu Arg Glu Ala Phe
65 70 75 80
Leu Ala Thr Arg His Glu Asn Ile Ile Asp Thr Val Phe Asp Leu Val
85 90 95
Ile Asn Leu Phe Gly Asn His Asp Asn Ser Lys Ser Gly Val Ala Gly
100 105 110
Asp I1e Met Arg Arg Arg Lys Glu Ile Arg Val Lys His Pro Glu Leu
115 120 125
Ala Val Gln His Phe Ala Arg Phe His Gln Ala Arg Glu Gly Leu Glu
130 135 140
His Leu Ile Val Glu Tyr Phe Glu Lys Trp Pro Gly Ser Gln His Leu
145 150 155 160
Asp Glu Pro Ala Asp Arg Glu Ala Ile Ala Ile Val Gly Leu Leu Ile
165 170 175
Ser Val Met Leu Gln Gly Ser Arg Glu Trp His Asp Met Pro Gln Gly
180 185 190
Thr Gln Ala Asp Phe Gln Ala Cys Cys Arg Lys Ala Ile Lys Asn Thr
195 200 205
Phe Leu Leu Arg Gly Gly Phe Ser Glu
210 215
<210>25
<211>2212
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(2182)
<223>RXA01573
<400>25
agccccagct caccgaattc tccattcgtt ttaattgctt cgttaattaa aacgccatat 60
aaaaaccggc gcattgccgg tatttttcca ggagaattta atg aag agg ctt tcc 115
Met Lys Arg Leu Ser
l 5
cgt gca gcc ctc gca gtg gtc gcc acc acc gca gtt agc ttc agc gca 163
Arg Ala Ala Leu Ala Val Val Ala Thr Thr Ala Val Ser Phe Ser Ala
10 15 20
ctc gca gtt cca gct ttc gca gac gaa gca agc aat gtt gag ctc aac 211
Leu Ala Val Pro Ala Phe Ala Asp Glu Ala Ser Asn Val Glu Leu Asn
25 30 35
atc ctc ggt gtc acc gac ttc cac gga cac atc gag cag aag gct gtt 259
Ile Leu Gly Val Thr Asp Phe His Gly His Ile Glu Gln Lys Ala Val
40 45 50
aaa gat gat aag gga gta atc acc ggt tac tca gaa atg ggt gcc agt 307
Lys Asp Asp Lys Gly Val Ile Thr Gly Tyr Ser Glu Met Gly Ala Ser
55 60 65
ggc gtt gcc tgc tac gtc gac gct gaa cgc gcg gac aac cca aac acc 355
Gly Val Ala Cys Tyr Val Asp Ala Glu Arg Ala Asp Asn Pro Asn Thr
70 75 80 85
cgc ttc atc acc gtt ggt gac aac att ggt gga tcc cca ttc gtg tcc 403
Arg Phe Ile Thr Val Gly Asp Asn Ile Gly Gly Ser Pro Phe Val Ser
90 95 100
tcc atc ctg aag gat gag cca acc ttg caa gcc ctc agc gcc atc ggt 451
Ser Ile Leu Lys Asp Glu Pro Thr Leu Gln Ala Leu Ser Ala Ile Gly
105 110 115
gtt gac gca tcc gca ctg ggc aat cac gaa ttc gac cag ggc tac tca 499
Val Asp Ala Ser Ala Leu Gly Asn His Glu Phe Asp Gln Gly Tyr Ser
120 125 130
gac ctg gtg aac cgc gtt tcc ctc gac ggc tcc ggc agc gca aag ttc 547
Asp Leu Val Asn Arg Val Ser Leu Asp Gly Ser Gly Ser Ala Lys Phe
135 140 145
cca tac ctc ggc gca aac gtt gaa ggt ggc acc cca gca cct gca aag 595
Pro Tyr Leu Gly Ala Asn Val Glu Gly Gly Thr Pro Ala Pro Ala Lys
150 155 160 165
tct gaa atc atc gag atg gac ggc gtc aag atc gct tac gtc ggc gca 643
Ser Glu Ile Ile Glu Met Asp Gly Val Lys Ile Ala Tyr Val Gly Ala
170 175 180
gta acc gag gag acc gca acc ttg gtc tcc cca gca ggc atc gaa ggc 691
Val Thr Glu Glu Thr Ala Thr Leu Val Ser Pro Ala Gly Ile Glu Gly
185 190 195
atc acc ttc acc ggc gac atc gac gct atc aac gca gaa gca gat cgc 739
Ile Thr Phe Thr Gly Asp Ile Asp Ala Ile Asn Ala Glu Ala Asp Arg
200 205 210
gtc att gag gca ggc gaa gca gac gta gtc atc gca ttg atc cac gct 787
Val Ile Glu Ala Gly Glu Ala Asp Val Val Ile Ala Leu Ile His Ala
215 220 225
gaa gcc gct cca acc gat cta ttc tcc aac aac gtt gac gtt gta ttc 835
Glu Ala Ala Pro Thr Asp Leu Phe Ser Asn Asn Val Asp Val Val Phe
230 235 240 245
tcc gga cac acc cac ttc gac tac gtt gct gaa ggc gaa gca cgt ggc 883
Ser Gly His Thr His Phe Asp Tyr Val Ala Glu Gly Glu Ala Arg Gly
250 255 260
gac aag cag cca ctc gtt gtc atc cag ggc cac gaa tac ggc aag gtc 931
Asp Lys Gln Pro Leu Val Val Ile Gln Gly His Glu Tyr Gly Lys Val
265 270 275
atc tcc gac gtg gag atc tcc tac gac cgc gaa gca ggc aag atc acc 979
Ile Ser Asp Val Glu Ile Ser Tyr Asp Arg Glu Ala Gly Lys Ile Thr
280 285 290
aac att gag gcg aag aat gtc tct gct act gac gtt gtg gaa aac tgt 1027
Asn Ile Glu Ala Lys Asn Val Ser Ala Thr Asp Val Val Glu Asn Cys
295 300 305
gag act cca aac aca gca gtc gac gca atc gtt gca gct gct gtt gag 1075
Glu Thr Pro Asn Thr Ala Val Asp Ala Ile Val Ala Ala Ala Val Glu
310 315 320 325
gcc gct gaa gaa gca ggt aat gaa gtt gtt gca acc att gac aac ggc 1123
Ala Ala Glu Glu Ala Gly Asn Glu Val Val Ala Thr Ile Asp Asn Gly
330 335 340
ttc tac cgt ggg gcg gat gaa gag ggt acg acc ggc tcc aac cgt ggt 117l
Phe Tyr Arg Gly Ala Asp Glu Glu Gly Thr Thr Gly Ser Asn Arg Gly
345 350 355
gtt gag tct tcc ctg agc aac ctc atc gca gaa gct gga ctg tgg gca 1219
Val Glu Ser Ser Leu Ser Asn Leu Ile Ala Glu Ala Gly Leu Trp Ala
360 365 370
gtc aac gac gcg acc atc ctg aac gct gac atc ggc atc atg aac gca 1267
Val Asn Asp Ala Thr Ile Leu Asn Ala Asp Ile Gly Ile Met Asn Ala
375 380 385
ggc ggc gtg cgt gcg gac ctc gaa gca ggc gaa gtt acc ttc gca gat 1315
Gly Gly Val Arg Ala Asp Leu Glu Ala Gly Glu Val Thr Phe Ala Asp
390 395 400 405
gca tac gca acc cag aac ttc tcc aac acc tac ggc gta cgt gaa gtg 1363
Ala Tyr Ala Thr Gln Asn Phe Ser Asn Thr Tyr Gly Val Arg Glu Val
410 415 420
tct ggt gcg cag ttc aaa gaa gca ctg gaa cag cag tgg aag gaa acc 1411
Ser Gly Ala Gln Phe Lys Glu Ala Leu Glu Gln Gln Trp Lys Glu Thr
425 430 435
ggc gac cgc cca cgt ctg gca ttg gga ctg tcc agc aac gtc cag tac 1459
Gly Asp Arg Pro Arg Leu Ala Leu Gly Leu Ser Ser Asn Val Gln Tyr
440 445 450
tcc tac gac gag acc cgc gaa tac ggc gac cgc atc acc cac atc acc 1507
Ser Tyr Asp Glu Thr Arg Glu Tyr Gly Asp Arg Ile Thr His Ile Thr
455 460 465
ttc aac ggt gag cca atg gat atg aag gag acc tac cgc gtc aca gga 1555
Phe Asn Gly Glu Pro Met Asp Met Lys Glu Thr Tyr Arg Val Thr Gly
470 475 480 485
tca tcc ttc ctg ctc gca ggt ggc gac tcc ttc act gca ttc gct gaa 1603
Ser Ser Phe Leu Leu Ala Gly Gly Asp Ser Phe Thr Ala Phe Ala Glu
490 495 500
ggc ggc cca atc gct gaa acc ggc atg gtt gac att gac ctg ttc aac 1651
Gly Gly Pro Ile Ala Glu Thr Gly Met Val Asp Ile Asp Leu Phe Asn
505 510 515
aac tac atc gca gct cac cca gat gca cca att cgt gca aat cag agc 1699
Asn Tyr Ile Ala Ala His Pro Asp Ala Pro Ile Arg Ala Asn Gln Ser
520 525 530
tca gta ggc atc gcc ctt tcc ggc ccg gca gtt gca gaa gac gga act 1747
Ser Val Gly Ile Ala Leu Ser Gly Pro Ala Val Ala Glu Asp Gly Thr
535 540 545
ttg gtc cct ggt gaa gag ctg acc gtc gat ctt tct tcc ctc tcc tac 1795
Leu Val Pro Gly Glu Glu Leu Thr Val Asp Leu Ser Ser Leu Ser Tyr
550 555 560 565
acc gga cct gaa gct aag cca acc acc gtt gag gtg acc gtt ggt act 1843
Thr Gly Pro Glu Ala Lys Pro Thr Thr Val Glu Val Thr Val Gly Thr
570 575 580
gag aag aag act gcg gac gtc gat aac acc atc gtt cct cag ttt gac 1891
Glu Lys Lys Thr Ala Asp Val Asp Asn Thr Ile Val Pro Gln Phe Asp
585 590 595
agc acc ggc aag gca act gtc acc ctg act gtt cct gag gga gct acc 1939
Ser Thr Gly Lys Ala Thr Val Thr Leu Thr Val Pro Glu Gly Ala Thr
600 605 610
tct gtc aag atc gca act gac aat ggc act acc ttt gaa ctg cca gta 1987
Ser Val Lys Ile Ala Thr Asp Asn Gly Thr Thr Phe Glu Leu Pro Val
615 620 625
acc gta aac ggt gaa ggc aac aat gat gac gat gat gat aag gag cag 2035
Thr Val Asn Gly Glu Gly Asn Asn Asp Asp Asp Asp Asp Lys Glu Gln
630 635 640 645
cag tcc tcc gga tcc tcc gac gcc ggt tcc ctt gta gca gtt ctc ggt 2083
Gln Ser Ser Gly Ser Ser Asp Ala Gly Ser Leu Val Ala Val Leu Gly
650 655 660
gtt ctt gga gca ctc ggt ggc ctg gtg gcg ttc ttc ctg aac tct gcg 2131
Val Leu Gly Ala Leu Gly Gly Leu Val Ala Phe Phe Leu Asn Ser Ala
665 670 675
cag ggc gca cca ttc ttg gct cag ctt cag gct atg ttt gcg cag ttc 2179
Gln Gly Ala Pro Phe Leu Ala Gln Leu Gln Ala Met Phe Ala Gln Phe
680 685 690
atg taataacttg tagtaaataa atcgggcctt 2212
Met
<210>26
<211>694
<212>PRT
<213>谷氨酸棒状杆菌
<400>26
Met Lys Arg Leu Ser Arg Ala Ala Leu Ala Val Val Ala Thr Thr Ala
1 5 10 15
Val Ser Phe Ser Ala Leu Ala Val Pro Ala Phe Ala Asp Glu Ala Ser
20 25 30
Asn Val Glu Leu Asn Ile Leu Gly Val Thr Asp Phe His Gly His Ile
35 40 45
Glu Gln Lys Ala Val Lys Asp Asp Lys Gly Val Ile Thr Gly Tyr Ser
50 55 60
Glu Met Gly Ala Ser Gly Val Ala Cys Tyr Val Asp Ala Glu Arg Ala
65 70 75 80
Asp Asn Pro Asn Thr Arg Phe Ile Thr Val Gly Asp Asn Ile Gly Gly
85 90 95
Ser Pro Phe Val Ser Ser Ile Leu Lys Asp Glu Pro Thr Leu Gln Ala
100 105 110
Leu Ser Ala Ile Gly Val Asp Ala Ser Ala Leu Gly Asn His Glu Phe
115 120 125
Asp Gln Gly Tyr Ser Asp Leu Val Asn Arg Val Ser Leu Asp Gly Ser
130 135 140
Gly Ser Ala Lys Phe Pro Tyr Leu Gly Ala Asn Val Glu Gly Gly Thr
145 150 155 160
Pro Ala Pro Ala Lys Ser Glu Ile Ile Glu Met Asp Gly Val Lys Ile
165 170 175
Ala Tyr Val Gly Ala Val Thr Glu Glu Thr Ala Thr Leu Val Ser Pro
180 185 190
Ala Gly Ile Glu Gly Ile Thr Phe Thr Gly Asp Ile Asp Ala Ile Asn
195 200 205
Ala Glu Ala Asp Arg Val Ile Glu Ala Gly Glu Ala Asp Val Val Ile
210 215 220
Ala Leu Ile His Ala Glu Ala Ala Pro Thr Asp Leu Phe Ser Asn Asn
225 230 235 240
Val Asp Val Val Phe Ser Gly His Thr His Phe Asp Tyr Val Ala Glu
245 250 255
Gly Glu Ala Arg Gly Asp Lys Gln Pro Leu Val Val Ile Gln Gly His
260 265 270
Glu Tyr Gly Lys Val Ile Ser Asp Val Glu Ile Ser Tyr Asp Arg Glu
275 280 285
Ala Gly Lys Ile Thr Asn Ile Glu Ala Lys Asn Val Ser Ala Thr Asp
290 295 300
Val Val Glu Asn Cys Glu Thr Pro Asn Thr Ala Val Asp Ala Ile Val
305 310 315 320
Ala Ala Ala Val Glu Ala Ala Glu Glu Ala Gly Asn Glu Val Val Ala
325 330 335
Thr Ile Asp Asn Gly Phe Tyr Arg Gly Ala Asp Glu Glu Gly Thr Thr
340 345 350
Gly Ser Asn Arg Gly Val Glu Ser Ser Leu Ser Asn Leu Ile Ala Glu
355 360 365
Ala Gly Leu Trp Ala Val Asn Asp Ala Thr Ile Leu Asn Ala Asp Ile
370 375 380
Gly Ile Met Asn Ala Gly Gly Val Arg Ala Asp Leu Glu Ala Gly Glu
385 390 395 400
Val Thr Phe Ala Asp Ala Tyr Ala Thr Gln Asn Phe Ser Asn Thr Tyr
405 410 415
Gly Val Arg Glu Val Ser Gly Ala Gln Phe Lys Glu Ala Leu Glu Gln
420 425 430
Gln Trp Lys Glu Thr Gly Asp Arg Pro Arg Leu Ala Leu Gly Leu Ser
435 440 445
Ser Asn Val Gln Tyr Ser Tyr Asp Glu Thr Arg Glu Tyr Gly Asp Arg
450 455 460
Ile Thr His Ile Thr Phe Asn Gly Glu Pro Met Asp Met Lys Glu Thr
465 470 475 480
Tyr Arg Val Thr Gly Ser Ser Phe Leu Leu Ala Gly Gly Asp Ser Phe
485 490 495
Thr Ala Phe Ala Glu Gly Gly Pro Ile Ala Glu Thr Gly Met Val Asp
500 505 510
Ile Asp Leu Phe Asn Asn Tyr Ile Ala Ala His Pro Asp Ala Pro Ile
515 520 525
Arg Ala Asn Gln Ser Ser Val Gly Ile Ala Leu Ser Gly Pro Ala Val
530 535 540
Ala Glu Asp Gly Thr Leu Val Pro Gly Glu Glu Leu Thr Val Asp Leu
545 550 555 560
Ser Ser Leu Ser Tyr Thr Gly Pro Glu Ala Lys Pro Thr Thr Val Glu
565 570 575
Val Thr Val Gly Thr Glu Lys Lys Thr Ala Asp Val Asp Asn Thr Ile
580 585 590
Val Pro Gln Phe Asp Ser Thr Gly Lys Ala Thr Val Thr Leu Thr Val
595 600 605
Pro Glu Gly Ala Thr Ser Val Lys Ile Ala Thr Asp Asn Gly Thr Thr
610 615 620
Phe Glu Leu Pro Val Thr Val Asn Gly Glu Gly Asn Asn Asp Asp Asp
625 630 635 640
Asp Asp Lys Glu Gln Gln Ser Ser Gly Ser Ser Asp Ala Gly Ser Leu
645 650 655
Val Ala Val Leu Gly Val Leu Gly Ala Leu Gly Gly Leu Val Ala Phe
660 665 670
Phe Leu Asn Ser Ala Gln Gly Ala Pro Phe Leu Ala Gln Leu Gln Ala
675 680 685
Met Phe Ala Gln Phe Met
690
<210>27
<211>760
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(730)
<223>RXA01607
<400>27
gggctgaagg gctgggcgga acaataatta ttgaatctac aatcggatcg ggaactggaa 60
tttccgcccg ttttccctat ccacaaaagg accaagataa gtg atc cgt att ctg 115
Val Ile Arg Ile Leu
1 5
ttg gct gat gat cat ccc gtt gtt cgc gca ggc ctt gcc tcc ttg ctg 163
Leu Ala Asp Asp His Pro Val Val Arg Ala Gly Leu Ala Ser Leu Leu
10 15 20
gtg agt gaa gat gat ttt gag ata gtg gac atg gtg ggc acc cca gat 211
Val Ser Glu Asp Asp Phe Glu Ile Val Asp Met Val Gly Thr Pro Asp
25 30 35
gat gcc gtt gcg cgc gcc gcg gaa ggc ggg gtg gat gtg gtg ttg atg 259
Asp Ala Val Ala Arg Ala Ala Glu Gly Gly Val Asp Val Val Leu Met
40 45 50
gat ctg cgt ttt ggt gat caa cca ggc atc gag gtc gcc ggc ggg gta 307
Asp Leu Arg Phe Gly Asp Gln Pro Gly Ile Glu Val Ala Gly Gly Val
55 60 65
gag gca acg cgt cgc atc cgt gcg ctg gac aac ccg cca cag gta ctg 355
Glu Ala Thr Arg Arg Ile Arg Ala Leu Asp Asn Pro Pro Gln Val Leu
70 75 80 85
gtg gtg acc aac tac tcc aca gac ggc gat gtg gtg ggc gca gta tct 403
Val Val Thr Asn Tyr Ser Thr Asp Gly Asp Val Val Gly Ala Val Ser
90 95 100
gct ggt gcc gtg ggg tat ttg ctc aaa gat agc tcc cca gaa gat ctc 451
Ala Gly Ala Val Gly Tyr Leu Leu Lys Asp Ser Ser Pro Glu Asp Leu
105 110 115
att gcc ggt gtt cgc gat gcc gcg cgg gga gaa tca gtg ctt tca aag 499
Ile Ala Gly Val Arg Asp Ala Ala Arg Gly Glu Ser Val Leu Ser Lys
120 125 130
cag gtc gcc agc aag atc atg ggg cgg atg aac aac ccc atg act gct 547
Gln Val Ala Ser Lys Ile Met Gly Arg Met Asn Asn Pro Met Thr Ala
135 140 145
ctc agt gcc aga gaa att gaa gtg ctg tcc ttg gtg gcg caa ggg caa 595
Leu Ser Ala Arg Glu Ile Glu Val Leu Ser Leu Val Ala Gln Gly Gln
150 155 160 165
agc aat aga gaa atc ggc aag aaa ctt ttc ctc act gag gcc acg gtg 643
Ser Asn Arg Glu Ile Gly Lys Lys Leu Phe Leu Thr Glu Ala Thr Val
170 175 180
aaa agt cac atg ggg cat gtg ttc aac aag ctg gat gtc acc tct aga 691
Lys Ser His Met Gly His Val Phe Asn Lys Leu Asp Val Thr Ser Arg
185 190 195
aca gct gcg gta gct gaa gcc aga cag cgc gga att atc tagacgcaca 740
Thr Ala Ala Val Ala Glu Ala Arg Gln Arg Gly Ile Ile
200 205 210
cgtgttggta accgatcaca 760
<210>28
<211>210
<212>PRT
<213>谷氨酸棒状杆菌
<400>28
Val Ile Arg Ile Leu Leu Ala Asp Asp His Pro Val Val Arg Ala Gly
1 5 10 15
Leu Ala Ser Leu Leu Val Ser Glu Asp Asp Phe Glu Ile Val Asp Met
20 25 30
Val Gly Thr Pro Asp Asp Ala Val Ala Arg Ala Ala Glu Gly Gly Val
35 40 45
Asp Val Val Leu Met Asp Leu Arg Phe Gly Asp Gln Pro Gly Ile Glu
50 55 60
Val Ala Gly Gly Val Glu Ala Thr Arg Arg Ile Arg Ala Leu Asp Asn
65 70 75 80
Pro Pro Gln Val Leu Val Val Thr Asn Tyr Ser Thr Asp Gly Asp Val
85 90 95
Val Gly Ala Val Ser Ala Gly Ala Val Gly Tyr Leu Leu Lys Asp Ser
100 105 110
Ser Pro Glu Asp Leu Ile Ala Gly Val Arg Asp Ala Ala Arg Gly Glu
115 120 125
Ser Val Leu Ser Lys Gln Val Ala Ser Lys Ile Met Gly Arg Met Asn
130 135 140
Asn Pro Met Thr Ala Leu Ser Ala Arg Glu Ile Glu Val Leu Ser Leu
145 150 155 160
Val Ala Gln Gly Gln Ser Asn Arg Glu Ile Gly Lys Lys Leu Phe Leu
165 170 175
Thr Glu Ala Thr Val Lys Ser His Met Gly His Val Phe Asn Lys Leu
180 185 190
Asp Val Thr Ser Arg Thr Ala Ala Val Ala Glu Ala Arg Gln Arg Gly
195 200 205
Ile Ile
210
<210>29
<211>892
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(862)
<223>RXA01759
<400>29
cactaaagaa cttctgagcg cgctatcgtt ggtcgatgct attggtctgg gtacttctcc 60
ggtagaccat cactctgaat aagggggata acatatagtt atg acc aaa cgg ctc 115
Met Thr Lys Arg Leu
l 5
agc ctt gaa ggg ctc cgc tat gcg cag gcc gtc gca gaa act cac tca 163
Ser Leu Glu Gly Leu Arg Tyr Ala Gln Ala Val Ala Glu Thr His Ser
10 15 20
ttc agc gca gca gcc cgc gaa tac gga gtc acc caa cct gcg cta tcc 211
Phe Ser Ala Ala Ala Arg Glu Tyr Gly Val Thr Gln Pro Ala Leu Ser
25 30 35
aac ggc atc gcc aaa ctg gaa gat cgg ctc ggt gaa caa ctc ttc gat 259
Asn Gly Ile Ala Lys Leu Glu Asp Arg Leu Gly Glu Gln Leu Phe Asp
40 45 50
cga tct act caa ggc gtc acc ccg acg tcc ttt ggc ctc cac atc ctc 307
Arg Ser Thr Gln Gly Val Thr Pro Thr Ser Phe Gly Leu His Ile Leu
55 60 65
ccc ctg atc caa cgc gcg ctg act gaa atc gac gca atc acc gcg gaa 355
Pro Leu Ile Gln Arg Ala Leu Thr Glu Ile Asp Ala Ile Thr Ala Glu
70 75 80 85
gcg cac cgt ttg att aac tca gaa gca cgc agc att cga gtt gga atc 403
Ala His Arg Leu Ile Asn Ser Glu Ala Arg Ser Ile Arg Val Gly Ile
90 95 100
tcc cca ctt atc aac cct caa ctg gtt gca cga aca tat acc gcg gtt 451
Ser Pro Leu Ile Asn Pro Gln Leu Val Ala Arg Thr Tyr Thr Ala Val
105 110 115
cgt gag ctt ccc aca gca cac gac cta gta ctc cgc gaa gca aac atg 499
Arg Glu Leu Pro Thr Ala His Asp Leu Val Leu Arg Glu Ala Asn Met
120 125 130
aaa gaa cta cat gaa gga ctt ctt gca ggt gaa ctt aat gta att ctc 547
Lys Glu Leu His Glu Gly Leu Leu Ala Gly Glu Leu Asn Val Ile Leu
135 140 145
att ccc gca gtg aaa cca cta ccc cat ttt gaa cac cgc atc att gac 595
Ile Pro Ala Val Lys Pro Leu Pro His Phe Glu His Arg Ile Ile Asp
150 155 160 165
tcc gaa cca gtc gtt atc gtc gaa tcc acc cag gac agc acc gac ccc 643
Ser Glu Pro Val Val Ile Val Glu Ser Thr Gln Asp Ser Thr Asp Pro
170 175 180
ata gaa ctt cgc gag act cag cac gaa ccg ttc att ctg gta ccc gac 691
Ile Glu Leu Arg Glu Thr Gln His Glu Pro Phe Ile Leu Val Pro Asp
185 190 195
aca tgc ggt tta acc act ttc acc aat caa ctg ttt gaa aca aat gac 739
Thr Cys Gly Leu Thr Thr Phe Thr Asn Gln Leu Phe Glu Thr Asn Asp
200 205 210
ctg gca tta aac gcc tat tcc ggc gaa gca gcc agc tac caa gta ctc 787
Leu Ala Leu Asn Ala Tyr Ser Gly Glu Ala Ala Ser Tyr Gln Val Leu
215 220 225
gaa cag tgg gcc aca ctt gga ctc gga tct gca atg ctt cca ctt tct 835
Glu Gln Trp Ala Thr Leu Gly Leu Gly Ser Ala Met Leu Pro Leu Ser
230 235 240 245
aaa ctc agc tcc cct aca gca ccc cat tgaccactcc gcgaacaagg 882
Lys Leu Ser Ser Pro Thr Ala Pro His
250
cctcgacgtg 892
<210>30
<211>254
<212>PRT
<213>谷氨酸棒状杆菌
<400>30
Met Thr Lys Arg Leu Ser Leu Glu Gly Leu Arg Tyr Ala Gln Ala Val
1 5 10 15
Ala Glu Thr His Ser Phe Ser Ala Ala Ala Arg Glu Tyr Gly Val Thr
20 25 30
Gln Pro Ala Leu Ser Asn Gly Ile Ala Lys Leu Glu Asp Arg Leu Gly
35 40 45
Glu Gln Leu Phe Asp Arg Ser Thr Gln Gly Val Thr Pro Thr Ser Phe
50 55 60
Gly Leu His Ile Leu Pro Leu Ile Gln Arg Ala Leu Thr Glu Ile Asp
65 70 75 80
Ala Ile Thr Ala Glu Ala His Arg Leu Ile Asn Ser Glu Ala Arg Ser
85 90 95
Ile Arg Val Gly Ile Ser Pro Leu Ile Asn Pro Gln Leu Val Ala Arg
100 105 110
Thr Tyr Thr Ala Val Arg Glu Leu Pro Thr Ala His Asp Leu Val Leu
115 120 125
Arg Glu Ala Asn Met Lys Glu Leu His Glu Gly Leu Leu Ala Gly Glu
130 135 140
Leu Asn Val Ile Leu Ile Pro Ala Val Lys Pro Leu Pro His Phe Glu
145 150 155 160
His Arg Ile Ile Asp Ser Glu Pro Val Val Ile Val Glu Ser Thr Gln
165 170 175
Asp Ser Thr Asp Pro Ile Glu Leu Arg Glu Thr Gln His Glu Pro Phe
180 185 190
Ile Leu Val Pro Asp Thr Cys Gly Leu Thr Thr Phe Thr Asn Gln Leu
195 200 205
Phe Glu Thr Asn Asp Leu Ala Leu Asn Ala Tyr Ser Gly Glu Ala Ala
210 215 220
Ser Tyr Gln Val Leu Glu Gln Trp Ala Thr Leu Gly Leu Gly Ser Ala
225 230 235 240
Met Leu Pro Leu Ser Lys Leu Ser Ser Pro Thr Ala Pro His
245 250
<210>31
<211>2068
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(2038)
<223>RXA01826
<400>31
ctacacctag gacgagtgct agcgttccag ttgagactaa tgcaccggct gatgatttaa 60
tcgacgccgt aaatggccta ttggatgtag gaggagcgca gtg acc ttc gtg atc 115
Val Thr Phe Val Ile
1 5
gct gat cgc tat gaa ctg gat gcc gtc atc ggc tcc ggt ggc atg agc 163
Ala Asp Arg Tyr Glu Leu Asp Ala Val Ile Gly Ser Gly Gly Met Ser
10 15 20
gag gtg ttc gcg gcc acc gac acg ctc att ggt cgg gag gtc gcg gta 211
Glu Val Phe Ala Ala Thr Asp Thr Leu Ile Gly Arg Glu Val Ala Val
25 30 35
aag atg ctg cgc atc gac ctt gcg aaa gat ccc aat ttc cga gaa cgc 259
Lys Met Leu Arg Ile Asp Leu Ala Lys Asp Pro Asn Phe Arg Glu Arg
40 45 50
ttc cgc agg gaa gcc caa aac tcc gga agg ttg agc cac tct tcg atc 307
Phe Arg Arg Glu Ala Gln Asn Ser Gly Arg Leu Ser His Ser Ser Ile
55 60 65
gtc gct gtt ttt gac acc ggc gaa gta gac aaa gac ggc acc tct gtt 355
Val Ala Val Phe Asp Thr Gly Glu Val Asp Lys Asp Gly Thr Ser Val
70 75 80 85
ccc tac att gtg atg gaa cgc gtg cag ggt cga aac ctg cgc gaa gtt 403
Pro Tyr Ile Val Met Glu Arg Val Gln Gly Arg Asn Leu Arg Glu Val
90 95 100
gtc acc gaa gac ggc gta ttc acc cca gtt gag gca gcc aac atc ctc 45l
Val Thr Glu Asp Gly Val Phe Thr Pro Val Glu Ala Ala Asn Ile Leu
105 110 115
atc cct gtg tgt gaa gcg ctg cag gca tcc cat gac gcc ggc att att 499
