CN1756847A

CN1756847A - 发酵产生含硫精细化学品的方法(metY)

Info

Publication number: CN1756847A
Application number: CNA038204525A
Authority: CN
Inventors: B·克勒格尔; O·策尔德尔; C·克洛普罗格; H·施罗德; S·哈夫纳
Original assignee: BASF SE
Priority date: 2002-08-26
Filing date: 2003-08-26
Publication date: 2006-04-05
Anticipated expiration: 2023-08-26
Also published as: BR0313758A; WO2004024933A3; KR20050034747A; US7381549B2; US20050260721A1; AU2003264109A1; EP1537225A2; JP2005537025A; DE10239082A1; WO2004024933A2; CN100523208C

Abstract

本发明涉及通过使用表达编码甲硫氨酸合酶(metH)基因的核苷酸序列的细菌，发酵产生含硫精细化学品尤其是L－甲硫氨酸的方法。

Description

发酵产生含硫精细化学品的方法(metY)

描述

本发明涉及通过使用表达编码O-乙酰基高丝氨酸硫化氢解酶(metY)基因的核苷酸序列的细菌，发酵产生含硫精细化学品，尤其是L-甲硫氨酸的方法。

现有技术

含硫精细化学品如甲硫氨酸、高半胱氨酸、S-腺苷甲硫氨酸、谷胱甘肽、半胱氨酸、生物素、硫胺素、硫辛酸通过天然代谢过程在细胞中产生并且用于许多工业领域，包括食品、动物饲料、化妆品和制药工业。这些统称为“含硫精细化学品”的物质包括有机酸、蛋白原性(proteinogenic)和非蛋白原性氨基酸、维生素和辅因子。通过培养细菌可以极其方便地大规模生产这些物质，已经开发这些细菌以产生并大量分泌在每种情况下所希望的物质。尤其适于该目的的生物是棒状细菌，它们是革兰氏阳性非病原性细菌。

公知通过棒状细菌，尤其是谷氨酸棒杆菌(Corynebacteriumglutamicum)的发酵生产氨基酸。由于非常重要，所以生产方法不断改进。方法改进可涉及测定相关的发酵技术方面如搅拌和氧供给，或者涉及营养培养基组分如发酵过程中的糖浓度，或者涉及得到产物的操作(work-up)，例如通过离子交换层析，或者涉及微生物自身的内在性能特性。

通过菌株选择已经开发了从含硫精细化学品产生各种所希望的化合物的许多突变菌株。通过应用诱变、选择和突变选择的方法，在特定分子的产生方面所述微生物的性能特性得到提高。然而，这是一种费时而且困难的方法。以这种方式获得了例如对下述抗代谢物具有抗性或者对于调节重要的代谢物为营养缺陷型的并产生含硫精细化学品如L-甲硫氨酸的菌株，所述抗代谢物如甲硫氨酸类似物α-甲基甲硫氨酸、乙硫氨酸、正亮氨酸、n-乙酰基正亮氨酸、S-三氟甲基高半胱氨酸、2-氨基-5-heprenoitic acid、硒代蛋氨酸、甲硫氨酸亚砜胺(methioninesulfoximine)、methoxine、1-氨基环戊烷羧酸。

重组DNA技术的方法通过扩增单个氨基酸生物合成基因并研究其对氨基酸产生的影响数年来也已经被用于改良产生L-氨基酸的棒杆菌菌株。

WO-A-02/18613描述了谷氨酸棒杆菌中metY的核酸序列和氨基酸序列以及其用于产生L-赖氨酸的用途。

发明简述

本发明的一个目的是提供含硫精细化学品尤其是L-甲硫氨酸的改良的发酵生产的新方法。

我们已经发现通过提供一种含硫精细化学品的发酵生产方法实现了该目的，该方法包括在棒状细菌中表达编码具有metY活性的蛋白质的异源核苷酸序列。

本发明首先涉及用于发酵产生至少一种含硫精细化学品的方法，其包括下面的步骤：

a)发酵产生目的含硫精细化学品的棒状细菌培养物，该棒状细菌表达至少一种这种的异源核苷酸序列，该序列编码具有O-乙酰基高丝氨酸硫化氢解酶(metY)活性的蛋白质；

b)浓缩培养基或细菌细胞中的含硫精细化学品；和

c)分离含硫精细化学品，其优选含有L-甲硫氨酸。

上面的异源编码metY的核苷酸序列与谷氨酸棒杆菌ATCC 13032的编码metY的序列优选具有100％以下的同源性，如70％以上的同源性，诸如75、80、85、90或95％的同源性，或具有小于70％的同源性，如不超过60、50、40、30、20或10％的同源性。编码metY的序列优选来自下面表I生物中的任一种。

表I

白喉棒杆菌(Corynebacterium diphteriae)	ATCC 14779
白喉棒杆菌(Corynebacterium diphteriae)	ATCC 14779	结核分枝杆菌(Mycobacterium tuberculosis)CDC1551	ATCC 25584
丙酮丁醇梭菌(Clostridium acetobutylicum)	ATCC 824	结核分枝杆菌(Mycobacterium tuberculosis)CDC1551	ATCC 25584
丙酮丁醇梭菌(Clostridium acetobutylicum)	ATCC 824	嗜碱芽孢杆菌(Bacillus halodurans)	ATCC 21591
嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)	ATCC 12980	嗜碱芽孢杆菌(Bacillus halodurans)	ATCC 21591
嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)	ATCC 12980	微温绿菌(Chlorobium tepidum)	ATCC 49652
聚球藻属(Synechococcus)中的种	ATCC 27104	微温绿菌(Chlorobium tepidum)	ATCC 49652
聚球藻属(Synechococcus)中的种	ATCC 27104	构巢裸孢壳(Emericella nidulans)	ATCC 36104
脆弱拟杆菌(Bacteroides fragilis)	ATCC 25285	构巢裸孢壳(Emericella nidulans)	ATCC 36104
脆弱拟杆菌(Bacteroides fragilis)	ATCC 25285	乳酸乳球菌(Lactococcus lactis)	ATCC 7962
支气管炎博德特氏菌(Bordetella bronchiseptica)	ATCC 19395	乳酸乳球菌(Lactococcus lactis)	ATCC 7962
支气管炎博德特氏菌(Bordetella bronchiseptica)	ATCC 19395	铜绿假单胞菌(Pseudomonas aeruginosa)	ATCC 17933
欧洲亚硝化单胞菌(Nitrosomonas europaea)	ATCC 19718	铜绿假单胞菌(Pseudomonas aeruginosa)	ATCC 17933
欧洲亚硝化单胞菌(Nitrosomonas europaea)	ATCC 19718	苜蓿中华根瘤菌(Sinorhizobium meliloti)	ATCC 4399
海栖热袍菌(Thermotoga maritima)	ATCC 43589	苜蓿中华根瘤菌(Sinorhizobium meliloti)	ATCC 4399
海栖热袍菌(Thermotoga maritima)	ATCC 43589	变异链球菌(Streptococcus mutans)	ATCC 25175
洋葱伯克霍尔德氏菌(Burkholderia cepacia)	ATCC 25416	变异链球菌(Streptococcus mutans)	ATCC 25175
洋葱伯克霍尔德氏菌(Burkholderia cepacia)	ATCC 25416	耐辐射奇异球菌(Deinococcus radiodurans)	ATCC 13939
荚膜红细菌(Rhodobacter capsulatus)	ATCC 11166	耐辐射奇异球菌(Deinococcus radiodurans)	ATCC 13939
荚膜红细菌(Rhodobacter capsulatus)	ATCC 11166	多杀巴斯德氏菌(Pasteurella multocida)	ATCC 6530
艰难梭菌(Clostridium difficile)	ATCC 9689	多杀巴斯德氏菌(Pasteurella multocida)	ATCC 6530
艰难梭菌(Clostridium difficile)	ATCC 9689	空肠弯曲杆菌(Campylobacter jejuni)	ATCC 33560
肺炎链球菌(Streptococcus pneumoniae)	ATCC 6308	空肠弯曲杆菌(Campylobacter jejuni)	ATCC 33560
肺炎链球菌(Streptococcus pneumoniae)	ATCC 6308	酿酒酵母(Saccharomyces cerevisiae)	ATCC 2704
乳酸克鲁维酵母(Kluyveromyces lactis)	ATCC 8585	酿酒酵母(Saccharomyces cerevisiae)	ATCC 2704
乳酸克鲁维酵母(Kluyveromyces lactis)	ATCC 8585	白假丝酵母(Candida albicans)	ATCC 10231
粟酒裂殖酵母(Schizosaccharomyces pombe)	ATCC 24969	白假丝酵母(Candida albicans)	ATCC 10231

ATCC：美国典型培养物保藏中心，美国Rockville，MD。

本发明使用的metY-编码序列优选含有根据SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、41、43、45、47、49、51和53的编码序列或者与它们同源的编码具有metY活性蛋白质的核苷酸序列。

此外，本发明使用的metY-编码序列优选编码具有metY活性的蛋白质，所述蛋白质含有根据SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52和54的氨基酸序列或与它们同源的代表具有metY活性的蛋白质的氨基酸序列。

编码metY的序列优选为可以在棒状细菌中复制或者被稳定整合到染色体中的DNA或RNA。

根据一个优选的实施方案，本发明的方法通过下面的步骤实施：

a)使用质粒载体转化的细菌菌株，该质粒载体携带处于调节序列控制下的至少一份编码metY序列的拷贝，或者

b)使用这样的菌株，该菌株中编码metY的序列已经被整合到细菌染色体中。

此外，发酵优选过量表达编码metY的序列。

还希望发酵这样的细菌，其中目的含硫精细化学品的生物合成途径的至少另一基因已经被扩增；和/或其中至少一条代谢途径已经至少部分被关闭，其中所述该代谢途径降低目的含硫精细化学品的产生。

还希望发酵这样的细菌，其中额外地目的含硫精细化学品的生物合成途径的至少另一基因不被代谢的代谢物不利地影响。

因此，根据本发明方法的另一实施方案，发酵这样的棒状细菌，其中同时存在选自：

a)基因lysC，其编码天冬氨酸激酶，

b)基因asd，其编码天冬氨酸-半醛脱氢酶，

c)甘油醛-3-磷酸脱氢酶编码基因gap，

d)3-磷酸甘油酸激酶编码基因pgk，

e)丙酮酸羧化酶编码基因pyc，

f)磷酸丙糖异构酶编码基因tpi，

g)高丝氨酸O-乙酰转移酶编码基因metA，

h)γ胱硫醚合酶编码基因metB，

i)γ胱硫醚裂合酶编码基因metC，

j)丝氨酸羟甲基转移酶编码基因glyA，

k)甲硫氨酸合酶编码基因metH，

l)亚甲基四氢叶酸还原酶编码基因metF，

m)磷酸丝氨酸氨基转移酶编码基因serC，

n)磷酸丝氨酸磷酸酶编码基因serB，

o)丝氨酸乙酰转移酶编码基因cysE，

p)高丝氨酸脱氢酶编码基因hom

的至少一种基因被过量表达。

根据本发明方法的另一实施方案，发酵这样的棒杆菌，其中同时有选自上面的组a)到p)的基因的至少一种基因以某种方式突变使得相应蛋白质的活性与未突变蛋白质相比，被所代谢的代谢物影响程度较小(如果有)，并且尤其是精细化学品的发明性生产不被不利地影响。