Ile Pro Val Cys Glu Ala Leu Gln Ala Ser His Asp Ala Gly Ile Ile
120 125 130
cac cgc gat gtg aaa ccc gcc aac atc atg atc acc aac acc ggt ggc 547
His Arg Asp Val Lys Pro Ala Asn Ile Met Ile Thr Asn Thr Gly Gly
135 140 145
gtg aaa gtc atg gac ttc ggc atc gcc cgc gcg gtc aac gat tcc acc 595
Val Lys Val Met Asp Phe Gly Ile Ala Arg Ala Val Asn Asp Ser Thr
150 155 160 165
tcc gcc atg act caa acc tcc gca gtc atc ggc acc gcc cag tac ctc 643
Ser Ala Met Thr Gln Thr Ser Ala Val Ile Gly Thr Ala Gln Tyr Leu
170 175 180
tcc cct gag cag gcc cgc ggc aaa ccc gcc gat gcg cgt tcc gat att 691
Ser Pro Glu Gln Ala Arg Gly Lys Pro Ala Asp Ala Arg Ser Asp Ile
185 190 195
tac gcc acc ggc tgc gtc atg tac gaa tta gtc acc ggt aag cca cct 739
Tyr Ala Thr Gly Cys Val Met Tyr Glu Leu Val Thr Gly Lys Pro Pro
200 205 210
ttt gaa ggc gag tcc cct ttc gcc gtg gcc tac caa cac gtc cag gaa 787
Phe Glu Gly Glu Ser Pro Phe Ala Val Ala Tyr Gln His Val Gln Glu
215 220 225
gac ccc acc cct cct tcg gat ttc atc gcg gac ctc acc ccg acc tct 835
Asp Pro Thr Pro Pro Ser Asp Phe Ile Ala Asp Leu Thr Pro Thr Ser
230 235 240 245
gct gtc aac gtg gat gcc gtg gta ctc acc gcc atg gca aaa cac ccc 883
Ala Val Asn Val Asp Ala Val Val Leu Thr Ala Met Ala Lys His Pro
250 255 260
gcc gac cgc tac caa aca gcc tcc gaa atg gcc gct gac ctg ggc cgg 931
Ala Asp Arg Tyr Gln Thr Ala Ser Glu Met Ala Ala Asp Leu Gly Arg
265 270 275
cta tcc cgc aat gca gtc tcc cat gcc gca cgc gcg cat gta gaa aca 979
Leu Ser Arg Asn Ala Val Ser His Ala Ala Arg Ala His Val Glu Thr
280 285 290
gaa gaa acc cca gaa gag ccc gaa act cgc ttc tcg acg cgc acc tcc 1027
Glu Glu Thr Pro Glu Glu Pro Glu Thr Arg Phe Ser Thr Arg Thr Ser
295 300 305
acc caa gtg gcc ccc gcc gca ggc gtg gct gcg gcc agt acg ggg tca 1075
Thr Gln Val Ala Pro Ala Ala Gly Val Ala Ala Ala Ser Thr Gly Ser
310 315 320 25
ggg tct tct tcg cgt aaa cgt gga tcc aga ggc ctc acc gcc ctg gcc 1123
Gly Ser Ser Ser Arg Lys Arg Gly Ser Arg Gly Leu Thr Ala Leu Ala
330 335 340
atc gtg tta tcc cta ggt gtc gtc ggc gtt gcc ggt gcc ttc acc tac 1171
Ile Val Leu Ser Leu Gly Val Val Gly Val Ala Gly Ala Phe Thr Tyr
345 350 355
gac tac ttt gcc aac agc tcc tcc act gca acc agc gcg atc ccc aat 1219
Asp Tyr Phe Ala Asn Ser Ser Ser Thr Ala Thr Ser Ala Ile Pro Asn
360 365 370
gtg gaa ggc ctc ccg cag caa gaa gct ctc aca gaa ctt caa gca gca 1267
Val Glu Gly Leu Pro Gln Gln Glu Ala Leu Thr Glu Leu Gln Ala Ala
375 380 385
gga ttt gtt gtc aac atc gtc gaa gaa gcc agc gcc gac gtc gcc gaa 1315
Gly Phe Val Val Asn Ile Val Glu Glu Ala Ser Ala Asp Val Ala Glu
390 395 400 405
ggc ctc gtc atc cga gca aac cca agc gtt gga tcc gaa atc cgc caa 1363
Gly Leu Val Ile Arg Ala Asn Pro Ser Val Gly Ser Glu Ile Arg Gln
410 415 420
ggg gcc acc gtc acc atc acc gtg tcc acc ggc cga gaa atg atc aac 141l
Gly Ala Thr Val Thr Ile Thr Val Ser Thr Gly Arg Glu Met Ile Asn
425 430 435
atc cca gac gtc tcc ggc atg aca ctt gag gac gcc gcc cgc gcc ctc 1459
Ile Pro Asp Val Ser Gly Met Thr Leu Glu Asp Ala Ala Arg Ala Leu
440 445 450
gaa gac gtt ggt ctc ata ctc aac caa aac gtt cgg gaa gaa acc tcc 1507
Glu Asp Val Gly Leu Ile Leu Asn Gln Asn Val Arg Glu Glu Thr Ser
455 460 465
gac gac gtc gaa tct ggc ctc gtc atc gac caa aac ccc gaa gcc ggc 1555
Asp Asp Val Glu Ser Gly Leu Val Ile Asp Gln Asn Pro Glu Ala Gly
470 475 480 485
caa gaa gta gtc gtg ggt tcc tct gta tct cta acc atg tct tca ggc 1603
Gln Glu Val Val Val Gly Ser Ser Val Ser Leu Thr Met Ser Ser Gly
490 495 500
acc gag agc atc cga gtg ccc aac ctc acc ggc atg aac tgg tca caa 1651
Thr Glu Ser Ile Arg Val Pro Asn Leu Thr Gly Met Asn Trp Ser Gln
505 510 515
gca gaa caa aac ctc atc tcc atg ggc ttt aac ccc aca gct tcc tac 1699
Ala Glu Gln Asn Leu Ile Ser Met Gly Phe Asn Pro Thr Ala Ser Tyr
520 525 530
tta gac agc agc gaa cca gaa ggc gaa gtc ctc tca gtt tcc agc caa 1747
Leu Asp Ser Ser Glu Pro Glu Gly Glu Val Leu Ser Val Ser Ser Gln
535 540 545
gga act gaa cta ccc aag ggt tca tcc atc aca gtg gaa gtc tcc aac 1795
Gly Thr Glu Leu Pro Lys Gly Ser Ser Ile Thr Val Glu Val Ser Asn
550 555 560 565
ggc atg ctc atc caa gcc ccc gat ctc gcc cgc atg tcc acc gaa cag 1843
Gly Met Leu Ile Gln Ala Pro Asp Leu Ala Arg Met Ser Thr Glu Gln
570 575 580
gcc atc agt gcc ctc cgc gct gct ggc tgg acc gcc cca gat caa tcc 1891
Ala Ile Ser Ala Leu Arg Ala Ala Gly Trp Thr Ala Pro Asp Gln Ser
585 590 595
ctg atc gtc ggc gac ccc atc cac acc gca gcc ctc gtg gat caa aac 1939
Leu Ile Val Gly Asp Pro Ile His Thr Ala Ala Leu Val Asp Gln Asn
600 605 610
aaa atc gga ttc caa tcc cca acc cct gca acc ctc ttc cgc aaa gac 1987
Lys Ile Gly Phe Gln Ser Pro Thr Pro Ala Thr Leu Phe Arg Lys Asp
615 620 625
gcc caa gtg caa gtg cga ctc ttc gaa ttc gat ctc gct gca ctc gtg 2035
Ala Gln Val Gln Val Arg Leu Phe Glu Phe Asp Leu Ala Ala Leu Val
630 635 640 645
caa tagccaacaa ggaaaccgtc aaggtagctg 2068
Gln
<210>32
<211>646
<212>PRT
<213>谷氨酸棒状杆菌
<400>32
Val Thr Phe Val Ile Ala Asp Arg Tyr Glu Leu Asp Ala Val Ile Gly
1 5 10 15
Ser Gly Gly Met Ser Glu Val Phe Ala Ala Thr Asp Thr Leu Ile Gly
20 25 30
Arg Glu Val Ala Val Lys Met Leu Arg Ile Asp Leu Ala Lys Asp Pro
35 40 45
Asn Phe Arg Glu Arg Phe Arg Arg Glu Ala Gln Asn Ser Gly Arg Leu
50 55 60
Ser His Ser Ser Ile Val Ala Val Phe Asp Thr Gly Glu Val Asp Lys
65 70 75 80
Asp Gly Thr Ser Val Pro Tyr Ile Val Met Glu Arg Val Gln Gly Arg
85 90 95
Asn Leu Arg Glu Val Val Thr Glu Asp Gly Val Phe Thr Pro Val Glu
100 105 110
Ala Ala Asn Ile Leu Ile Pro Val Cys Glu Ala Leu Gln Ala Ser His
115 120 125
Asp Ala Gly Ile Ile His Arg Asp Val Lys Pro Ala Asn Ile Met Ile
130 135 140
Thr Asn Thr Gly Gly Val Lys Val Met Asp Phe Gly Ile Ala Arg Ala
145 150 155 160
Val Asn Asp Ser Thr Ser Ala Met Thr Gln Thr Ser Ala Val Ile Gly
165 170 175
Thr Ala Gln Tyr Leu Ser Pro Glu Gln Ala Arg Gly Lys Pro Ala Asp
180 185 190
Ala Arg Ser Asp Ile Tyr Ala Thr Gly Cys Val Met Tyr Glu Leu Val
195 200 205
Thr Gly Lys Pro Pro Phe Glu Gly Glu Ser Pro Phe Ala Val Ala Tyr
210 215 220
Gln His Val Gln Glu Asp Pro Thr Pro Pro Ser Asp Phe Ile Ala Asp
225 230 235 240
Leu Thr Pro Thr Ser Ala Val Asn Val Asp Ala Val Val Leu Thr Ala
245 250 255
Met Ala Lys His Pro Ala Asp Arg Tyr Gln Thr Ala Ser Glu Met Ala
260 265 270
Ala Asp Leu Gly Arg Leu Ser Arg Asn Ala Val Ser His Ala Ala Arg
275 280 285
Ala His Val Glu Thr Glu Glu Thr Pro Glu Glu Pro Glu Thr Arg Phe
290 295 300
Ser Thr Arg Thr Ser Thr Gln Val Ala Pro Ala Ala Gly Val Ala Ala
305 310 315 320
Ala Ser Thr Gly Ser Gly Ser Ser Ser Arg Lys Arg Gly Ser Arg Gly
325 330 335
Leu Thr Ala Leu Ala Ile Val Leu Ser Leu Gly Val Val Gly Val Ala
340 345 350
Gly Ala Phe Thr Tyr Asp Tyr Phe Ala Asn Ser Ser Ser Thr Ala Thr
355 360 365
Ser Ala Ile Pro Asn Val Glu Gly Leu Pro Gln Gln Glu Ala Leu Thr
370 375 380
Glu Leu Gln Ala Ala Gly Phe Val Val Asn Ile Val Glu Glu Ala Ser
385 390 395 400
Ala Asp Val Ala Glu Gly Leu Val Ile Arg Ala Asn Pro Ser Val Gly
405 410 415
Ser Glu Ile Arg Gln Gly Ala Thr Val Thr Ile Thr Val Ser Thr Gly
420 425 430
Arg Glu Met Ile Asn Ile Pro Asp Val Ser Gly Met Thr Leu Glu Asp
435 440 445
Ala Ala Arg Ala Leu Glu Asp Val Gly Leu Ile Leu Asn Gln Asn Val
450 455 460
Arg Glu Glu Thr Ser Asp Asp Val Glu Ser Gly Leu Val Ile Asp Gln
465 470 475 480
Asn Pro Glu Ala Gly Gln Glu Val Val Val Gly Ser Ser Val Ser Leu
485 490 495
Thr Met Ser Ser Gly Thr Glu Ser Ile Arg Val Pro Asn Leu Thr Gly
500 505 510
Met Asn Trp Ser Gln Ala Glu Gln Asn Leu Ile Ser Met Gly Phe Asn
515 520 525
Pro Thr Ala Ser Tyr Leu Asp Ser Ser Glu Pro Glu Gly Glu Val Leu
530 535 540
Ser Val Ser Ser Gln Gly Thr Glu Leu Pro Lys Gly Ser Ser Ile Thr
545 550 555 560
Val Glu Val Ser Asn Gly Met Leu Ile Gln Ala Pro Asp Leu Ala Arg
565 570 575
Met Ser Thr Glu Gln Ala Ile Ser Ala Leu Arg Ala Ala Gly Trp Thr
580 585 590
Ala Pro Asp Gln Ser Leu Ile Val Gly Asp Pro Ile His Thr Ala Ala
595 600 605
Leu Val Asp Gln Asn Lys Ile Gly Phe Gln Ser Pro Thr Pro Ala Thr
610 615 620
Leu Phe Arg Lys Asp Ala Gln Val Gln Val Arg Leu Phe Glu Phe Asp
625 630 635 640
Leu Ala Ala Leu Val Gln
645
<210>33
<211>3502
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(3472)
<223>RXA02097
<400>33
ttggctacag atgtttcagc gcttgcagtg gggtagtgtg tttaaacatc acaattagtt 60
ctagaggaaa acgcattttt tgcgcgggga gagtgtatac atg ccg gct ggc atc 115
Met Pro Ala Gly Ile
1 5
gca gac atg aca gat tca ttg ctc gga tgg gca tca caa act gag ctg 163
Ala Asp Met Thr Asp Ser Leu Leu Gly Trp Ala Ser Gln Thr Glu Leu
10 15 20
gat ctg aac cag cgt ctt gca ggg gta gag tac ttt cca caa att cag 211
Asp Leu Asn Gln Arg Leu Ala Gly Val Glu Tyr Phe Pro Gln Ile Gln
25 30 35
ctg cga cac gat gag ctc gag cgc att cat cgg ttt tac ggc acc ttt 259
Leu Arg His Asp Glu Leu Glu Arg Ile His Arg Phe Tyr Gly Thr Phe
40 45 50
ttg tcc cgc cag gta ggc gcg ggc gca agc ctt ggg gat ctt ttt gaa 307
Leu Ser Arg Gln Val Gly Ala Gly Ala Ser Leu Gly Asp Leu Phe Glu
55 60 65
atg acc cca tgc ctg aca gtc acc acc ttg gtg tct cgg gcg tca cgg 355
Met Thr Pro Cys Leu Thr Val Thr Thr Leu Val Ser Arg Ala Ser Arg
70 75 80 85
atc agc gat cca gca gat ttc ttc ggt gaa tac atc gga gga ctg gga 403
Ile Ser Asp Pro Ala Asp Phe Phe Gly Glu Tyr Ile Gly Gly Leu Gly
90 95 100
ctt agc gca gaa cac gca gca gtt gtt gaa ggg ttg acc gaa aag ctc 451
Leu Ser Ala Glu His Ala Ala Val Val Glu Gly Leu Thr Glu Lys Leu
105 110 115
ttc gca cag gct ggc ctg ctc gtt cct gag gga att gca tct cca ttg 499
Phe Ala Gln Ala Gly Leu Leu Val Pro Glu Gly Ile Ala Ser Pro Leu
120 125 130
gag ttg tta tcc atc cac gca ggc att agt aac cac gaa gtg gcc gca 547
Glu Leu Leu Ser Ile His Ala Gly Ile Ser Asn His Glu Val Ala Ala
135 140 145
gtg ctg acc gaa gtg gaa aac ggc acc acc gaa tat cca ttc atg ttc 595
Val Leu Thr Glu Val Glu Asn Gly Thr Thr Glu Tyr Pro Phe Met Phe
150 155 160 165
gac gct gtc ctg cgc cta acc cct gag tgg gca cag acc ctt atc ggc 643
Asp Ala Val Leu Arg Leu Thr Pro Glu Trp Ala Gln Thr Leu Ile Gly
170 175 180
gga gtt caa gaa ctc att gaa ttt gcc acc acc cac cga act tct tgg 691
Gly Val Gln Glu Leu Ile Glu Phe Ala Thr Thr His Arg Thr Ser Trp
185 190 195
tca gac cgc cag cgc gaa tcc tca ctg cca gcc atg atc gat gag atc 739
Ser Asp Arg Gln Arg Glu Ser Ser Leu Pro Ala Met Ile Asp Glu Ile
200 205 210
gtt gtg gcg gaa ctt cgg gaa cgc cca gtt ggt act gcc gac cgt gaa 787
Val Val Ala Glu Leu Arg Glu Arg Pro Val Gly Thr Ala Asp Arg Glu
215 220 225
aac tcc gtt ggt gtg gca ctt cgt gag ctt cgc cca cgc ctc atc ctg 835
Asn Ser Val Gly Val Ala Leu Arg Glu Leu Arg Pro Arg Leu Ile Leu
230 235 240 45
gat gca gaa cgc cgc aaa gtc tgc ctg cgt cta cct gaa cag cgc gtc 883
Asp Ala Glu Arg Arg Lys Val Cys Leu Arg Leu Pro Glu Gln Arg Val
250 255 260
agc gac gat gaa atc aac tgg cga gtc agc cta gaa ggc acc acc cgg 931
Ser Asp Asp Glu Ile Asn Trp Arg Val Ser Leu Glu Gly Thr Thr Arg
265 270 275
att ttc tcc acc cgc cga gca tgg ggc gat act tct gga tac tcc gaa 979
Ile Phe Ser Thr Arg Arg Ala Trp Gly Asp Thr Ser Gly Tyr Ser Glu
280 285 290
gcc ctc gac atc act gtc gag cgt caa atc cgc gaa acc acc gtc acc 1027
Ala Leu Asp Ile Thr Val Glu Arg Gln Ile Arg Glu Thr Thr Val Thr
295 300 305
gac acc tca aac caa atc acc tgg gtt gtc cca gtc gtg gac ttc aac 1075
Asp Thr Ser Asn Gln Ile Thr Trp Val Val Pro Val Val Asp Phe Asn
310 315 320 325
gac cca gtg ctg gtg ttt tcc gcg cgc ggt gaa aac ctc acc gac aag 1123
Asp Pro Val Leu Val Phe Ser Ala Arg Gly Glu Asn Leu Thr Asp Lys
330 335 340
gtc tcc ctg cac cat caa gag att tac gtt ctc gcg cca gcg gaa gca 1171
Val Ser Leu His His Gln Glu Ile Tyr Val Leu Ala Pro Ala Glu Ala
345 350 355
aaa ctc gaa gac atg gtc act ggc cag cca gta cca gtt att gag caa 1219
Lys Leu Glu Asp Met Val Thr Gly Gln Pro Val Pro Val Ile Glu Gln
360 365 370
ttc ctc gta gag ggc tgg aac tca tgg gtg tgc tcc cgc gtg gac gcc 1267
Phe Leu Val Glu Gly Trp Asn Ser Trp Val Cys Ser Arg Val Asp Ala
375 380 385
cgt ggc ctg tcc tct ctg aag gtc aac aaa gaa gtc cga tgc att gac 1315
Arg Gly Leu Ser Ser Leu Lys Val Asn Lys Glu Val Arg Cys Ile Asp
390 395 400 405
cca cgt cga cgc gtt gcc ttc cac cac cca gcc gaa ttg gtc cct cac 1363
Pro Arg Arg Arg Val Ala Phe His His Pro Ala Glu Leu Val Pro His
410 415 420
gta cga tcc att tcc gga ctc ccc gta cac gcg cag tcc ctg atc gcc 1411
Val Arg Ser Ile Ser Gly Leu Pro Val His Ala Gln Ser Leu Ile Ala
425 430 435
gag ttc cca cca acc ctg agc gga caa gac gaa acc tgg atg ctc tcc 1459
Glu Phe Pro Pro Thr Leu Ser Gly Gln Asp Glu Thr Trp Met Leu Ser
440 445 450
atc tcg gct ttc gca ggt gta ggc gct gct ggt gaa gaa atc gcc gag 1507
Ile Ser Ala Phe Ala Gly Val Gly Ala Ala Gly Glu Glu Ile Ala Glu
455 460 465
cca gag cct ttg gaa gtc cct gcc gac ggt ggc ctt ttc gcc atc ttc 1555
Pro Glu Pro Leu Glu Val Pro Ala Asp Gly Gly Leu Phe Ala Ile Phe
470 475 480 485
gac cca gaa ata tac gac gcc cca tgg gtg ggt gaa tac ctg gtc cga 1603
Asp Pro Glu Ile Tyr Asp Ala Pro Trp Val Gly Glu Tyr Leu Val Arg
490 495 500
ctc cgc ggc cca cgc aat gaa tcc ttc cga ccc gaa ttc gcc atc gtc 1651
Leu Arg Gly Pro Arg Asn Glu Ser Phe Arg Pro Glu Phe Ala Ile Val
505 510 515
gaa gac atg acc acc gaa ttc gaa gtc gcc tca ggt gca tca ttt cga 1699
Glu Asp Met Thr Thr Glu Phe Glu Val Ala Ser Gly Ala Ser Phe Arg
520 525 530
atc cca acc acc act ggt ctc agc gaa gcc agc cta cgc gtg cgt tcc 1747
Ile Pro Thr Thr Thr Gly Leu Ser Glu Ala Ser Leu Arg Val Arg Ser
535 540 545
ggt gaa aag cac ttc acc gca gag cca cgc ctg gtc acc gtt gaa gca 1795
Gly Glu Lys His Phe Thr Ala Glu Pro Arg Leu Val Thr Val Glu Ala
550 555 560 565
acc gac ccc aac gca tca ttc gtg gtc acc acc gat gaa ggc gat caa 1843
Thr Asp Pro Asn Ala Ser Phe Val Val Thr Thr Asp Glu Gly Asp Gln
570 575 580
atg cca ttg cga ttt gtg cca cca caa atc gcc atc gaa ctt cca ctg 1891
Met Pro Leu Arg Phe Val Pro Pro Gln Ile Ala Ile Glu Leu Pro Leu
585 590 595
acc acc gag cca cca acc tgg cgc gtc acc cgt act gtc tgt gga cca 1939
Thr Thr Glu Pro Pro Thr Trp Arg Val Thr Arg Thr Val Cys Gly Pro
600 605 610
cgc gac ctc gac ggt gca ggc gaa ctc cgc atc cgc acc ggt gtc gat 1987
Arg Asp Leu Asp Gly Ala Gly Glu Leu Arg Ile Arg Thr Gly Val Asp
615 620 625
gtc ggc gat cca aag gtc agt gtg cgc aac cac cac ggt tca cca ctg 2035
Val Gly Asp Pro Lys Val Ser Val Arg Asn His His Gly Ser Pro Leu
630 635 640 645
cga acc gtg aaa atg gtc acc cct gac aac ggc cgt acc tgg att gcc 2083
Arg Thr Val Lys Met Val Thr Pro Asp Asn Gly Arg Thr Trp Ile Ala
650 655 660
agc atg aag gaa atc gca gcc agt acc ttt gtg atg cca cgc gga tcc 2131
Ser Met Lys Glu Ile Ala Ala Ser Thr Phe Val Met Pro Arg Gly Ser
665 670 675
atc gaa ttt gag tgg act gac cgc aag gtt gac cgt cgc gtt tcc gtg 2179
Ile Glu Phe Glu Trp Thr Asp Arg Lys Val Asp Arg Arg Val Ser Val
680 685 690
acg att gct gtc att gac aaa act gag aac ttt act ggc atc acc atc 2227
Thr Ile Ala Val Ile Asp Lys Thr Glu Asn Phe Thr Gly Ile Thr Ile
695 700 705
gaa gat gga aag ctc gta ttc gaa gaa ctc gca gcc ggt cgc caa ctc 2275
Glu Asp Gly Lys Leu Val Phe Glu Glu Leu Ala Ala Gly Arg Gln Leu
710 715 720 725
gct gca tgg gtg tgg cca caa acc gca ccg tgg gta agc gca gtg gaa 2323
Ala Ala Trp Val Trp Pro Gln Thr Ala Pro Trp Val Ser Ala Val Glu
730 735 740
ctt gct gtc acc gga cca gag ctg gaa ctc cct gaa gtt ctc gtc ggc 2371
Leu Ala Val Thr Gly Pro Glu Leu Glu Leu Pro Glu Val Leu Val Gly
745 750 755
gca ggc aac ctg att gtt caa ctc cac acc gct gac cca ttc act acc 2419
Ala Gly Asn Leu Ile Val Gln Leu His Thr Ala Asp Pro Phe Thr Thr
760 765 770
tcc gtg acc cca ctg tca cca gga aaa gct gcg gtc acc gtt gag caa 2467
Ser Val Thr Pro Leu Ser Pro Gly Lys Ala Ala Val Thr Val Glu Gln
775 780 785
gaa ggc tac tac tca gca caa acc gaa gaa tat gca cag ctt tca gca 2515
Glu Gly Tyr Tyr Ser Ala Gln Thr Glu Glu Tyr Ala Gln Leu Ser Ala
790 795 800 805
ttc ttc ggt ggg gaa gta gaa gaa cca cca atc agt gac gct gtg gtc 2563
Phe Phe Gly Gly Glu Val Glu Glu Pro Pro Ile Ser Asp Ala Val Val
810 815 820
ccc gca ctt tgg gat gtt tcc cat atc tgg acc gaa cag gga aac acc 2611
Pro Ala Leu Trp Asp Val Ser His Ile Trp Thr Glu Gln Gly Asn Thr
825 830 835
gag cat ctt cca gta gtc cat gcc gcc ctg cgc tcc tca cca gcc gca 2659
Glu His Leu Pro Val Val His Ala Ala Leu Arg Ser Ser Pro Ala Ala
840 845 850
gca ctg aag ggt ctg tcc gct tcg ctg gtt ccc gca cag gca cta cct 2707
Ala Leu Lys Gly Leu Ser Ala Ser Leu Val Pro Ala Gln Ala Leu Pro
855 860 865
gga aaa gtc att tcc tcc gga ctg gca gcc tca ccg ttc acc acg gaa 2755
Gly Lys Val Ile Ser Ser Gly Leu Ala Ala Ser Pro Phe Thr Thr Glu
870 875 880 885
tca cca gca aca gaa gtg cac cgc acc gca tgg atc gga acc ctg caa 2803
Ser Pro Ala Thr Glu Val His Arg Thr Ala Trp Ile Gly Thr Leu Gln
890 895 900
ctc ctg ggt gca ctg cca agc gca ttc aag gaa gcc gaa gag ctt ggc 2851
Leu Leu Gly Ala Leu Pro Ser Ala Phe Lys Glu Ala Glu Glu Leu Gly
905 910 915
aac cgc aca cca ctg ctg cca atc ctc gga caa ctt gag gaa gtc gcc 2899
Asn Arg Thr Pro Leu Leu Pro Ile Leu Gly Gln Leu Glu Glu Val Ala
920 925 930
ggc aag aac atc ctg tcc acc ctt gca act ggc cgt gac tcc act ttg 2947
Gly Lys Asn Ile Leu Ser Thr Leu Ala Thr Gly Arg Asp Ser Thr Leu
935 940 945
gac acc gca tgc atc gac caa tcc acc gtt gcg att gcc ggc atg aac 2995
Asp Thr Ala Cys Ile Asp Gln Ser Thr Val Ala Ile Ala Gly Met Asn
950 955 960 965
gaa acc cag caa aaa gcc ctg ctg gac atg ttc ttc agc aac gcc gac 3043
Glu Thr Gln Gln Lys Ala Leu Leu Asp Met Phe Phe Ser Asn Ala Asp
970 975 980
atc gtt cct gga cca cta atg gaa gac aac acc cgc ctc atg gca gtg 3091
Ile Val Pro Gly Pro Leu Met Glu Asp Asn Thr Arg Leu Met Ala Val
985 990 995
ttc gaa acc ttc aag aag cgc gat gca ctc cgt gag gtt ctc cag act 3139
Phe Glu Thr Phe Lys Lys Arg Asp Ala Leu Arg Glu Val Leu Gln Thr
1000 1005 1010
gaa ggc ttg att aag acc gct gta gaa ctt ctt cgt gcc atg cgt gga 3187
Glu Gly Leu Ile Lys Thr Ala Val Glu Leu Leu Arg Ala Met Arg Gly
1015 1020 1025
acc cag cgt cag ctg tat tcc tcc gca cgt att cga ttc gac aag ctc 3235
Thr Gln Arg Gln Leu Tyr Ser Ser Ala Arg Ile Arg Phe Asp Lys Leu
1030 1035 1040 1045
gat ggt gtc aac act gac aac cca gaa aac atg tgg gca ctc acc cca 3283
Asp Gly Val Asn Thr Asp Asn Pro Glu Asn Met Trp Ala Leu Thr Pro
1050 1055 1060
gtt gtg tca ctg gtg ttc gcg ttg tca tcc cgt ttg cat gca cac gaa 3331
Val Val Ser Leu Val Phe Ala Leu Ser Ser Arg Leu His Ala His Glu
1065 1070 1075
ttg atc ggc aag acc cga act ctc gat cgt gca tct gcc ggt tgg ggt 3379
Leu Ile Gly Lys Thr Arg Thr Leu Asp Arg Ala Ser Ala Gly Trp Gly
1080 1085 1090
cga atc gct gat ctg gtg cca gac ctt gtc acc ggt gac ttg atc tcc 3427
Arg Ile Ala Asp Leu Val Pro Asp Leu Val Thr Gly Asp Leu Ile Ser
1095 1100 1105
gcg gag gca atg gtt ttg gga gct cga aac cca gga ctc gtc gat 3472
Ala Glu Ala Met Val Leu Gly Ala Arg Asn Pro Gly Leu Val Asp
1110 1115 1120
tagtccctga tttcatcgga gggcacctag 3502
<210>34
<211>1124
<212>PRT
<213>谷氨酸棒状杆菌
<400>34
Met Pro Ala Gly Ile Ala Asp Met Thr Asp Ser Leu Leu Gly Trp Ala
1 5 10 15
Ser Gln Thr Glu Leu Asp Leu Asn Gln Arg Leu Ala Gly Val Glu Tyr
20 25 30
Phe Pro Gln Ile Gln Leu Arg His Asp Glu Leu Glu Arg Ile His Arg
35 40 45
Phe Tyr Gly Thr Phe Leu Ser Arg Gln Val Gly Ala Gly Ala Ser Leu
50 55 60
Gly Asp Leu Phe Glu Met Thr Pro Cys Leu Thr Val Thr Thr Leu Val
65 70 75 80
Ser Arg Ala Ser Arg Ile Ser Asp Pro Ala Asp Phe Phe Gly Glu Tyr
85 90 95
Ile Gly Gly Leu Gly Leu Ser Ala Glu His Ala Ala Val Val Glu Gly
100 105 110
Leu Thr Glu Lys Leu Phe Ala Gln Ala Gly Leu Leu Val Pro Glu Gly
115 120 125
Ile Ala Ser Pro Leu Glu Leu Leu Ser Ile His Ala Gly Ile Ser Asn
130 135 140
His Glu Val Ala Ala Val Leu Thr Glu Val Glu Asn Gly Thr Thr Glu
145 150 155 160
Tyr Pro Phe Met Phe Asp Ala Val Leu Arg Leu Thr Pro Glu Trp Ala
165 170 175
Gln Thr Leu Ile Gly Gly Val Gln Glu Leu Ile Glu Phe Ala Thr Thr
180 185 190
His Arg Thr Ser Trp Ser Asp Arg Gln Arg Glu Ser Ser Leu Pro Ala
195 200 205
Met Ile Asp Glu Ile Val Val Ala Glu Leu Arg Glu Arg Pro Val Gly
210 215 220
Thr Ala Asp Arg Glu Asn Ser Val Gly Val Ala Leu Arg Glu Leu Arg
225 230 235 240
Pro Arg Leu Ile Leu Asp Ala Glu Arg Arg Lys Val Cys Leu Arg Leu
245 250 255
Pro Glu Gln Arg Val Ser Asp Asp Glu Ile Asn Trp Arg Val Ser Leu
260 265 270
Glu Gly Thr Thr Arg Ile Phe Ser Thr Arg Arg Ala Trp Gly Asp Thr
275 280 285
Ser Gly Tyr Ser Glu Ala Leu Asp Ile Thr Val Glu Arg Gln Ile Arg
290 295 300
Glu Thr Thr Val Thr Asp Thr Ser Asn Gln Ile Thr Trp Val Val Pro
305 310 315 320
Val Val Asp Phe Asn Asp Pro Val Leu Val Phe Ser Ala Arg Gly Glu
325 330 335
Asn Leu Thr Asp Lys Val Ser Leu His His Gln Glu Ile Tyr Val Leu
340 345 350
Ala Pro Ala Glu Ala Lys Leu Glu Asp Met Val Thr Gly Gln Pro Val
355 360 365
Pro Val Ile Glu Gln Phe Leu Val Glu Gly Trp Asn Ser Trp Val Cys
370 375 380
Ser Arg Val Asp Ala Arg Gly Leu Ser Ser Leu Lys Val Asn Lys Glu
385 390 395 400
Val Arg Cys Ile Asp Pro Arg Arg Arg Val Ala Phe His His Pr0 Ala
405 410 415
Glu Leu Val Pro His Val Arg Ser Ile Ser Gly Leu Pro Val His Ala
420 425 430
Gln Ser Leu Ile Ala Glu Phe Pro Pro Thr Leu Ser Gly Gln Asp Glu
435 440 445
Thr Trp Met Leu Ser Ile Ser Ala Phe Ala Gly Val Gly Ala Ala Gly
450 455 460
Glu Glu Ile Ala Glu Pro Glu Pro Leu Glu Val Pro Ala Asp Gly Gly
465 470 475 480
Leu Phe Ala Ile Phe Asp Pro Glu Ile Tyr Asp Ala Pro Trp Val Gly
485 490 495
Glu Tyr Leu Val Arg Leu Arg Gly Pro Arg Asn Glu Ser Phe Arg Pro
500 505 510
Glu Phe Ala Ile Val Glu Asp Met Thr Thr Glu Phe Glu Val Ala Ser
515 520 525
Gly Ala Ser Phe Arg Ile Pro Thr Thr Thr Gly Leu Ser Glu Ala Ser
530 535 540
Leu Arg Val Arg Ser Gly Glu Lys His Phe Thr Ala Glu Pro Arg Leu
545 550 555 560
Val Thr Val Glu Ala Thr Asp Pro Asn Ala Ser Phe Val Val Thr Thr
565 570 575
Asp Glu Gly Asp Gln Met Pro Leu Arg Phe Val Pro Pro Gln Ile Ala
580 585 590
Ile Glu Leu Pro Leu Thr Thr Glu Pro Pro Thr Trp Arg Val Thr Arg
595 600 605
Thr Val Cys Gly Pro Arg Asp Leu Asp Gly Ala Gly Glu Leu Arg Ile
610 615 620
Arg Thr Gly Val Asp Val Gly Asp Pro Lys Val Ser Val Arg Asn His
625 630 635 640
His Gly Ser Pro Leu Arg Thr Val Lys Met Val Thr Pro Asp Asn Gly
645 650 655
Arg Thr Trp Ile Ala Ser Met Lys Glu Ile Ala Ala Ser Thr Phe Val
660 665 670
Met Pro Arg Gly Ser Ile Glu Phe Glu Trp Thr Asp Arg Lys Val Asp
675 680 685
Arg Arg Val Ser Val Thr Ile Ala Val Ile Asp Lys Thr Glu Asn Phe
690 695 700
Thr Gly Ile Thr Ile Glu Asp Gly Lys Leu Val Phe Glu Glu Leu Ala
705 710 715 720
Ala Gly Arg Gln Leu Ala Ala Trp Val Trp Pro Gln Thr Ala Pro Trp
725 730 735
Val Ser Ala Val Glu Leu Ala Val Thr Gly Pro Glu Leu Glu Leu Pro
740 745 750
Glu Val Leu Val Gly Ala Gly Asn Leu Ile Val Gln Leu His Thr Ala
755 760 765
Asp Pro Phe Thr Thr Ser Val Thr Pro Leu Ser Pro Gly Lys Ala Ala
770 775 780
Val Thr Val Glu Gln Glu Gly Tyr Tyr Ser Ala Gln Thr Glu Glu Tyr
785 790 795 800
Ala Gln Leu Ser Ala Phe Phe Gly Gly Glu Val Glu Glu Pro Pro Ile
805 810 815
Ser Asp Ala Val Val Pro Ala Leu Trp Asp Val Ser His Ile Trp Thr
820 825 830
Glu Gln Gly Asn Thr Glu His Leu Pro Val Val His Ala Ala Leu Arg
835 840 845
Ser Ser Pro Ala Ala Ala Leu Lys Gly Leu Ser Ala Ser Leu Val Pro
850 855 860
Ala Gln Ala Leu Pro Gly Lys Val Ile Ser Ser Gly Leu Ala Ala Ser
865 870 875 880
Pro Phe Thr Thr Glu Ser Pro Ala Thr Glu Val His Arg Thr Ala Trp
885 890 895
Ile Gly Thr Leu Gln Leu Leu Gly Ala Leu Pro Ser Ala Phe Lys Glu
900 905 910
Ala Glu Glu Leu Gly Asn Arg Thr Pro Leu Leu Pro Ile Leu Gly Gln
915 920 925
Leu Glu Glu Val Ala Gly Lys Asn Ile Leu Ser Thr Leu Ala Thr Gly
930 935 940
Arg Asp Ser Thr Leu Asp Thr Ala Cys Ile Asp Gln Ser Thr Val Ala
945 950 955 960
Ile Ala Gly Met Asn Glu Thr Gln Gln Lys Ala Leu Leu Asp Met Phe
965 970 975
Phe Ser Asn Ala Asp Ile Val Pro Gly Pro Leu Met Glu Asp Asn Thr
980 985 990
Arg Leu Met Ala Val Phe Glu Thr Phe Lys Lys Arg Asp Ala Leu Arg
995 1000 1005
Glu Val Leu Gln Thr Glu Gly Leu Ile Lys Thr Ala Val Glu Leu Leu
1010 1015 1020
Arg Ala Met Arg Gly Thr Gln Arg Gln Leu Tyr Ser Ser Ala Arg Ile
1025 1030 1035 1040
Arg Phe Asp Lys Leu Asp Gly Val Asn Thr Asp Asn Pro Glu Asn Met
1045 1050 1055
Trp Ala Leu Thr Pro Val Val Ser Leu Val Phe Ala Leu Ser Ser Arg
1060 1065 1070
Leu His Ala His Glu Leu Ile Gly Lys Thr Arg Thr Leu Asp Arg Ala
1075 1080 1085
Ser Ala Gly Trp Gly Arg Ile Ala Asp Leu Val Pro Asp Leu Val Thr
1090 1095 1100
Gly Asp Leu Ile Ser Ala Glu Ala Met Val Leu Gly Ala Arg Asn Pro
1105 1110 1115 1120
Gly Leu Val Asp
<210>35
<211>694
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(664)
<223>RXA02210
<400>35
ttcatggcca atgtggtggg gtcctttcta atacgccaaa tttttcaaac ccatcctctt 60
ccatgcgtgt tttcaccgct attttccata ggagtacatt gtg tcc gta gcg gca 115
Val Ser Val Ala Ala
1 5
ggc gac aaa cca aca aat agc cgt caa gaa atc ctc gaa ggt gcc cga 163
Gly Asp Lys Pro Thr Asn Ser Arg Gln Glu Ile Leu Glu Gly Ala Arg
10 15 20
cgg tgc ttc gct gag cac ggc tat gaa ggc gca acc gta cgc cga ctg 211
Arg Cys Phe Ala Glu His Gly Tyr Glu Gly Ala Thr Val Arg Arg Leu
25 30 35
gaa gaa gca aca ggt aaa tca cgc gga gcg atc ttt cat cac ttc ggt 259
Glu Glu Ala Thr Gly Lys Ser Arg Gly Ala Ile Phe His His Phe Gly
40 45 50
gac aaa gaa aac ctg ttc cta gcc ctc gcg cgg gaa gat gca gcc cgc 307
Asp Lys Glu Asn Leu Phe Leu Ala Leu Ala Arg Glu Asp Ala Ala Arg
55 60 65
atg gcg gag gtg gtg tct gaa aat ggc ctc gtt gaa gtg atg cga gga 355
Met Ala Glu Val Val Ser Glu Asn Gly Leu Val Glu Val Met Arg Gly
70 75 80 85
atg ctg gaa gat cct gaa cga tat gac tgg atg tca gta cgc ctg gag 403
Met Leu Glu Asp Pro Glu Arg Tyr Asp Trp Met Ser Val Arg Leu Glu
90 95 100
atc tcc aag cag ctg cgc acc gac ccg gta ttc cgc gca aaa tgg att 451
Ile Ser Lys Gln Leu Arg Thr Asp Pro Val Phe Arg Ala Lys Trp Ile
105 110 115
gat cac caa agt gtt cta gac gaa gct gtc cgc gtg cgt ttg tcc cgc 499
Asp His Gln Ser Val Leu Asp Glu Ala Val Arg Val Arg Leu Ser Arg
120 125 130
aac gtg gat aag gga caa atg cgc act gac gtc ccg atc gaa gtg ctg 547
Asn Val Asp Lys Gly Gln Met Arg Thr Asp Val Pro Ile Glu Val Leu
135 140 145
cac acc ttc tta gag act gtt ctc gac ggt ttc atc tcc cgt ctt gct 595
His Thr Phe Leu Glu Thr Val Leu Asp Gly Phe Ile Ser Arg Leu Ala
150 155 160 165
acc ggc gca tcc aca gaa gga ctg tcc gaa gta ttg gat ctg gtc gag 643
Thr Gly Ala Ser Thr Glu Gly Leu Ser Glu Val Leu Asp Leu Val Glu
170 175 180
gga act gtc cgt aaa cgc gac taaacgaccc ctgattcaca ctttcagact 694
Gly Thr Val Arg Lys Arg Asp
185
<210>36
<211>188
<212>PRT
<213>谷氨酸棒状杆菌
<400>36
Val Ser Val Ala Ala Gly Asp Lys Pro Thr Ash Ser Arg Gln Glu Ile
1 5 10 15
Leu Glu Gly Ala Arg Arg Cys Phe Ala Glu His Gly Tyr Glu Gly Ala
20 25 30
Thr Val Arg Arg Leu Glu Glu Ala Thr Gly Lys Ser Arg Gly Ala Ile
35 40 45
Phe His His Phe Gly Asp Lys Glu Asn Leu Phe Leu Ala Leu Ala Arg
50 55 60
Glu Asp Ala Ala Arg Met Ala Glu Val Val Ser Glu Asn Gly Leu Val
65 70 75 80
Glu Val Met Arg Gly Met Leu Glu Asp Pro Glu Arg Tyr Asp Trp Met
85 90 95
Ser Val Arg Leu Glu Ile Ser Lys Gln Leu Arg Thr Asp Pro Val Phe
100 105 110
Arg Ala Lys Trp Ile Asp His Gln Ser Val Leu Asp Glu Ala Val Arg
115 120 125
Val Arg Leu Ser Arg Asn Val Asp Lys Gly Gln Met Arg Thr Asp Val
130 135 140
Pro Ile Glu Val Leu His Thr Phe Leu Glu Thr Val Leu Asp Gly Phe
145 150 155 160
Ile Ser Arg Leu Ala Thr Gly Ala Ser Thr Glu Gly Leu Ser Glu Val
165 170 175
Leu Asp Leu Val Glu Gly Thr Val Arg Lys Arg Asp
180 185
<210>37
<211>3829
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(3799)
<223>RXA02362
<400>37
acaattgttc ccattcgcat atctcgtgtc caccagagtc gctacagtaa cgagaaactt60
aattatttga tccgattctt ccgttcaaag gcctcattcc gtg act atc tct cgc 115
Val Thr Ile Ser Arg
1 5
cga ctc aaa caa gag cgc agt ttc gct gac gat ctt caa gat ctc aaa 163
Arg Leu Lys Gln Glu Arg Ser Phe Ala Asp Asp Leu Gln Asp Leu Lys
10 15 20
act ctc aat gat caa ctg cgg ttt aca aac gcc aaa ttg caa gct cgc 211
Thr Leu Asn Asp Gln Leu Arg Phe Thr Asn Ala Lys Leu Gln Ala Arg
25 30 35
atc agt ggt att ggc aat gat gga aag aaa atc acg cgc cct acc cca 259
Ile Ser Gly Ile Gly Asn Asp Gly Lys Lys Ile Thr Arg Pro Thr Pro
40 45 50
ctc ctt gcg ctg gat ttt cag ctg acc gtt gaa gaa tac gaa acg atc 307
Leu Leu Ala Leu Asp Phe Gln Leu Thr Val Glu Glu Tyr Glu Thr Ile
55 60 65
att gca atc ttg gtg gaa gca gtt ggc gga aat caa tcc aag cca gcg 355
Ile Ala Ile Leu Val Glu Ala Val Gly Gly Asn Gln Ser Lys Pro Ala
70 75 80 85
att ctt aaa gat ctg ttt ata gaa tat cca ctc gtc ttc ctg gca gcg 403
Ile Leu Lys Asp Leu Phe Ile Glu Tyr Pro Leu Val Phe Leu Ala Ala
90 95 100
ctt tct gga acc gcc atg ctc gat gct caa gaa ggt ttc tgg cct gcg 451
Leu Ser Gly Thr Ala Met Leu Asp Ala Gln Glu Gly Phe Trp Pro Ala
105 110 115
ttc tgg aaa cgc act cag gtg tca gtt cea gag cat gta tac gac gcg 499
Phe Trp Lys Arg Thr Gln Val Ser Val Pro Glu His Val Tyr Asp Ala
120 125 130
atc cgt aaa gaa cta gtt aat agc atc cgc aaa aat ggc cta gaa act 547
Ile Arg Lys Glu Leu Val Asn Ser Ile Arg Lys Asn Gly Leu Glu Thr
135 140 145
ttt tct ctc gct gac ctc aat cga cgc gaa tat gtc gga ctc atc caa 595
Phe Ser Leu Ala Asp Leu Asn Arg Arg Glu Tyr Val Gly Leu Ile Gln
150 155 160 165
ctt cac agt ggc ctt tct gca aaa gac atg ctc gcc ttg gtc aaa ttt 643
Leu His Ser Gly Leu Ser Ala Lys Asp Met Leu Ala Leu Val Lys Phe
170 175 180
atc gat cac act cga gca gaa aac caa gga tgg gat tct ggt gag gac 691
Ile Asp His Thr Arg Ala Glu Asn Gln Gly Trp Asp Ser Gly Glu Asp
185 190 195
ttt gca tca tat gcg aag agt gtc ttc tcc tcc ggg gac aac cta tta 739
Phe Ala Ser Tyr Ala Lys Ser Val Phe Ser Ser Gly Asp Asn Leu Leu
200 205 210
acc acg gag tcg ctc aag caa tta gtc acc cac atc cct gcg cgt tcc 787
Thr Thr Glu Ser Leu Lys Gln Leu Val Thr His Ile Pro Ala Arg Ser
215 220 225
gtc gac ttc atc gcc aga gtc tat gaa cta acc aat tgg tac cgc gac 835
Val Asp Phe Ile Ala Arg Val Tyr Glu Leu Thr Asn Trp Tyr Arg Asp
230 235 240 245
ctc aaa gac ctc aat gaa gta gaa gcc ttc gta ggt act cat ggg ctg 883
Leu Lys Asp Leu Asn Glu Val Glu Ala Phe Val Gly Thr His Gly Leu
250 255 260
ccg gaa ttg tct ttc aaa ttt ctt ctg gag tgt ctg agc ggc gaa gct 931
Pro Glu Leu Ser Phe Lys Phe Leu Leu Glu Cys Leu Ser Gly Glu Ala
265 270 275
gaa caa att gcc gaa aag acg aaa gca gca cca gca agc ctg gaa aac 979
Glu Gln Ile Ala Glu Lys Thr Lys Ala Ala Pro Ala Ser Leu Glu Asn
280 285 290
ctg gaa cct ccg cat ctc tat ctg gat cca cag agt ttt gaa ctc agt 1027
Leu Glu Pro Pro His Leu Tyr Leu Asp Pro Gln Ser Phe Glu Leu Ser
295 300 305
ctt gtt ttc cca gcg atc tct aaa act gca gca ctt cag att cca gca 1075
Leu Val Phe Pro Ala Ile Ser Lys Thr Ala Ala Leu Gln Ile Pro Ala
310 315 320 325
cca gaa tgg aca gtg att tat gac gga aac tcc att aaa gtt cgt ccc 1123
Pro Glu Trp Thr Val Ile Tyr Asp Gly Asn Ser Ile Lys Val Arg Pro
330 335 340
gaa cag gac tgg tcc tac gga ggt ttc gcc gaa tac cgt ttg cct tta 1171
Glu Gln Asp Trp Ser Tyr Gly Gly Phe Ala Glu Tyr Arg Leu Pro Leu
345 350 355
gac aaa ccg ctc tcc agc ttg aga gtc atc act cca aca gag aaa tcc 1219
Asp Lys Pro Leu Ser Ser Leu Arg Val Ile Thr Pro Thr Glu Lys Ser
360 365 370
cta att ctg att gaa gga ttt ggc cac aag aat ccc att atg ttc ttt 1267
Leu Ile Leu Ile Glu Gly Phe Gly His Lys Asn Pro Ile Met Phe Phe
375 380 385
aag aac aac ggt cag cca tat gca aac caa gaa atg ctc agt ggc aac 1315
Lys Asn Asn Gly Gln Pro Tyr Ala Asn Gln Glu Met Leu Ser Gly Asn
390 395 400 405
gct gtc aca gct ata gtc cca gct gca gca atc att cgt gca cgt atg 1363
Ala Val Thr Ala Ile Val Pro Ala Ala Ala Ile Ile Arg Ala Arg Met
410 415 420
cga gct tcc aag act ttc aac tat caa gac ttg ggt ccc ttg tcc gga 1411
Arg Ala Ser Lys Thr Phe Asn Tyr Gln Asp Leu Gly Pro Leu Ser Gly
425 430 435
tgg aac aag tgg gtc att cgt tcg atc cca ctc aaa cga gct gaa tcg 1459
Trp Asn Lys Trp Val Ile Arg Ser Ile Pro Leu Lys Arg Ala Glu Ser
440 445 450
atc aca gtc tcc cac ggt ggc ttc aga aaa gaa ctc cca gtt cga cgc 1507
Ile Thr Val Ser His Gly Gly Phe Arg Lys Glu Leu Pro Val Arg Arg
455 460 465
aaa gtt gat gtt caa tgg att act gag gat ctc acg atc gag aat ctt 1555
Lys Val Asp Val Gln Trp Ile Thr Glu Asp Leu Thr Ile Glu Asn Leu
470 475 480 485
caa ggt ctc gat cat gag ccc gtt ttc cac acg agt ccc cgc atc gaa 1603
Gln Gly Leu Asp His Glu Pro Val Phe His Thr Ser Pro Arg Ile Glu
490 495 500
ttc ccc acc tct gga tca aac tgg gta att cag tat tca cag att ctt 1651
Phe Pro Thr Ser Gly Ser Asn Trp Val Ile Gln Tyr Ser Gln Ile Leu
505 510 515
cca gat ggc agc ctc atc gaa atg gaa gat tac cca gtc gaa cct gaa 1699
Pro Asp Gly Ser Leu Ile Glu Met Glu Asp Tyr Pro Val Glu Pro Glu
520 525 530
aac ttc gga tac gaa cta gac ctc ttc gaa gaa tcc gac gac cct tgg 1747
Asn Phe Gly Tyr Glu Leu Asp Leu Phe Glu Glu Ser Asp Asp Pro Trp
535 540 545
gtc ggg caa ttt tta gta act ctg ctc aag gat gaa aaa gtc tat gaa 1795
Val Gly Gln Phe Leu Val Thr Leu Leu Lys Asp Glu Lys Val Tyr Glu
550 555 560 565
acc cgc aaa ttc aat ctc gcg gaa ggc ctc gat ctt tcc cta aca ttc 1843
Thr Arg Lys Phe Asn Leu Ala Glu Gly Leu Asp Leu Ser Leu Thr Phe
570 575 580
agc gga ggc gga cct gaa aat cga ttt agg tac ccc agc atc aat cag 1891
Ser Gly Gly Gly Pro Glu Asn Arg Phe Arg Tyr Pro Ser Ile Asn Gln
585 590 595
gga caa act ggc tta aca aag act ttc gcc cgt ttt agt tcc aat tct 1939
Gly Gln Thr Gly Leu Thr Lys Thr Phe Ala Arg Phe Ser Ser Asn Ser
600 605 610
gaa aag cac atc agg ttc cca gat gag atc atc ggg ctt gat gca ttc 1987
Glu Lys His Ile Arg Phe Pro Asp Glu Ile Ile Gly Leu Asp Ala Phe
615 620 625
acc tct caa aaa gcg ttt aac atc gca agc ggt gat ttc cct gag gac 2035
Thr Ser Gln Lys Ala Phe Asn Ile Ala Ser Gly Asp Phe Pro Glu Asp
630 635 640 645
tac aac ctc gac gtt ttc atc acg cct ccg caa ctt cac tac caa gta 2083
Tyr Asn Leu Asp Val Phe Ile Thr Pro Pro Gln Leu His Tyr Gln Val
650 655 660
cct gtc aca cac agc caa aca aag tgg gaa agc aca aag acg aca cta 2131
Pro Val Thr His Ser Gln Thr Lys Trp Glu Ser Thr Lys Thr Thr Leu
665 670 675
gat ttc aat gac ttt gcc gat gga aac ctc cag atc aga ttc cct aat 2179
Asp Phe Asn Asp Phe Ala Asp Gly Asn Leu Gln Ile Arg Phe Pro Asn
680 685 690
gaa gtc tat gat cca aac ttg aaa atc att aaa atg gtg gca tac aag 2227
Glu Val Tyr Asp Pro Asn Leu Lys Ile Ile Lys Met Val Ala Tyr Lys
695 700 705
aaa cct gag tcc agt gag cct aaa tac tta agc aaa att ggt tca agc 2275
Lys Pro Glu Ser Ser Glu Pro Lys Tyr Leu Ser Lys Ile Gly Ser Ser
710 715 720 725
aaa gtg tgg tct atc cct atg gat cgc atc aag gaa ctc atg gat gat 2323
Lys Val Trp Ser Ile Pro Met Asp Arg Ile Lys Glu Leu Met Asp Asp
730 735 740
gat gcc caa ttc ctt ttg atc gcg gag tgg ttc gct gaa agt aaa gac 2371
Asp Ala Gln Phe Leu Leu Ile Ala Glu Trp Phe Ala Glu Ser Lys Asp
745 750 755
cag cac cga gag aag atc att agc gaa gct aag cga act gga aaa atc 2419
Gln His Arg Glu Lys Ile Ile Ser Glu Ala Lys Arg Thr Gly Lys Ile
760 765 770
tcc aat gca gcg ctt aag agt gct cgt cct caa cct caa gca agt tcc 2467
Ser Asn Ala Ala Leu Lys Ser Ala Arg Pro Gln Pro Gln Ala Ser Ser
775 780 785
cac att gca aca att gag aaa aag ccc cta cta gct gcg gct gaa att 2515
His Ile Ala Thr Ile Glu Lys Lys Pro Leu Leu Ala Ala Ala Glu Ile
790 795 800 805
aag ctt tct acc gtg gag ttg gaa ctt ggt cgg cac act tct aag aga 2563
Lys Leu Ser Thr Val Glu Leu Glu Leu Gly Arg His Thr Ser Lys Arg
810 815 820
ctg gaa ggc tgg gca tgg tct gcg ctc aac ccg ctt gat cca cca atc 2611
Leu Glu Gly Trp Ala Trp Ser Ala Leu Asn Pro Leu Asp Pro Pro Ile
825 830 835
aaa gtc gat ttc caa gga acc tca ggc tca ctt cca gac acc cac ttc 2659
Lys Val Asp Phe Gln Gly Thr Ser Gly Ser Leu Pro Asp Thr His Phe
840 845 850
gtc gtt ggc cct tta atc gtg gaa gtg aga gaa aaa gag ttt ctc tcc 2707
Val Val Gly Pro Leu Ile Val Glu Val Arg Glu Lys Glu Phe Leu Ser
855 860 865
caa tgg cag cca aaa gtt ccc tca gtt aaa gcc gtg gtt gca aat gat 2755
Gln Trp Gln Pro Lys Val Pro Ser Val Lys Ala Val Val Ala Asn Asp
870 875 880 885
ccc tca ttt gaa ttg gac cct caa ttt gat cct ttc ctc aca cac cga 2803
Pro Ser Phe Glu Leu Asp Pro Gln Phe Asp Pro Phe Leu Thr His Arg
890 895 900
tgg atg ttc gct cca cga agt ggg aag gtc tta ctc cca caa gaa atc 2851
Trp Met Phe Ala Pro Arg Ser Gly Lys Val Leu Leu Pro Gln Glu Ile
905 910 915
cgc aca gtg tgg gac gcc cga ttc aat atg cgc cat gtc tta gcg cag 2899
Arg Thr Val Trp Asp Ala Arg Phe Asn Met Arg His Val Leu Ala Gln
920 925 930
cgt gaa aac ctt cat gtg aaa tcg att caa gat ttt gac gat gcc acc 2947
Arg Glu Asn Leu His Val Lys Ser Ile Gln Asp Phe Asp Asp Ala Thr
935 940 945
agt acc tat ctc acc agt gat cct cgg gtg gca tta gat gaa ttg gat 2995
Ser Thr Tyr Leu Thr Ser Asp Pro Arg Val Ala Leu Asp Glu Leu Asp
950 955 960 965
aag agc tca att ccg tct aat tcc cac ttt gaa tca ttc atc cga tcc 3043
Lys Ser Ser Ile Pro Ser Asn Ser His Phe Glu Ser Phe Ile Arg Ser
970 975 980
gga tta gct gag ctt tct ttc gaa gtt gac gac aca gcc gga gat atc 3091
Gly Leu Ala Glu Leu Ser Phe Glu Val Asp Asp Thr Ala Gly Asp Ile
985 990 995
cat cgc gtt ccc tgg atc ggc ctg atc cag gaa atg aac gac ctc aga 3139
His Arg Val Pro Trp Ile Gly Leu Ile Gln Glu Met Asn Asp Leu Arg
1000 1005 1010
att ctg cag ata caa ggc tat gaa aca gaa gaa cga gcc atc gaa cgc 3187
Ile Leu Gln Ile Gln Gly Tyr Glu Thr Glu Glu Arg Ala Ile Glu Arg
1015 1020 1025
cga aat tcg cag agc tac atc cgt gag ata gga ggc agt gaa ttg tgg 3235
Arg Asn Ser Gln Ser Tyr Ile Arg Glu Ile Gly Gly Ser Glu Leu Trp
1030 1035 1040 1045
aat atc cta aaa gga aat tca gag gga ttg tct ctt gct caa aaa tgc 3283
Asn Ile Leu Lys Gly Asn Ser Glu Gly Leu Ser Leu Ala Gln Lys Cys
1050 1055 1060
gca cca caa gcc act gag att aat gtg att cgt aat tca ggc ttg gaa 3331
Ala Pro Gln Ala Thr Glu Ile Asn Val Ile Arg Asn Ser Gly Leu Glu
1065 1070 1075
gct atg cgc aat ggg ctg ggc gcc gat cag ttc agc gcc gag ttt att 3379
Ala Met Arg Asn Gly Leu Gly Ala Asp Gln Phe Ser Ala Glu Phe Ile
1080 1085 1090
tca gca gac tca cgc cta cga gct cag ctt gaa tgg ttg gaa aac cgc 3427
Ser Ala Asp Ser Arg Leu Arg Ala Gln Leu Glu Trp Leu Glu Asn Arg
1095 1100 1105
cga gag ctc aat gat ctc ggc cag ctc cca acg ctc ttc gat ttc gcc 3475
Arg Glu Leu Asn Asp Leu Gly Gln Leu Pro Thr Leu Phe Asp Phe Ala
1110 1115 1120 1125
gag aaa tac gag tac ctc atc gat cac tta ggt gat gat cgc atc aag 3523
Glu Lys Tyr Glu Tyr Leu Ile Asp His Leu Gly Asp Asp Arg Ile Lys
1130 1135 1140
gtc act gca cgt gag ctg tct act ctt gcg tcg gaa cac cgt cgc ggc 3571
Val Thr Ala Arg Glu Leu Ser Thr Leu Ala Ser Glu His Arg Arg Gly
1145 1150 1155
aac gct gaa aac tgg ctt tat gca cca tat gtg tca ttc att tac agc 3619
Asn Ala Glu Asn Trp Leu Tyr Ala Pro Tyr Val Ser Phe Ile Tyr Ser
1160 1165 1170
ttg ctt aac cga atg atc gct cat gaa gta ata cgt ccg atc gct cag 3667