根据本发明的方法的另一实施方案，发酵这样的棒杆菌，其中同时存在选自：

q)高丝氨酸激酶编码基因thrB，

r)苏氨酸脱水酶编码基因ilvA，

s)苏氨酸合酶编码基因thrC，

t)内消旋-二氨基庚二酸D-脱氢酶编码基因ddh，

u)磷酸烯醇丙酮酸羧激酶编码基因pck，

v)葡萄糖-6-磷酸6-异构酶编码基因pgi，

w)丙酮酸氧化酶编码基因poxB，

x)二氢吡啶二羧酸合酶编码基因dapA，

y)二氢吡啶二羧酸还原酶编码基因dapB；或

z)二氨基吡啶甲酸脱羧酶编码基因lysA的至少一种基因被弱化，尤其通过降低相应基因的表达速率而被弱化。

根据本发明的另一实施方案，发酵这样的棒杆菌，其中同时存在至少一种选自上面组q)到z)的基因以某种方式突变使得相应蛋白质的酶活性被部分或完全降低。

在本发明的方法中，优选谷氨酸棒杆菌种的微生物。

本发明还涉及从发酵液产生含L-甲硫氨酸的动物饲料添加剂的方法，该方法包括下面的步骤：

a)在发酵培养基中培养和发酵产生L-甲硫氨酸的微生物；

b)从含L-甲硫氨酸的发酵液除去水；

c)除去发酵过程中形成的生物量重量的0到100％；和

d)干燥根据b)和/或c)所得发酵液，以得到所希望的粉剂或粒剂形式的动物饲料添加剂。

本发明同样涉及第一次从上面的微生物分离的编码metY的序列，涉及由其编码的O-乙酰基高丝氨酸硫化氢解酶，还分别涉及这些多核苷酸和蛋白质的功能同系物。

发明详述

a)一般术语

具有O-乙酰基高丝氨酸硫化氢解酶活性的蛋白质，也称作metY(EC4.2.99.10)，被描述为使用辅因子磷酸吡哆醛(pyrodoxal phosphate)能够将O-乙酰基高丝氨酸和硫化物转化成高半胱氨酸的蛋白质。技术人员能够区分O-乙酰基高丝氨酸硫化氢解酶的活性和O-琥珀酰高丝氨酸硫化氢解酶的活性，O-琥珀酰高丝氨酸硫化氢解酶在文献中也被称为metz。对于后一种酶，以O-琥珀酰高丝氨酸而不是O-乙酰高丝氨酸作为反应的底物。技术人员可以通过酶测定法检测mety的酶活性，该酶测定法的方案可以是：Shimizu H.Yamagata S.Masui R.Inoue Y.Shibata T.Yokoyama S.Kuramitsu S.Iwama T.Biochimica et Biophysica Acta.1549(1)：61-72，2001，Yamagata S.Isaji M.Nakamura K.Fujisaki S.Doi K.Bawden S.D’Andrea R.Applied Microbiology & Biotechnology.42(1)：92-9，1994。

在本发明的范围中，术语“含硫精细化学品”包括含有至少一个共价结合的硫原子并且可通过本发明的发酵方法得到的化合物。它们的非限制性实例为甲硫氨酸、高半胱氨酸、S-腺苷甲硫氨酸，尤其是甲硫氨酸和S-腺苷甲硫氨酸。

在本发明的范围内，术语“L-甲硫氨酸”、“甲硫氨酸”、高半胱氨酸和S-腺苷甲硫氨酸还包括相应的盐如甲硫氨酸盐酸盐或甲硫氨酸硫酸盐。

“多核苷酸”通常指多核糖核苷酸(RNA)和多脱氧核糖核苷酸(DNA)，其可以分别是未修饰的RNA和DNA，或者分别是修饰的RNA和DNA。

根据本发明，“多肽”指含有通过肽键连接的两个或多个氨基酸的肽或蛋白质。

术语“代谢的代谢物”指在生物体的代谢中发生的作为中间产物或者作为终产物并且，它们除了作为化学结构单元的性质，还可以对酶和对它们的催化活性具有调节作用的化合物。从文献中已知这些代谢的代谢物可以以抑制和刺激的方式作用于酶活性(Biochemistry，Stryer，Lubert，1995W.H.Freeman & Company，New York，纽约)。在文献中还描述了可能在生物体中产生酶，其中代谢的代谢物的影响已经被一些措施改变，这些措施为例如通过紫外辐射、电离辐射或诱变而突变基因组DNA，随后选择特定表型(Sahm H.，Eggeling L.，de Graaf AA.，Biological Chemistry381(9-10)：899-910，2000；Eikmanns BJ.，Eggeling L.，Sahm H.，Antonie vanLeeuwenhoek.，64：145-63，1993-94)。这些改变的特性也可以通过特定测量实现。技术人员公知可能以如此方法特异修饰编码蛋白质的DNA的酶基因中特定核苷酸从而由表达的DNA序列得到的蛋白质具有某些新的性质，例如，代谢的代谢物对未修饰的蛋白质的调节作用被改变。

酶的活性可以以某种方式被影响从而反应速率被减小或者对底物的亲和性被改变或者反应速率被改变。

术语“表达”和“扩增”或“过量表达”在本发明的上下文中描述了微生物中相应DNA编码的一种或多种酶的产生或细胞内活性增加。为此，例如，可将基因导入生物体以通过另一基因替换现有基因，增加该一种基因或几种基因的拷贝数，使用强启动子或使用编码具有高活性的相应酶的基因，并且适当时可以组合这些措施。

b)本发明的metY蛋白质

本发明同样包括上面的表I中具体公开生物的metY酶的“功能等同物”。

在本发明范围内，具体公开的多肽的“功能等同物”或类似物是与其不同的多肽，该多肽还具有所希望的生物学活性如底物特异性。

根据本发明，“功能等同物”指特定突变体，其在上面提到的序列位置的至少一个位置具有不同于特定提到的氨基酸的氨基酸，但是仍然具有上面提到的生物学活性之一。从而“功能等同物”还包括通过一个或多个氨基酸添加、替换、缺失和/或倒位可以得到的突变，所述修饰可能在该序列的任何位置发生，只要它们导致具有本发明特性的突变体。尤其当突变体和未修饰多肽的反应模式定性地匹配，即，例如相同的底物以不同速率被转化时，则存在功能等同物。

“功能等同物”自然也包括从其他生物可以得到的多肽，和天然存在的变体。例如，通过序列比较可以发现同源序列区，按照本发明的特定指导方针可以建立等同酶。

“功能等同物”同样包括本发明多肽的片段、优选单个结构域或序列基序，它们具有例如目的生物学功能。

“功能等同物”还包括融合蛋白质，其具有上面提到的多肽序列之一或者衍生自该序列的功能等同物以及在N-或C-末端功能性连接的与该序列功能不同的至少一种其他异源序列(即，融合蛋白部分的功能的可忽略功能损失)。这些异源序列的非限制性实例为，例如，信号肽、酶、免疫球蛋白、表面抗原、受体或受体配体。

根据本发明，“功能等同物”包括具体公开的蛋白质的同系物。这些同系物与具体公开的序列之一具有至少30％，或者约40％、50％，优选至少约60％、65％、70％或75％，尤其至少85％，如90％、95％或99％的同源性，该同源性通过Pearson和Lipman(Proc.Natl.Acad.，Sci.(USA)85(8)，1988，2444-2448)的算法计算。

通过诱变，例如通过蛋白质的点突变或截短可以产生本发明的蛋白质或多肽的同系物。如此处所用的术语“同系物”涉及蛋白质的变体形式，其作为蛋白质活性的激动剂或拮抗剂。

通过筛选突变体组合文库如截短突变体组合文库，可以鉴定本发明蛋白质的同系物。可例如通过核酸水平的组合诱变，例如，通过合成的寡核苷酸混合物的酶促连接产生蛋白质变体的多样化文库。有多种方法可用于从简并寡核苷酸序列制备潜在同系物的文库。简并基因序列的化学合成可以在自动DNA合成仪中进行，然后合成的基因可以被连接到适宜的表达载体中。一组简并基因的使用使得可能在一种混合物中提供编码一组目的潜在蛋白质序列的全部序列。合成简并寡核苷酸的方法是技术人员公知的(例如，Narang，S.A.，(1983)Tetrahedron 39：3；Itakura等，(1984)Annu.Rev.Biochem.53：323；Itakura等，(1984)Science 198：1056；Ike等，(1983)Nucleic Acids Res.11：477)。

此外，蛋白质密码子片段的文库可用于产生蛋白质片段的多样化群体，该群体用于筛选和随后选择本发明蛋白质的同系物。在一个实施方案中，可以如下产生编码序列片段的文库，这可通过用核酸酶在一定条件下处理编码序列的双链PCR片段，在该条件下切开发生仅仅约为每个分子一次，变性双链DNA，复性该DNA形成双链DNA，其可含有不同切口产物的有意/反义对，通过S1核酸酶处理重新形成的双链体除去单链部分并将所得片段文库连接到表达载体而实现。可通过该方法设计编码本发明蛋白质的N-末端、C-末端和内部片段的表达文库，这些片段具有不同大小。

在现有技术中公知一些技术用于从已经通过点突变或截短产生的组合文库筛选基因产物和筛选DNA文库以得到具有所选择特性的基因产物。这些技术可适于快速筛选通过本发明的同系物的组合诱变所产生的基因文库。用于筛选经历高通量分析的大基因文库的最经常使用的技术包括将基因文库克隆到可复制的表达载体中，用所得载体文库转化适宜的细胞并在一定条件下表达组合基因，其中在该条件下对目的活性的检测方便了这样的载体的分离，该载体的基因编码的产物已经被检测。递归整体诱变(Recursive ensemble mutagenesis，REM)一增加文库中功能突变体频率的一种技术一可以与筛选试验组合使用以鉴定同系物(Arkin und Yourvan(1992)PNAS 89：7811-7815；Delgrave等(1993)，Protein Engineering 6(3)：327-331。

c)本发明的多核苷酸

本发明还涉及编码上面的metY酶之一的核酸序列(单链和双链DNA和RNA如cDNA和mRNA)及其功能等同物，其也可以通过例如使用人工核苷酸类似物得到。

本发明涉及分离的核酸分子，其编码本发明的多肽或蛋白质或者其生物学活性部分，还涉及这样的核酸片段，该片段可用作例如用于鉴定或扩增本发明的编码核酸的杂交探针或引物。

此外，本发明的核酸分子可以含有基因编码区的3’和/或5’端的非翻译序列。

“分离的”核酸分子分离自存在于该核酸的天然来源的其他核酸分子并且还可以基本上无其他细胞物质或培养基(如果其通过重组技术制备)，或者无化学前体或其他化学品(如果其通过化学合成)。

本发明还包括与具体描述的核苷酸序列或其部分互补的核酸分子。

本发明的核苷酸序列使得可产生可用于鉴定和/或克隆其他细胞型或生物中的同源序列的探针和引物。这些探针和引物通常组成这样的核苷酸序列区，其在严格条件下与本发明核酸序列的有意链或者相应的反义链的至少约12、优选至少约25，如40、50或75个连续核苷酸杂交。

本发明的其他核酸序列来自SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、41、43、45、47、49、51或53并且通过添加、替换、插入或缺失一个或多个核苷酸而与它们不同，但是仍然编码具有目的特性的多肽。这些可以是在至少约50％、55％、60％、65％、70％、80％或90％，优选在至少约95％、96％、97％、98％或99％的序列位置中与上面的序列相同的多核苷酸。

本发明还包括通过与特定提到的序列比较，按照特定来源或宿主生物的密码子使用，含有“沉默”突变或被修饰的那些核酸序列，以及天然存在的变体如剪接变体或等位基因变体。本发明还涉及通过保守核苷酸替换(即，相关氨基酸被具有相同电荷、大小、极性和/或溶解性的氨基酸替换)可以得到的序列。

本发明还涉及通过序列多态性从具体公开的核酸衍生的分子。这些遗传多态性可以由于群体内个体间的天然变异而存在。这些天然变异通常导致基因的核苷酸序列中1到5％的变化。

本发明还包括与上面提到的编码序列杂交或者与它们互补的核酸序列。这些多核苷酸可以在筛选基因组或cDNA文库时发现，并且适宜时，通过PCR使用适宜的引物从它们扩增，然后，例如，用适宜的探针分离。另一可能性是用本发明的多核苷酸或载体转化适宜的微生物，繁殖该微生物从而增殖该多核苷酸，然后分离这些多核苷酸。另一可能性是通过化学途径合成本发明的多核苷酸。

能够“杂交”多核苷酸的性质指多核苷酸或寡核苷酸能够在严格条件下结合几乎互补的序列，而非互补序列在这些条件下没有非特异结合。为此，序列应该70-100％，优选90-100％互补，互补序列能够特异地相互结合的性质被例如用于RNA印迹技术或DNA印迹技术或者PCR或者RT-PCR(对于引物结合的情况)中。具有长为30个碱基对或更多碱基对的寡核苷酸通常用于该目的。严格条件指，例如，在RNA印迹技术中，使用50-70℃，优选60-65℃的洗涤溶液，例如，含有0.1％SDS的0.1×SSC缓冲液(20×SSC；3M NaCl，0.3M柠檬酸钠，pH7.0)用于洗脱非特异杂交的cDNA探针或寡核苷酸。在该情况下，如上面提到的，仅仅具有高度互补性的核酸保持相互结合。严格条件的设置是技术人员公知的并且在例如，Ausubel等，Current Protocols in Molecular Biology，John Wiley & Sons，N.Y.(1989)，6.3.1-6.3.6中描述。

d)编码metY基因的分离

可以以本身公知的方法从上面表I的生物分离编码O-乙酰基高丝氨酸硫化氢解酶的metY基因。

为了分离上面表I的生物的metY基因或其他基因，首先在大肠杆菌(E.coli)中产生该生物的基因文库。基因文库的产生在一般已知的教科书和手册中详细描述。可以提及的实例是Winnacker：Gene und Klone，EineEinführung in die Gentechnologie(Verlag Chemie，Weinheim，德国，1990)的教科书，和Sambrook等：分子克隆实验指南(冷泉港，1989)。一种非常熟知的基因文库是大肠杆菌K-12菌株W3110的基因文库，其由Kohara等人(Cell50，495-508(1980))在λ载体中产生。

为了在大肠杆菌中产生来自表I中生物的基因文库，可以使用粘粒如粘粒载体SuperCos I(Wahl等人，1987，Proceedings of the NationalAcademy of Sciences USA，84：2160-2164)或者质粒如pBR322(BoliVal；Life Sciences，25，807-818(1979))或pUC9(Vieira等人，1982，Gene，19：259-268)。适宜的宿主尤其是限制性和重组缺陷的大肠杆菌菌株。该菌株的一个实例是菌株DH5αmcr，其已经由Grant等人(Proceedings of theNational Academy of Sciences USA，87(1990)4645-4649)描述。用粘粒克隆的长DNA片段然后又可以亚克隆到适于测序的通用载体中并随后被测序，如在Sanger等人(proceedings of the National Academy of Sciences of theUnited States of America，74：5463-5467，1977)中所描述的。

然后所得DNA序列可以使用公知的算法或序列分析程序研究，这些算法或序列分析程序为如Staden(Nucleic Acids Research 14，217-232(1986))的算法、Marck(Nucleic Acids Research 16，1829-1836(1988))的算法或者Butler(Methods of Biochemical Analysis 39，74-97(1998))的GCG程序。

发现了来自上面表I生物的编码metY的DNA序列。具体地，发现了根据SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、41、43、45、47、49、51和53的DNA序列。此外，使用上述方法，从存在的所述DNA序列得到了相应蛋白质的氨基酸序列。SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52和54描述了metY基因产物的所得氨基酸序列。

由于遗传密码的简并性从根据SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、41、43、45、47、49、51和53的序列得到的编码DNA序列也是本发明的主题。同样，本发明涉及与所述序列或从它们衍生的序列的部分杂交的DNA序列。

通过杂交鉴定DNA序列的教导可以由技术人员在例如来自Boehringer Mannheim GmbH的手册《滤膜杂交的DIG系统用户指南》(Mannheim，德国，1993)和在Liebl等人(International Journal ofSystematic Bacteriology(1991)41：255-260)中发现。利用聚合酶链式反应(PCR)扩增DNA序列的教导尤其可以由技术人员在Gait编著的手册：Oligonucleotide synthesis：A Practical Approach(IRL Press，Oxford，UK，1984)以及Newton和Graham：PCR(Spektrum Akademischer Verlag，Heidelberg，德国，1994)中找到。

还公知蛋白质的N-和/或C-末端的改变不实质性地损害其功能或者甚至可稳定所述功能。关于此的信息可以由技术人员尤其在Ben-Bassat等人(Journal of Bacteriology 169：751-757(1987))、O′Regan等人(Gene 77：237-251(1989)、Sahin-Toth等人(Protein Sciences 3：240-247(1994))、Hochuli等人(Biotechnology 6：1321-1325(1988))以及在遗传学和分子生物学的公知教科书中找到。

因此从SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52和54获得的氨基酸序列同样是本发明的部分。

e)根据本发明使用的宿主细胞

本发明还涉及作为宿主细胞的微生物，尤其是棒细菌，该微生物含有载体，尤其是穿梭载体或质粒载体，携带本发明定义的至少一种metY基因或者其中本发明的metY基因被表达或扩增。

这些微生物可以从葡萄糖、蔗糖、乳糖、果糖、麦芽糖、糖蜜、淀粉、纤维素或从甘油和乙醇产生含硫精细化学品，尤其是L-甲硫氨酸。所述微生物优选为棒状细菌，尤其是棒杆菌属的细菌。对于棒杆菌属，必须提及的是谷氨酸棒杆菌，在文献中已知其能够产生L-氨基酸。

可以提及的棒状细菌的适宜菌株的实例是棒杆菌属的菌株，尤其是谷氨酸棒杆菌(C.glutamicum)种的菌株，如

谷氨酸棒杆菌ATCC 13032、

醋谷氨酸棒杆菌(Corynebacterium acetoglutamicum)ATCC 15806、

嗜乙酰乙酸棒杆菌(Corynebacterium acetoacidophilum)ATCC13870、

热产氨棒杆菌(Corynebacterium thermoaminogenes)FERMBP-1539、

Corynebacterium melassecola ATCC 17965

或者短杆菌属(Brevibacterium)的菌株，如

黄色短杆菌(Brevibacterium flavum)ATCC 14067、

乳发酵短杆菌(Brevibacterium lactofermentum)ATCC 13869和

叉开短杆菌(Brevibacterium divaricatum)ATCC 14020；

后者从中衍生的菌株，如

谷氨酸棒杆菌KFCC10065、

谷氨酸棒杆菌ATCC21608，

其同样产生目的精细化学品或者其前体。

缩写KFCC指韩国培养物保藏联合会(Korean Federation of CultureCollection)，缩写ATCC指美国典型菌株培养物保藏中心，缩写FERM BP指日本工业科学和技术局的国立生命科学和人体技术研究所的保藏中心。

f)实施本发明的发酵

根据本发明，发现棒状细菌过量表达来自表I生物的metY基因后，以有利的方式产生含硫精细化学品，尤其是L-甲硫氨酸。

为了实现过量表达，技术人员可以采用单独的或联合的不同措施。从而可能增加适宜基因的拷贝数或者突变启动子和调节区或者位于结构基因上游的核糖体结合位点。掺入结构基因上游的表达盒以同样的方式作用。可诱导的启动子使得还可能在发酵性L-甲硫氨酸产生过程中增加表达。通过延长mRNA寿命的措施也可以提高表达。此外，通过防止酶蛋白质的降解也可以增强酶活性。基因或基因构建体可以或者以不同的拷贝数存在于质粒中或者被整合到染色体并在染色体中扩增。另一可能的备选方案是通过改变培养基组分和操纵培养实现相关基因的过量表达。

过量表达的教导可以由技术人员在Martin等人(Biontechnology 5，137-146(1987))、Guerrero等人(Gene 138，35-41(1994))、Tsuchiya和Morinaga(Bio/Technology 6，428-430(1988))、Eikmanns等人(Gene 102，93-98(1991))、欧洲专利0472869、美国专利4,601,893、Schwarzer和Pühler(Biotechnology 9，84-87(1991)、Remscheid等人(Applied andEnvironmental Microbiology 60，126-132(1994)、LaBarre等人(Journal ofBacteriology 175，1001-1007(1993))、专利申请WO 96/15246、Malumbres等人(Gene 134，15-24(1993))、日本公开的说明书JP-A-10-229891、Jensen和Hammer(Biotechnology and Bioengineering 58，191-195(1998))、Makrides(Microbiological Reviews 60：512-538(1996)以及遗传学和分子生物学的公知教科书中找到。

本发明因此还涉及含有处于调节性核酸序列的遗传控制下的编码本发明多肽的核酸序列的表达构建体，还涉及含有至少一种那所述表达构建体的载体。本发明的这类构建体优选包括特定编码序列5’上游的启动子和3’下游的终止子序列和适当时包括其他调节元件，在每种情况下它们均可操作地连接到编码序列。“可操作地连接”指启动子、编码序列、终止子序列和适宜时其他调节元件的顺序排列从而每种调节元件可以在编码序列的表达中正确发挥其功能。可操作地连接的序列的实例为活化序列和增强子等。其他调节元件包括可选择标记、扩增信号、复制起点等。适宜的调节序列在例如Goeddel，基因表达技术：酶学方法185，Academic Press，SanDiego，CA(1990)中描述。

除了人工调节序列外，天然调节序列仍然可以存在于实际的结构基因的上游。遗传修饰可以在适宜时关闭该天然调节并且增加或减少该基因的表达。基因构建体也可以具有更简单的设计，即没有额外的调节信号被插入结构基因的上游并且天然启动子与其调节没有被除去。取而代之的是，天然调节序列被突变从而调节不再发生并且基因表达被增强或减弱。基因构建体可以含有核酸序列的一份或多份拷贝。

有用的启动子的实例为来自谷氨酸棒杆菌启动子的ddh、amy、lysC、dapA、lysA，以及革兰氏阳性启动子SPO2，如在《枯草芽孢杆菌及其最接近的菌株》，Sonenshein，Abraham L.，Hoch，James A.，Losick，Richard；ASM Press，华盛顿哥伦比亚特区以及Patek M.Eikmanns BJ.，Patek J.，Sahm H.，Microbiology.142 1297-309，1996中所描述的，或者优选有利地用于革兰氏阴性细菌中的cos、tac、trp、tet、trp-tet、lpp、lac、lpp-lac、lacIq、T7、T5、T3、gal、trc、ara、SP6、λ-PR和λ-PL启动子。还优选使用可诱导的启动子如光可诱导的启动子，尤其是温度可诱导的启动子如P_rP_L启动子。原则上可以使用具有调节序列的所有天然启动子。此外，还可以有利地使用合成的启动子。

所提及的调节序列旨在使得核酸序列的特异表达成为可能。根据宿主生物，这可以指例如基因仅仅在诱导后被表达或过量表达，或者其被立即表达和/或过量表达。

关于这一点，调节序列和因子可以优选对表达具有有益影响，并能从而增加或减少表达。从而，可能并有利地通过使用强转录信号如启动子和/或增强子增强转录水平上的调节元件。然而，除了这之外还可通过例如提高mRNA的稳定性增强翻译。

通过将适宜的启动子、适宜的SD序列融合到metA核苷酸序列和适宜的终止信号制备表达盒。为此，使用常规重组和克隆技术，如在CurrentProtocols in Molecular Biology，1993，John Wiley & Sons，Incorporated，New York，纽约；PCR Methods，Gelfand，David H.，Innis，Michael A.，Sninsky，John J.，1999，Academic Press，Incorporated，California，SanDiego；PCR Cloning Protocols，Methods in Molecular Biology Ser.，192卷，第二版，Humana Press，New Jersey；Totowa.T.Maniatis，E.F.Fritsch和J.Sambrook，分子克隆实验指南，冷泉港实验室，冷泉港，NY(1989)；以及T.J.Silhavy，M.L.Berman和L.W.Enquist，Experiments with GeneFusions，Cold Spring Harbor Laboratory，Cold Spring Harbor，NY(1984)；以及Ausubel，F.M.等人，Current Protocols in Molecular Biology，Greene Publishing Assoc.and Wiley Interscience(1987)中描述的那些技术。

通过将重组核酸构建体或基因构建体有利地插入宿主特异的载体而实现在适宜的宿主生物中表达所述重组核酸构建体或基因构建体，其中所述载体使得可能在宿主中最优表达这些基因。载体是本领域技术人员熟知的并且可以在例如，“Cloning Vectors”(Pouwels P.H.等人，Hrsg，Elsevier，Amsterdam-New York-Oxford，1985)中找到。术语“载体”除了质粒，还指技术人员公知的所有其他载体，如噬菌体、转座子、IS元件、质粒、粘粒和线性或环状DNA。这些载体可以在宿主生物中自主复制或者随染色体复制。

通过例如利用游离型质粒过量表达本发明的metY基因而扩增这些基因。适宜的质粒为在棒状细菌中复制的那些质粒。许多公知的质粒载体如pZ1(Menkel等人，Applied and Environmental Microbiology(1989)64：549-554)、pEKEx1(Eikmanns等人，Gene 102：93-98(1991))或pHS2-1(Sonnen等人，Gene 107：69-74(1991))是基于隐性质粒(cryptic plasmid)pHM1519、pBL1或pGA1。其他质粒载体如pCLiK5MCS或者基于pCG4(US-A 4,489,160)或pNG2(Serwold-Davis等人，FEMS MicrobiologyLetters 66，119-124(1990))或pAG1(US-A 5,158,891)的那些质粒可以以相同方式使用。