Leu Leu Asn Arg Met Ile Ala His Glu Val Ile Arg Pro Ile Ala Gln
1175 1180 1185
atc aat tac tca cgg cac gat tgg gca aac gct gct cgg ctg att cct 3715
Ile Asn Tyr Ser Arg His Asp Trp Ala Asn Ala Ala Arg Leu Ile Pro
1190 1195 1200 1205
cgt ctc aca gga ttt gac ctg gtg agt gcc gaa gcg aaa gtg ctc agc 3763
Arg Leu Thr Gly Phe Asp Leu Val Ser Ala Glu Ala Lys Val Leu Ser
1210 1215 1220
gca ata aac aac aac aat ata atc cca act gca att taaggatcac 3809
Ala Ile Asn Asn Asn Asn Ile Ile Pro Thr Ala Ile
1225 1230
tatgtccaac gcacctaaaa 3829
<210>38
<211>1233
<212>PRT
<213>谷氨酸棒状杆菌
<400>38
Val Thr Ile Ser Arg Arg Leu Lys Gln Glu Arg Ser Phe Ala Asp Asp
1 5 10 15
Leu Gln Asp Leu Lys Thr Leu Asn Asp Gln Leu Arg Phe Thr Asn Ala
20 25 30
Lys Leu Gln Ala Arg Ile Ser Gly Ile Gly Asn Asp Gly Lys Lys Ile
35 40 45
Thr Arg Pro Thr Pro Leu Leu Ala Leu Asp Phe Gln Leu Thr Val Glu
50 55 60
Glu Tyr Glu Thr Ile Ile Ala Ile Leu Val Glu Ala Val Gly Gly Asn
65 70 75 80
Gln Ser Lys Pro Ala Ile Leu Lys Asp Leu Phe Ile Glu Tyr Pro Leu
85 90 95
Val Phe Leu Ala Ala Leu Ser Gly Thr Ala Met Leu Asp Ala Gln Glu
100 105 110
Gly Phe Trp Pro Ala Phe Trp Lys Arg Thr Gln Val Ser Val Pro Glu
115 120 125
His Val Tyr Asp Ala Ile Arg Lys Glu Leu Val Asn Ser Ile Arg Lys
130 135 140
Asn Gly Leu Glu Thr Phe Ser Leu Ala Asp Leu Asn Arg Arg Glu Tyr
145 150 155 160
Val Gly Leu Ile Gln Leu His Ser Gly Leu Ser Ala Lys Asp Met Leu
165 170 175
Ala Leu Val Lys Phe Ile Asp His Thr Arg Ala Glu Asn Gln Gly Trp
180 185 190
Asp Ser Gly Glu Asp Phe Ala Ser Tyr Ala Lys Ser Val Phe Ser Ser
195 200 205
Gly Asp Asn Leu Leu Thr Thr Glu Ser Leu Lys Gln Leu Val Thr His
210 215 220
Ile Pro Ala Arg Ser Val Asp Phe Ile Ala Arg Val Tyr Glu Leu Thr
225 230 235 240
Asn Trp Tyr Arg Asp Leu Lys Asp Leu Asn Glu Val Glu Ala Phe Val
245 250 255
Gly Thr His Gly Leu Pro Glu Leu Ser Phe Lys Phe Leu Leu Glu Cys
260 265 270
Leu Ser Gly Glu Ala Glu Gln Ile Ala Glu Lys Thr Lys Ala Ala Pro
275 280 285
Ala Ser Leu Glu Asn Leu Glu Pro Pro His Leu Tyr Leu Asp Pro Gln
290 295 300
Ser Phe Glu Leu Ser Leu Val Phe Pro Ala Ile Ser Lys Thr Ala Ala
305 310 315 320
Leu Gln Ile Pro Ala Pro Glu Trp Thr Val Ile Tyr Asp Gly Asn Ser
325 330 335
Ile Lys Val Arg Pro Glu Gln Asp Trp Ser Tyr Gly Gly Phe Ala Glu
340 345 350
Tyr Arg Leu Pro Leu Asp Lys Pro Leu Ser Ser Leu Arg Val Ile Thr
355 360 365
Pro Thr Glu Lys Ser Leu Ile Leu Ile Glu Gly Phe Gly His Lys Asn
370 375 380
Pro Ile Met Phe Phe Lys Asn Asn Gly Gln Pro Tyr Ala Asn Gln Glu
385 390 395 400
Met Leu Ser Gly Asn Ala Val Thr Ala Ile Val Pro Ala Ala Ala Ile
405 410 415
Ile Arg Ala Arg Met Arg Ala Ser Lys Thr Phe Asn Tyr Gln Asp Leu
420 425 430
Gly Pro Leu Ser Gly Trp Asn Lys Trp Val Ile Arg Ser Ile Pro Leu
435 440 445
Lys Arg Ala Glu Ser Ile Thr Val Ser His Gly Gly Phe Arg Lys Glu
450 455 460
Leu Pro Val Arg Arg Lys Val Asp Val Gln Trp Ile Thr Glu Asp Leu
465 470 475 480
Thr Ile Glu Asn Leu Gln Gly Leu Asp His Glu Pro Val Phe His Thr
485 490 495
Ser Pro Arg Ile Glu Phe Pro Thr Ser Gly Ser Asn Trp Val Ile Gln
500 505 510
Tyr Ser Gln Ile Leu Pro Asp Gly Ser Leu Ile Glu Met Glu Asp Tyr
515 520 525
Pro Val Glu Pro Glu Asn Phe Gly Tyr Glu Leu Asp Leu Phe Glu Glu
530 535 540
Ser Asp Asp Pro Trp Val Gly Gln Phe Leu Val Thr Leu Leu Lys Asp
545 550 555 560
Glu Lys Val Tyr Glu Thr Arg Lys Phe Asn Leu Ala Glu Gly Leu Asp
565 570 575
Leu Ser Leu Thr Phe Ser Gly Gly Gly Pro Glu Asn Arg Phe Arg Tyr
580 585 590
Pro Ser Ile Asn Gln Gly Gln Thr Gly Leu Thr Lys Thr Phe Ala Arg
595 600 605
Phe Ser Ser Asn Ser Glu Lys His Ile Arg Phe Pro Asp Glu Ile Ile
610 615 620
Gly Leu Asp Ala Phe Thr Ser Gln Lys Ala Phe Asn Ile Ala Ser Gly
625 630 635 640
Asp Phe Pro Glu Asp Tyr Asn Leu Asp Val Phe Ile Thr Pro Pro Gln
645 650 655
Leu His Tyr Gln Val Pro Val Thr His Ser Gln Thr Lys Trp Glu Ser
660 665 670
Thr Lys Thr Thr Leu Asp Phe Asn Asp Phe Ala Asp Gly Asn Leu Gln
675 680 685
Ile Arg Phe Pro Asn Glu Val Tyr Asp Pro Asn Leu Lys Ile Ile Lys
690 695 700
Met Val Ala Tyr Lys Lys Pro Glu Ser Ser Glu Pro Lys Tyr Leu Ser
705 710 715 720
Lys Ile Gly Ser Ser Lys Val Trp Ser Ile Pro Met Asp Arg Ile Lys
725 730 735
Glu Leu Met Asp Asp Asp Ala Gln Phe Leu Leu Ile Ala Glu Trp Phe
740 745 750
Ala Glu Ser Lys Asp Gln His Arg Glu Lys Ile Ile Ser Glu Ala Lys
755 760 765
Arg Thr Gly Lys Ile Ser Asn Ala Ala Leu Lys Ser Ala Arg Pro Gln
770 775 780
Pro Gln Ala Ser Ser His Ile Ala Thr Ile Glu Lys Lys Pro Leu Leu
785 790 795 800
Ala Ala Ala Glu Ile Lys Leu Ser Thr Val Glu Leu Glu Leu Gly Arg
805 810 815
His Thr Ser Lys Arg Leu Glu Gly Trp Ala Trp Ser Ala Leu Asn Pro
820 825 830
Leu Asp Pro Pro Ile Lys Val Asp Phe Gln Gly Thr Ser Gly Ser Leu
835 840 845
Pro Asp Thr His Phe Val Val Gly Pro Leu Ile Val Glu Val Arg Glu
850 855 860
Lys Glu Phe Leu Ser Gln Trp Gln Pro Lys Val Pro Ser Val Lys Ala
865 870 875 880
Val Val Ala Asn Asp Pro Ser Phe Glu Leu Asp Pro Gln Phe Asp Pro
885 890 895
Phe Leu Thr His Arg Trp Met Phe Ala Pro Arg Ser Gly Lys Val Leu
900 905 910
Leu Pro Gln Glu Ile Arg Thr Val Trp Asp Ala Arg Phe Asn Met Arg
915 920 925
His Val Leu Ala Gln Arg Glu Asn Leu His Val Lys Ser Ile Gln Asp
930 935 940
Phe Asp Asp Ala Thr Ser Thr Tyr Leu Thr Ser Asp Pro Arg Val Ala
945 950 955 960
Leu Asp Glu Leu Asp Lys Ser Ser Ile Pro Ser Asn Ser His Phe Glu
965 970 975
Ser Phe Ile Arg Ser Gly Leu Ala Glu Leu Ser Phe Glu Val Asp Asp
980 985 990
Thr Ala Gly Asp Ile His Arg Val Pro Trp Ile Gly Leu Ile Gln Glu
995 1000 1005
Met Asn Asp Leu Arg Ile Leu Gln Ile Gln Gly Tyr Glu Thr Glu Glu
1010 1015 1020
Arg Ala Ile Glu Arg Arg Asn Ser Gln Ser Tyr Ile Arg Glu Ile Gly
1025 1030 1035 1040
Gly Ser Glu Leu Trp Asn Ile Leu Lys Gly Asn Ser Glu Gly Leu Ser
1045 1050 1055
Leu Ala Gln Lys Cys Ala Pro Gln Ala Thr Glu Ile Asn Val Ile Arg
1060 1065 1070
Asn Ser Gly Leu Glu Ala Met Arg Asn Gly Leu Gly Ala Asp Gln Phe
1075 1080 1085
Ser Ala Glu Phe Ile Ser Ala Asp Ser Arg Leu Arg Ala Gln Leu Glu
1090 1095 1100
Trp Leu Glu Asn Arg Arg Glu Leu Asn Asp Leu Gly Gln Leu Pro Thr
1105 1110 1115 1120
Leu Phe Asp Phe Ala Glu Lys Tyr Glu Tyr Leu Ile Asp His Leu Gly
1125 1130 1135
Asp Asp Arg Ile Lys Val Thr Ala Arg Glu Leu Ser Thr Leu Ala Ser
1140 1145 1150
Glu His Arg Arg Gly Asn Ala Glu Asn Trp Leu Tyr Ala Pro Tyr Val
1155 1160 1165
Ser Phe Ile Tyr Ser Leu Leu Asn Arg Met Ile Ala His Glu Val Ile
1170 1175 1180
Arg Pro Ile Ala Gln Ile Asn Tyr Ser Arg His Asp Trp Ala Asn Ala
1185 1190 1195 1200
Ala Arg Leu Ile Pro Arg Leu Thr Gly Phe Asp Leu Val Ser Ala Glu
1205 1210 1215
Ala Lys Val Leu Ser Ala Ile Asn Asn Asn Asn Ile Ile Pro Thr Ala
1220 1225 1230
Ile
<210>39
<211>1633
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1603)
<223>RXA02376
<400>39
actggtagga tcgaagaacc ttggcgccca cacatccccc tttatttttc tacttatacg 60
tcggcgcccg ttttatcccg atcaggagtg acaccgtttt atg aac cga ttt att 115
Met Asn Arg Phe Ile
1 5
gac cgc gtt gtg cta cac ctc gcc gca ggt gac ggc ggc aat ggt tgt 163
Asp Arg Val Val Leu His Leu Ala Ala Gly Asp Gly Gly Asn Gly Cys
10 15 20
gtc tcg gtg cac cgc gaa aaa ttc aag cca ctt ggt gga cca gac ggc 211
Val Ser Val His Arg Glu Lys Phe Lys Pro Leu Gly Gly Pro Asp Gly
25 30 35
ggc aac ggc ggc cac ggt gga gac atc atc ttg gaa gtc acc gca cag 259
Gly Asn Gly Gly His Gly Gly Asp Ile Ile Leu Glu Val Thr Ala Gln
40 45 50
gtc cac acc ctg ctt gac ttc cac ttc cac cca cac gtg aag gcc gag 307
Val His Thr Leu Leu Asp Phe His Phe His Pro His Val Lys Ala Glu
55 60 65
cgc ggc gct aac ggc gct ggc gat cat cgc aac ggt gcc cga ggc aag 355
Arg Gly Ala Asn Gly Ala Gly Asp His Arg Asn Gly Ala Arg Gly Lys
70 75 80 85
gac ctt gtc ttg gaa gtt cca cca gga act gtc gtg ctt aat gaa aag 403
Asp Leu Val Leu Glu Val Pro Pro Gly Thr Val Val Leu Asn Glu Lys
90 95 100
ggc gag act ctg gca gac ctg acc agc gtg ggc atg aag ttc atc gct 451
Gly Glu Thr Leu Ala Asp Leu Thr Ser Val Gly Met Lys Phe Ile Ala
105 110 115
gct gct ggc ggt aac ggc ggt ttg ggt aac gca gcg ctt gcc tcc aag 499
Ala Ala Gly Gly Asn Gly Gly Leu Gly Asn Ala Ala Leu Ala Ser Lys
120 125 130
gct cgt aag gcc cca ggc ttc gcc ctg atc ggt gag cca ggc gag gcc 547
Ala Arg Lys Ala Pro Gly Phe Ala Leu Ile Gly Glu Pro Gly Glu Ala
135 140 145
cac gac ttg att ctt gaa ctc aaa tcc atg gca gat gtg gga ttg gtg 595
His Asp Leu Ile Leu Glu Leu Lys Ser Met Ala Asp Val Gly Leu Val
150 155 160 165
ggc ttc cca tca gcc ggc aaa tca tca ctg att tct gtg atg tct gca 643
Gly Phe Pro Ser Ala Gly Lys Ser Ser Leu Ile Ser Val Met Ser Ala
170 175 180
gca aag cca aag atc ggt gat tac cca ttc acc acc ctg cag cca aac 691
Ala Lys Pro Lys Ile Gly Asp Tyr Pro Phe Thr Thr Leu Gln Pro Asn
185 190 195
ctc ggc gta gtt aac gtt ggt cat gag act ttc acc atg gca gac gtg 739
Leu Gly Val Val Asn Val Gly His Glu Thr Phe Thr Met Ala Asp Val
200 205 210
cct ggt ttg atc cct ggt gct tct gag ggc aag ggc ttg ggt ctg gat 787
Pro Gly Leu Ile Pro Gly Ala Ser Glu Gly Lys Gly Leu Gly Leu Asp
215 220 225
ttc ttg cgc cac att gag cgc acc tcc gtg ctg gtt cac gtt gtc gat 835
Phe Leu Arg His Ile Glu Arg Thr Ser Val Leu Val His Val Val Asp
230 235 240 245
acc gca acg atg gat cca ggc cgc gat ccg atc tct gat att gag gct 883
Thr Ala Thr Met Asp Pro Gly Arg Asp Pro Ile Ser Asp Ile Glu Ala
250 255 260
ttg gaa gca gaa ctt gcc gcc tac cag tcg gct ttg gat gaa gac acc 931
Leu Glu Ala Glu Leu Ala Ala Tyr Gln Ser Ala Leu Asp Glu Asp Thr
265 270 275
gga ctt ggt gac ttg agc cag cgc cct cgc ctt gtt gtg ttg aac aag 979
Gly Leu Gly Asp Leu Ser Gln Arg Pro Arg Leu Val Val Leu Asn Lys
280 285 290
gct gat gtc cct gag gct gaa gag ctt gct gag ttc ctc aaa gaa gat 1027
Ala Asp Val Pro Glu Ala Glu Glu Leu Ala Glu Phe Leu Lys Glu Asp
295 300 305
att gag aag caa ttc gga tgg ccc gtg ttc att atc tcc gca gtg gca 1075
Ile Glu Lys Gln Phe Gly Trp Pro Val Phe Ile Ile Ser Ala Val Ala
310 315 320 325
cgc aag ggc ttg gat cct ttg aag tac aag ctg ctg gaa atc gtc cag 1123
Arg Lys Gly Leu Asp Pro Leu Lys Tyr Lys Leu Leu Glu Ile Val Gln
330 335 340
gat gcc cga aag aag cgt cca aag gag aag gct gag tct gtc atc att 1171
Asp Ala Arg Lys Lys Arg Pro Lys Glu Lys Ala Glu Ser Val Ile Ile
345 350 355
aag cct aag gct gtt gat cac cgc act aag ggg cag ttc cag atc aag 1219
Lys Pro Lys Ala Val Asp His Arg Thr Lys Gly Gln Phe Gln Ile Lys
360 365 370
cct gac cca gag gtt cag ggc gga ttc atc atc acc ggc gaa aag cca 1267
Pro Asp Pro Glu Val Gln Gly Gly Phe Ile Ile Thr Gly Glu Lys Pro
375 380 385
gag cgc tgg att ttg cag acc gac ttt gaa aac gac gaa gca gtt ggc 1315
Glu Arg Trp Ile Leu Gln Thr Asp Phe Glu Asn Asp Glu Ala Val Gly
390 395 400 405
tac ctg gct gac cgt ctg gcc aag ttg ggc att gag gac ggg ctt cgt 1363
Tyr Leu Ala Asp Arg Leu Ala Lys Leu Gly Ile Glu Asp Gly Leu Arg
410 415 420
aag gca gga gca cat gtg ggt gca aac gtc acc atc gga ggc att tcc 1411
Lys Ala Gly Ala His Val Gly Ala Asn Val Thr Ile Gly Gly Ile Ser
425 430 435
ttc gag tgg gag cca atg acc acc gct ggc gac gat cca gtc ctt acc 1459
Phe Glu Trp Glu Pro Met Thr Thr Ala Gly Asp Asp Pro Val Leu Thr
440 445 450
gga cgt ggc acc gat gtg cgc ctt gaa cag acc tct cgt atc tct gct 1507
Gly Arg Gly Thr Asp Val Arg Leu Glu Gln Thr Ser Arg Ile Ser Ala
455 460 465
gca gag cgt aaa cgc gca tct cag gta cgt cgt ggc ctc atc gat gag 1555
Ala Glu Arg Lys Arg Ala Ser Gln Val Arg Arg Gly Leu Ile Asp Glu
470 475 480 485
ttg gat tat ggc gag gac caa gag gct tcc cgc gaa cgc tgg gaa gga 1603
Leu Asp Tyr Gly Glu Asp Gln Glu Ala Ser Arg Glu Arg Trp Glu Gly
490 495 500
taaaaccgag cacttttcag gtctacgtgt 1633
<210>40
<211>501
<212>PRT
<213>谷氨酸棒状杆菌
<400>40
Met Asn Arg Phe Ile Asp Arg Val Val Leu His Leu Ala Ala Gly Asp
1 5 10 15
Gly Gly Asn Gly Cys Val Ser Val His Arg Glu Lys Phe Lys Pro Leu
20 25 30
Gly Gly Pro Asp Gly Gly Asn Gly Gly His Gly Gly Asp Ile Ile Leu
35 40 45
Glu Val Thr Ala Gln Val His Thr Leu Leu Asp Phe His Phe His Pro
50 55 60
His Val Lys Ala Glu Arg Gly Ala Ash Gly Ala Gly Asp His Arg Asn
65 70 75 80
Gly Ala Arg Gly Lys Asp Leu Val Leu Glu Val Pro Pro Gly Thr Val
85 90 95
Val Leu Asn Glu Lys Gly Glu Thr Leu Ala Asp Leu Thr Ser Val Gly
100 105 110
Met Lys Phe Ile Ala Ala Ala Gly Gly Asn Gly Gly Leu Gly Asn Ala
115 120 125
Ala Leu Ala Ser Lys Ala Arg Lys Ala Pro Gly Phe Ala Leu Ile Gly
130 135 140
Glu Pro Gly Glu Ala His Asp Leu Ile Leu Glu Leu Lys Ser Met Ala
145 150 155 160
Asp Val Gly Leu Val Gly Phe Pro Ser Ala Gly Lys Ser Ser Leu Ile
165 170 175
Ser Val Met Ser Ala Ala Lys Pro Lys Ile Gly Asp Tyr Pro Phe Thr
180 185 190
Thr Leu Gln Pro Asn Leu Gly Val Val Asn Val Gly His Glu Thr Phe
195 200 205
Thr Met Ala Asp Val Pro Gly Leu Ile Pro Gly Ala Ser Glu Gly Lys
210 215 220
Gly Leu Gly Leu Asp Phe Leu Arg His Ile Glu Arg Thr Ser Val Leu
225 230 235 240
Val His Val Val Asp Thr Ala Thr Met Asp Pro Gly Arg Asp Pro Ile
245 250 255
Ser Asp Ile Glu Ala Leu Glu Ala Glu Leu Ala Ala Tyr Gln Ser Ala
260 265 270
Leu Asp Glu Asp Thr Gly Leu Gly Asp Leu Ser Gln Arg Pro Arg Leu
275 280 285
Val Val Leu Asn Lys Ala Asp Val Pro Glu Ala Glu Glu Leu Ala Glu
290 295 300
Phe Leu Lys Glu Asp Ile Glu Lys Gln Phe Gly Trp Pro Val Phe Ile
305 310 315 320
Ile Ser Ala Val Ala Arg Lys Gly Leu Asp Pro Leu Lys Tyr Lys Leu
325 330 335
Leu Glu Ile Val Gln Asp Ala Arg Lys Lys Arg Pro Lys Glu Lys Ala
340 345 350
Glu Ser Val Ile Ile Lys Pro Lys Ala Val Asp His Arg Thr Lys Gly
355 360 365
Gln Phe Gln Ile Lys Pro Asp Pro Glu Val Gln Gly Gly Phe Ile Ile
370 375 380
Thr Gly Glu Lys Pro Glu Arg Trp Ile Leu Gln Thr Asp Phe Glu Asn
385 390 395 400
Asp Glu Ala Val Gly Tyr Leu Ala Asp Arg Leu Ala Lys Leu Gly Ile
405 410 415
Glu Asp Gly Leu Arg Lys Ala Gly Ala His Val Gly Ala Asn Val Thr
420 425 430
Ile Gly Gly Ile Ser Phe Glu Trp Glu Pro Met Thr Thr Ala Gly Asp
435 440 445
Asp Pro Val Leu Thr Gly Arg Gly Thr Asp Val Arg Leu Glu Gln Thr
450 455 460
Ser Arg Ile Ser Ala Ala Glu Arg Lys Arg Ala Ser Gln Val Arg Arg
465 470 475 480
Gly Leu Ile Asp Glu Leu Asp Tyr Gly Glu Asp Gln Glu Ala Ser Arg
485 490 495
Glu Arg Trp Glu Gly
500
<210>41
<211>1144
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1114)
<223>RXA02627
<400>41
tcgcctattg ggggttattt tcgatgcccg atcacagtaa tctcgaacag gttaggtaag 60
gtttgcatac tttattttta tcctgaaggg acgggcgcca atg aaa aag cgt gtc 115
Met Lys Lys Arg Val
1 5
agt tta ttt aag tca gct gca tcg att gtg gtg gtt gcg gga atg gcg 163
Ser Leu Phe Lys Ser Ala Ala Ser Ile Val Val Val Ala Gly Met Ala
10 15 20
ttg ggt atg gtg ggt tgc tca tcg acc gaa tcg gag agt gaa tct agc 211
Leu Gly Met Val Gly Cys Ser Ser Thr Glu Ser Glu Ser Glu Ser Ser
25 30 35
gct gac gtg aca tca tct gcc gat ggg gca tat cca gtg tcc atc gcg 259
Ala Asp Val Thr Ser Ser Ala Asp Gly Ala Tyr Pro Val Ser Ile Ala
40 45 50
acc aaa ttc ggt gat gtc act gtg gaa agc caa cca gaa cgc gtc gtt 307
Thr Lys Phe Gly Asp Val Thr Val Glu Ser Gln Pro Glu Arg Val Val
55 60 65
gcc ctg ggt tgg gga gat gct gag gct gcg ctg gaa ttc ggt gtg cag 355
Ala Leu Gly Trp Gly Asp Ala Glu Ala Ala Leu Glu Phe Gly Val Gln
70 75 80 85
cct gtg ggt gca tca gat tgg ctc gca ttc ggt ggt gaa ggc gtg gga 403
Pro Val Gly Ala Ser Asp Trp Leu Ala Phe Gly Gly Glu Gly Val Gly
90 95 100
ccg tgg att gag gat tct gcc tac gat gaa gcg cca gaa ata atc gga 451
Pro Trp Ile Glu Asp Ser Ala Tyr Asp Glu Ala Pro Glu Ile Ile Gly
105 110 115
acc atg gaa ccg gag tat gaa aag att gca gcg ctt gaa ccg gat ctg 499
Thr Met Glu Pro Glu Tyr Glu Lys Ile Ala Ala Leu Glu Pro Asp Leu
120 125 130
att ttg gac gtg cgc agc tct ggc gac cag gaa cgc tat gac aag ttg 547
Ile Leu Asp Val Arg Ser Ser Gly Asp Gln Glu Arg Tyr Asp Lys Leu
135 140 145
tct tca atc gca ctg acc atc ggc gtt cca gaa ggt ggc gat agc tac 595
Ser Ser Ile Ala Leu Thr Ile Gly Val Pro Glu Gly Gly Asp Ser Tyr
150 155 160 165
ctc acc cca cgc gct gag cag gta acc atg atc gcc act gct ctg ggg 643
Leu Thr Pro Arg Ala Glu Gln Val Thr Met Ile Ala Thr Ala Leu Gly
170 175 180
cag gct gaa cgt ggt gaa gaa gtg aac gct gaa tac gag cag ctc act 691
Gln Ala Glu Arg Gly Glu Glu Val Asn Ala Glu Tyr Glu Gln Leu Thr
185 190 195
gct gat att cgt gca gct cac ccg ggc tgg cct gag aag acc gcg gct 739
Ala Asp Ile Arg Ala Ala His Pro Gly Trp Pro Glu Lys Thr Ala Ala
200 205 210
gct gta tct gca acg gca acc agc tgg ggt gca tac atc aag ggc tcc 787
Ala Val Ser Ala Thr Ala Thr Ser Trp Gly Ala Tyr Ile Lys Gly Ser
215 220 225
aac cgt gta gat act ttg ctg gac ctg ggc ttc cag gaa aac cct gag 835
Asn Arg Val Asp Thr Leu Leu Asp Leu Gly Phe Gln Glu Asn Pro Glu
230 235 240 245
ctg gct aaa cag caa cct ggc gat acg ggt ttc tcc atc aaa ttc agt 883
Leu Ala Lys Gln Gln Pro Gly Asp Thr Gly Phe Ser Ile Lys Phe Ser
250 255 260
gaa gag act ttc ggc gtt gtg gat tcc gac ctg gtt gtc ggc ttt gcc 931
Glu Glu Thr Phe Gly Val Val Asp Ser Asp Leu Val Val Gly Phe Ala
265 270 275
ate ggt atg act cct gag gaa atg gca gag cag gtt cca tgg cag atg 979
Ile Gly Met Thr Pro Glu Glu Met Ala Glu Gln Val Pro Trp Gln Met
280 285 290
ttg acc gcc act cgt gac ggc cgt tcc ttt gtg atg ccc cgt gag att 1027
Leu Thr Ala Thr Arg Asp Gly Arg Ser Phe Val Met Pro Arg Glu Ile
295 300 305
tcc aat gcg ttt tct ttg ggt tcc ccg cag tcc act cgg ttc gcg tta 1075
Ser Asn Ala Phe Ser Leu Gly Ser Pro Gln Ser Thr Arg Phe Ala Leu
310 315 320 325
gac gcc ttg gtg cca ctt ctg gag gag cat gca ggg gag tagtggtccg 1124
Asp Ala Leu Val Pro Leu Leu Glu Glu His Ala Gly Glu
330 335
gtggtgcggg cagggggtag 1144
<210>42
<211>338
<212>PRT
<213>谷氨酸棒状杆菌
<400>42
Met Lys Lys Arg Val Ser Leu Phe Lys Ser Ala Ala Ser Ile Val Val
1 5 10 15
Val Ala Gly Met Ala Leu Gly Met Val Gly Cys Ser Ser Thr Glu Ser
20 25 30
Glu Ser Glu Ser Ser Ala Asp Val Thr Ser Ser Ala Asp Gly Ala Tyr
35 40 45
Pro Val Ser Ile Ala Thr Lys Phe Gly Asp Val Thr Val Glu Ser Gln
50 55 60
Pro Glu Arg Val Val Ala Leu Gly Trp Gly Asp Ala Glu Ala Ala Leu
65 70 75 80
Glu Phe Gly Val Gln Pro Val Gly Ala Ser Asp Trp Leu Ala Phe Gly
85 90 95
Gly Glu Gly Val Gly Pro Trp Ile Glu Asp Ser Ala Tyr Asp Glu Ala
100 105 110
Pro Glu Ile Ile Gly Thr Met Glu Pro Glu Tyr Glu Lys Ile Ala Ala
115 120 125
Leu Glu Pro Asp Leu Ile Leu Asp Val Arg Ser Ser Gly Asp Gln Glu
130 135 140
Arg Tyr Asp Lys Leu Ser Ser Ile Ala Leu Thr Ile Gly Val Pro Glu
145 150 155 160
Gly Gly Asp Ser Tyr Leu Thr Pro Arg Ala Glu Gln Val Thr Met Ile
165 170 175
Ala Thr Ala Leu Gly Gln Ala Glu Arg Gly Glu Glu Val Asn Ala Glu
180 185 190
Tyr Glu Gln Leu Thr Ala Asp Ile Arg Ala Ala His Pro Gly Trp Pro
195 200 205
Glu Lys Thr AlaAla Ala Val Ser Ala Thr Ala Thr Ser Trp Gly Ala
210 215 220
Tyr Ile Lys Gly Ser Asn Arg Val Asp Thr Leu Leu Asp Leu Gly Phe
225 230 235 240
Gln Glu Asn Pro Glu Leu Ala Lys Gln Gln Pro Gly Asp Thr Gly Phe
245 250 255
Ser Ile Lys Phe Ser Glu Glu Thr Phe Gly Val Val Asp Ser Asp Leu
260 265 270
Val Val Gly Phe Ala Ile Gly Met Thr Pro Glu Glu Met Ala Glu Gln
275 280 285
Val Pro Trp Gln Met Leu Thr Ala Thr Arg Asp Gly Arg Ser Phe Val
290 295 300
Met Pro Arg Glu Ile Ser Asn Ala Phe Ser Leu Gly Ser Pro Gln Ser
305 310 315 320
Thr Arg Phe Ala Leu Asp Ala Leu Val Pro Leu Leu Glu Glu His Ala
325 330 335
Gly Glu
<210>43
<211>793
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(763)
<223>RXA02667
<400>43
gttacgttaa atatggtaga attccatagg ttaaagcctg ccccgaattc gattgaaatt 60