适宜的质粒载体还包括通过它们可以应用通过整合到染色体扩增基因的方法的那些质粒载体，如Remscheid等人(Applied and EnvironmentalMicrobiology 60，126-132(1994))已经描述的用于复制和扩增hom-thrB操纵子的那些质粒载体。在该方法中，完整基因被克隆到质粒载体中，该质粒载体可以在宿主(一般为大肠杆菌)但是不能在谷氨酸棒杆菌中复制。适宜的载体为例如pSUP301(Sirnon等人，Bio/Technology 1，784-791(1983))、pK18mob或pK19mob(Sch_fer等人，Gene 145，69-73(1994))，Bernard等人，Journal of Molecular Biology，234：534-541(1993))、pEM1(Schrumpf等人，1991，Journal of Bacteriology 173：4510-4516)或pBGS8(Spratt等人，1986，Gene 41：337-342)。含有待扩增基因的质粒载体然后通过转化被转移到目的谷氨酸棒杆菌菌株中。转化方法在例如Thierbach等人(Applied Microbiology and Biotechnology 29，356-362(1988))、Dunican和Shivnan(Biotechnology 7，1067-1070(1989))和Tauch等人(FEMSMicrobiological Letters 123，343-347(1994))中描述。

酶的活性可以被相应基因中的突变影响从而使得酶反应的速率被部分或完全降低。这些突变的实例是技术人员公知的(Motoyama H.，Yano H.，Terasaki Y.，Anazawa H.，Applied & Environmental Microbiology.67：3064-70，2001，Eikmanns BJ.，Eggeling L.，Sahm H.，Antonie vanLeeuwenhoek.64：145-63，1993-94)。

此外，对于含硫精细化学品，尤其是L-甲硫氨酸的产生有利的是，除了表达和扩增本发明的metY基因，还扩增各自生物合成途径、半胱氨酸途径、天冬氨酸-半醛合成、糖酵解、回补、磷酸戊糖代谢、柠檬酸循环或者氨基酸输出的一种或多种酶。

从而，可以扩增一种或多种下面的基因以产生含硫精细化学品，尤其是L-甲硫氨酸：

-基因lysC，其编码天冬氨酸激酶(EP 1 108 790 A2；DNA-SEQ NO.281)，

-基因asd，其编码天冬氨酸-半醛脱氢酶(EP 1 108 790 A2；DNA-SEQNO.282)，

-甘油醛-3-磷酸脱氢酶编码基因gap(Eikmanns(1992)，Journal ofBacteriology 174：6076-6086)，

-3-磷酸甘油酸激酶编码基因pgk(Eikmanns(1992)，Journal ofBacteriology 174：6076-6086)，

-丙酮酸羧化酶编码基因pyc(Eikmanns(1992)，Journal ofBacteriology 174：6076-6086)，

-磷酸丙糖异构酶编码基因tpi(Eikmanns(1992)，Journal ofBacteriology 174：6076-6086)，

-高丝氨酸O-乙酰转移酶编码基因metA(EP 1 108 790 A2；DNA-SEQ NO.725)，

-γ胱硫醚合酶编码基因metB(EP 1 108 790 A2；DNA-SEQ NO.3491)，

-γ胱硫醚裂合酶编码基因metC(EP 1 108 790 A2；DNA-SEQ NO.3061)，

-丝氨酸羟甲基转移酶编码基因glyA(EP 1 108 790 A2；DNA-SEQNO.1110)，

-甲硫氨酸合酶编码基因metH(EP 1 108 790 A2)，

-亚甲基四氢叶酸还原酶编码基因metF(EP 1 108 790 A2；DNA-SEQNO.2379)，

-磷酸丝氨酸氨基转移酶编码基因serC(EP 1 108 790 A2；DNA-SEQNO.928)，

-磷酸丝氨酸磷酸酶编码基因serB(EP 1 108 790 A2；DNA-SEQ NO.334，DNA-SEQ NO.467，DNA-SEQ NO.2767)，

-基因cysE，其编码丝氨酸乙酰转移酶(EP 1 108 790 A2；DNA-SEQNO.2818)，

-基因hom，其编码高丝氨酸脱氢酶(EP 1 108 790 A2；DNA-SEQ NO.1306)

从而，对在棒状细菌中产生含硫精细化学品尤其是L-甲硫氨酸有利的是同时突变至少一种下面的基因，从而相应蛋白质的活性与未突变的蛋白质的相比，受代谢的代谢物影响程度较小或不受影响：

-基因lysC，其编码天冬氨酸激酶(EP 1 108 790 A2；DNA-SEQ NO.281)，

-γ胱硫醚合酶编码基因metB(EP 1 108 790 A2；DNA-SEQ NO.3491)，

-γ胱硫醚裂合酶编码基因metC(EP 1 108 790 A2；DNA-SEQ NO.3061)，

-甲硫氨酸合酶编码基因metH(EP 1 108 790 A2)，

-丝氨酸乙酰转移酶编码基因cysE(EP 1 108 790 A2；DNA-SEQ NO.2818)，

-基因hom，其编码高丝氨酸脱氢酶(EP 1 108 790 A2；DNA-SEQ NO.1306)

另外对产生含硫精细化学品尤其是L-甲硫氨酸有利的是，除了表达和扩增本发明的metY基因之一外，还弱化一种或多种下面的基因，尤其是减少它们的表达，或者将它们关闭：

-高丝氨酸激酶编码基因thrB(EP 1 108 790 A2；DNA-SEQ NO.3453)，

-苏氨酸脱水酶编码基因ilvA(EP 1 108 790 A2；DNA-SEQ NO.2328)，

-苏氨酸合酶编码基因thrC(EP 1 108 790 A2；DNA-SEQ NO.3486)，

-内消旋二氨基庚二酸D-脱氢酶编码基因ddh(EP 1 108 790 A2；DNA-SEQ NO.3494)，

-磷酸烯醇丙酮酸羧激酶编码基因pck(EP 1 108 790 A2；DNA-SEQNO.3157)，

-葡萄糖-6-磷酸6-异构酶编码基因pgi(EP 1 108 790 A2；DNA-SEQNO.950)，

-丙酮酸氧化酶编码基因poxB(EP 1 108 790 A2；DNA-SEQ NO.2873)，

-二氢吡啶二羧酸合酶编码基因dapA(EP 1 108 790 A2；DNA-SEQNO.3476)，

-二氢吡啶二羧酸还原酶编码基因dapB(EP 1 108 790 A2；DNA-SEQNO.3477)

-二氨基吡啶甲酸脱羧酶编码基因lysA(EP 1 108 790 A2；DNA-SEQNO.3451)。

另外对含硫精细化学品尤其是L-甲硫氨酸的产生有利的是，除了在棒状细菌中表达和扩增本发明的metY基因之一外，同时突变至少一种下面的基因使得相应蛋白质的酶活性部分或完全降低：

-高丝氨酸激酶编码基因thrB(EP 1 108 790 A2；DNA-SEQ NO.3453)，

-苏氨酸脱水酶编码基因ilvA(EP 1 108 790 A2；DNA-SEQ NO.2328)，

-苏氨酸合酶编码基因thrC(EP 1 108 790 A2；DNA-SEQ NO.3486)，

-丙酮酸氧化酶编码基因poxB(EP 1 108 790 A2；DNA-SEQ NO.2873)，

另外对含硫精细化学品尤其是L-甲硫氨酸的产生有利的是，除了表达和扩增本发明的一种metY基因，还消除不需要的副反应(Nakayama：微生物产物的过量产生中的“产氨基酸微生物的喂饲”，Krumphanzl，Sikyta，Vanek(eds.)，Academic Press，伦敦，UK，1982)。

根据本发明产生的微生物可以连续地或者分批地或者补料分批或者反复补料分批方法培养以产生含硫精细化学品，尤其是L-甲硫氨酸。公知的培养方法的概述可以在Chmiel的教科书(Bioprozeβtechnik 1.Einführungin die Bioverfahrenstechnik(Gustav Fischer Verlag，Stuttgart，1991))或者Storhas的教科书(Bioreaktoren und periphere Einrichtungen(ViewegVerlag，Braunschweig/Wiesbaden，1994))中找到。

所用的培养基必须以适当的方式满足特定菌株的要求。美国细菌学协会(the American Society for Bacteriology)的教科书″Manual of Methodsfür General Bacteriology″包含各种微生物培养基的描述。

可以根据本发明使用的所述培养基通常含有一种或多种碳源、氮源、无机盐、维生素和/或微量元素。

优选的碳源为糖如单糖、二糖或多糖。非常好的碳源的实例为葡萄糖、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。也可通过复杂化合物如糖蜜或其他糖精炼的副产物将糖加入培养基。还有利的是加入不同碳源的混合物。其他可能的碳源为油和脂肪如大豆油、向日葵油、花生油和椰子脂，脂肪酸如棕榈酸、硬脂酸和亚油酸，醇如甘油、甲醇和乙醇以及有机酸如乙酸和乳酸。

氮源通常为有机或无机氮化合物或含有所述化合物的物质。氮源的实例包括氨气或铵盐如硫酸铵、氯化铵、磷酸铵、碳酸铵和硝酸铵、硝酸盐、尿素、氨基酸和复杂氮源如玉米浆、大豆粉、大豆蛋白、酵母提取物、肉膏等。氮源可以单独地或者作为混合物使用。

可以包括在培养基的无机盐化合物包括钙、镁、钠、钴、钼、钾、锰、锌、铜和铁的氯化物、磷酸盐或硫酸盐。

无机含硫化合物如硫酸盐、亚硫酸盐、连二亚硫酸盐、连四硫酸盐、硫代硫酸盐、硫化物或者有机含硫化合物如硫醇类也可用作产生含硫精细化学品尤其是甲硫氨酸的硫源。

磷酸、磷酸二氢钾和磷酸氢二钾或相应的含钠盐可用作磷源。

可向培养基中加入螯合剂以保持溶液中的金属离子。尤其适宜的螯合剂包括二羟基酚类如儿茶酚或原儿茶酸以及有机酸如柠檬酸。

根据本发明使用的发酵培养基通常还含有其他生长因子如维生素或生长促进剂，其包括例如生物素、核黄素、硫胺素、叶酸、烟酸、泛酸和吡哆醇。生长因子和盐经常来自复杂的培养基组分如酵母提取物、糖蜜、玉米浆等。还可向培养基中加入适宜的前体。培养基的精确组分很大程度上取决于特定实验并且对于每种特定情况单独决定。优化培养基的信息可以在教科书″Applied Microbiol.Physiology，A Practical Approach″(编者P.M.Rhodes，P.F.Stanbury，IRL Press(1997)53-73页，ISBN 0 19 9635773)中发现。还可以从供应商得到生长培养基，例如Standard 1(Merck)或BHI(脑心浸液，DIFCO)等。

所有培养基组分通过热(1.5巴下20分钟，121℃)或通过无菌过滤除菌。各组分可以一起或者，如果需要，分开灭菌。所有培养基组分可以在培养开始时存在或者根据需要连续或者分批加入。

培养温度通常为15℃到45℃，优选25℃到40℃，并且可以保持恒定或者在实验过程中改变。培养基的pH应该为5到8.5，优选约7.0。培养的pH可以在培养中通过加入碱性化合物如氢氧化钠、氢氧化钾、氨和氨水或者酸性化合物如磷酸或硫酸控制。通过使用防沫剂如脂肪酸聚乙二醇酯控制起泡沫。为了保持质粒的稳定，可向培养基加入具有选择作用的适宜物质，例如抗生素。通过将氧气或者含氧的气体混合物如空气导入培养基可以保持需氧条件。培养温度通常为20℃到45℃。连续培养直到目的产物达到最大量。该目标通常在10到160小时内实现。

以这种方法得到的发酵液，尤其是含有L-甲硫氨酸的培养基，通常含有按重量计7.5到25％的干生物量。

另一额外的益处是至少在末尾，但是优选在至少30％的发酵期间实施限糖发酵。这表示在该时间内发酵培养基中可利用糖的浓度保持在或者减小到≥0到3g/l。

然后进一步处理发酵液。生物量可以根据需要通过分离方法如离心、过滤、倒出或这些方法的组合从发酵液完全或部分除去或者完全保留在所述发酵液中。

随后，使用公知的方法如利用旋转蒸发器、薄膜蒸发器、降膜蒸发器、反向渗透或者通过纳过滤(nanofiltration)增稠或者浓缩发酵液。该浓缩的发酵液然后可以通过冷冻干燥、喷雾干燥、喷雾粒化或其他方法处理。