tgaggggtag cgcagcaggg tgaagtatta aataggtcgc ttg cat gga caa tgc 115
Leu His Gly Gln Cys
1 5
ggg cgc cac cgg ctg gtt cta gca aaa atg gaa ccg cca tta gaa gga 163
Gly Arg His Arg Leu Val Leu Ala Lys Met Glu Pro Pro Leu Glu Gly
10 15 20
gtg gga atg gaa ttc aag gtc gga gat acc gtc gtt tac ccg cac cac 211
Val Gly Met Glu Phe Lys Val Gly Asp Thr Val Val Tyr Pro His His
25 30 35
gga gct gca att att tca gcc ctg gag cag cgt gaa atg aat ggt gag 259
Gly Ala Ala Ile Ile Ser Ala Leu Glu Gln Arg Glu Met Asn Gly Glu
40 45 50
acg gtg gac tac ctg gtt ctc cag atc aat cat tcc gat ctc gtc gtt 307
Thr Val Asp Tyr Leu Val Leu Gln Ile Asn His Ser Asp Leu Val Val
55 60 65
cgc gtt cca gca aag aac gct gaa ctc gtt ggc gtg cgt gac gtt gtc 355
Arg Val Pro Ala Lys Asn Ala Glu Leu Val Gly Val Arg Asp Val Val
70 75 80 85
ggc gag gag ggc ctg cag aag gtt ttc tct gtt ctt cgt gaa att gac 403
Gly Glu Glu Gly Leu Gln Lys Val Phe Ser Val Leu Arg Glu Ile Asp
90 95 100
gtc gaa gaa gcc ggc aac tgg tcc cgc cgt tac aag gct aac cag gag 451
Val Glu Glu Ala Gly Asn Trp Ser Arg Arg Tyr Lys Ala Asn Gln Glu
105 110 115
cgt ttg gct tcc ggt gac gtg aac aag gtc gct gag gtt gtc cgt gac 499
Arg Leu Ala Ser Gly Asp Val Asn Lys Val Ala Glu Val Val Arg Asp
120 125 130
ctg tgg cgt cgt gat cag gat cgt ggc ctt tcc gct ggt gag aag cgc 547
Leu Trp Arg Arg Asp Gln Asp Arg Gly Leu Ser Ala Gly Glu Lys Arg
135 140 145
atg ctc tcc aag gcg cgt cag gtt ctt gtt ggt gag ctc gcg ctc gcc 595
Met Leu Ser Lys Ala Arg Gln Val Leu Val Gly Glu Leu Ala Leu Ala
150 155 160 165
gaa acc gtg gac gat gag aag gcg gat gct ttc ctc agc cag gtc gat 643
Glu Thr Val Asp Asp Glu Lys Ala Asp Ala Phe Leu Ser Gln Val Asp
170 175 180
gag acc att gct cgc cac cgc gct gac ctg ctc ggc gac gag gaa gag 691
Glu Thr Ile Ala Arg His Arg Ala Asp Leu Leu Gly Asp Glu Glu Glu
185 190 195
aag aag gac gca ttc gac gac ttc gac gat tcc gac gtg gat ctt gac 739
Lys Lys Asp Ala Phe Asp Asp Phe Asp Asp Ser Asp Val Asp Leu Asp
200 205 210
gat ctg agc ttc gac gac gaa gat tagacgccca tgtcgtctac acgaatcccc 793
Asp Leu Ser Phe Asp Asp Glu Asp
215 220
<210>44
<211>221
<212>PRT
<213>谷氨酸棒状杆菌
<400>44
Leu His Gly Gln Cys Gly Arg His Arg Leu Val Leu Ala Lys Met Glu
1 5 10 15
Pro Pro Leu Glu Gly Val Gly Met Glu Phe Lys Val Gly Asp Thr Val
20 25 30
Val Tyr Pro His His Gly Ala Ala Ile Ile Ser Ala Leu Glu Gln Arg
35 40 45
Glu Met Asn Gly Glu Thr Val Asp Tyr Leu Val Leu Gln Ile Asn His
50 55 60
Ser Asp Leu Val Val Arg Val Pro Ala Lys Asn Ala Glu Leu Val Gly
65 70 75 80
Val Arg Asp Val Val Gly Glu Glu Gly Leu Gln Lys Val Phe Ser Val
85 90 95
Leu Arg Glu Ile Asp Val Glu Glu Ala Gly Asn Trp Ser Arg Arg Tyr
100 105 110
Lys Ala Asn Gln Glu Arg Leu Ala Ser Gly Asp Val Asn Lys Val Ala
115 120 125
Glu Val Val Arg Asp Leu Trp Arg Arg Asp Gln Asp Arg Gly Leu Ser
130 135 140
Ala Gly Glu Lys Arg Met Leu Ser Lys Ala Arg Gln Val Leu Val Gly
145 150 155 160
Glu Leu Ala Leu Ala Glu Thr Val Asp Asp Glu Lys Ala Asp Ala Phe
165 170 175
Leu Ser Gln Val Asp Glu Thr Ile Ala Arg His Arg Ala Asp Leu Leu
180 185 190
Gly Asp Glu Glu Glu Lys Lys Asp Ala Phe Asp Asp Phe Asp Asp Ser
195 200 205
Asp Val Asp Leu Asp Asp Leu Ser Phe Asp Asp Glu Asp
210 215 220
<210>45
<211>1468
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1438)
<223>RXA02758
<400>45
ggctggtgct tttcatattc cgacttgggg cacccctgaa tacatctcac ccaattcccc 60
ataactagac aattgcccag caacgactga taagtctcca atg tcg tgt tcc gcg 115
Met Ser Cys Ser Ala
1 5
ctc aga cat gag aca att gtt gcc gtg act gaa ctc atc cag aat gaa 163
Leu Arg His Glu Thr Ile Val Ala Val Thr Glu Leu Ile Gln Asn Glu
10 15 20
tcc caa gaa atc gct gag ctg gaa gcc ggc cag cag gtt gca ttg cgt 211
Ser Gln Glu Ile Ala Glu Leu Glu Ala Gly Gln Gln Val Ala Leu Arg
25 30 35
gaa ggt tat ctt cct gcg gtg atc aca gtg agc ggt aaa gac cgc cca 259
Glu Gly Tyr Leu Pro Ala Val Ile Thr Val Ser Gly Lys Asp Arg Pro
40 45 50
ggt gtg act gcc gcg ttc ttt agg gtc ttg tcc gct aat cag gtt cag 307
Gly Val Thr Ala Ala Phe Phe Arg Val Leu Ser Ala Asn Gln Val Gln
55 60 65
gtc ttg gac gtt gag cag tca atg ttc cgt ggc ttt ttg aac ttg gcg 355
Val Leu Asp Val Glu Gln Ser Met Phe Arg Gly Phe Leu Asn Leu Ala
70 75 80 85
gcg ttt gtg ggt atc gca cct gag cgt gtc gag acc gtc acc aca ggc 403
Ala Phe Val Gly Ile Ala Pro Glu Arg Val Glu Thr Val Thr Thr Gly
90 95 100
ctg act gac acc ctc aag gtg cat gga cag tcc gtg gtg gtg gag ctg 451
Leu Thr Asp Thr Leu Lys Val His Gly Gln Ser Val Val Val Glu Leu
105 110 115
cag gaa act gtg cag tcg tcc cgt cct cgt tct tcc cat gtt gtt gtg 499
Gln Glu Thr Val Gln Ser Ser Arg Pro Arg Ser Ser His Val Val Val
120 125 130
gtg ttg ggt gat ccg gtt gat gcg ttg gat att tcc cgc att ggt cag 547
Val Leu Gly Asp Pro Val Asp Ala Leu Asp Ile Ser Arg Ile Gly Gln
135 140 145
acc ctg gcg gat tac gat gcc aac att gac acc att cgt ggt att tcg 595
Thr Leu Ala Asp Tyr Asp Ala Asn Ile Asp Thr Ile Arg Gly Ile Ser
150 155 160 165
gat tac cct gtg acc ggc ctg gag ctg aag gtg act gtg ccg gat gtc 643
Asp Tyr Pro Val Thr Gly Leu Glu Leu Lys Val Thr Val Pro Asp Val
170 175 180
agc cct ggt ggt ggt gaa gcg atg cgt aag gcg ctt gct gct ctt acc 691
Ser Pro Gly Gly Gly Glu Ala Met Arg Lys Ala Leu Ala Ala Leu Thr
185 190 195
tct gag ctg aat gtg gat att gcg att gag cgt tct ggt ttg ctg cgt 739
Ser Glu Leu Asn Val Asp Ile Ala Ile Glu Arg Ser Gly Leu Leu Arg
200 205 210
cgt tct aag cgt ctg gtg tgc ttc gat tgt gat tcc acg ttg atc act 787
Arg Ser Lys Arg Leu Val Cys Phe Asp Cys Asp Ser Thr Leu Ile Thr
215 220 225
ggt gag gtc att gag atg ctg gcg gct cac gcg ggc aag gaa gct gaa 835
Gly Glu Val Ile Glu Met Leu Ala Ala His Ala Gly Lys Glu Ala Glu
230 235 240 246
gtt gcg gca gtt act gag cgt gcg atg cgc ggt gag ctc gat ttc gag 883
Val Ala Ala Val Thr Glu Arg Ala Met Arg Gly Glu Leu Asp Phe Glu
250 255 260
gag tct ctg cgt gag cgt gtg aag gcg ttg gct ggt ttg gat gcg tcg 931
Glu Ser Leu Arg Glu Arg Val Lys Ala Leu Ala Gly Leu Asp Ala Ser
265 270 275
gtg atc gat gag gtc gct gcc gct att gag ctg acc cct ggt gcg cgc 979
Val Ile Asp Glu Val Ala Ala Ala Ile Glu Leu Thr Pro Gly Ala Arg
280 285 290
acc acg atc cgt acg ctg aac cgc atg ggt tac cag acc gct gtt gtt 1027
Thr Thr Ile Arg Thr Leu Asn Arg Met Gly Tyr Gln Thr Ala Val Val
295 300 305
tcc ggt ggt ttc atc cag gtg ttg gaa ggt ttg gct gag gag ttg gag 1075
Ser Gly Gly Phe Ile Gln Val Leu Glu Gly Leu Ala Glu Glu Leu Glu
310 315 320 325
ttg gat tat gtc cgc gcc aac act ttg gaa atc gtt gat ggc aag ctg 1123
Leu Asp Tyr Val Arg Ala Asn Thr Leu Glu Ile Val Asp Gly Lys Leu
330 335 340
acc ggc aac gtc acc gga aag atc gtt gac cgc gct gcg aag gct gag 1171
Thr Gly Asn Val Thr Gly Lys Ile Val Asp Arg Ala Ala Lys Ala Glu
345 350 355
ttc ctc cgt gag ttc gct gcg gat tct ggc ctg aag atg tac cag act 1219
Phe Leu Arg Glu Phe Ala Ala Asp Ser Gly Leu Lys Met Tyr Gln Thr
360 365 370
gtc gct gtc ggt gat ggc gct aat gac atc gat atg ctc tcc gct gcg 1267
Val Ala Val Gly Asp Gly Ala Asn Asp Ile Asp Met Leu Ser Ala Ala
375 380 385
ggt ctg ggt gtt gct ttc aac gcg aag cct gcg ctg aag gag att gcg 1315
Gly Leu Gly Val Ala Phe Asn Ala Lys Pro Ala Leu Lys Glu Ile Ala
390 395 400 405
gat act tcc gtg aac cac cca ttc ctc gac gag gtt ttg cac atc atg 1363
Asp Thr Ser Val Asn His Pro Phe Leu Asp Glu Val Leu His Ile Met
410 415 420
ggc att tcc cgc gac gag atc gat ctg gcg gat cag gaa gac ggc act 1411
Gly Ile Ser Arg Asp Glu Ile Asp Leu Ala Asp Gln Glu Asp Gly Thr
425 430 435
ttc cac cgc gtt cca ttg acc aat gcc taaagattcg cttctcgacg 1458
Phe His Arg Val Pro Leu Thr Asn Ala
440 445
cccacctcct 1468
<210>46
<211>446
<212>PRT
<213>谷氨酸棒状杆菌
<400>46
Met Ser Cys Ser Ala Leu Arg His Glu Thr Ile Val Ala Val Thr Glu
1 5 10 15
Leu Ile Gln Asn Glu Ser Gln Glu Ile Ala Glu Leu Glu Ala Gly Gln
20 25 30
Gln Val Ala Leu Arg Glu Gly Tyr Leu Pro Ala Val Ile Thr Val Ser
35 40 45
Gly Lys Asp Arg Pro Gly Val Thr Ala Ala Phe Phe Arg Val Leu Ser
50 55 60
Ala Asn Gln Val Gln Val Leu Asp Val Glu Gln Ser Met Phe Arg Gly
65 70 75 80
Phe Leu Asn Leu Ala Ala Phe Val Gly Ile Ala Pro Glu Arg Val Glu
85 90 95
Thr Val Thr Thr Gly Leu Thr Asp Thr Leu Lys Val His Gly Gln Ser
100 105 110
Val Val Val Glu Leu Gln Glu Thr Val Gln Ser Ser Arg Pro Arg Ser
115 120 125
Ser His Val Val Val Val Leu Gly Asp Pro Val Asp Ala Leu Asp Ile
130 135 140
Ser Arg Ile Gly Gln Thr Leu Ala Asp Tyr Asp Ala Asn Ile Asp Thr
145 150 155 160
Ile Arg Gly Ile Ser Asp Tyr Pro Val Thr Gly Leu Glu Leu Lys Val
165 170 175
Thr Val Pro Asp Val Ser Pro Gly Gly Gly Glu Ala Met Arg Lys Ala
180 185 190
Leu Ala Ala Leu Thr Ser Glu Leu Asn Val Asp Ile Ala Ile Glu Arg
195 200 205
Ser Gly Leu Leu Arg Arg Ser Lys Arg Leu Val Cys Phe Asp Cys Asp
210 215 220
Ser Thr Leu Ile Thr Gly Glu Val Ile Glu Met Leu Ala Ala His Ala
225 230 235 240
Gly Lys Glu Ala Glu Val Ala Ala Val Thr Glu Arg Ala Met Arg Gly
245 250 255
Glu Leu Asp Phe Glu Glu Ser Leu Arg Glu Arg Val Lys Ala Leu Ala
260 265 270
Gly Leu Asp Ala Ser Val Ile Asp Glu Val Ala Ala Ala Ile Glu Leu
275 280 285
Thr Pro Gly Ala Arg Thr Thr Ile Arg Thr Leu Asn Arg Met Gly Tyr
290 295 300
Gln Thr Ala Val Val Ser Gly Gly Phe Ile Gln Val Leu Glu Gly Leu
305 310 315 320
Ala Glu Glu Leu Glu Leu Asp Tyr Val Arg Ala Asn Thr Leu Glu Ile
325 330 335
Val Asp Gly Lys Leu Thr Gly Asn Val Thr Gly Lys Ile Val Asp Arg
340 345 350
Ala Ala Lys Ala Glu Phe Leu Arg Glu Phe Ala Ala Asp Ser Gly Leu
355 360 365
Lys Met Tyr Gln Thr Val Ala Val Gly Asp Gly Ala Asn Asp Ile Asp
370 375 380
Met Leu Ser Ala Ala Gly Leu Gly Val Ala Phe Asn Ala Lys Pro Ala
385 390 395 400
Leu Lys Glu Ile Ala Asp Thr Ser Val Asn His Pro Phe Leu Asp Glu
405 410 415
Val Leu His Ile Met Gly Ile Ser Arg Asp Glu Ile Asp Leu Ala Asp
420 425 430
Gln Glu Asp Gly Thr Phe His Arg Val Pro Leu Thr Asn Ala
435 440 445
<210>47
<211>844
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(814)
<223>RXA02910
<400>47
tagcggacca acctaggccg atacccatgt ggaaatctcg acgtcttaaa tggacgattg 60
gagctaaaac cacgaacagc tgggattttc cacgatagga ttg ggt ctc gtg gag 115
Leu Gly Leu Val Glu
1 5
att cgt tgg ttg gaa ggc ttt atc gcg gtc gcg gaa gaa ttg cac ttt 163
Ile Arg Trp Leu Glu Gly Phe Ile Ala Val Ala Glu Glu Leu His Phe
10 15 20
agt aat gct gcg att cgt ttg ggg atg ccg caa tcg ccg ttg agt cag 211
Ser Asn Ala Ala Ile Arg Leu Gly Met Pro Gln Ser Pro Leu Ser Gln
25 30 35
ttg atc cgg cgg ttg gag tcg gag ttg ggg cag aag ctt ttt gat cgc 259
Leu Ile Arg Arg Leu Glu Ser Glu Leu Gly Gln Lys Leu Phe Asp Arg
40 45 50
agt acc cgg tcg gtg gag tta act gcc gcg ggt cgg gcg ttt ttg cca 307
Ser Thr Arg Ser Val Glu Leu Thr Ala Ala Gly Arg Ala Phe Leu Pro
55 60 65
cat gcc agg ggg att gtg gcg agc gct gcg gtg gcg agg gaa gct gtg 355
His Ala Arg Gly Ile Val Ala Ser Ala Ala Val Ala Arg Glu Ala Val
70 75 80 85
aat gct gcc gag ggg gag atc gtt ggt gtt gtt cgc att ggt ttt tct 403
Asn Ala Ala Glu Gly Glu Ile Val Gly Val Val Arg Ile Gly Phe Ser
90 95 100
ggt gtg ctg aac tat tcc acg ctg ccg ctt ttg acc agt gag gtg cat 451
Gly Val Leu Asn Tyr Ser Thr Leu Pro Leu Leu Thr Ser Glu Val His
105 110 115
aaa cgg ctt cct aat gtg gag ttg gag ctc gtt ggt cag aag ttg acg 499
Lys Arg Leu Pro Asn Val Glu Leu Glu Leu Val Gly Gln Lys Leu Thr
120 125 130
agg gaa gcg gta agt ttg ctg cgc ttg ggg gcg ttg gat att acg ttg 547
Arg Glu Ala Val Ser Leu Leu Arg Leu Gly Ala Leu Asp Ile Thr Leu
135 140 145
atg ggt ttg ccc att gag gat cca gag att gag act cgg ctg att agt 595
Met Gly Leu Pro Ile Glu Asp Pro Glu Ile Glu Thr Arg Leu Ile Ser
150 155 160 165
ttg gaa gag ttt tgc gtg gtg ttg ccg aag gat cat cgt ctt gcg ggg 643
Leu Glu Glu Phe Cys Val Val Leu Pro Lys Asp His Arg Leu Ala Gly
170 175 180
gaa gga gtg gtg gat ttg gtg gat ctg gct aaa gat ggg ttt gtg acg 691
Glu Gly Val Val Asp Leu Val Asp Leu Ala Lys Asp Gly Phe Val Thr
185 190 195
acg ccg gag ttt gcg ggg tct gtg ttt agg aat tcc acc ttt cag ttg 739
Thr Pro Glu Phe Ala Gly Ser Val Phe Arg Asn Ser Thr Phe Gln Leu
200 205 210
tgt gct gag gct ggt ttt gtg ccg agg atc agc cag caa gtt aat gat 787
Cys Ala Glu Ala Gly Phe Val Pro Arg Ile Ser Gln Gln Val Asn Asp
215 220 225
cct tac atg gcg ctg ttg ttg gcg cgg tagtcaatca tgggggagta 834
Pro Tyr Met Ala Leu Leu Leu Ala Arg
230 235
tcccaccgta 844
<210>48
<211>238
<212>PRT
<213>谷氨酸棒状杆菌
<400>48
Leu Gly Leu Val Glu Ile Arg Trp Leu Glu Gly Phe Ile Ala Val Ala
1 5 10 15
Glu Glu Leu His Phe Ser Asn Ala Ala Ile Arg Leu Gly Met Pro Gln
20 25 30
Ser Pro Leu Ser Gln Leu Ile Arg Arg Leu Glu Ser Glu Leu Gly Gln
35 40 45
Lys Leu Phe Asp Arg Ser Thr Arg Ser Val Glu Leu Thr Ala Ala Gly
50 55 60
Arg Ala Phe Leu Pro His Ala Arg Gly Ile Val Ala Ser Ala Ala Val
65 70 75 80
Ala Arg Glu Ala Val Asn Ala Ala Glu Gly Glu Ile Val Gly Val Val
85 90 95
Arg Ile Gly Phe Ser Gly Val Leu Asn Tyr Ser Thr Leu Pro Leu Leu
100 105 110
Thr Ser Glu Val His Lys Arg Leu Pro Asn Val Glu Leu Glu Leu Val
115 120 125
Gly Gln Lys Leu Thr Arg Glu Ala Val Ser Leu Leu Arg Leu Gly Ala
130 135 140
Leu Asp Ile Thr Leu Met Gly Leu Pro Ile Glu Asp Pro Glu Ile Glu
145 150 155 160
Thr Arg Leu Ile Ser Leu Glu Glu Phe Cys Val Val Leu Pro Lys Asp
165 170 175
His Arg Leu Ala Gly Glu Gly Val Val Asp Leu Val Asp Leu Ala Lys
180 185 190
Asp Gly Phe Val Thr Thr Pro Glu Phe Ala Gly Ser Val Phe Arg Asn
195 200 205
Ser Thr Phe Gln Leu Cys Ala Glu Ala Gly Phe Val Pro Arg Ile Ser
210 215 220
Gln Gln Val Asn Asp Pro Tyr Met Ala Leu Leu Leu Ala Arg
225 230 235
<210>49
<211>1348
<212>DNA
<21 3>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1318)
<223>RXA03100
<400>49
aaactctaaa aactatagag ctatagaaac tttaacttcg gaggtattca tgtcccgtcc 60
aatcgttaaa caagcattca ccgtcaccgc agtcaccgcg atg gct ttt gcc ctg 115
Met Ala Phe Ala Leu
1 5
gca tca tgc acc cgc gca gtg gat gca acc tcc gca gat gga acc gcg 163
Ala Ser Cys Thr Arg Ala Val Asp Ala Thr Ser Ala Asp Gly Thr Ala
10 15 20
agc aac acc gca gct tcc tgt gtg gat aca tcc ggc gac tcc atc aaa 211
Ser Asn Thr Ala Ala Ser Cys Val Asp Thr Ser Gly Asp Ser Ile Lys
25 30 35
atc ggc ttc atc aac tcc ctg tcc ggc acg atg gct atc tct gaa acc 259
Ile Gly Phe Ile Asn Ser Leu Ser Gly Thr Met Ala Ile Ser Glu Thr
40 45 50
acg gtg aac caa tcc ctg cac atg gca gcc gat gaa atc aac gca gcc 307
Thr Val Asn Gln Ser Leu His MetAla Ala Asp Glu Ile Asn Ala Ala
55 60 65
ggc ggc gtt ttg ggc aag cag ttg gag atc tct gaa gaa gac ggc gcc 355
Gly Gly Val Leu Gly Lys Gln Leu Glu Ile Ser Glu Glu Asp Gly Ala
70 75 80 85
agc gaa ccc gcc acc ttc gcc gaa cgc tcc caa cgc ctc atc cag cag 403
Ser Glu Pro Ala Thr Phe Ala Glu Arg Ser Gln Arg Leu Ile Gln Gln
90 95 100
gaa tgc gtt gca gcc gtg ttt ggt gga tgg acc tcc gcc tcc cgc aaa 451
Glu Cys Val Ala Ala Val Phe Gly Gly Trp Thr Ser Ala Ser Arg Lys
105 110 115
gca atg ctc ccc gtc ttt gag ggc aat aac tcc ctg ctg ttc tac ccg 499
Ala Met Leu Pro Val Phe Glu Gly Asn Asn Ser Leu Leu Phe Tyr Pro
120 125 130
gtg cag tac gag ggc atg gaa tcc tcg ccg aat att ttc tac acc ggc 547
Val Gln Tyr Glu Gly Met Glu Ser Ser Pro Asn Ile Phe Tyr Thr Gly
135 140 145
gcc acc acc aac cag cag atc atc ccg gct ctt gat tac ctg cgt gaa 595
Ala Thr Thr Asn Gln Gln Ile Ile Pro Ala Leu Asp Tyr Leu Arg Glu
150 155 160 165
aac ggc ctg aac cgc ctt ttc ctt gtc ggt tcc gat tat gtt ttc cca 643
Asn Gly Leu Asn Arg Leu Phe Leu Val Gly Ser Asp Tyr Val Phe Pro
170 175 180
cgc act gca aat tcc atc atc aag gac tac gcc gaa gcc aat ggt atg 691
Arg Thr Ala Asn Ser Ile Ile Lys Asp Tyr Ala Glu Ala Asn Gly Met
185 190 195
gaa atc gtc ggc gaa gac tac gcg ccg ttg gga tcc acc gac ttc acc 739
Glu Ile Val Gly Glu Asp Tyr Ala Pro Leu Gly Ser Thr Asp Phe Thr
200 205 210
acc atc gcc aac cgc atg cgt gac tcc aac gca gat gcc gtg ttc aac 787
Thr Ile Ala Asn Arg Met Arg Asp Ser Asn Ala Asp Ala Val Phe Asn
215 220 225
act ttg aat ggc gat tcc aac gtg gcg ttc ttc cgc cag tac aac agc 835
Thr Leu Asn Gly Asp Ser Asn Val Ala Phe Phe Arg Gln Tyr Asn Ser
230 235 240 245
ctc ggc ttc aat gca gac acc ctt ccg gtg atg tca gta tcc att gcg 883
Leu Gly Phe Asn Ala Asp Thr Leu Pro Val Met Ser Val Ser Ile Ala
250 255 260
gaa gaa gaa gtc gga ggc atc ggc acc gca aat att gag ggc cag ctg 931
Glu Glu Glu Val Gly Gly Ile Gly Thr Ala Asn Ile Glu Gly Gln Leu
265 270 275
gtg gcg tgg gac tac tac caa acc atc gac acc cca gaa aac gag acc 979
Val Ala Trp Asp Tyr Tyr Gln Thr Ile Asp Thr Pro Glu Asn Glu Thr
280 285 290
ttc gtg gag aat ttc aag gac ctc tac ggc cag gac aaa gtg acc tcc 1027
Phe Val Glu Asn Phe Lys Asp Leu Tyr Gly Gln Asp Lys Val Thr Ser
295 300 305
gat ccg atg gaa gct gct tac act agc ctc tac ctc tgg aaa gaa atg 1075
Asp Pro Met Glu Ala Ala Tyr Thr Ser Leu Tyr Leu Trp Lys Glu Met
310 315 320 325
gta gag aag gcc gat tcc ttt gat gtc gcc gca att caa gca gcc gcc 1123
Val Glu Lys Ala Asp Ser Phe Asp Val Ala Ala Ile Gln Ala Ala Ala
330 335 340
gac gga acc act ttt gat gca cca gaa gga acc gtg gtg gtt gac ggc 1171
Asp Gly Thr Thr Phe Asp Ala Pro Glu Gly Thr Val Val Val Asp Gly
345 350 355
gat aac cac cac atc tcc aaa aca ccg cgc atc ggt cga atc cgc ccg 1219
Asp Asn His His Ile Ser Lys Thr Pro Arg Ile Gly Arg Ile Arg Pro
360 365 370
gat gga ttg atc gac acc att tgg gaa acc gat tcc cca gtt gat ccg 1267
Asp Gly Leu Ile Asp Thr Ile Trp Glu Thr Asp Ser Pro Val Asp Pro
375 380 385
gac cca tac ttg tct tcc tat gac tgg gcc aag acc acc gct gcg act 1315
Asp Pro Tyr Leu Ser Ser Tyr Asp Trp Ala Lys Thr Thr Ala Ala Thr
390 395 400 405
tcc taagagataa aaatcatgga cattttactc 1348
Ser
<210>50
<211>406
<212>PRT
<213>谷氨酸棒状杆菌
<400>50
Met Ala Phe Ala Leu Ala Ser Cys Thr Arg Ala Val Asp Ala Thr Ser
1 5 10 15
Ala Asp Gly Thr Ala Ser Asn Thr Ala Ala Ser Cys Val Asp Thr Ser
20 25 30
Gly Asp Ser Ile Lys Ile Gly Phe Ile Asn Ser Leu Ser Gly Thr Met
35 40 45
Ala Ile Ser Glu Thr Thr Val Asn Gln Ser Leu His Met Ala Ala Asp
50 55 60
Glu Ile Asn Ala Ala Gly Gly Val Leu Gly Lys Gln Leu Glu Ile Ser
65 70 75 80
Glu Glu Asp Gly Ala Ser Glu Pro Ala Thr Phe Ala Glu Arg Ser Gln
85 90 95
Arg Leu Ile Gln Gln Glu Cys Val Ala Ala Val Phe Gly Gly Trp Thr
100 105 110
Ser Ala Ser Arg Lys Ala Met Leu Pro Val Phe Glu Gly Asn Asn Ser
115 120 125
Leu Leu Phe Tyr Pro Val Gln Tyr Glu Gly Met Glu Ser Ser Pro Asn
130 135 140
Ile Phe Tyr Thr Gly Ala Thr Thr Asn Gln Gln Ile Ile Pro Ala Leu
145 150 155 160
Asp Tyr Leu Arg Glu Asn Gly Leu Asn Arg Leu Phe Leu Val Gly Ser
165 170 175
Asp Tyr Val Phe Pro Arg Thr Ala Asn Ser Ile Ile Lys Asp Tyr Ala
180 185 190
Glu Ala Asn Gly Met Glu Ile Val Gly Glu Asp Tyr Ala Pro Leu Gly
195 200 205
Ser Thr Asp Phe Thr Thr Ile Ala Asn Arg Met Arg Asp Ser Asn Ala
210 215 220
Asp Ala Val Phe Asn Thr Leu Asn Gly Asp Ser Asn Val Ala Phe Phe
225 230 235 240
Arg Gln Tyr Asn Ser Leu Gly Phe Asn Ala Asp Thr Leu Pro Val Met
245 250 255
Ser Val Ser Ile Ala Glu Glu Glu Val Gly Gly Ile Gly Thr Ala Asn
260 265 270
Ile Glu Gly Gln Leu Val Ala Trp Asp Tyr Tyr Gln Thr Ile Asp Thr
275 280 285
Pro Glu Asn Glu Thr Phe Val Glu Asn Phe Lys Asp Leu Tyr Gly Gln
290 295 300
Asp Lys Val Thr Ser Asp Pro Met Glu Ala Ala Tyr Thr Ser Leu Tyr
305 310 315 320
Leu Trp Lys Glu Met Val Glu Lys Ala Asp Ser Phe Asp Val Ala Ala
325 330 335
Ile Gln Ala Ala Ala Asp Gly Thr Thr Phe Asp Ala Pro Glu Gly Thr
340 345 350