然而，还可进一步纯化含硫精细化学品，尤其是L-甲硫氨酸。为此，含产物的发酵液，在除去生物量后，使用适宜的树脂进行层析，目的产物或者杂质完全或部分保留在层析树脂上。如果需要，可以使用相同的或者不同的层析树脂重复这些层析步骤。技术人员熟悉适宜的层析树脂的选择和它们最有效的应用。纯化的产物可以通过过滤或者超滤浓缩并保持在某一温度下，在该温度下产物的稳定性最大。

通过本领域技术可以确定所分离的一种或几种化合物的身份和纯度。这些技术包括高效液相层析(HPLC)、光谱方法、染色方法、薄层层析、NIRS、酶测定法或微生物学测定法。这些分析方法在Patek等人(1994)Appl.Environ.Microbiol.60：133-140；Malakhova 等人(1996)Biotekhnologiya 1127-32；和Schmidt等人(1998)Bioprocess Engineer.19：67-70.Ulmann′s Encyclopedia of Industrial Chemistry(1996)Bd.A27，VCH：Weinheim，89-90页，521-540页，540-547页，559-566页，575-581和581-587页；Michal，G.，(1999)Biochemical Pathways：An Atlas ofBiochemistry and Molecular Biology，John Wiley and Sons；Fallon，A.等人(1987)，在Laboratory Techniques in Biochemistry and Molecular Biology，17卷的HPLC在生物化学中的应用中概述。

下面的非限制性实施例和附图更详细时描述本发明：

图1显示了质粒pClysC的质粒图；

图2质粒pCISlysCthr311ile的质粒图；

图3质粒pCPhsdhmetY_Mt的质粒图；

限制性切割位点及它们各自的位置(在括号中)在质粒图中显示。必需的序列片段以粗体印刷。KanR指卡那霉素抗性基因；ask指天冬氨酸激酶基因。

实施例1：pCLiK5MCS的构建

首先，使用寡核苷酸p1.3(SEQ ID NO：55)和p2.3(SEQ ID NO：56)，利用聚合酶链式反应(PCR)扩增载体pBR322的氨苄青霉素抗性和复制起点。

p1.3(SEQ ID NO：55)

5‘-CCCGGGATCCGCTAGCGGCGCGCCGGCCGGCCCGGTGTGAAATACCGCACAG-3‘

p2.3(SEQ ID NO：56)

5‘-TCTAGACTCGAGCGGCCGCGGCCGGCCTTTAAATTGAAGACGAAAGGGCCTCG-3‘

寡核苷酸p1.3(SEQ ID NO：55)除了含有与pBR322互补的序列，还含有5’-3’方向限制性核酸酶SmaI、BamHI、NheI和AscI的切割位点，寡核苷酸p2.3(SEQ ID NO：56)含有5’-3’方向限制性核酸内切酶XbaI、XhoI、NotI和DraI的切割位点。根据标准方法如Innis等(PCR Protocols.A Guideto Methods and Applications，Academic Press(1990))使用PfuTurbo聚合酶(Stratagene，La Jolla，USA)实施PCR反应。得到的大小约2.1kb的DNA片段用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明纯化。使用快速DNA连接试剂盒(RocheDiagnostics，Mannheim)根据生产商的使用说明将DNA片段的钝端相互连接并根据标准方法，如Sambrook等人(分子克隆实验指南，冷泉港实验室，(1989))中描述方法将连接混合物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过将细胞涂在含有氨苄青霉素(50μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板选择携带质粒的细胞。

使用Qiaprep spin miniprep试剂盒(Qiagen，Hilden)根据生产商的使用说明书分离单个克隆的质粒DNA并将它们通过限制性消化检查。以这种方法得到的质粒称为pCLiK1。

以质粒pWLT1(Liebl等，1992)作为PCR反应的模板开始，使用寡核苷酸neo1(SEQ ID NO：57)和neo2(SEQ ID NO：58)扩增卡那霉素抗性盒。

neo1(SEQ ID NO：57)：

5‘-GAGATCTAGACCCGGGGATCCGCTAGCGGGCTGCTAAAGGAAGCGGA-3‘

neo2(SEQ ID NO：58)：

5‘-GAGAGGCGCGCCGCTAGCGTGGGCGAAGAACTCCAGCA-3‘

寡核苷酸neo1除了含有与pWLT1互补的序列外，还含有5’-3’方向限制性内切酶XbaI、SmaI、BamHI、NheI的切割位点，寡核苷酸neo2(SEQID NO：58)含有5’-3’方向限制性核酸内切酶AscI和NheI的切割位点。使用PfuTurbo聚合酶(Stratagene，La Jolla，USA)根据标准方法如Innis等(PCR Protocols.A Guide to Methods and Applications，Academic Press(1990))的方法实施PCR反应。得到的约1.3kb大小的DNA片段用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书纯化。DNA片段用限制性内切酶XbaI和AscI(New England Biolabs，Beverly，USA)切割并且，之后，再次用用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书纯化。载体pCLiK1也用限制性内切酶XbaI和AscI切割并使用碱性磷酸酶(Roche Diagnostics，Mannheim)根据生产商的使用说明书去磷酸。在0.8％强度的琼脂糖凝胶中电泳后，线性化载体(约2.1kb)使用GFX^TMPCR、DNA和凝胶带纯化试剂盒(AmershamPharmacia，Freiburg)根据生产商的使用说明书分离。该载体片段使用快速DNA连接试剂盒(Roche Diagnostics，Mannheim)根据生产商的使用说明用切割的PCR片段连接并将根据标准方法，如Sambrook等(分子克隆实验指南，冷泉港，(1989))中描述的方法将连接混合物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过涂在含有氨苄青霉素(50μg/ml)和卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板上选择携带质粒的细胞。

使用Qiaprep spin miniprep试剂盒(Qiagen，Hilden)根据生产商的使用说明书分离单个克隆的质粒DNA并将它们通过限制性消化检查。以这种方法得到的质粒称为pCLiK2。

载体pCLiK2用限制性内切酶DraI(New England Biolabs，Beverly，USA)切割。在0.8％强度的琼脂糖凝胶中电泳后，使用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书分离约2.3kb载体片段。该载体片段使用快速DNA连接试剂盒(Roche Diagnostics，Mannheim)根据生产商的使用说明重新连接并将根据标准方法，如Sambrook等(分子克隆实验指南，冷泉港，(1989))中描述的方法将连接混合物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过涂在含有卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板上选择携带质粒的细胞。

使用Qiaprep spin miniprep试剂盒(Qiagen，Hilden)根据生产商的使用说明书分离单个克隆的质粒DNA并将它们通过限制性消化检查。以这种方法得到的质粒称为pCLiK3。

以质粒pWLQ2(Liebl等，1992)作为PCR反应的模板开始，使用寡核苷酸cg1(SEQ ID NO：59)和cg2(SEQ ID NO：60)扩增复制起点pHM1519。

cg1(SEQ ID NO：59)：

5‘-GAGAGGGCGGCCGCGCAAAGTCCCGCTTCGTGAA-3‘

cg2(SEQ ID NO：60)：

5‘-GAGAGGGCGGCCGCTCAAGTCGGTCAAGCCACGC-3‘

寡核苷酸cg1(SEQ ID NO：59)和cg2(SEQ ID NO：60)除了包含与pWLQ2互补的序列外，还含有限制性内切酶NotI的切割位点。使用PfuTurbo聚合酶(Stratagene，La Jolla，USA)根据标准方法如Innis等(PCR Protocols.A Guide to Methods and Applications，Academic Press(1990))的方法实施PCR反应。得到DNA片段大小约2.7kb并用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书纯化。DNA片段用限制性内切酶NotI(NewEngland Biolabs，Beverly，USA)切割，并且之后再次用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书纯化。载体pCLiK3也用限制性内切酶NotI切割并使用碱性磷酸酶(Roche Diagnostics，Mannheim)根据生产商的使用说明书去磷酸。在0.8％强度的琼脂糖凝胶中电泳后，线性化载体(约2.3kb)使用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书分离。该载体片段使用快速DNA连接试剂盒(Roche Diagnostics，Mannheim)根据生产商的使用说明用切割的PCR片段连接并根据标准方法，如Sambrook等(分子克隆实验指南，冷泉港，(1989))中描述的方法将连接混合物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过涂在含有卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板上选择携带质粒的细胞。

使用Qiaprep spin miniprep试剂盒(Qiagen，Hilden)根据生产商的使用说明书分离单个克隆的质粒DNA并将它们通过限制性消化检查。以这种方法得到的质粒称为pCLiK5。

通过组合两种合成的基本互补的寡核苷酸HS445((SEQ ID NO：61)和HS446(SEQ ID NO：62))通过多克隆位点(MCS)延伸PCLik5，HS445和HS446含有限制性内切酶SwaI、XhoI、AatI、ApaI、Asp718、MluI、NdeI、SpeI、EcoRV、SalI、ClaI、BamHI、XbaI和SmaI的切割位点，延伸后通过将它们一起加热到95℃，然后缓慢冷却得到双链DNA片段。

HS445(SEQ ID NO：61)：

5‘-TCGAATTTAAATCTCGAGAGGCCTGACGTCGGGCCCGGTACCACGCGTCATATGACTAGTTCGGACCTAGGGATATCGTCGACATCGATGCTCTTCTGCGTTAATTAACAATTGGGATCCTCTAGACCCGGGATTTAAAT-3‘

HS446(SEQ ID NO：62)：

5‘-GATCATTTAAATCCCGGGTCTAGAGGATCCCAATTGTTAATTAACGCAGAAGAGCATCGATGTCGACGATATCCCTAGGTCCGAACTAGTCATATGACGCGTGGTACCGGGCCCGACGTCAGGCCTCTCGAGATTTAAAT-3‘

载体pCLiK5用限制性内切酶XhoI和BamHI(New England Biolabs，Beverly，USA)切割并用碱性磷酸酶(I(Roche Diagnostics，Mannheim))根据生产商的使用说明书去磷酸。在0.8％强度的琼脂糖凝胶中电泳后，线性化载体(约5.0kb)使用GFX^TMPCR、DNA和凝胶带纯化试剂盒(AmershamPharmacia，Freiburg)根据生产商的使用说明书分离。该载体片段使用快速DNA连接试剂盒(Roche Diagnostics，Mannheim)根据生产商的使用说明用合成的双链DNA片段连接，并根据如Sambrook等(分子克隆实验指南，冷泉港，(1989))中描述的标准方法将连接混合物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过涂在含有卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板上选择携带质粒的细胞。

使用Qiaprep spin miniprep试剂盒(Qiagen，Hilden)根据生产商的使用说明书分离单个克隆的质粒DNA并将它们通过限制性消化检查。以这种方法得到的质粒称为pCLiK5MCS。

根据Sanger等人(1977)所述(Proceedings of the National Academy ofSciences USA 74：5463-5467)实施测序反应。分段进行测序反应并通过ABIPrism 377(PE Applied Biosystems，Weiterstadt)分析。

所得质粒pCLiK5MCS如SEQ ID NO：65所示。

实施例2：pCLiK5MCS integrativ sacB的构建

以质粒pK19mob(Sch_fer等人，Gene 145，69-73(1994))作为模板开始PCR反应，使用寡核苷酸BK1732和BK1733扩增枯草芽孢杆菌sacB基因(编码果聚糖蔗糖酶)。

BK1732(SEQ ID NO：63)：

5‘-GAGAGCGGCCGCCGATCCTTTTTAACCCATCAC-3‘

BK1733(SEQ ID NO：64)：

5‘-AGGAGCGGCCGCCATCGGCATTTTCTTTTGCG-3‘

寡核苷酸BK1732和BK1733除了含有与pEK19mobsac互补的序列外，还含有限制性内切酶NotI的切割位点。使用PfuTurbo聚合酶(Stratagene，La Jolla，USA)根据标准方法如Innis等(PCR Protocols.AGuide to Methods and Applications，Academic Press(1990))的方法实施PCR反应。得到大小约1.9kb的DNA片段用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书纯化。DNA片段用限制性内切酶NotI(New England Biolabs，Beverly，USA)切割并且，之后再次用GFX^TMPCR、DNA和凝胶带纯化试剂盒(AmershamPharmacia，Freiburg)根据生产商的使用说明书纯化。