Val Val Val Asp Gly Asp Asn His His Ile Ser Lys Thr Pro Arg Ile
355 360 365
Gly Arg Ile Arg Pro Asp Gly Leu Ile Asp Thr Ile Trp Glu Thr Asp
370 375 380
Ser Pro Val Asp Pro Asp Pro Tyr Leu Ser Set Tyr Asp Trp Ala Lys
385 390 395 400
Thr Thr Ala Ala Thr Ser
405
<210>51
<211>850
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(820)
<223>RXA03 127
<400>51
aggtagaaca tctgaaagca cttcagataa gacggatggg gtctgatgga aacgacagtc 60
gatatgatca gaaccatctc cagattagga agtgaacaca atg gaa agc tcc aaa 115
Met Glu Ser Ser Lys
1 5
aag act tcg cga tca agg tcc act act caa gaa gca gtg cgc gac att 163
Lys Thr Ser Arg Ser Arg Ser Thr Thr Gln Glu Ala Val Arg Asp Ile
10 15 20
aaa aaa tac att cgg gac aac cgg ctg cgt acg gga gac ctt ctt cct 211
Lys Lys Tyr Ile Arg Asp Asn Arg Leu Arg Thr Gly Asp Leu Leu Pro
25 30 35
tcc gaa gcg ttc tta tgt gag gaa ttg ggt tgt tcc cgt tct gcg atc 259
Ser Glu Ala Phe Leu Cys Glu Glu Leu Gly Cys Ser Arg Ser Ala Ile
40 45 50
agg gag gcg atc cgc gcg ctc gtg acc ttg gac atc gtc gag gtt cgc 307
Arg Glu Ala Ile Arg Ala Leu Val Thr Leu Asp Ile Val Glu Val Arg
55 60 65
cac ggc tac ggc act ttc gtg tcc agg atg tcc ctc gag ccc ctg atc 355
His Gly Tyr Gly Thr Phe Val Ser Arg Met Ser Leu Glu Pro Leu Ile
70 75 80 85
aac ggg atg gtg ttc cgc acg gtg ttg gac aat gac acc tcg gtg gaa 403
Asn Gly Met Val Phe Arg Thr Val Leu Asp Asn Asp Thr Ser Val Glu
90 95 100
aac ctt ttc tac gtg gtg gat acc cgc gaa atc ctt gac ctt tca ctt 451
Asn Leu Phe Tyr Val Val Asp Thr Arg Glu Ile Leu Asp Leu Ser Leu
105 110 115
ggc gaa gag ctg atc gag gtg ttc acc gac gat gac cgc gag cta ctc 499
Gly Glu Glu Leu Ile Glu Val Phe Thr Asp Asp Asp Arg Glu Leu Leu
120 125 130
ctt gat ctg gtg gac aag atg cgc gag cac aac gat cag ggc gaa tcc 547
Leu Asp Leu Val Asp Lys Met Arg Glu His Asn Asp Gln Gly Glu Ser
135 140 145
ttt gtg gtg gag gat caa aaa ttc cac cga gca ctc cta gcg cga acg 595
Phe Val Val Glu Asp Gln Lys Phe His Arg Ala Leu Leu Ala Arg Thr
150 155 160 165
aaa aac ccg ctg att aga gag ctc aac gat gcg ttt tgg cag atc caa 643
Lys Asn Pro Leu Ile Arg Glu Leu Asn Asp Ala Phe Trp Gln Ile Gln
170 175 180
acc gag gcg cag ccc atg ctc aat ctg gct atg ccc gca gac atc gac 691
Thr Glu Ala Gln Pro Met Leu Asn Leu Ala Met Pro Ala Asp Ile Asp
185 190 195
gaa acc atc aaa gct cac agc gac atc gtc gaa gcg ctc tcc agc ggg 739
Glu Thr Ile Lys Ala His Ser Asp Ile Val Glu Ala Leu Ser Ser Gly
200 205 210
aac atc gac gat tat cgc agc gcc gtg ctc gct cac tac gcg ccg ttt 787
Asn Ile Asp Asp Tyr Arg Ser Ala Val Leu Ala His Tyr Ala Pro Phe
215 220 225
cgc cgc atg att tcc aac atg ctc gat gcg cac tagcctcatt gcgcgcgggt840
Arg Arg Met Ile Ser Asn Met Leu Asp Ala His
230 235 240
tgtaccgcat 850
<210>52
<211>240
<212>PRT
<213>谷氨酸棒状杆菌
<400>52
Met Glu Ser Ser Lys Lys Thr Ser Arg Ser Arg Ser Thr Thr Gln Glu
1 5 10 15
Ala Val Arg Asp Ile Lys Lys Tyr Ile Arg Asp Asn Arg Leu Arg Thr
20 25 30
Gly Asp Leu Leu Pro Ser Glu Ala Phe Leu Cys Glu Glu Leu Gly Cys
35 40 45
Ser Arg Ser Ala Ile Arg Glu Ala Ile Arg Ala Leu Val Thr Leu Asp
50 55 60
Ile Val Glu Val Arg His Gly Tyr Gly Thr Phe Val Ser Arg Met Ser
65 70 75 80
Leu Glu Pro Leu Ile Asn Gly Met Val Phe Arg Thr Val Leu Asp Asn
85 90 95
Asp Thr Ser Val Glu Asn Leu Phe Tyr Val Val Asp Thr Arg Glu Ile
100 105 110
Leu Asp Leu Ser Leu Gly Glu Glu Leu Ile Glu Val Phe Thr Asp Asp
115 120 125
Asp Arg Glu Leu Leu Leu Asp Leu Val Asp Lys Met Arg Glu His Asn
130 135 140
Asp Gln Gly Glu Ser Phe Val Val Glu Asp Gln Lys Phe His Arg Ala
145 150 155 160
Leu Leu Ala Arg Thr Lys Asn Pro Leu Ile Arg Glu Leu Asn Asp Ala
165 170 175
Phe Trp Gln Ile Gln Thr Glu Ala Gln Pro Met Leu Asn Leu Ala Met
180 185 190
Pro Ala Asp Ile Asp Glu Thr Ile Lys Ala His Ser Asp Ile Val Glu
195 200 205
Ala Leu Ser Ser Gly Asn Ile Asp Asp Tyr Arg Ser Ala Val Leu Ala
210 215 220
His Tyr Ala Pro Phe Arg Arg Met Ile Ser Asn Met Leu Asp Ala His
225 230 235 240
<210>53
<211>2749
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(2719)
<223>RXA03136
<400>53
ggtacaagtc ccttttttcg tctagtattg atttatggtt gcctcagtct ctttcacccc 60
gtcatcgtcc caaacgttcc taccgcgggc gtcgatcatc gtg caa gaa gtc agc 115
Val Gln Glu Val Ser
1 5
aca cag atc aaa aga tta gaa tcc ggc gaa ggc aaa ctc ttc cac atc 163
Thr Gln Ile Lys Arg Leu Glu Ser Gly Glu Gly Lys Leu Phe His Ile
10 15 20
agt ggc cag ccc ggc gcg ggt aaa aca gag ttc ggc atg cag ctt atc 211
Ser Gly Gln Pro Gly Ala Gly Lys Thr Glu Phe Gly Met Gln Leu Ile
25 30 35
gac gag ctc caa ggc tgg acc gtg gta cgt gcc act tcc ctg tcg tgg 259
Asp Glu Leu Gln Gly Trp Thr Val Val Arg Ala Thr Ser Leu Ser Trp
40 45 50
ctg aaa aac agc ccc cga aac ctc ctt gga cac atc gcc cac aaa ttg 307
Leu Lys Asn Ser Pro Arg Asn Leu Leu Gly His Ile Ala His Lys Leu
55 60 65
ggt gcg cac tcc gcc aac tcc atc cgt ggt gtg atc gac cgt ctc gat 355
Gly Ala His Ser Ala Asn Ser Ile Arg Gly Val Ile Asp Arg Leu Asp
70 75 80 85
gcc tcc acc gtg gtg atc gtt gac gat gtc cac tgg gcc gac gtg gaa 403
Ala Ser Thr Val Val Ile Val Asp Asp Val His Trp Ala Asp Val Glu
90 95 100
tcc atg caa aaa ctc atc gaa tat tcc atg cgc atg gtt tct ggc cgt 451
Ser Met Gln Lys Leu Ile Glu Tyr Ser Met Arg Met Val Ser Gly Arg
105 110 115
ttc gca ctc atc atg att ggc ctt gat gaa gag aac tta gtg ttc cac 499
Phe Ala Leu Ile Met Ile Gly Leu Asp Glu Glu Asn Leu Val Phe His
120 125 130
gat gag gtg gtc tcg ctc ccc tcc atc gca gac tcc acc tac gta ttg 547
Asp Glu Val Val Ser Leu Pro Ser Ile Ala Asp Ser Thr Tyr Val Leu
135 140 145
ccg ccg atg agt att gaa gaa atc cgc cag ctt gcg ctt acc gat gtc 595
Pro Pro Met Ser Ile Glu Glu Ile Arg Gln Leu Ala Leu Thr Asp Val
150 155 160 165
cgc ggc cgc atc agc acc acc acc gcc aca gac atc cag cgc atc acc 643
Arg Gly Arg Ile Ser Thr Thr Thr Ala Thr Asp Ile Gln Arg Ile Thr
170 175 180
ggc ggc atc tac ggg cga gtc aaa gaa gtc ctc cac tcg gaa tcc ccc 691
Gly Gly Ile Tyr Gly Arg Val Lys Glu Val Leu His Ser Glu Ser Pro
185 190 195
gat cac tgg cga atg ccc aac cca aat att ccc atc cca caa agc tgg 739
Asp His Trp Arg Met Pro Asn Pro Asn Ile Pro Ile Pro Gln Ser Trp
200 205 210
cat gcc aac ctg ttg aga cgc atc acc aac gaa gaa gtc tgg cat gta 787
His Ala Asn Leu Leu Arg Arg Ile Thr Asn Glu Glu Val Trp His Val
215 220 225
cta ctc gcc gtc gct gtc ctt ccc tcc gga ggc ccc att gac ctg gta 835
Leu Leu Ala Val Ala Val Leu Pro Ser Gly Gly Pro Ile Asp Leu Val
230 235 240 245
aaa ctc ata ggc aac gac ccc acg ggc atg ctt tgc gac gac gcc gtc 883
Lys Leu Ile Gly Asn Asp Pro Thr Gly Met Leu Cys Asp Asp Ala Val
250 255 260
cgc tca ggc ctg ctc cgc gtg ctg ccg tct gac ggc caa cca caa gtg 931
Arg Ser Gly Leu Leu Arg Val Leu Pro Ser Asp Gly GIn Pro Gln Val
265 270 275
gat ttg gtc ctg ccg atc gac cgc gcc gta ctg caa tca cgc act ccg 979
Asp Leu Val Leu Pro Ile Asp Arg Ala Val Leu Gln Ser Arg Thr Pro
280 285 290
ctc aac att ctg gcg cag ttg cac cac aag gca gcc gaa tat tac ggc 1027
Leu Asn Ile Leu Ala Gln Leu His His Lys Ala Ala Glu Tyr Tyr Gly
295 300 305
aag tgg aat caa aaa gat gcc caa ctg gag cac gaa gca ttt gct gca 1075
Lys Trp Asn Gln Lys Asp Ala Gln Leu Glu His Glu Ala Phe Ala Ala
310 315 320 325
att gat cca aat gat cca gca gtg cga gcc cta gcg cag cgc gga tat 1123
Ile Asp Pro Asn Asp Pro Ala Val Arg Ala Leu Ala Gln Arg Gly Tyr
330 335 340
gcg ttg ggt agg act ggc cac tgg atg gaa tcg gca cac gcc cta tct 1171
Ala Leu Gly Arg Thr Gly His Trp Met Glu Ser Ala His Ala Leu Ser
345 350 355
ctt gcc gcg aac cgc act gca cac caa gaa gaa tca aat aag tac ttg 1219
Leu Ala Ala Asn Arg Thr Ala His Gln Glu Glu Ser Asn Lys Tyr Leu
360 365 370
ctg gag tcc atc gat tca ctg atc gcc gcc gcc gat ctc ccc caa gct 1267
Leu Glu Ser Ile Asp Ser Leu Ile Ala Ala Ala Asp Leu Pro Gln Ala
375 380 385
cga tcc aga gca tcc acc ctt gat ctt gga gaa acc ggc att caa caa 1315
Arg Ser Arg Ala Ser Thr Leu Asp Leu Gly Glu Thr Gly Ile Gln Gln
390 395 400 405
gac tca atg ctg ggc tac ctg gca atc cac gaa ggc cgg cgc ctc gaa 1363
Asp Ser Met Leu Gly Tyr Leu Ala Ile His Glu Gly Arg Arg Leu Glu
410 415 420
gca cgc aat ctc ctt cat cgt gct tct gaa gaa ttg ctg gcg cag cac 1411
Ala Arg Asn Leu Leu His Arg Ala Ser Glu Glu Leu Leu Ala Gln His
425 430 435
ccg att gat ccg atc cac ggc ccc cgc atg gct cag cgc aaa gta ctg 1459
Pro Ile Asp Pro Ile His Gly Pro Arg Met Ala Gln Arg Lys Val Leu
440 445 450
tta aac tta gtg gac tgg aat cca gaa gaa ctc ctg gtg tgg gct gat 1507
Leu Asn Leu Val Asp Trp Asn Pro Glu Glu Leu Leu Val Trp Ala Asp
455 460 465
aga gca gtc gca tgg act gaa gag gat gct ggc gaa aag gtt gag gcc 1555
Arg Ala Val Ala Trp Thr Glu Glu Asp Ala Gly Glu Lys Val Glu Ala
470 475 480 485
caa gct att tcc ctc att gga caa tcc atc ctc gat ggc tgc ctc ccc 1603
Gln Ala Ile Ser Leu Ile Gly Gln Ser Ile Leu Asp Gly Cys Leu Pro
490 495 500
gaa gat aaa ccc atc ccc ggt gaa acc acc ctt cac gca caa cgc cgc 1651
Glu Asp Lys Pro Ile Pro Gly Glu Thr Thr Leu His Ala Gln Arg Arg
505 510 515
cac atg gca atg ggc tgg ctt tcc atg gtt cac gat gat cca gta act 1699
His Met Ala Met Gly Trp Leu Ser Met Val His Asp Asp Pro Val Thr
520 525 530
gca cgt caa aag ctt gaa cgt cgc aca tcc atc aat ggt tca gaa cgc 1747
Ala Arg Gln Lys Leu Glu Arg Arg Thr Ser Ile Asn Gly Ser Glu Arg
535 540 545
atc agt ttg tgg caa gac gga tgg ctg gct cgg tcc cta ctg ctg ctc 1795
Ile Ser Leu Trp Gln Asp Gly Trp Leu Ala Arg Ser Leu Leu Leu Leu
550 555 560 565
ggc gaa tgg gag tcc gca gca cgc acc gta gaa atc ggt ctg gcc cgc 1843
Gly Glu Trp Glu Ser Ala Ala Arg Thr Val Glu Ile Gly Leu Ala Arg
570 575 580
gcc gaa cag ttt ggc atc cgc ttc ctc gaa cca ctg tta ctg tgg tcg 1891
Ala Glu Gln Phe Gly Ile Arg Phe Leu Glu Pro Leu Leu Leu Trp Ser
585 590 595
ggc gcc aca att gca aca gcc cgc gga aac tct gac ttg gca cga aat 1939
Gly Ala Thr Ile Ala Thr Ala Arg Gly Asn Ser Asp Leu Ala Arg Asn
600 605 610
tac atg agc aga ctg tcc acc gat caa gac tcc ttc atc gtc caa tct 1987
Tyr Met Ser Arg Leu Ser Thr Asp Gln Asp Ser Phe Ile Val Gln Ser
615 620 625
atg cca tct gcg atg tgt cgc atg tgg gtc cac cgc cat aga aat gaa 2035
Met Pro Ser Ala Met Cys Arg Met Trp Val His Arg His Arg Asn Glu
630 635 640 645
atc ccc ggt gcg atc gtg gcc gga gaa caa ttg gaa aaa atc gcc gca 2083
Ile Pro Gly Ala Ile Val Ala Gly Glu Gln Leu Glu Lys Ile Ala Ala
650 655 660
cac aaa cac gtc aac gca cct gga ttc tgg cca tgg caa gac gtc cac 2131
His Lys His Val Asn Ala Pro Gly Phe Trp Pro Trp Gln Asp Val His
665 670 675
gca acg cat ctc atc cgc atc ggc gaa act gag cgc gcc cag gag tta 2179
Ala Thr His Leu Ile Arg Ile Gly Glu Thr Glu Arg Ala Gln Glu Leu
680 685 690
gtg aac tcc acg ctt gag gag ctc aga ggc tcc gat atc atg tct gcc 2227
Val Asn Ser Thr Leu Glu Glu Leu Arg Gly Ser Asp Ile Met Ser Ala
695 700 705
cac gca aaa att gcc gtt ccc gac gcc atg ttg atg atc cac cac gga 2275
His Ala Lys Ile Ala Val Pro Asp Ala Met Leu Met Ile His His Gly
710 715 720 725
gat gtg aaa aag gga ttt aag cgt ttc gac gac gcc ctc gat atg ctc 2323
Asp Val Lys Lys Gly Phe Lys Arg Phe Asp Asp Ala Leu Asp Met Leu
730 735 740
gat ccc ctc acc ctc ccc tac tat cgg gca cgc atc tgc ttt gaa tac 2371
Asp Pro Leu Thr Leu Pro Tyr Tyr Arg Ala Arg Ile Cys Phe Glu Tyr
745 750 755
ggc cag gcc ctg aga cgc cag ggg caa cgt cga cgt gct gat gaa caa 2419
Gly Gln Ala Leu Arg Arg Gln Gly Gln Arg Arg Arg Ala Asp Glu Gln
760 765 770
ttt gcc cgt gca gct tcc cta ttc caa gac atg ggc gcc gac gcg atg 2467
Phe Ala Arg Ala Ala Ser Leu Phe Gln Asp Met Gly Ala Asp Ala Met
775 780 785
gtc acc cta gcc aac cga gaa cgc cgg gtg ggt ggc ctt ggt caa cga 2515
Val Thr Leu Ala Asn Arg Glu Arg Arg Val Gly Gly Leu Gly Gln Arg
790 795 800 805
tcc gag caa gcc ggt ggg ctc acc cct cag gaa tat gaa att gcc cga 2563
Ser Glu Gln Ala Gly Gly Leu Thr Pro Gln Glu Tyr Glu Ile Ala Arg
810 815 820
tta gtg tca tct ggg cat gcc aac cga gag gtc gca cag gag ctt ttc 2611
Leu Val Ser Ser Gly His Ala Asn Arg Glu Val Ala Gln Glu Leu Phe
825 830 835
ctc tcg cct aag acc gtg gaa tac cat ctc acc cgg gtg tac aaa aag 2659
Leu Ser Pro Lys Thr Val Glu Tyr His Leu Thr Arg Val Tyr Lys Lys
840 845 850
ctc gga ata cgc aat cgg atg gaa ctt gcc gag gct ttg aag aag tac 2707
Leu Gly Ile Arg Asn Arg Met Glu Leu Ala Glu Ala Leu Lys Lys Tyr
855 860 865
tca cac gac gcc tagcagcgga tatgtttgcg gacaagtatc 2749
Ser His Asp Ala
870
<210>54
<211>873
<212>PRT
<213>谷氨酸棒状杆菌
<400>54
Val Gln Glu Val Ser Thr Gln Ile Lys Arg Leu Glu Ser Gly Glu Gly
1 5 10 15
Lys Leu Phe His Ile Ser Gly Gln Pro Gly Ala Gly Lys Thr Glu Phe
20 25 30
Gly Met Gln Leu Ile Asp Glu Leu Gln Gly Trp Thr Val Val Arg Ala
35 40 45
Thr Ser Leu Ser Trp Leu Lys Asn Ser Pro Arg Asn Leu Leu Gly His
50 55 60
Ile Ala His Lys Leu Gly Ala His Ser Ala Asn Ser Ile Arg Gly Val
65 70 75 80
Ile Asp Arg Leu Asp Ala Ser Thr Val Val Ile Val Asp Asp Val His
85 90 95
Trp Ala Asp Val Glu Ser Met Gln Lys Leu Ile Glu Tyr Ser Met Arg
100 105 110
Met Val Ser Gly Arg Phe Ala Leu Ile Met Ile Gly Leu Asp Glu Glu
115 120 125
Asn Leu Val Phe His Asp Glu Val Val Ser Leu Pro Ser Ile Ala Asp
130 135 140
Ser Thr Tyr Val Leu Pro Pro Met Ser Ile Glu Glu Ile Arg Gln Leu
145 150 155 160
Ala Leu Thr Asp Val Arg Gly Arg Ile Ser Thr Thr Thr Ala Thr Asp
165 170 175
Ile Gln Arg Ile Thr Gly Gly Ile Tyr Gly Arg Val Lys Glu Val Leu
180 185 190
His Ser Glu Ser Pro Asp His Trp Arg Met Pro Asn Pro Asn Ile Pro
195 200 205
Ile Pro Gln Ser Trp His Ala Asn Leu Leu Arg Arg Ile Thr Asn Glu
210 215 220
Glu Val Trp His Val Leu Leu Ala Val Ala Val Leu Pro Ser Gly Gly
225 230 235 240
Pro Ile Asp Leu Val Lys Leu Ile Gly Asn Asp Pro Thr Gly Met Leu
245 250 255
Cys Asp Asp Ala Val Arg Ser Gly Leu Leu Arg Val Leu Pro Ser Asp
260 265 270
Gly Gln Pro Gln Val Asp Leu Val Leu Pro Ile Asp Arg Ala Val Leu
275 280 285
Gln Ser Arg Thr Pro Leu Asn Ile Leu Ala Gln Leu His His Lys Ala
290 295 300
Ala Glu Tyr Tyr Gly Lys Trp Asn Gln Lys Asp Ala Gln Leu Glu His
305 310 315 320
Glu Ala Phe Ala Ala Ile Asp Pro Asn Asp Pro Ala Val Arg Ala Leu
325 330 335
Ala Gln Arg Gly Tyr Ala Leu Gly Arg Thr Gly His Trp Met Glu Ser
340 345 350
Ala His Ala Leu Ser Leu Ala Ala Asn Arg Thr Ala His Gln Glu Glu
355 360 365
Ser Asn Lys Tyr Leu Leu Glu Ser Ile Asp Ser Leu Ile Ala Ala Ala
370 375 380
Asp Leu Pro Gln Ala Arg Ser Arg Ala Ser Thr Leu Asp Leu Gly Glu
385 390 395 400
Thr Gly Ile Gln Gln Asp Ser Met Leu Gly Tyr Leu Ala Ile His Glu
405 410 415
Gly Arg Arg Leu Glu Ala Arg Asn Leu Leu His Arg Ala Ser Glu Glu
420 425 430
Leu Leu Ala Gln His Pro Ile Asp Pro Ile His Gly Pro Arg Met Ala
435 440 445
Gln Arg Lys Val Leu Leu Asn Leu Val Asp Trp Asn Pro Glu Glu Leu
450 455 460
Leu Val Trp Ala Asp Arg Ala Val Ala Trp Thr Glu Glu Asp Ala Gly
465 470 475 480
Glu Lys Val Glu Ala Gln Ala Ile Ser Leu Ile Gly Gln Ser Ile Leu
485 490 495
Asp Gly Cys Leu Pro Glu Asp Lys Pro Ile Pro Gly Glu Thr Thr Leu
500 505 510
His Ala Gln Arg Arg His Met Ala Met Gly Trp Leu Ser Met Val His
515 520 525
Asp Asp Pro Val Thr Ala Arg Gln Lys Leu Glu Arg Arg Thr Ser Ile
530 535 540
Asn Gly Ser Glu Arg Ile Ser Leu Trp Gln Asp Gly Trp Leu Ala Arg
545 550 555 560
Ser Leu Leu Leu Leu Gly Glu Trp Glu Ser Ala Ala Arg Thr Val Glu
565 570 575
Ile Gly Leu Ala Arg Ala Glu Gln Phe Gly Ile Arg Phe Leu Glu Pro
580 585 590
Leu Leu Leu Trp Ser Gly Ala Thr Ile Ala Thr Ala Arg Gly Asn Ser
595 600 605
Asp Leu Ala Arg Asn Tyr Met Ser Arg Leu Ser Thr Asp Gln Asp Ser
610 615 620
Phe Ile Val Gln Ser Met Pro Ser Ala Met Cys Arg Met Trp Val His
625 630 635 640
Arg His Arg Asn Glu Ile Pro Gly Ala Ile Val Ala Gly Glu Gln Leu
645 650 655
Glu Lys Ile Ala Ala His Lys His Val Asn Ala Pro Gly Phe Trp Pro
660 665 670
Trp Gln Asp Val His Ala Thr His Leu Ile Arg Ile Gly Glu Thr Glu
675 680 685
Arg Ala Gln Glu Leu Val Asn Ser Thr Leu Glu Glu Leu Arg Gly Ser
690 695 700
Asp Ile Met Ser Ala His Ala Lys Ile Ala Val Pro Asp Ala Met Leu
705 710 715 720
Met Ile His His Gly Asp Val Lys Lys Gly Phe Lys Arg Phe Asp Asp
725 730 735
Ala Leu Asp Met Leu Asp Pro Leu Thr Leu Pro Tyr Tyr Arg Ala Arg
740 745 750
Ile Cys Phe Glu Tyr Gly Gln Ala Leu Arg Arg Gln Gly Gln Arg Arg
755 760 765
Arg Ala Asp Glu Gln Phe Ala Arg Ala Ala Ser Leu Phe Gln Asp Met
770 775 780
Gly Ala Asp Ala Met Val Thr Leu Ala Asn Arg Glu Arg Arg Val Gly
785 790 795 800
Gly Leu Gly Gln Arg Ser Glu Gln Ala Gly Gly Leu Thr Pro Gln Glu
805 810 815
Tyr Glu Ile Ala Arg Leu Val Ser Ser Gly His Ala Asn Arg Glu Val
820 825 830
Ala Gln Glu Leu Phe Leu Ser Pro Lys Thr Val Glu Tyr His Leu Thr
835 840 845
Arg Val Tyr Lys Lys Leu Gly Ile Arg Asn Arg Met Glu LeuAla Glu
850 855 860
Ala Leu Lys Lys Tyr Ser His Asp Ala
865 870
<210>55
<211>1117
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(1087)
<223>RXA03201
<400>55
gcagggccgg gtcgtgatca cccacaacgg ctccagcgta gaactgattg acgatcttgc 60
acatgcagtc cgcgtagaaa aggttgaggg ctaacacacg atg aag ctt ctg gtt 115
Met Lys Leu Leu Val
1 5
acc ggt ggc gct ggc tat gtt ggc agc gtt gct gcc gct gtt ctt cta 163
Thr Gly Gly Ala Gly Tyr Val Gly Ser Val Ala Ala Ala Val Leu Leu
10 15 20
gaa cat ggg cac gat gtc aca atc att gac aat ttc agt acc ggc aac 211
Glu His Gly His Asp Val Thr Ile Ile Asp Asn Phe Ser Thr Gly Asn
25 30 35
cgt gaa gca gtc cct gct gac gct cgc ctg ata gaa ggt gac gtc aac 259
Arg Glu Ala Val Pro Ala Asp Ala Arg Leu Ile Glu Gly Asp Val Asn
40 45 50
gat gtc gtt gag gaa gtc ctt tct gaa ggc gga ttc gag ggt gtc gtc 307
Asp Val Val Glu Glu Val Leu Ser Glu Gly Gly Phe Glu Gly Val Val
55 60 65
cac ttc gct gct cgc tca ttg gta ggc gaa tca gtg gaa aag ccc aat 355
His Phe Ala Ala Arg Ser Leu Val Gly Glu Ser Val Glu Lys Pro Asn
70 75 80 85
gaa tac tgg cac gac aat gtg gtc acg gcg ctc acc ttg ctc gat gca 403
Glu Tyr Trp His Asp Asn Val Val Thr Ala Leu Thr Leu Leu Asp Ala
90 95 100
atg cgt gcc cat ggg gtg aac aac ctt gtg ttc tcc tcc act gct gct 451
Met Arg Ala His Gly Val Asn Asn Leu Val Phe Ser Ser Thr Ala Ala
105 110 115
acc tac ggt gaa cca gac gtt gtt ccc atc acc gag gac atg ccc acc 499
Thr Tyr Gly Glu Pro Asp Val Val Pro Ile Thr Glu Asp Met Pro Thr
120 125 130
cag ccc acc aat gct tat ggc gca acg aag ctg tcc atc gac tac gca 547
Gln Pro Thr Asn Ala Tyr Gly Ala Thr Lys Leu Ser Ile Asp Tyr Ala
135 140 145
att acg tcc tac gct gct gcc ttt ggt ctg gct gca acc agc ttg cgc 595
Ile Thr Ser Tyr Ala Ala Ala Phe Gly Leu Ala Ala Thr Ser Leu Arg
150 155 160 165
tac ttc aac gtt gct ggc gca tac gga aac atc ggt gaa aat cgt gaa 643
Tyr Phe Asn Val Ala Gly Ala Tyr Gly Asn Ile Gly Glu Asn Arg Glu
170 175 180
gtt gaa act cac ctc att cca cta gtg ctg cag gta gca acc ggt cac 691
Val Glu Thr His Leu Ile Pro Leu Val Leu Gln Val Ala Thr Gly His
185 190 195
cgc gag aaa acc ttc atg ttc ggc gat gac tgg cca act ccc gat gga 739
Arg Glu Lys Thr Phe Met Phe Gly Asp Asp Trp Pro Thr Pro Asp Gly
200 205 210
acc gcg gtc cgc gat tac atc cac att ctt gat ctt gcc aaa gca cac 787
Thr Ala Val Arg Asp Tyr Ile His Ile Leu Asp Leu Ala Lys Ala His
215 220 225
gtg ttg gct ttg gaa tcc aac gaa gcc ggt aag cac cgc att ttc aac 835
Val Leu Ala Leu Glu Ser Asn Glu Ala Gly Lys His Arg Ile Phe Asn
230 235 240 245
ctt ggt tcc gga gat ggc tac agc gtg aaa cag gtt gtg gag atg tgc 883
Leu Gly Ser Gly Asp Gly Tyr Ser Val Lys Gln Val Val Glu Met Cys
250 255 260
cgt gaa gta acc gga cat ccc atc ccg gca gag gtc gca ccc agg cgt 931
Arg Glu Val Thr Gly His Pro Ile Pro Ala Glu Val Ala Pro Arg Arg
265 270 275
gca gga gat cca gca aca cta att gca tct tct gag aag gca aag caa 979
Ala Gly Asp Pro Ala Thr Leu Ile Ala Ser Ser Glu Lys Ala Lys Gln
280 285 290
gaa ctg ggt tgg acc cct gag cac act gat ctg cgc acc atc gtt gag 1027
Glu Leu Gly Trp Thr Pro Glu His Thr Asp Leu Arg Thr Ile Val Glu
295 300 305
gac gct tgg gcg ttt aca tcc gca ctg ggc gat cgt tcc cat gct gca 1075
Asp Ala Trp Ala Phe Thr Ser Ala Leu Gly Asp Arg Ser His Ala Ala
310 315 320 325
aag aag aaa gct taagctttcg gatcggggac gtgaaagtcc 1117
Lys Lys Lys Ala
<210>56
<211>329
<212>PRT
<213>谷氨酸棒状杆菌
<400>56
Met Lys Leu Leu Val Thr Gly Gly Ala Gly Tyr Val Gly Ser Val Ala
1 5 10 15
Ala Ala Val Leu Leu Glu His Gly His Asp Val Thr Ile Ile Asp Asn
20 25 30
Phe Ser Thr Gly Asn Arg Glu Ala Val Pro Ala Asp Ala Arg Leu Ile
35 40 45
Glu Gly Asp Val Asn Asp Val Val Glu Glu Val Leu Ser Glu Gly Gly
50 55 60
Phe Glu Gly Val Val His Phe Ala Ala Arg Ser Leu Val Gly Glu Ser
65 70 75 80
Val Glu Lys Pro Asn Glu Tyr Trp His Asp Asn Val Val Thr Ala Leu
85 90 95
Thr Leu Leu Asp Ala Met Arg Ala His Gly Val Asn Asn Leu Val Phe
100 105 110
Ser Ser Thr Ala Ala Thr Tyr Gly Glu Pro Asp Val Val Pro Ile Thr
115 120 125
Glu Asp Met Pro Thr Gln Pro Thr Asn Ala Tyr Gly Ala Thr Lys Leu
130 135 140
Ser Ile Asp Tyr Ala Ile Thr Ser Tyr Ala Ala Ala Phe Gly Leu Ala
145 150 155 160
Ala Thr Ser Leu Arg Tyr Phe Asn Val Ala Gly Ala Tyr Gly Asn Ile
165 170 175
Gly Glu Asn Arg Glu Val Glu Thr His Leu Ile Pro Leu Val Leu Gln
180 185 190
Val Ala Thr Gly His Arg Glu Lys Thr Phe Met Phe Gly Asp Asp Trp
195 200 205
Pro Thr Pro Asp Gly Thr Ala Val Arg Asp Tyr Ile His Ile Leu Asp
210 215 220
Leu Ala Lys Ala His Val Leu Ala Leu Glu Ser Asn Glu Ala Gly Lys
225 230 235 240
His Arg Ile Phe Asn Leu Gly Ser Gly Asp Gly Tyr Ser Val Lys Gln
245 250 255
Val Val Glu Met Cys Arg Glu Val Thr Gly His Pro Ile Pro Ala Glu
260 265 270
Val Ala Pro Arg Arg Ala Gly Asp Pro Ala Thr Leu Ile Ala Ser Ser
275 280 285
Glu Lys Ala Lys Gln Glu Leu Gly Trp Thr Pro Glu His Thr Asp Leu
290 295 300
Arg Thr Ile Val Glu Asp Ala Trp Ala Phe Thr Ser Ala Leu Gly Asp
305 310 315 320
Arg Ser His Ala Ala Lys Lys Lys Ala
325
<210>57
<211>463
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(433)
<223>RXA03407
<400>57
gcccgaaaag tcctcaaagc atggaaagcc aaacagtgac cattgataga atatgtgtca 60
aaacatatat ttggatttgt gcttttactt gtctcgcgaa ttg gca ata gga gtc 115
Leu Ala Ile Gly Val
1 5
agc aac atg tca tta aag agt ttt agt tct cta gca caa tca gag aag 163
Ser Asn Met Ser Leu Lys Ser Phe Ser Ser Leu Ala Gln Ser Glu Lys
10 15 20
tca aca tgg agt aaa gaa gct aag caa gat tac gag gac gct tcc gcg 211
Ser Thr Trp Ser Lys Glu Ala Lys Gln Asp Tyr Glu Asp Ala Ser Ala
25 30 35
cta atc act caa gaa ctc gcc tca cga aaa gct ctt ggc caa aaa cta 259
Leu Ile Thr Gln Glu Leu Ala Ser Arg Lys Ala Leu Gly Gln Lys Leu
40 45 50
gtc aag gct cgg aaa caa cga gga gtc acc caa gtt caa ctt gct gaa 307
Val Lys Ala Arg Lys Gln Arg Gly Val Thr Gln Val Gln Leu Ala Glu
55 60 65
gct tct gga gtc caa cag gca gaa ata agc aag att gaa cgt ggc ctc 355
Ala Ser Gly Val Gln Gln Ala Glu Ile Ser Lys Ile Glu Arg Gly Leu
70 75 80 85
gcc aat ccc act ttt tcc aca ctt gaa agc ctc gca tcc cac cta gga 403
Ala Asn Pro Thr Phe Ser Thr Leu Glu Ser Leu Ala Ser His Leu Gly
90 95 100
ctt caa ttc act ttt aca gaa tca gcc gca tagcatccaa cccttaataa 453
Leu Gln Phe Thr Phe Thr Glu Ser Ala Ala
105 110
ataactccaa 463
<210>58
<211>111
<212>PRT
<213>谷氨酸棒状杆菌
<400>58
Leu Ala Ile Gly Val Ser Asn Met Ser Leu Lys Ser Phe Ser Ser Leu
1 5 10 15
Ala Gln Ser Glu Lys Ser Thr Trp Ser Lys Glu Ala Lys Gln Asp Tyr
20 25 30
Glu Asp Ala Ser Ala Leu Ile Thr Gln Glu Leu Ala Ser Arg Lys Ala
35 40 45
Leu Gly Gln Lys Leu Val Lys Ala Arg Lys Gln Arg Gly Val Thr Gln
50 55 60
Val Gln Leu Ala Glu Ala Ser Gly Val Gln Gln Ala Glu Ile Ser Lys
65 70 75 80
Ile Glu Arg Gly Leu Ala Asn Pro Thr Phe Ser Thr Leu Glu Ser Leu
85 90 95
Ala Ser His Leu Gly Leu Gln Phe Thr Phe Thr Glu Ser Ala Ala
100 105 110
<210>59
<211>487
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(457)
<223>RXA03629
<400>59
tgatggatca ccagtagatt aattcgtctt aacatcaacg aactaccccc attggcgagt 60
ttttagaaaa aattgaatga gaggatataa ggtaaaactc atg acg cac agt aag 115
Met Thr His Ser Lys
1 5
aaa gcg atg aaa gct aaa gct aaa gct ctt ctg gct tcc cag cgt gaa 163
Lys Ala Met Lys Ala Lys Ala Lys Ala Leu Leu Ala Ser Gln Arg Glu
10 15 20
ttc ctt gac tca ctt gta gct ttg aga aag aaa gct ggc att agc caa 211
Phe Leu Asp Ser Leu Val Ala Leu Arg Lys Lys Ala Gly Ile Ser Gln
25 30 35
gac gag gtg gct aat cgg atg ggc gtt tcc cag agt gct att tct caa 259
Asp Glu Val Ala Asn Arg Met Gly Val Ser Gln Ser Ala Ile Ser Gln
40 45 50
ttt gag cac tac gat gca aat ccc act ctg tct acg atc cga cgc tat 307
Phe Glu His Tyr Asp Ala Asn Pro Thr Leu Ser Thr Ile Arg Arg Tyr
55 60 65
gcg cta gcg gta gat gct tct ata tct tat agg gtt agc tct tct gcc 355
Ala Leu Ala Val Asp Ala Ser Ile Ser Tyr Arg Val Ser Ser Ser Ala
70 75 80 85
acc ctc tat caa gag tat gag tcg agg aca aac aat cac gta tcc gtg 403
Thr Leu Tyr Gln Glu Tyr Glu Ser Arg Thr Asn Asn His Val Ser Val
90 95 100
tcc ggc cag gaa gct gcc ccg ccc tac gtt gat tgg gaa aag ccc att 451
Ser Gly Gln Glu Ala Ala Pro Pro Tyr Val Asp Trp Glu Lys Pro Ile
105 ll0 115
ttg gcg tagtgtttcg ggttttatag gttgagattg 487
Leu Ala
<210>60
<211>119
<212>PRT
<213>谷氨酸棒状杆菌
<400>60
Met Thr His Ser Lys Lys Ala Met Lys Ala Lys Ala Lys Ala Leu Leu
1 5 10 15
Ala Ser Gln Arg Glu Phe Leu Asp Ser Leu Val Ala Leu Arg Lys Lys
20 25 30
Ala Gly Ile Ser Gln Asp Glu Val Ala Asn Arg Met Gly Val Ser Gln
35 40 45
Ser Ala Ile Ser Gln Phe Glu His Tyr Asp Ala Asn Pro Thr Leu Ser
50 55 60
Thr Ile Arg Arg Tyr Ala Leu Ala Val Asp Ala Ser Ile Ser Tyr Arg
65 70 75 80
Val Ser Ser Ser Ala Thr Leu Tyr Gln Glu Tyr Glu Ser Arg Thr Asn
85 90 95
Asn His Val Ser Val Ser Gly Gln Glu Ala Ala Pro Pro Tyr Val Asp
100 105 110
Trp Glu Lys Pro Ile Leu Ala
115
<210>61
<211>892
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(862)
<223>RXA03928
<400>61
tggtggcggc gggctgcgtc gaaaagcgaa aatcaacaag tttgcaacac ctcagtgcca 60
agagtggtta aggtgatggt gatcacgcta tagttgcgcc atg gga aag aca tat 115
Met Gly Lys Thr Tyr
1 5
gtg ggg tcc agg ctg cgc caa ctg cgc cgc gaa aga gac ctg agc cag 163
Val Gly Ser Arg Leu Arg Gln Leu Arg Arg Glu Arg Asp Leu Ser Gln
10 15 20
gca tcc tta gca gca acc ctt ggc tta tct gca agt tat gta aat cag 211
Ala Ser Leu Ala Ala Thr Leu Gly Leu Ser Ala Ser Tyr Val Asn Gln
25 30 35
att gag cac gac gta cgc ccg ctc acc gta ccg gtg tta ttg cgc atc 259
Ile Glu His Asp Val Arg Pro Leu Thr Val Pro Val Leu Leu Arg Ile
40 45 50
acc gag gcg ttc ggc gta gac gca acg ttt ttc tcc cgc gac gat gac 307
Thr Glu Ala Phe Gly Val Asp Ala Thr Phe Phe Ser Arg Asp Asp Asp
55 60 65
tcc cgc ctg ctc gcc gag gtc caa gac gtc atg ctg gac cgg gag atc 355
Ser Arg Leu Leu Ala Glu Val Gln Asp Val Met Leu Asp Arg Glu Ile
70 75 80 85
aat cct gcg aac gtg gag ctg caa gag ctt tcg gag atg gtg tac aac 403
Asn Pro Ala Asn Val Glu Leu Gln Glu Leu Ser Glu Met Val Tyr Asn
90 95 100
cac ccc caa cta gcg cgc gcg atg gtg gaa atg cac cag cgt tac cga 451
His Pro Gln Leu Ala Arg Ala Met Val Glu Met His Gln Arg Tyr Arg
105 110 115
aac gtg cgc gat aag ttc tcc atc gca gtg gat aat cgc acc aac acg 499
Asn Val Arg Asp Lys Phe Ser Ile Ala Val Asp Asn Arg Thr Asn Thr
120 125 130
cct gag gaa cgc cgt ccc atc gcg gag gcc gtg agc atg ccg cac gaa 547
Pro Glu Glu Arg Arg Pro Ile Ala Glu Ala Val Ser Met Pro His Glu
135 140 145
gag gtc cgc gat ttc att tac gcc cgc caa aac tac ttc gat gcc ctt 595
Glu Val Arg Asp Phe Ile Tyr Ala Arg Gln Asn Tyr Phe Asp Ala Leu
150 155 160 165
gac cgc cgc gcc gaa gcc atg cgc cgc gca act ggg ctg gca gcc gta 643
Asp Arg Arg Ala Glu Ala Met Arg Arg Ala Thr Gly Leu Ala Ala Val
170 175 180
cga ttc ccg cgc cat gga aga ttc gat cgc ccg ccg cct gca aat gga 691
Arg Phe Pro Arg His Gly Arg Phe Asp Arg Pro Pro Pro Ala Asn Gly
185 190 195
tca cga tgt cac cat cac ctc ctc caa aga gga atc cgg cac gct gca 739
Ser Arg Cys His His His Leu Leu Gln Arg Gly Ile Arg His Ala Ala
200 205 210
cca ctt cga ccc cga gac gcg tct gct gac aat cca cgc acg cct caa 787
Pro Leu Arg Pro Arg Asp Ala Ser Ala Asp Asn Pro Arg Thr Pro Gln
215 220 225
ccc cgg gca acg cgc ctt ccg cat ggc cac cga act cgg cta cct aga 835
Pro Arg Ala Thr Arg Leu Pro His Gly His Arg Thr Arg Leu Pro Arg
230 235 240 245
agc caa cga cct cat cga agg tat cgt tgacgacggc atctggtcca 882
Ser Gln Arg Pro His Arg Arg Tyr Arg
250
cccccgaagc 892
<210>62
<211>254
<212>PRT
<213>谷氨酸棒状杆菌
<400>62
Met Gly Lys Thr Tyr Val Gly Ser Arg Leu Arg Gln Leu Arg Arg Glu
1 5 10 15
Arg Asp Leu Ser Gln Ala Ser Leu Ala Ala Thr Leu Gly Leu Ser Ala
20 25 30
Ser Tyr Val Asn Gln Ile Glu His Asp Val Arg Pro Leu Thr Val Pro
35 40 45
Val Leu Leu Arg Ile Thr Glu Ala Phe Gly Val Asp Ala Thr Phe Phe
50 55 60
Ser Arg Asp Asp Asp Ser Arg Leu Leu Ala Glu Val Gln Asp Val Met
65 70 75 80
Leu Asp Arg Glu Ile Asn Pro Ala Asn Val Glu Leu Gln Glu Leu Ser
85 90 95
Glu Met Val Tyr Asn His Pro Gln Leu Ala Arg Ala Met Val Glu Met
100 105 110
His Gln Arg Tyr Arg Asn Val Arg Asp Lys Phe Ser Ile Ala Val Asp
115 120 125
Asn Arg Thr Asn Thr Pro Glu Glu Arg Arg Pro Ile Ala Glu Ala Val
130 135 140
Ser Met Pro His Glu Glu Val Arg Asp Phe Ile Tyr Ala Arg Gln Asn
145 150 155 160
Tyr Phe Asp Ala Leu Asp Arg Arg Ala Glu Ala Met Arg Arg Ala Thr
165 170 175
Gly Leu Ala Ala Val Arg Phe Pro Arg His Gly Arg Phe Asp Arg Pro
180 185 190
Pro Pro Ala Asn Gly Ser Arg Cys His His His Leu Leu Gln Arg Gly
195 200 205
Ile Arg His Ala Ala Pro Leu Arg Pro Arg Asp Ala Ser Ala Asp Asn
210 215 220
Pro Arg Thr Pro Gln Pro Arg Ala Thr Arg Leu Pro His Gly His Arg
225 230 235 240
Thr Arg Leu Pro Arg Ser Gln Arg Pro His Arg Arg Tyr Arg
245 250
<210>63
<211>499
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(469)
<223>RXA04129
<400>63
ctattttttc atttccctca taaaaggttt atatagaagg taaaatagca agcgtgctag 60
atccattcca gacctagcct aaggacggaa ggacttcccc gtg gct caa cac tca 115
Val Ala Gln His Ser
1 5
aat cgg aat ttc gtc acc cct gca gaa gag tcc gaa tcc aca gca tca 163
Asn Arg Asn Phe Val Thr Pro Ala Glu Glu Ser Glu Ser Thr Ala Ser
10 15 20
gct gag ctt cag aaa ctt gca aca gca aaa aac atc aaa gcg atc agc 211
Ala Glu Leu Gln Lys Leu Ala Thr Ala Lys Asn Ile Lys Ala Ile Ser
25 30 35
ctg ctg att agg gca cta gat tcc ccc tta aga atc gaa atc atc ctc 259
Leu Leu Ile Arg Ala Leu Asp Ser Pro Leu Arg Ile Glu Ile Ile Leu
40 45 50
gcc ctc aat gaa agg ccc cac tac gtc cac gaa ttg gtc aag cta gta 307
Ala Leu Asn Glu Arg Pro His Tyr Val His Glu Leu Val Lys Leu Val
55 60 65
aaa agt tcg caa cca cta gtg agc cag cac ctc aaa gtc ctt aaa act 355
Lys Ser Ser Gln Pro Leu Val Ser Gln His Leu Lys Val Leu Lys Thr
70 75 80 85
gca ggt atc gtc gac gca gaa cgt caa ggc cgg caa atg act tat tca 403
Ala Gly Ile Val Asp Ala Glu Arg Gln Gly Arg Gln Met Thr Tyr Ser
90 95 100
ctt gcc gaa cca ctc gtc ctc gac ata ctt ctc ctg gca cta aac gca 451
Leu Ala Glu Pro Leu Val Leu Asp Ile Leu Leu Leu Ala Leu Asn Ala
105 110 115
ggg gtt gac aca tct ggg tagactatcg aagtacattt tgtgtcattg 499
Gly Val Asp Thr Ser Gly
120
<210>64
<211>123
<212>PRT
<213>谷氨酸棒状杆菌
<400>64
Val Ala Gln His Ser Asn Arg Asn Phe Val Thr Pro Ala Glu Glu Ser
1 5 10 15
Glu Ser Thr Ala Ser Ala Glu Leu Gln Lys Leu Ala Thr Ala Lys Asn
20 25 30
Ile Lys Ala Ile Ser Leu Leu Ile Arg Ala Leu Asp Ser Pro Leu Arg
35 40 45
Ile Glu Ile Ile Leu Ala Leu Asn Glu Arg Pro His Tyr Val His Glu
50 55 60
Leu Val Lys Leu Val Lys Ser Ser Gln Pro Leu Val Ser Gln His Leu
65 70 75 80
Lys Val Leu Lys Thr Ala Gly Ile Val Asp Ala Glu Arg Gln Gly Arg
85 90 95
Gln Met Thr Tyr Ser Leu Ala Glu Pro Leu Val Leu Asp Ile Leu Leu
100 105 110
Leu Ala Leu Asn Ala Gly Val Asp Thr Ser Gly
115 120
<210>65
<211>784
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(754)
<223>RXA04350
<400>65
catcggggga aacggtgttg caacgcacca tgcgggtgtt gcactgcttt gatgcagaaa 60
acgcgaggtt ggggcctcgg aaatcgcgcg tcgcagtggt ttg cct agc tcg acg 115
Leu Pro Ser Ser Thr
1 5
gcg cac cgt ttg gtc gtg gaa ttg acg tcg gtg aag ttg ctg gag cgc 163
Ala His Arg Leu Val Val Glu Leu Thr Ser Val Lys Leu Leu Glu Arg
10 15 20
ttt tct gac ggc aaa tat ggc atc ggc acg cat gcg tgg gag acg ttt 211
Phe Ser Asp Gly Lys Tyr Gly Ile Gly Thr His Ala Trp Glu Thr Phe
25 30 35
gtg agg gcg aat cct ttg gag cgc atg agg ctg caa gct cag acc atc 259
Val Arg Ala Asn Pro Leu Glu Arg Met Arg Leu Gln Ala Gln Thr Ile
40 45 50
ttg agt gat gtg cgt gaa gag ctg ggg caa tac gtg tgt ttg gcg gtg 307
Leu Ser Asp Val Arg Glu Glu Leu Gly Gln Tyr Val Cys Leu Ala Val
55 60 65
ccg gat ttt gtg gat cgt acg att ttg tat gtg gag cgt ttc gat tcg 355
Pro Asp Phe Val Asp Arg Thr Ile Leu Tyr Val Glu Arg Phe Asp Ser
70 75 80 85
acg gat agt cag ctg cgt ctt ttg ggc agg cat gcg ggg cgt ttg gat 403
Thr Asp Ser Gln Leu Arg Leu Leu Gly Arg His Ala Gly Arg Leu Asp
90 95 100
ttg cac acc aca tca ttg ggg ttg gtg atg ttg gcg ttt gct tcg ccg 451
Leu His Thr Thr Ser Leu Gly Leu Val Met Leu Ala Phe Ala Ser Pro
105 110 115
caa acc att aag gca gtg acg agt tcg ccg ttt aag gat tcc att tcg 499
Gln Thr Ile Lys Ala Val Thr Ser Ser Pro Phe Lys Asp Ser Ile Ser
120 125 130
ggg gat att acg gat ggt gct gct gta gct gcg atg ctc ccg gaa att 547
Gly Asp Ile Thr Asp Gly Ala Ala Val Ala Ala Met Leu Pro Glu Ile
135 140 145
cgc gcg acg ggg cat ttt gta ttt gtg ggt ggt ctg att ccg gag aat 595
Arg Ala Thr Gly His Phe Val Phe Val Gly Gly Leu Ile Pro Glu Asn
150 155 160 165
acg gcg gtt gcg gct ccg gtt ttt gat cga aaa ggg gtt gtt cgg gca 643
Thr Ala Val Ala Ala Pro Val Phe Asp Arg Lys Gly Val Val Arg Ala
170 175 180
tca gtg ggg gta gtg gcg agg aac gat gaa att gat gtg aat cag gcg 691
Ser Val Gly Val Val Ala Arg Asn Asp Glu Ile Asp Val Asn Gln Ala
185 190 195
gtt tct gtg gtg ctt gat gca tgc caa aag ctc agc gta agg cta caa 739
Val Ser Val Val Leu Asp Ala Cys Gln Lys Leu Ser Val Arg Leu Gln
200 205 210
aaa gac tct cta tat taagttcgca tttcgaacgc gcccatttgg 784
Lys Asp Ser Leu Tyr
215
<210>66
<211>218
<212>PRT
<213>谷氨酸棒状杆菌
<400>66
Leu Pro Ser Ser Thr Ala His Arg Leu Val Val Glu Leu Thr Ser Val
1 5 10 15
Lys Leu Leu Glu Arg Phe Ser Asp Gly Lys Tyr Gly Ile Gly Thr His
20 25 30
Ala Trp Glu Thr Phe Val Arg Ala Asn Pro Leu Glu Arg Met Arg Leu
35 40 45
Gln Ala Gln Thr Ile Leu Ser Asp Val Arg Glu Glu Leu Gly Gln Tyr
50 55 60
Val Cys Leu Ala Val Pro Asp Phe Val Asp Arg Thr Ile Leu Tyr Val
65 70 75 80
Glu Arg Phe Asp Ser Thr Asp Ser Gln Leu Arg Leu Leu Gly Arg His
85 90 95
Ala Gly Arg Leu Asp Leu His Thr Thr Ser Leu Gly Leu Val Met Leu
100 105 110
Ala Phe Ala Ser Pro Gln Thr Ile Lys Ala Val Thr Ser Ser Pro Phe
115 120 125
Lys Asp Ser Ile Ser Gly Asp Ile Thr Asp Gly Ala Ala Val Ala Ala
130 135 140
Met Leu Pro Glu Ile Arg Ala Thr Gly His Phe Val Phe Val Gly Gly
145 150 155 160
Leu Ile Pro Glu Asn Thr Ala Val Ala Ala Pro Val Phe Asp Arg Lys
165 170 175
Gly Val Val Arg Ala Ser Val Gly Val Val Ala Arg Asn Asp Glu Ile
180 185 190
Asp Val Asn Gln Ala Val Ser Val Val Leu Asp Ala Cys Gln Lys Leu
195 200 205
Ser Val Arg Leu Gln Lys Asp Ser Leu Tyr
210 215
<210>67
<211>904
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(874)
<223>RXA04363
<400>67
cacaagtcct cccgattctg gtgcgtgtga tgtcataaac acataatcga ggtgagccaa 60
gttacaatca aagggattcc gctatctgga aagagtgatt atg gat aac gtc gcc 115
Met Asp Asn Val Ala
1 5
ccc acc cag ggg tta ccc ccg aaa gaa ttt cta agc tct gtc gac att 163
Pro Thr Gln Gly Leu Pro Pro Lys Glu Phe Leu Ser Ser Val Asp Ile
10 15 20
gca ttg cag ctc att ttg ctg ctc cgt gac tct gga agt ttg acc att 211
Ala Leu Gln Leu Ile Leu Leu Leu Arg Asp Ser Gly Ser Leu Thr Ile
25 30 35
tcc ggt gcc gcc gaa acc ctt ggg gtg ggc gcg tct acg atc cat cga 259
Ser Gly Ala Ala Glu Thr Leu Gly Val Gly Ala Ser Thr Ile His Arg
40 45 50
tcc atg tca atg ctg gtc tac cgg ggt ttt gca gtc aga agc gag tcc 307
Ser Met Ser Met Leu Val Tyr Arg Gly Phe Ala Val Arg Ser Glu Ser
55 60 65
cgc acc tac ctt cca ggc tct gca ttg gcg acc tcc gcg ctg cag cca 355
Arg Thr Tyr Leu Pro Gly Ser Ala Leu Ala Thr Ser Ala Leu Gln Pro
70 75 80 85
ggc ctt ggc gct gac ttg acg aaa aaa tgc agc cac tac atg gaa tca 403
Gly Leu Gly Ala Asp Leu Thr Lys Lys Cys Ser His Tyr Met Glu Ser
90 95 100
atc ggc aag gaa act ggc gaa aca acc cac ttg gtg att ctg cag gga 451
Ile Gly Lys Glu Thr Gly Glu Thr Thr His Leu Val Ile Leu Gln Gly
105 110 115
gat agc gtt cac ttt att cac agt gtt gaa ggt tcc ctg ccg gtg cgc 499
Asp Ser Val His Phe Ile His Ser Val Glu Gly Ser Leu Pro Val Arg
120 125 130
gtg ggc aat cgc cga ggt caa gtc atg ccc gcc atc caa aat tca ggt 547
Val Gly Asn Arg Arg Gly Gln Val Met Pro Ala Ile Gln Asn Ser Gly
135 140 145
gga tta gtg atg ctt gca gag atg tca gcc cgg gag ctt cgg gca ctg 595
Gly Leu Val Met Leu Ala Glu Met Ser Ala Arg Glu Leu Arg Ala Leu
150 155 160 165
tat tcc agc ctg ggc gat gag gaa ttt gag aat tta aga aag cgt ctt 643
Tyr Ser Ser Leu Gly Asp Glu Glu Phe Glu Asn Leu Arg Lys Arg Leu
170 175 180
cgc agg acc cgg gat cga ggc cat ggc gca aac ttt ggc ttc ttt gag 691
Arg Arg Thr Arg Asp Arg Gly His Gly Ala Asn Phe Gly Phe Phe Glu
185 190 195
cag gac gtt agt gca gtt gcg gag cct tta ctc aac gat gtg ggt gat 739
Gln Asp Val Ser Ala Val Ala Glu Pro Leu Leu Asn Asp Val Gly Asp
200 205 210
gtt tta ggt gca att aca gtg gct gtg ccg tcg aat cgg ttc cgg gag 787
Val Leu Gly Ala Ile Thr Val Ala Val Pro Ser Asn Arg Phe Arg Glu
215 220 225
gtc tat ccg aag gcg gtg cag gtt ttg gaa cga cat atg cgg gat ctg 835
Val Tyr Pro Lys Ala Val Gln Val Leu Glu Arg His Met Arg Asp Leu
230 235 240 245
aac aag gct tta gct gat tac cga gtg ccc gaa aaa ggg tgaatggggg 884
Asn Lys Ala Leu Ala Asp Tyr Arg Val Pro Glu Lys Gly
250 255
gaattttcag agctgtcaca 904
<210>68
<211>258
<212>PRT
<213>谷氨酸棒状杆菌
<400>68
Met Asp Asn Val Ala Pro Thr Gln Gly Leu Pro Pro Lys Glu Phe Leu
1 5 10 15
Ser Ser Val Asp Ile Ala Leu Gln Leu Ile Leu Leu Leu Arg Asp Ser
20 25 30
Gly Ser Leu Thr Ile Ser Gly Ala Ala Glu Thr Leu Gly Val Gly Ala
35 40 45
Ser Thr Ile His Arg Ser Met Ser Met Leu Val Tyr Arg Gly Phe Ala
50 55 60
Val Arg Ser Glu Ser Arg Thr Tyr Leu Pro Gly Ser Ala Leu Ala Thr
65 70 75 80
Ser Ala Leu Gln Pro Gly Leu Gly Ala Asp Leu Thr Lys Lys Cys Ser
85 90 95
His Tyr Met Glu Ser Ile Gly Lys Glu Thr Gly Glu Thr Thr His Leu
100 105 110
Val Ile Leu Gln Gly Asp Ser Val His Phe Ile His Ser Val Glu Gly
115 120 125
Ser Leu Pro Val Arg Val Gly Asn Arg Arg Gly Gln Val Met Pro Ala
130 135 140
Ile Gln Asn Ser Gly Gly Leu Val Met Leu Ala Glu Met Ser Ala Arg
145 150 155 160
Glu Leu Arg Ala Leu Tyr Ser Ser Leu Gly Asp Glu Glu Phe Glu Asn
165 170 175
Leu Arg Lys Arg Leu Arg Arg Thr Arg Asp Arg Gly His Gly Ala Asn
180 185 190
Phe Gly Phe Phe Glu Gln Asp Val Ser Ala Val Ala Glu Pro Leu Leu
195 200 205
Asn Asp Val Gly Asp Val Leu Gly Ala Ile Thr Val Ala Val Pro Ser
210 215 220
Asn Arg Phe Arg Glu Val Tyr Pro Lys Ala Val Gln Val Leu Glu Arg
225 230 235 240
His Met Arg Asp Leu Asn Lys Ala Leu Ala Asp Tyr Arg Val Pro Glu
245 250 255
Lys Gly
<210>69
<211>973
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(943)