载体pCLiK5MCS(根据实施例1制备)也用限制性内切酶NotI切割并使用碱性磷酸酶(I(Roche Diagnostics，Mannheim))根据生产商的使用说明书去磷酸。在0.8％强度的琼脂糖凝胶中电泳后，使用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)根据生产商的使用说明书分离约2.4kb大小的载体。该载体片段使用快速DNA连接试剂盒(Roche Diagnostics，Mannheim)根据生产商的使用说明与切割的PCR片段连接并根据标准方法，如Sambrook等(分子克隆实验指南，冷泉港，(1989))中描述的方法将连接混合物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过涂在含有卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板上选择携带质粒的细胞。

使用Qiaprep spin miniprep试剂盒(Qiagen，Hilden)根据生产商的使用说明书分离单个克隆的质粒DNA并将它们通过限制性消化检查。以这种方法得到的质粒称为pCLiK5MCS integrativ sacB。

所得质粒pCLiK5MCS integrativ sacB如SEQ ID NO：66所示。

可以类似方式制备适于metY基因的发明性表达或过量产生的其他载体。

实施例3：从谷氨酸棒杆菌菌株LU1479分离lysC基因

菌株构建的第一步计划为谷氨酸棒杆菌ATCC13032(以下称为LU1479)中编码天冬氨酸激酶的lysC野生型基因的等位基因替换。计划在lysC基因中实施核苷酸替换从而在所得蛋白质中，311位的氨基酸Thr改变为氨基酸Ile。

以LU1479染色体DNA作为模板开始PCR，用寡核苷酸引物SEQ IDNO：67和SEQ ID NO：68 lysC，利用Pfu-Turbo PCR系统(Stratagene USA)按照生产商的使用说明书实施扩增。如Tauch等人(1995)Plasmid33：168-179或Eikmanns等人(1994)Microbiology 140：1817-1828描述的制备谷氨酸棒杆菌ATCC 13032的染色体DNA。所扩增片段的5’端侧翼位为SalI限制性切割，其3’端侧翼位为MluI限制性切割。克隆步骤前，扩增的片段用这两种限制酶消化并用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)纯化。

SEQ ID NO：67

5‘-GAGAGAGAGACGCGTCCCAGTGGCTGAGACGCATC-3‘

SEQ ID NO：68

5‘-CTCTCTCTGTCGACGAATTCAATCTTACGGCCTG-3‘

所得多核苷酸通过SalI和MluI切割被克隆到pCLIK5MCS integrativSacB(此后称为pCIS；实施例2的SEQ ID NO：66)并被转化到大肠杆菌XL-1 blue中。通过涂含有卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板实现对携带质粒的细胞的选择。分离质粒并通过测序验证预期的核苷酸序列。通过Quiagen的方法并使用来自Quiagen的材料制备质粒DNA。如Sanger等人(1977)Proceedings ofthe National Academyof Sciences USA 74：5463-5467描述的实施测序反应。通过ABI Prism 377(PE Applied Biosystems，Weiterstadt)分离测序反应物并对其评定。所得质粒pCIS lysC如SEQ ID NO：69所示。相应的质粒图在图1中显示。

序列SEQ ID NO：69包括下面的必需部分-区域：

基因座 pCIS\lysC 5860bp DNA 环状

特征定位/定义 (Qualifiers)

CDS¹⁾ 155..1420

/vntifkey＝″4″

/label＝lysC

CDS 互补的²⁾(3935..5356)

/vntifkey＝″4″

/label＝sacB\(枯草芽孢杆菌)

启动子互补的(5357..5819)

/vntifkey＝″30″

/label＝启动子\sacB

C_region 互补的(3913..3934)

/vntifkey＝″2″

/label＝sacB\下游区

CDS 1974..2765

/vntifkey＝″4″

/label＝Kan\R

CDS 互补的(3032..3892)

/vntifkey＝″4″

/label＝Ori\-EC\(pMB)

¹⁾编码序列

²⁾在互补链上

实施例4：谷氨酸棒杆菌lysC基因的诱变

使用QuickChange试剂盒(Stratagene/USA)按照生产商的使用说明书实施谷氨酸棒杆菌lysC基因(实施例3)的位点专一诱变。在质粒pCIS lysC，SEQ ID NO：69中实施诱变。利用Quickchange方法(Stratagene)合成了下面的寡核苷酸引物，其用于将thr311变为311ile。

SEQ ID NO：70

5‘-CGGCACCACCGACATCATCTTCACCTGCCCTCGTTCCG-3‘

SEQ ID NO：71

5‘-CGGAACGAGGGCAGGTGAAGATGATGTCGGTGGTGCCG-3‘

Quickchange反应中这些寡核苷酸引物的使用导致lysC基因中932位核苷酸的替换(用T代替C)(参考SEQ ID NO：72)和相应酶中311位中氨基酸的替换(Thr→Ile)(参考SEQ ID NO：73)。LysC基因中所得氨基酸替换Thr311Ile通过转化到大肠杆菌XL1-blue和质粒制备后测序来检验。该质粒被命名为pCIS lysC thr311ile并且如SEQ ID NO：74所示。相应的质粒图在图2中显示。

序列SEQ ID NO：74包括下面的必需部分区域：

基因座 pCIS\lysC\thr311ile 5860bp DNA环状

特征定位/定义

CDS¹⁾ 155..1420

/vntifkey＝″4″

/label＝lysC

CDS 互补的²⁾(3935..5356)

/vntifkey＝″4″

/label＝sacB\(枯草芽孢杆菌)

启动子互补的(5357..5819)

/vntifkey＝″30″

/label＝启动子\sacB

C_region 互补的(3913..3934)

/vntifkey＝″2″

/label＝sacB\下游区

CDS 1974..2765

/vntifkey＝″4″

/label＝Kan\R

CDS 互补的(3032..3892)

/vntifkey＝″4″

/label＝Ori\-EC\(pMB)

¹⁾编码区

²⁾在互补链上

通过如Liebl，等人(1989)FEMS Microbiology Letters 53：299-303描述的电穿孔法将质粒pCIS lysC thr311ile转化到谷氨酸棒杆菌LU1479中。方案的修改在DE-A-10046870中描述。使用标准方法如Sambrook等人((1989)，分子克隆实验指南，冷泉港)中描述的DNA印迹和杂交检查单个转化体的lysC基因座的染色体排列。从而确保转化体为具有通过同源重组整合在lysC基因座的被转化质粒的转化体。这些菌落在没有抗生素的培养基中生长过夜后，将细胞涂布在蔗糖-CM琼脂培养基(10％蔗糖)的平板上并在30℃孵育24小时。

因为存在于载体pCIS lysC thr311ile中的sacB基因将蔗糖转化成毒性产物，所以仅仅那些通过另一同源重组步骤将野生型lysC基因和突变的基因lysC thr311ile之间的sacB基因缺失的菌落能够生长。同源重组过程中，野生型基因或者突变基因与sacB基因一起可以被缺失。如果sacB基因与野生型基因一起被除去，则产生突变的转化体。

挑选生长菌落并检查卡那霉素敏感表型。缺失SacB基因的克隆必须同时表现出卡那霉素-敏感的生长行为。在摇瓶中研究这种Kan-敏感克隆的赖氨酸产量(见实施例6)。生长未处理的菌株LU1479用于比较目的。选择赖氨酸产量比对照增加的克隆，得到染色体DNA，并通过PCR反应扩增lysC基因的相应区域并测序。具有增加的赖氨酸合成和具有lysC中932位经证实突变的这种克隆之一称为LU1479 lysC 311ile。

实施例5：乙硫氨酸抗性谷氨酸棒杆菌菌株的产生

在第二个菌株构建步骤，处理所得菌株LU1479 lysC 311ile(实施例4)以诱导对乙硫氨酸的抗性(Kase，H.Nakayama K.Agr.Biol.Chem.39，153-106，1975)，通过谷氨酸棒杆菌的甲硫氨酸类似物抗性突变株产生L-甲硫氨酸)：BHI培养基(Difco)中的过夜培养物用柠檬酸盐缓冲液(50mMpH5.5)洗涤并在30℃用N-甲基亚硝基胍(50mM柠檬酸盐pH5.5中10mg/ml)处理20分钟。用化学诱变剂N-甲基亚硝基胍处理后，洗涤(柠檬酸盐缓冲液50mM pH5.5)细胞并将其涂布在含有下面组分的培养基平板上，在500ml中含有：10g(NH₄)₂SO₄、0.5g KH₂PO₄、0.5g K₂HPO₄、0.125gMgSO₄·7H₂O、21g MOPS、50mg CaCl₂、15mg原儿茶酸(proteocatechuate)、0.5mg生物素、1mg硫胺素、5g/l D，L-乙硫氨酸(SigmaChemicals Deutschland)，pH7.0。此外，培养基含有0.5ml微量盐溶液，其由10g/l FeSO₄·7H₂O、1g/l MnSO₄·H₂O、0.1g/l ZnSO₄·7H₂O、0.02g/lCuSO₄、0.002g/l NiCl₂·6H₂O组成，所有盐溶于0.1M HCl中。所完成的培养基通过过滤除菌，并且加入40ml无菌50％葡萄糖溶液后，加入液体无菌琼脂，其终浓度为1.5％琼脂，并将混合物倒入培养皿中。

将已经经历诱变处理的细胞应用于含有上述培养基的平板并在30℃孵育3-7天。分离所得克隆，单个克隆在选择培养基上分离至少一次然后在装有培养基II(见实施例6)的摇瓶中分析它们的甲硫氨酸产量。

实施例6：使用菌株LU1479 lysC 311ile ET-16制备甲硫氨酸

实施例5中产生的菌株在含有CM培养基的琼脂板上于30℃生长2天。

CM琼脂：

10.0g/l D-葡萄糖、2.5g/l NaCl、2.0g/l尿素、10.0g/l细菌培养用蛋白胨(Difco)、5.0g/l酵母提取物(Difco)、5.0g/l牛肉膏(Difco)、22.0g/l琼脂(Difco)，高压灭菌(20分钟，121℃)

细胞随后从平板刮下并重悬在盐水中。对于主培养，向100ml锥形烧瓶中的10ml培养基II和0.5g高压灭菌的CaCO₃(Riedel de Haen)接种细胞悬浮物至OD 600nm为1.5并在有轨摇床上以200转/分钟在30℃下孵育72小时。

培养基II：

40g/l 蔗糖

60g/l 糖蜜(基于100％糖含量)

10g/l (NH₄)₂SO₄

0.4g/l MgSO₄·7H₂O

0.6g/l KH₂PO₄

0.3mg/l 硫胺素·HCl

1mg/l 生物素(来自用NH₄OH调节至pH8.0的1mg/ml过滤除菌的母液)

2mg/l FeSO₄

2mg/l MnSO₄

用NH₄OH建立7.8的pH并将混合物高压灭菌(121℃，20分钟)。此外，加入来自母液(200μg/ml，过滤除菌的)的维生素B12(羟钴胺素，SigmaChemicals)至终浓度100μg/l。

使用Agilent氨基酸确定方法在Agilent 1100 Series LC System HPLC上确定培养液中形成的甲硫氨酸以及其他氨基酸。用正-邻苯二醛(ortho-phtalaldehyde)柱前衍生使得可以确定所形成的氨基酸量。氨基酸混合物在柱上分离。氨基酸混合物在Hypersil AA柱(Agilent)上分离。

分离甲硫氨酸产量比原始菌株LU1479 lysC 311ile的产量高至少2倍的克隆。一个这种克隆用于进一步的实验中，并命名为LU1479 lysC 311ileET-16。

实施例7：从结核分枝杆菌克隆metY并克隆到质粒pC PhsdhmetY_Mt中

结核分枝杆菌的染色体DNA来自菌株ATCC 25584，该菌株获自美国典型菌株培养物保藏中心(ATCC，Atlanta-USA)。通过Tauch等人(1995)Plasmid 33：168-179或Eikmanns等人(1994)Microbiology 140：1817-1828描述的方法制备谷氨酸棒杆菌ATCC 13032的染色体DNA。

使用寡核苷酸引物SEQ ID NO：75和SEQ ID NO：76、谷氨酸棒杆菌染色体DNA作为模板和Pfu Turbo聚合酶(Stratagene)，通过聚合酶链式反应(PCR)，按照标准方法如Innis等人(1990)(PCR Protocols.A Guide toMethods and Applications，Academic Press)所述从高丝氨酸脱氢酶(HsDH)的5’非编码区(启动子区)扩增了约180个碱基对的DNA片段。扩增的片段在其5’端侧翼为BamHI限制性切割位点，其3’端侧翼为与结核分枝杆菌的metY同源的区域并且已通过寡核苷酸引物导入。

SEQ ID NO：75

5‘-GAGAGGATCCGGAAGGTGAATCGAATTTCGG-3‘

和

SEQ ID NO：76

5‘-CTATTGCTGTCGGCGCTCATGATTCTCCAAAAATAATCGC-3‘

所得DNA片段用GFX^TMPCR、DNA和凝胶带纯化试剂盒(AmershamPharmacia，Freiburg)按照生产商的使用说明书纯化。

从作为PCR反应模板的来自结核分枝杆菌染色体DNA开始，通过富含GC的PCR系统(Roche Diagnostics，Mannheim)按照生产商的使用说明，使用寡核苷酸引物SEQ ID NO：77和SEQ ID NO：78扩增metY。被扩增的片段在其3’端侧翼为Xbal限制性切割位点，其已通过寡核苷酸引物导入。

SEQ ID NO：77

5‘-ATGAGCGCCGACAGCAATAG-3‘

和

SEQ ID NO：78

5‘-GAACTCTAGATCAGAACGCCGCCACGGAC-3‘

所得约1.4kb DNA片段用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)按照生产商的使用说明书纯化。

在进一步的PCR反应中，上面所得两种片段联合用作模板。由于存在与metY片段同源并已经通过寡核苷酸引物SEQ ID NO：76导入的区域，这两种片段在PCR反应中相互退火并被所用聚合酶延伸到连续的DNA链。修改了标准方法，其中将所用的寡核苷酸引物SEQ ID NO：75和SEQID NO：78仅在第二个循环开始时加至反应物。

扩增的DNA片段大小约为1.6kb，该DNA片段用GFX^TMPCR、DNA和凝胶带纯化试剂盒按照生产商的使用说明纯化。此后，将其用限制酶BamHI和Xbal(Roche Diagnostics，Mannheim)切割并通过凝胶电泳分离。使用GFX^TMPCR、DNA和凝胶带纯化试剂盒(Amersham Pharmacia，Freiburg)从琼脂糖分离约1.6kb DNA片段。

载体pCIik5MCS SEQ ID NO：65，此后称为pC，用限制酶BamHI和Xbal(Roche Diagnostics，Mannheim)切割，并且，通过电泳分离后，用GFX^TMPCR、DNA和凝胶带纯化试剂盒分离5kb片段。

利用快速DNA连接试剂盒(Roche Diagnostics，Mannheim)按照生产商的使用说明书将载体片段与被切割并分离的PCR片段连接，使用如Sambrook等人(分子克隆实验指南，冷泉港，(1989))描述的标准方法将连接反应物转化到感受态大肠杆菌XL-1Blue(Stratagene，La Jolla，USA)中。通过涂布在含有卡那霉素(20μg/ml)的LB琼脂(Lennox，1955，Virology，1：190)板上实现含有质粒的细胞的选择。

使用Quiagen的方法和来自Quiagen的材料制备质粒DNA。如Sanger等人(1977)Proceedings of the National Academy of Sciences USA74：5463-5467描述的实施测序反应。通过ABI Prism 377(PE AppliedBiosystems，Weiterstadt)分离和评价测序反应物。

所得质粒pC Phsdh metY_Mt(结核分枝杆菌)如SEQ ID NO：79所列。相应的质粒图在图3中显示。

SEQ ID NO：79含有下面的必需部分-区域：

基因座 pC\Phsdh\metY_Mt 6591 bp DNA 环状

2003年7月21日

特征定位/定义

CDS 156..1505

/vntifkey＝″4″

/label＝metY\aus\M\结核分枝杆菌

CDS 1855..2646

/vntifkey＝″4″

/label＝Kan\R

CDS 4927..6048

/vntifkey＝″4″

/label＝Rep\蛋白质

CDS 3919..4593

/vntifkey＝″4″

/label＝ORF\1

CDS 互补的(2913..3773)

/vntifkey＝″4″

/label＝Ori\-EC\(pMB)

实施例8：用质粒pC Phsdh metY_Mt转化菌株LU1479 lysC 311ileET-16

通过上述方法(Liebl，等人(1989)FEMS Microbiology Letters53：299-303)用质粒pC Phsdh metY_Mt转化菌株LU1479 lysC 311ileET-16。将转化混合物涂布到额外含有20mg/l卡那霉素的CM板上以得到对含有质粒的细胞的选择。挑选所得卡那霉素-抗性克隆并分离单个克隆。在摇瓶实验中研究克隆的甲硫氨酸产量(见实施例6)。菌株LU1479 lysC311ile ET-16 pC Phsdh metY_Mt与LU1479 lysC 311ile ET-16相比产生明显更多的甲硫氨酸。

序列表

<110>巴斯福股份公司

<120>发酵产生含硫精细化学品的方法(metY)

<130>M/43128

<160>66

<170>PatentIn版本3.1

<210>1

<211>1317

<212>DNA

<213>白喉棒杆菌(Corynebacterium diphtheriae)

<220>

<221>CDS

<222>(1)..(1317)