<223>RXA04620
<400>69
cccaaagggg aagtacactg tacccttgtc gaatgattgt tactcgtgac gcgccctatg 60
ggtgtaccag cacgggtgta aagcaggagg aaatctgaag gtg gat acc cag cgg 115
Val Asp Thr Gln Arg
1 5
att aaa gat gac gaa gat gct att cgt tcg gcg ctg aca tcg ctg aaa 163
Ile Lys Asp Asp Glu Asp Ala Ile Arg Ser Ala Leu Thr Ser Leu Lys
10 15 20
acc gca aca ggc atc cca gtc acc atg ttc gcc act gtg ttg cag gac 211
Thr Ala Thr Gly Ile Pro Val Thr Met Phe Ala Thr Val Leu Gln Asp
25 30 35
aat cgc ctg caa att act cag tgg gtt ggg ttg cgt acc ccg gct ctg 259
Asn Arg Leu Gln Ile Thr Gln Trp Val Gly Leu Arg Thr Pro Ala Leu
40 45 50
cag aat ctg gtc att gaa cca ggt gtg ggc gtt ggt gga cgc gtc gtc 307
Gln Asn Leu Val Ile Glu Pro Gly Val Gly Val Gly Gly Arg Val Val
55 60 65
gca acc cgt cgt ccg gtt ggt gtg agt gat tac acc agg gca aat gtc 355
Ala Thr Arg Arg Pro Val Gly Val Ser Asp Tyr Thr Arg Ala Asn Val
70 75 80 85
att tca cat gag aag gat tcc gcg att cag gat gag ggc ctt cat tcc 403
Ile Ser His Glu Lys Asp Ser Ala Ile Gln Asp Glu Gly Leu His Ser
90 95 100
att gtc gca gtt ccc gtg atc gtg cac cgc gaa att cgt ggc gtt ttg 451
Ile Val Ala Val Pro Val Ile Val His Arg Glu Ile Arg Gly Val Leu
105 110 115
tat gtt ggc gtt cac tct gcg gtg cgt ctc ggc gac act gtt att gaa 499
Tyr Val Gly Val His Ser Ala Val Arg Leu Gly Asp Thr Val Ile Glu
120 125 130
gaa gtc acc atg act gcg cgc acg ttg gaa caa aac ctg gcg atc aac 547
Glu Val Thr Met Thr Ala Arg Thr Leu Glu Gln Asn Leu Ala Ile Asn
135 140 145
tcc gcg ctt cgc cgc aat ggc gtt cct gat ggt cgc ggt tcc ctc aaa 595
Ser Ala Leu Arg Arg Asn Gly Val Pro Asp Gly Arg Gly Ser Leu Lys
150 155 160 165
gct aac cgc gtg atg aat ggg gcg gag tgg gag cag gtt cgt tcc act 643
Ala Asn Arg Val Met Asn Gly Ala Glu Trp Glu Gln Val Arg Ser Thr
170 175 180
cat tcc aag ctg cgc atg ctg gca aat cgt gtg acc gat gag gat ctg 691
His Ser Lys Leu Arg Met Leu Ala Asn Arg Val Thr Asp Glu Asp Leu
185 190 195
cgc cgc gat ttg gaa gag ctt tgc gat cag atg gtc acc cca gtc cgc 739
Arg Arg Asp Leu Glu Glu Leu Cys Asp Gln Met Val Thr Pro Val Arg
200 205 210
atc aag cag acc acc aag ctg tcc gcg cgt gag ttg gac gtg ctg gct 787
Ile Lys Gln Thr Thr Lys Leu Ser Ala Arg Glu Leu Asp Val Leu Ala
215 220 225
tgt gtc gcg ctc ggt cac acc aac gtc gaa gct gct gaa gag atg ggc 835
Cys Val Ala Leu Gly His Thr Asn Val Glu Ala Ala Glu Glu Met Gly
230 235 240 245
atc ggc gcg gaa acc gtc aag agc tac ctg cgc tcg gtc atg cgc aag 883
Ile Gly Ala Glu Thr Val Lys Ser Tyr Leu Arg Ser Val Met Arg Lys
250 255 260
ctc ggc gcc cac acg cgc tac gag gca gtc aac gca gca cgc cgg atc 931
Leu Gly Ala His Thr Arg Tyr Glu Ala Val Asn Ala Ala Arg Arg Ile
265 270 275
ggc gca ctg cct taaaaagatt ttgctttacg acgccaccct 973
Gly Ala Leu Pro
280
<210>70
<211>281
<212>PRT
<213>谷氨酸棒状杆菌
<400>70
Val Asp Thr Gln Arg Ile Lys Asp Asp Glu Asp Ala Ile Arg Ser Ala
1 5 10 15
Leu Thr Ser Leu Lys Thr Ala Thr Gly Ile Pro Val Thr Met Phe Ala
20 25 30
Thr Val Leu Gln Asp Asn Arg Leu Gln Ile Thr Gln Trp Val Gly Leu
35 40 45
Arg Thr Pro Ala Leu Gln Asn Leu Val Ile Glu Pro Gly Val Gly Val
50 55 60
Gly Gly Arg Val Val Ala Thr Arg Arg Pro Val Gly Val Ser Asp Tyr
65 70 75 80
Thr Arg Ala Asn Val Ile Ser His Glu Lys Asp Ser Ala Ile Gln Asp
85 90 95
Glu Gly Leu His Ser Ile Val Ala Val Pro Val Ile Val His Arg Glu
100 105 110
Ile Arg Gly Val Leu Tyr Val Gly Val His Ser Ala Val Arg Leu Gly
115 120 125
Asp Thr Val Ile Glu Glu Val Thr Met Thr Ala Arg Thr Leu Glu Gln
130 135 140
Asn Leu Ala Ile Asn Ser Ala Leu Arg Arg Asn Gly Val Pro Asp Gly
145 150 155 160
Arg Gly Ser Leu Lys Ala Asn Arg Val Met Asn Gly Ala Glu Trp Glu
165 170 175
Gln Val Arg Ser Thr His Ser Lys Leu Arg Met Leu Ala Asn Arg Val
180 185 190
Thr Asp Glu Asp Leu Arg Arg Asp Leu Glu Glu Leu Cys Asp Gln Met
195 200 205
Val Thr Pro Val Arg Ile Lys Gln Thr Thr Lys Leu Ser Ala Arg Glu
210 215 220
Leu Asp Val Leu Ala Cys Val Ala Leu Gly His Thr Asn Val Glu Ala
225 230 235 240
Ala Glu Glu Met Gly Ile Gly Ala Glu Thr Val Lys Ser Tyr Leu Arg
245 250 255
Ser Val Met Arg Lys Leu Gly Ala His Thr Arg Tyr Glu Ala Val Asn
260 265 270
Ala Ala Arg Arg Ile Gly Ala Leu Pro
275 280
<210>71
<211>478
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(101)..(448)
<223>RXA06017
<400>71
cgatcacccc ttcgccggcc gccaagactt agaactatcc gccttagaag acctcgatct 60
gctgcttctc gacgacggac actgcctcca cgaccaaatt gtg gac ctg tgc cgc 115
Val Asp Leu Cys Arg
1 5
cgc gga gac atc aac ccc att agc tcc act act gct gtc acc cgc gca 163
Arg Gly Asp Ile Asn Pro Ile Ser Ser Thr Thr Ala Val Thr Arg Ala
10 15 20
tcc agc ctt acc acc gtc atg cag ctc gtc gtc gcc ggc ctt gga tcc 211
Ser Ser Leu Thr Thr Val Met Gln Leu Val Val Ala Gly Leu Gly Ser
25 30 35
acc ttg gtc cca atc agc gca atc cca tgg gaa tgc acc cga cca gga 259
Thr Leu Val Pro Ile Ser Ala Ile Pro Trp Glu Cys Thr Arg Pro Gly
40 45 50
ctg gca aca gcc aac ttc aac tct gat gtc acc gca aac cgc cgc att 307
Leu Ala Thr Ala Asn Phe Asn Ser Asp Val Thr Ala Asn Arg Arg Ile
55 60 65
gga ttg gtg tac cgt tcc tct tct tct cgc gcc gaa gag ttc gaa cag 355
Gly Leu Val Tyr Arg Ser Ser Ser Ser Arg Ala Glu Glu Phe Glu Gln
70 75 80 85
ttt gca ctc att ttg cag cgc gct ttc caa gaa gcc gtc gcg ctt gct 403
Phe Ala Leu Ile Leu Gln Arg Ala Phe Gln Glu Ala Val Ala Leu Ala
90 95 100
gcc tca act ggc atc acc ttg aag caa aat gtc gcg gta gcg cag 448
Ala Ser Thr Gly Ile Thr Leu Lys Gln Asn Val Ala Val Ala Gln
105 110 115
taagtttttc tagaggtttt ccagagtcag 478
<210>72
<211>116
<212>PRT
<213>谷氨酸棒状杆菌
<400>72
Val Asp Leu Cys Arg Arg Gly Asp Ile Asn Pro Ile Ser Ser Thr Thr
1 5 10 15
Ala Val Thr Arg Ala Ser Ser Leu Thr Thr Val Met Gln Leu Val Val
20 25 30
Ala Gly Leu Gly Ser Thr Leu Val Pro Ile Ser Ala Ile Pro Trp Glu
35 40 45
Cys Thr Arg Pro Gly Leu Ala Thr Ala Asn Phe Asn Ser Asp Val Thr
50 55 60
Ala Asn Arg Arg Ile Gly Leu Val Tyr Arg Ser Ser Ser Ser Arg Ala
65 70 75 80
Glu Glu Phe Glu Gln Phe Ala Leu Ile Leu Gln Arg Ala Phe Gln Glu
85 90 95
Ala Val Ala Leu Ala Ala Ser Thr Gly Ile Thr Leu Lys Gln Asn Val
100 105 110
Ala Val Ala Gln
115
<210>73
<211>1197
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(1)..(1197)
<223>RXA07002
<400>73
atg aat cac aga ccc ggc ctg acc ttc cgc ttc ctg gcc gcc cag gtg 48
Met Asn His Arg Pro Gly Leu Thr Phe Arg Phe Leu Ala Ala Gln Val
1 5 10 15
ttg gtc gtg gtg att agc ctg ctg gtg gcc gcg gcc gtg gcc acg atg 96
Leu Val Val Val Ile Ser Leu Leu Val Ala Ala Ala Val Ala Thr Met
20 25 30
gtg ggc ccg acc ctg ttc cat gat cat atg ttg atg acc ggc cgg gag 144
Val Gly Pro Thr Leu Phe His Asp His Met Leu Met Thr Gly Arg Glu
35 40 45
gac ccc tcg ctg gag ctg ttc cat gcc gag cag gcc tac cgg gac gcc 192
Asp Pro Ser Leu Glu Leu Phe His Ala Glu Gln Ala Tyr Arg Asp Ala
50 55 60
aac ctg atc acc ctg gcc gtc gcc ctg ccc acc gcc ttg atc agc gcc 240
Asn Leu Ile Thr Leu Ala Val Ala Leu Pro Thr Ala Leu Ile Ser Ala
65 70 75 80
ctg ctg gcc agc ctg tgg tta tcg cgt cgc ctg cgc acc ccc ctg cag 288
Leu Leu Ala Ser Leu Trp Leu Ser Arg Arg Leu Arg Thr Pro Leu Gln
85 90 95
gat ctc acc cgc gcc gct acc agc ctg acg gcc ggc aac tac cgt atc 336
Asp Leu Thr Arg Ala Ala Thr Ser Leu Thr Ala Gly Asn Tyr Arg Ile
100 105 110
cgc gtg ccc gcc gga gaa gca ggc ccc gag gtc acc acc ctg gcg cat 384
Arg Val Pro Ala Gly Glu Ala Gly Pro Glu Val Thr Thr Leu Ala His
115 120 125
gcc ttc aac acc atg gcc gac cgg ctg gaa cac acc gaa cag gtc cgc 432
Ala Phe Asn Thr Met Ala Asp Arg Leu Glu His Thr Glu Gln Val Arg
130 135 140
cgc cag atg ctc tct gat ctg gcc cac gaa atg ggc acc ccc tta tcg 480
Arg Gln Met Leu Ser Asp Leu Ala His Glu Met Gly Thr Pro Leu Ser
145 150 155 160
gtg ctc acg gtc tac ctc gat ggt ctc cag gac ggg gtc gtg gac tgg 528
Val Leu Thr Val Tyr Leu Asp Gly Leu Gln Asp Gly Val Val Asp Trp
165 170 175
aat aat gcc acc cac acg atc atg gct gac caa ctc acc cgc ctg acc 576
Asn Asn Ala Thr His Thr Ile Met Ala Asp Gln Leu Thr Arg Leu Thr
180 185 190
cgg ttg atg gaa gac atc gac gat gtc tcc cgg gcc cag gaa cac cgg 624
Arg Leu Met Glu Asp Ile Asp Asp Val Ser Arg Ala Gln Glu His Arg
195 200 205
atc gat ttg gac ctg gcg gag gaa ggg ctc ggg gat ctg ctc cat acc 672
Ile Asp Leu Asp Leu Ala Glu Glu Gly Leu Gly Asp Leu Leu His Thr
210 215 220
gcc gct gct gcc gcg ggg gaa gct tat gct gac aaa ggc gtc gat tta 720
Ala Ala Ala Ala Ala Gly Glu Ala Tyr Ala Asp Lys Gly Val Asp Leu
225 230 235 240
cag gtc gag acc att atg gac acc gtc cgg gtg ctc gtg gac cgg caa 768
Gln Val Glu Thr Ile Met Asp Thr Val Arg Val Leu Val Asp Arg Gln
245 250 255
cgc ttc ggc cag gtg atg agc aat ctc ctg tcg aac gcg cta cgg tac 816
Arg Phe Gly Gln Val Met Ser Asn Leu Leu Ser Asn Ala Leu Arg Tyr
260 265 270
acc ccg gcc ggc ggg cag gtc cgg atc agc gtc cac cga cag ggg gcg 864
Thr Pro Ala Gly Gly Gln Val Arg Ile Ser Val His Arg Gln Gly Ala
275 280 285
tcc acc gcg ctc atc cac gtc gcc gat gac ggc gag ggc atc cca cct 912
Ser Thr Ala Leu Ile His Val Ala Asp Asp Gly Glu Gly Ile Pro Pro
290 295 300
ggc cag ctc gga cac atc ttc gaa cgc ttc tac cgg ggg gat gcc gcc 960
Gly Gln Leu Gly His Ile Phe Glu Arg Phe Tyr Arg Gly Asp Ala Ala
305 310 315 320
cgc agc cgg gac aac ggc ggg gcc ggt atc ggt ctg acc atc tcc aag 1008
Arg Ser Arg Asp Asn Gly Gly Ala Gly Ile Gly Leu Thr Ile Ser Lys
325 330 335
gca ttg atc gag gcc cac ggc ggc act ctc acc gcc acc tct cct ggc 1056
Ala Leu Ile Glu Ala His Gly Gly Thr Leu Thr Ala Thr Ser Pro Gly
340 345 350
ccc ggt gcc gga tcg gtc ttc acc atc cgt ctt ccc ctg cac cag gaa 1104
Pro Gly Ala Gly Ser Val Phe Thr Ile Arg Leu Pro Leu His Gln Glu
355 360 365
aac gtg tcc ctg atg ctc agt gac ccc act ccc ggg gac aat aat tct 1152
Asn Val Ser Leu Met Leu Ser Asp Pro Thr Pro Gly Asp Asn Asn Ser
370 375 380
gat gac ctc ggc agc gat ccg tat caa ccc ctt gac aat acc cca 1197
Asp Asp Leu Gly Ser Asp Pro Tyr Gln Pro Leu Asp Asn Thr Pro
385 390 395
<210>74
<211>399
<212>PRT
<213>谷氨酸棒状杆菌
<400>74
Met Asn His Arg Pro Gly Leu Thr Phe Arg Phe Leu Ala Ala Gln Val
1 5 10 15
Leu Val Val Val Ile Ser Leu Leu Val Ala Ala Ala Val Ala Thr Met
20 25 30
Val Gly Pro Thr Leu Phe His Asp His Met Leu Met Thr Gly Arg Glu
35 40 45
Asp Pro Ser Leu Glu Leu Phe His Ala Glu Gln Ala Tyr Arg Asp Ala
50 55 60
Asn Leu Ile Thr Leu Ala Val Ala Leu Pro Thr Ala Leu Ile Ser Ala
65 70 75 80
Leu Leu Ala Ser Leu Trp Leu Ser Arg Arg Leu Arg Thr Pro Leu Gln
85 90 95
Asp Leu Thr Arg Ala Ala Thr Ser Leu Thr Ala Gly Asn Tyr Arg Ile
100 105 110
Arg Val Pro Ala Gly Glu Ala Gly Pro Glu Val Thr Thr Leu Ala His
115 120 125
Ala Phe Asn Thr Met Ala Asp Arg Leu Glu His Thr Glu Gln Val Arg
130 135 140
Arg Gln Met Leu Ser Asp Leu Ala His Glu Met Gly Thr Pro Leu Ser
145 150 155 160
Val Leu Thr Val Tyr Leu Asp Gly Leu Gln Asp Gly Val Val Asp Trp
165 170 175
Asn Asn Ala Thr His Thr Ile Met Ala Asp Gln Leu Thr Arg Leu Thr
180 185 190
Arg Leu Met Glu Asp Ile Asp Asp Val Ser Arg Ala Gln Glu His Arg
195 200 205
Ile Asp Leu Asp Leu Ala Glu Glu Gly Leu Gly Asp Leu Leu His Thr
210 215 220
Ala Ala Ala Ala Ala Gly Glu Ala Tyr Ala Asp Lys Gly Val Asp Leu
225 230 235 240
Gln Val Glu Thr Ile Met Asp Thr Val Arg Val Leu Val Asp Arg Gln
245 250 255
Arg Phe Gly Gln Val Met Ser Asn Leu Leu Ser Asn Ala Leu Arg Tyr
260 265 270
Thr Pro Ala Gly Gly Gln Val Arg Ile Ser Val His Arg Gln Gly Ala
275 280 285
Ser Thr Ala Leu Ile His Val Ala Asp Asp Gly Glu Gly Ile Pro Pro
290 295 300
Gly Gln Leu Gly His Ile Phe Glu Arg Phe Tyr Arg Gly Asp Ala Ala
305 310 315 320
Arg Ser Arg Asp Asn Gly Gly Ala Gly Ile Gly Leu Thr Ile Ser Lys
325 330 335
Ala Leu Ile Glu Ala His Gly Gly Thr Leu Thr Ala Thr Ser Pro Gly
340 345 350
Pro Gly Ala Gly Ser Val Phe Thr Ile Arg Leu Pro Leu His Gln Glu
355 360 365
Asn Val Ser Leu Met Leu Ser Asp Pro Thr Pro Gly Asp Asn Asn Ser
370 375 380
Asp Asp Leu Gly Ser Asp Pro Tyr Gln Pro Leu Asp Asn Thr Pro
385 390 395
<210>75
<211>951
<212>DNA
<213>谷氨酸棒状杆菌
<220>
<221>CDS
<222>(1)..(951)
<223>RXA07003
<400>75
gtg gcg gta gct ggg ctg agt gca agc gtt gag agc aaa atc gac acc 48
Val Ala Val Ala Gly Leu Ser Ala Ser Val Glu Ser Lys Ile Asp Thr
1 5 10 15
atc cgg gac acg ttg aag gaa act gcg tgc gca atc gca aac atg ccg 96
Ile Arg Asp Thr Leu Lys Glu Thr Ala Cys Ala Ile Ala Asn Met Pro
20 25 30
ccagcc aaa acg cag gaa ttc gat cca tca cga atc aag gaa ccc aat 144
Pro Ala Lys Thr Gln Glu Phe Asp Pro Ser Arg Ile Lys Glu Pro Asn
35 40 45
tca cct tcg gta atc acc aaa gcg gca acg ctc atg gat gtg ctc cgc 192
Ser Pro Ser Val Ile Thr Lys Ala Ala Thr Leu Met Asp Val Leu Arg
50 55 60
act gaa ggt ccc acc aac tcc gct cgc cta gct gag gtg ctg ggg gag 240
Thr Glu Gly Pro Thr Asn Ser Ala Arg Leu Ala Glu Val Leu Gly Glu
65 70 75 80
ccg atc agt tcg gtg tac aga atg ctc cac acc tta acc gcg atc ggg 288
Pro Ile Ser Ser Val Tyr Arg Met Leu His Thr Leu Thr Ala Ile Gly
85 90 95
tgg gtt gag cag gac gga aag cgc ggc tcc tat cgc gtt ggc cta gcc 336
Trp Val Glu Gln Asp Gly Lys Arg Gly Ser Tyr Arg Val Gly Leu Ala
100 105 110
atg ctc aca ctt gcc gaa ttg caa ttg cgc cat atg gat ctc cgc aag 384
Met Leu Thr Leu Ala Glu Leu Gln Leu Arg His Met Asp Leu Arg Lys
115 120 125
atc gcc gca cta aca atg cgg aaa att cat gca ctc act ggt gaa acc 432
Ile Ala Ala Leu Thr Met Arg Lys Ile His Ala Leu Thr Gly Glu Thr
130 135 140
aca ttt tta tgt gtt cgt cac ggc att cgc gcg gta tgc atc gag agg 480
Thr Phe Leu Cys Val Arg His Gly Ile Arg Ala Val Cys Ile Glu Arg
145 150 155 160
gtc gat ggc gat cgt gtg aat agt cga gtt ctc cag ctg gga acc tcg 528
Val Asp Gly Asp Arg Val Asn Ser Arg Val Leu Gln Leu Gly Thr Ser
165 170 175
ctg ccg ctc cat gtc ggt gcc gca cct cgt gcg ctt ctc gct ttt gag 576
Leu Pro Leu His Val Gly Ala Ala Pro Arg Ala Leu Leu Ala Phe Glu
180 185 190
gga cgc agg gcc tgg gaa aca tat gcc aca aac tta gga ttc gag ggc 624
Gly Arg Arg Ala Trp Glu Thr Tyr Ala Thr Asn Leu Gly Phe Glu Gly
195 200 205
cat aac tgg tct aaa gga ccg tcc aga ggc gag cta ttc caa cac ttg 672
His Asn Trp Ser Lys Gly Pro Ser Arg Gly Glu Leu Phe Gln His Leu
210 215 220
gac gaa gac cgc gat aag ggt ttt tgc ctg gta gac aat gaa att act 720
Asp Glu Asp Arg Asp Lys Gly Phe Cys Leu Val Asp Asn Glu Ile Thr
225 230 235 240
ccc ggg atc gcc gct gta gga gca ccg att tac aac cat cgt ggt gaa 768
Pro Gly Ile Ala Ala Val Gly Ala Pro Ile Tyr Asn His Arg Gly Glu
245 250 255
gtc gtg gca agc ctg tcc atg agt gga ctg cgg gac ggc atc ctc agc 816
Val Val Ala Ser Leu Ser Met Ser Gly Leu Arg Asp Gly Ile Leu Ser
260 265 270
gat acc acc gac tac tcg gcc gtg gaa ctg atc ctg cag ggg tct gcg 864
Asp Thr Thr Asp Tyr Ser Ala Val Glu Leu Ile Leu Gln Gly Ser Ala
275 280 285
gag att tcc caa gcg ctg gga gcg acc att gag cac aac ggg ggc aac 912
Glu Ile Ser Gln Ala Leu Gly Ala Thr Ile Glu His Asn Gly Gly Asn
290 295 300
caa aaa cta ccc cag gta acc cct ctg agc att gtg gtt 951
Gln Lys Leu Pro Gln Val Thr Pro Leu Ser Ile Val Val
305 310 315
<210>76
<211>317
<212>PRT
<213>谷氨酸棒状杆菌
<400>76
Val Ala Val Ala Gly Leu Ser Ala Ser Val Glu Ser Lys Ile Asp Thr
1 5 10 15
Ile Arg Asp Thr Leu Lys Glu Thr Ala Cys Ala Ile Ala Asn Met Pro
20 25 30
Pro Ala Lys Thr Gln Glu Phe Asp Pro Ser Arg Ile Lys Glu Pro Asn
35 40 45
Ser Pro Ser Val Ile Thr Lys Ala Ala Thr Leu Met Asp Val Leu Arg
50 55 60
Thr Glu Gly Pro Thr Asn Ser Ala Arg Leu Ala Glu Val Leu Gly Glu
65 70 75 80
Pro Ile Ser Ser Val Tyr Arg Met Leu His Thr Leu Thr Ala Ile Gly
85 90 95
Trp Val Glu Gln Asp Gly Lys Arg Gly Ser Tyr Arg Val Gly Leu Ala
100 105 110
Met Leu Thr Leu Ala Glu Leu Gln Leu Arg His Met Asp Leu Arg Lys
115 120 125
Ile Ala Ala Leu Thr Met Arg Lys Ile His Ala Leu Thr Gly Glu Thr
130 135 140
Thr Phe Leu Cys Val Arg His Gly Ile Arg Ala Val Cys Ile Glu Arg
145 150 155 160
Val Asp Gly Asp Arg Val Asn Ser Arg Val Leu Gln Leu Gly Thr Ser
165 170 175
Leu Pro Leu His Val Gly Ala Ala Pro Arg Ala Leu Leu Ala Phe Glu
180 185 190
Gly Arg Arg Ala Trp Glu Thr Tyr Ala Thr Asn Leu Gly Phe Glu Gly
195 200 205
His Asn Trp Ser Lys Gly Pro Ser Arg Gly Glu Leu Phe Gln His Leu
210 215 220
Asp Glu Asp Arg Asp Lys Gly Phe Cys Leu Val Asp Asn Glu Ile Thr
225 230 235 240
Pro Gly Ile Ala Ala Val Gly Ala Pro Ile Tyr Asn His Arg Gly Glu
245 250 255
Val Val Ala Ser Leu Ser Met Ser Gly Leu Arg Asp Gly Ile Leu Ser
260 265 270
Asp Thr Thr Asp Tyr Ser Ala Val Glu Leu Ile Leu Gln Gly Ser Ala
275 280 285
Glu Ile Ser Gln Ala Leu Gly Ala Thr Ile Glu His Asn Gly Gly Asn
290 295 300
Gln Lys Leu Pro Gln Val Thr Pro Leu Ser Ile Val Val
305 310 315
Claims (8)
1.分离的核酸分子,其编码的多肽所具有的氨基酸序列在每种情况下均涉及表1/第2列,其中在表1/第4列所示氨基酸位置处的核酸分子所编码的蛋白原氨基酸与表1/第5列相同行所示的特定氨基酸不同。
2.如权利要求1所述的分离的核酸分子,其中在表1/第4列中所示氨基酸位置处的核酸分子编码表1/第6列相同行所示的氨基酸。
3.载体,其包含至少一个如权利要求1所述的核酸序列。
4.宿主细胞,其用至少一种如权利要求3所述的载体转染。
5.如权利要求4所述的宿主细胞,其中所述核酸分子的表达调节所述细胞精细化学品的生产率。
6.制备精细化学品的方法,其包括培养已用至少一种如权利要求3所述的载体转染的细胞,从而产生精细化学品。
7.如权利要求6所述的方法,其中精细化学品是氨基酸。
8.如权利要求7所述的方法,其中所述氨基酸是赖氨酸。
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DE10154245A DE10154245A1 (de) | 2001-11-05 | 2001-11-05 | Gene die für regulatorische Proteine codieren |
DE10154245.3 | 2001-11-05 |
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US (1) | US20050014234A1 (zh) |
EP (1) | EP1444257A2 (zh) |
KR (1) | KR100861747B1 (zh) |
CN (1) | CN1596267A (zh) |
AU (1) | AU2002346805A1 (zh) |
BR (1) | BR0213775A (zh) |
DE (1) | DE10154245A1 (zh) |
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US6825030B2 (en) * | 2000-08-31 | 2004-11-30 | Degussa Ag | Nucleotide sequences encoding a sensor kinase, citA, from corynebacterium glutamicum |
DE10359661A1 (de) * | 2003-12-18 | 2005-07-28 | Basf Ag | Genvarianten die für Proteine aus dem Stoffwechselweg von Feinchemikalien codieren |
JP6519476B2 (ja) | 2013-10-23 | 2019-05-29 | 味の素株式会社 | 目的物質の製造法 |
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DE10039044A1 (de) * | 2000-08-10 | 2002-02-21 | Degussa | Neue für das IysR1-Gen kodierende Nukleotidsequenzen |
EP1311685B1 (en) * | 2000-08-26 | 2007-03-21 | Degussa GmbH | Nucleotide sequences which code for the ccpa1 gene |
-
2001
- 2001-11-05 DE DE10154245A patent/DE10154245A1/de not_active Withdrawn
-
2002
- 2002-10-31 BR BR0213775-5A patent/BR0213775A/pt not_active IP Right Cessation
- 2002-10-31 CN CNA028235843A patent/CN1596267A/zh active Pending
- 2002-10-31 KR KR1020047006801A patent/KR100861747B1/ko not_active IP Right Cessation
- 2002-10-31 EP EP02783044A patent/EP1444257A2/de not_active Withdrawn
- 2002-10-31 AU AU2002346805A patent/AU2002346805A1/en not_active Abandoned
- 2002-10-31 WO PCT/EP2002/012139 patent/WO2003040181A2/de not_active Application Discontinuation
- 2002-10-31 US US10/495,066 patent/US20050014234A1/en not_active Abandoned
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2004
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Also Published As
Publication number | Publication date |
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AU2002346805A1 (en) | 2003-05-19 |
WO2003040181A3 (de) | 2003-12-24 |
KR100861747B1 (ko) | 2008-10-29 |
KR20050042250A (ko) | 2005-05-06 |
DE10154245A1 (de) | 2003-06-05 |
US20050014234A1 (en) | 2005-01-20 |
BR0213775A (pt) | 2004-11-09 |
EP1444257A2 (de) | 2004-08-11 |
ZA200404417B (en) | 2006-05-31 |
WO2003040181A2 (de) | 2003-05-15 |
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