<223>

<400>1

atg cca aca aaa tac gat aat tcc aat gcc aac aaa tgg ggt ttc gag 48

Met Pro Thr Lys Tyr Asp Asn Ser Asn Ala Asn Lys Trp Gly Phe Glu

1 5 10 15

act cgc tcc atc cac gca gga caa agc gtc gat agt gat acc agt gcc 96

Thr Arg Ser Ile His Ala Gly Gln Ser Val Asp Ser Asp Thr Ser Ala

20 25 30

cgc aac cta ccg att tac ctg aca tca tcg tac gtt ttt aat gac gcc 144

Arg Asn Leu Pro Ile Tyr Leu Thr Ser Set Tyr Val Phe Asn Asp Ala

35 40 45

gaa cac gca gca aac cgc ttc aac ctt tcc gac gcc ggc ccg gtt tac 192

Glu His Ala Ala Asn Arg Phe Asn Leu Ser Asp Ala Gly Pro Val Tyr

50 55 60

tct cgc ctg acc aac cca act gtc gcg gca gtc gaa gaa cgc cta gcc 240

Ser Arg Leu Thr Asn Pro Thr Val Ala Ala Val Glu Glu Arg Leu Ala

65 70 75 80

aat ctt gaa ggt ggc gta cac gcc gta ctt ttc gct tcc gga atg gcc 288

Asn Leu Glu Gly Gly Val His Ala Val Leu Phe Ala Ser Gly Met Ala

85 90 95

gcc gaa acc gcc gca atc ctc aac atc gcc cgc gcg ggt tcc cac atc 336

Ala Glu Thr Ala Ala Ile Leu Asn Ile Ala Arg Ala Gly Ser His Ile

100 105 110

gtg tcc agt cct cgc att tac ggc ggc acc gaa aca ctc ttt gcc gtc 384

Val Ser Ser Pro Arg Ile Tyr Gly Gly Thr Glu Thr Leu Phe Ala Val

115 120 125

aca ttg gca cgc ctg ggc atc gaa acc act ttc gta gaa aat cct gac 432

Thr Leu Ala Arg Leu Gly Ile Glu Thr Thr Phe Val Glu Asn Pro Asp

130 135 140

gac cca gcc tca tgg gag gct gca gtt caa gac aac acg gta gct ctc 480

Asp Pro Ala Ser Trp Glu Ala Ala Val Gln Asp Asn Thr Val Ala Leu

145 150 155 160

tac gga gaa acc ttc gct aat cca caa gca gac gtg ctt gat att ccc 528

Tyr Gly Glu Thr Phe Ala Asn Pro Gln Ala Asp Val Leu Asp Ile Pro

165 170 175

gca atc gca gag gtt gcc cat aaa cat caa gta cca ctg atc gtc gac 576

Ala Ile Ala Glu Val Ala His Lys His Gln Val Pro Leu Ile Val Asp

180 185 190

aac acc ctc gca acc gca gcc ctt gta cgc ccc ctc gaa ctc ggt gca 624

Asn Thr Leu Ala Thr Ala Ala Leu Val Arg Pro Leu Glu Leu Gly Ala

195 200 205

gac gtc gtc gtg gca tcc cta acc aag ttc tac acc gga aat ggc tcc 672

Asp Val Val Val Ala Ser Leu Thr Lys Phe Tyr Thr Gly Asn Gly Ser

210 215 220

gga ctc ggc gga gtg ctt atc gac ggc gga aac ttc gac tgg acc gtc 720

Gly Leu Gly Gly Val Leu Ile Asp Gly Gly Asn Phe Asp Trp Thr Val

225 230 235 240

aca cgc aac ggc gaa ccg atc ttc ccc gac ttt gtc acc cca gat ccc 768

Thr Arg Asn Gly Glu Pro Ile Phe Pro Asp Phe Val Thr Pro Asp Pro

245 250 255

gcc tat cac ggt ctc aag tat tcc gat ctt ggt gcc ccc gcc ttc gga 816

Ala Tyr His Gly Leu Lys Tyr Ser Asp Leu Gly Ala Pro Ala Phe Gly

260 265 270

cta aag gct cgc gtc gga ctc ctg cgc gac acc ggc gca gcc cca tca 864

Leu Lys Ala Arg Val Gly Leu Leu Arg Asp Thr Gly Ala Ala Pro Ser

275 280 285

cca ctc aac gca tgg atc acc gca caa ggg ctc gac acc ctc tcg cta 912

Pro Leu Asn Ala Trp Ile Thr Ala Gln Gly Leu Asp Thr Leu Ser Leu

290 295 300

cga gta caa cgc cac aac gaa aac gca ctc gca gta gca caa ttc ctc 960

Arg Val Gln Arg His Asn Glu Asn Ala Leu Ala Val Ala Gln Phe Leu

305 310 315 320

gcc aac cac gag aaa gta gcc aag gtt aac tac gca ggc ctt ccc gac 1008

Ala Asn His Glu Lys Val Ala Lys Val Asn Tyr Ala Gly Leu Pro Asp

325 330 335

tcc cct tgg tac cca gtc aaa gaa aaa ctc gga ttc gac tac acc ggc 1056

Ser Pro Trp Tyr Pro Val Lys Glu Lys Leu Gly Phe Asp Tyr Thr Gly

340 345 350

tcc gta ctt tcc ttt gac gtt aaa ggt gga aaa aac gaa gca tgg cgc 1104

Ser Val Leu Ser Phe Asp Val Lys Gly Gly Lys Asn Glu Ala Trp Arg

355 360 365

ttt atc gac gca ctc aaa cta cac tcg aac ctc gcc aac gtc gga gac 1152

Phe Ile Asp Ala Leu Lys Leu His Ser Asn Leu Ala Asn Val Gly Asp

370 375 380

gta cgt tcc ctc gta gtc cac cca gcg acc acc acg cac tca caa tcg 1200

Val Arg Ser Leu Val Val His Pro Ala Thr Thr Thr His Ser Gln Ser

385 390 395 400

gaa gaa tcg gca ctt cta gcc gca gga att aat caa gca acc atc cga 1248

Glu Glu Ser Ala Leu Leu Ala Ala Gly Ile Asn Gln Ala Thr Ile Arg

405 410 415

ctc tcc gtc ggc atc gaa tcc atc gac gac atc atc gcc gac ctc aca 1296

Leu Ser Val Gly Ile Glu Ser Ile Asp Asp Ile Ile Ala Asp Leu Thr

420 425 430

gca ggt ttc gac gca atc taa 1317

Ala Gly Phe Asp Ala Ile

435

<210>2

<21l>438

<212>PRT

<213>白喉棒杆菌

<400>2

Met Pro Thr Lys Tyr Asp Asn Ser Asn Ala Asn Lys Trp Gly Phe Glu

1 5 10 15

Thr Arg Ser Ile His Ala Gly Gln Ser Val Asp Ser Asp Thr Ser Ala

20 25 30

Arg Asn Leu Pro Ile Tyr Leu Thr Ser Ser Tyr Val Phe Asn Asp Ala

35 40 45

Glu His Ala Ala Asn Arg Phe Asn Leu Ser Asp Ala Gly Pro Val Tyr

50 55 60

Ser Arg Leu Thr Asn Pro Thr Val Ala Ala Val Glu Glu Arg Leu Ala

65 70 75 80

Asn Leu Glu Gly Gly Val His Ala Val Leu Phe Ala Ser Gly Met Ala

85 90 95

Ala Glu Thr Ala Ala Ile Leu Asn Ile Ala Arg Ala Gly Ser His Ile

100 105 110

Val Ser Ser Pro Arg Ile Tyr Gly Gly Thr Glu Thr Leu Phe Ala Val

115 120 125

Thr Leu Ala Arg Leu Gly Ile Glu Thr Thr Phe Val Glu Asn Pro Asp

130 135 140

Asp Pro Ala Ser Trp Glu Ala Ala Val Gln Asp Asn Thr Val Ala Leu

145 150 155 160

Tyr Gly Glu Thr Phe Ala Asn Pro Gln Ala Asp Val Leu Asp Ile Pro

165 170 175

Ala Ile Ala Glu Val Ala His Lys His Gln Val Pro Leu Ile Val Asp

180 185 190

Asn Thr Leu Ala Thr Ala Ala Leu Val Arg Pro Leu Glu Leu Gly Ala

195 200 205

Asp Val Val Val Ala Ser Leu Thr Lys Phe Tyr Thr Gly Asn Gly Ser

210 215 220

Gly Leu Gly Gly Val Leu Ile Asp Gly Gly Asn Phe Asp Trp Thr Val

225 230 235 240

Thr Arg Asn Gly Glu Pro Ile Phe Pro Asp Phe Val Thr Pro Asp Pro

245 250 255

Ala Tyr His Gly Leu Lys Tyr Ser Asp Leu Gly Ala Pro Ala Phe Gly

260 265 270

Leu Lys Ala Arg Val Gly Leu Leu Arg Asp Thr Gly Ala Ala Pro Ser

275 280 285

Pro Leu Asn Ala Trp Ile Thr Ala Gln Gly Leu Asp Thr Leu Ser Leu

290 295 300

Arg Val Gln Arg His Asn Glu Asn Ala Leu Ala Val Ala Gln Phe Leu

305 310 315 320

Ala Asn His Glu Lys Val Ala Lys Val Asn Tyr Ala Gly Leu Pro Asp

325 330 335

Ser Pro Trp Tyr Pro Val Lys Glu Lys Leu Gly Phe Asp Tyr Thr Gly

340 345 350

Ser Val Leu Ser Phe Asp Val Lys Gly Gly Lys Asn Glu Ala Trp Arg

355 360 365

Phe Ile Asp Ala Leu Lys Leu His Ser Asn Leu Ala Asn Val Gly Asp

370 375 380

Val Arg Ser Leu Val Val His Pro Ala Thr Thr Thr His Ser Gln Ser

385 390 395 400

Glu Glu Ser Ala Leu Leu Ala Ala Gly Ile Asn Gln Ala Thr Ile Arg

405 410 415

Leu Ser Val Gly Ile Glu Ser Ile Asp Asp Ile Ile Ala Asp Leu Thr

420 425 430

Ala Gly Phe Asp Ala Ile

435

<210>3

<211>1350

<212>DNA

<213>结核分枝杆菌(Mycobacterium tuberculosis)

<220>

<221>CDS

<222>(1)..(1350)

<223>

<400>3

atg agc gcc gac agc aat agc acc gac gcc gat ccg acc gcg cat tgg 48

Met Ser Ala Asp Ser Asn Ser Thr Asp Ala Asp Pro Thr Ala His Trp

1 5 10 15

tcg ttc gaa acc aaa cag ata cac gct ggt cag cac cct gat ccg acc 96

Ser Phe Glu Thr Lys Gln Ile His Ala Gly Gln His Pro Asp Pro Thr

20 25 30

acc aac gcc cgg gct ctg ccg atc tat gcg acc acg tcg tac acc ttc 144

Thr Asn Ala Arg Ala Leu Pro Ile Tyr Ala Thr Thr Ser Tyr Thr Phe

35 40 45

gac gac acc gcg cac gcc gcc gcc ctg ttc gga ctg gaa att ccg ggc 192

Asp Asp Thr Ala His Ala Ala Ala Leu Phe Gly Leu Glu Ile Pro Gly

50 55 60

aat atc tac acc cgg atc ggc aac ccc acc acc gac gtc gtc gag cag 240

Asn Ile Tyr Thr Arg Ile Gly Asn Pro Thr Thr Asp Val Val Glu Gln

65 70 75 80

cgc atc gcc gcg ctc gag ggc ggt gtg gcc gcg ctg ttc ctg tcg tcg 288

Arg Ile Ala Ala Leu Glu Gly Gly Val Ala Ala Leu Phe Leu Ser Ser

85 90 95

ggg cag gcc gcg gag acg ttc gcc atc ttg aac ctg gcc ggc gcg ggc 336

Gly Gln Ala Ala Glu Thr Phe Ala Ile Leu Asn Leu Ala Gly Ala Gly

100 105 110

gat cac atc gtg tcc agc ccg cgc ctg tac ggc ggc acc tac aac ctg 384

Asp His Ile Val Ser Ser Pro Arg Leu Tyr Gly Gly Thr Tyr Asn Leu

115 120 125

ttc cac tat tcg ctg gcc aag ctc ggc atc gag gtc agc ttc gtc gac 432

Phe His Tyr Ser Leu Ala Lys Leu Gly Ile Glu Val Ser Phe Val Asp

130 135 140

gat ccg gac gat ctg gac acc tgg cag gcg gcg gta cgg ccc aac acc 480

Asp Pro Asp Asp Leu Asp Thr Trp Gln Ala Ala Val Arg Pro Asn Thr

145 150 155 160

aag gcg ttc ttc gcc gag acc atc tcc aac ccg cag atc gac ctg ctg 528

Lys Ala Phe Phe Ala Glu Thr Ile Ser Asn Pro Gln Ile Asp Leu Leu

165 170 175

gac acc ccg gcg gtt tcc gag gtc gcc cat cgc aac ggg gtg ccg ttg 576

Asp Thr Pro Ala Val Ser Glu Val Ala His Arg Asn Gly Val Pro Leu

180 185 190

atc gtc gac aac acc atc gcc acg cca tac ctg atc caa ccg ttg gcc 624

Ile Val Asp Asn Thr Ile Ala Thr Pro Tyr Leu Ile Gln Pro Leu Ala

195 200 205

cag ggc gcc gac atc gtc gtg cat tcg gcc acc aag tac ctg ggc ggg 672

Gln Gly Ala Asp Ile Val Val His Ser Ala Thr Lys Tyr Leu Gly Gly

210 215 220

cac ggt gcc gcc atc gcg ggt gtg atc gtc gac ggc ggc aac ttc gat 720

His Gly Ala Ala Ile Ala Gly Val Ile Val Asp Gly Gly Asn Phe Asp

225 230 235 240

tgg acc cag ggc cgc ttc ccc ggc ttc acc acc ccc gac ccc agc tac 768

Trp Thr Gln Gly Arg Phe Pro Gly Phe Thr Thr Pro Asp Pro Ser Tyr

245 250 255

cac ggc gtg gtg ttc gcc gag ctg ggt cca ccg gcg ttt gcg ctc aaa 816

His Gly Val Val Phe Ala Glu Leu Gly Pro Pro Ala Phe Ala Leu Lys

260 265 270

gct cga gtg cag ctg ctc cgt gac tac ggc tcg gcg gct tcg ccg ttc 864

Ala Arg Val Gln Leu Leu Arg Asp Tyr Gly Ser Ala Ala Ser Pro Phe

275 280 285

aac gcg ttc ttg gtg gcg cag ggt ctg gaa acg ctg agc ctg cgg atc 912

Asn Ala Phe Leu Val Ala Gln Gly Leu Glu Thr Leu Ser Leu Arg Ile

290 295 300

gag cgg cac gtc gcc aac gcg cag cgc gtc gcc gag ttc ctg gcc gcc 960

Glu Arg His Val Ala Asn Ala Gln Arg Val Ala Glu Phe Leu Ala Ala

305 310 315 320

cgc gac gac gtg ctt tcg gtc aac tat gcg ggg ctg ccc tcc tcg ccc 1008

Arg Asp Asp Val Leu Ser Val Asn Tyr Ala Gly Leu Pro Ser Ser Pro

325 330 335

tgg cat gag cgg gcc aag agg ctg gcg ccc aag gga acc ggg gcc gtg 1056

Trp His Glu Arg Ala Lys Arg Leu Ala Pro Lys Gly Thr Gly Ala Val

340 345 350

ctg tcc ttc gag ttg gcc ggc ggc atc gag gcc ggc aag gca ttc gtg 1104

Leu Ser Phe Glu Leu Ala Gly Gly Ile Glu Ala Gly Lys Ala Phe Val

355 360 365

aac gcg ttg aag ctg cac agc cac gtc gcc aac atc ggt gac gtg cgc 1152

Asn Ala Leu Lys Leu His Ser His Val Ala Asn Ile Gly Asp Val Arg

370 375 380

tcg ctg gtg atc cac ccg gca tcg acc act cat gcc cag ctg agc ccg 1200

Ser Leu Val Ile His Pro Ala Ser Thr Thr His Ala Gln Leu Ser Pro

385 390 395 400

gcc gag cag ctg gcg acc ggg gtc agc ccg ggc ctg gtg cgt ttg gct 1248

Ala Glu Gln Leu Ala Thr Gly Val Ser Pro Gly Leu Val Arg Leu Ala

405 410 415

gtg ggc atc gaa ggt atc gac gat atc ctg gcc gac ctg gag ctt ggc 1296

Val Gly Ile Glu Gly Ile Asp Asp Ile Leu Ala Asp Leu Glu Leu Gly

420 425 430

ttt gcc gcg gcc cgc aga ttc agc gcc gac ccg cag tcc gtg gcg gcg 1344

Phe Ala Ala Ala Arg Arg Phe Ser Ala Asp Pro Gln Ser Val Ala Ala

435 440 445

ttc tga 1350

Phe

<210>4

<211>449

<212>PRT

<213>结核分枝扦菌

<400>4

Met Ser Ala Asp Ser Asn Ser Thr Asp Ala Asp Pro Thr Ala His Trp

1 5 10 15

Ser Phe Glu Thr Lys Gln Ile His Ala Gly Gln His Pro Asp Pro Thr

20 25 30

Thr Asn Ala Arg Ala Leu Pro Ile Tyr Ala Thr Thr Ser Tyr Thr Phe

35 40 45

Asp Asp Thr Ala His Ala Ala Ala Leu Phe Gly Leu Glu Ile Pro Gly

50 55 60

Asn Ile Tyr Thr Arg Ile Gly Asn Pro Thr Thr Asp Val Val Glu Gln

65 70 75 80

Arg Ile Ala Ala Leu Glu Gly Gly Val Ala Ala Leu Phe Leu Ser Ser

85 90 95

Gly Gln Ala Ala Glu Thr Phe Ala Ile Leu Asn Leu Ala Gly Ala Gly

100 105 110

Asp His Ile Val Ser Ser Pro Arg Leu Tyr Gly Gly Thr Tyr Asn Leu

115 120 125

Phe His Tyr Ser Leu Ala Lys Leu Gly Ile Glu Val Ser Phe Val Asp

130 135 140

Asp Pro Asp Asp Leu Asp Thr Trp Gln Ala Ala Val Arg Pro Asn Thr

145 150 155 160

Lys Ala Phe Phe Ala Glu Thr Ile Ser Asn Pro Gln Ile Asp Leu Leu

165 170 175

Asp Thr Pro Ala Val Ser Glu Val Ala His Arg Asn Gly Val Pro Leu

180 185 190

Ile Val Asp Asn Thr Ile Ala Thr Pro Tyr Leu Ile Gln Pro Leu Ala

195 200 205

Gln Gly Ala Asp Ile Val Val His Ser Ala Thr Lys Tyr Leu Gly Gly

210 215 220

His Gly Ala Ala Ile Ala Gly Val Ile Val Asp Gly Gly Asn Phe Asp

225 230 235 240

Trp Thr Gln Gly Arg Phe Pro Gly Phe Thr Thr Pro Asp Pro Ser Tyr

245 250 255

His Gly Val Val Phe Ala Glu Leu Gly Pro Pro Ala Phe Ala Leu Lys

260 265 270

Ala Arg Val Gln Leu Leu Arg Asp Tyr Gly Ser Ala Ala Ser Pro Phe

275 280 285

Asn Ala Phe Leu Val Ala Gln Gly Leu Glu Thr Leu Ser Leu Arg Ile

290 295 300

Glu Arg His Val Ala Asn Ala Gln Arg Val Ala Glu Phe Leu Ala Ala

305 310 315 320

Arg Asp Asp Val Leu Ser Val Asn Tyr Ala Gly Leu Pro Ser Ser Pro

325 330 335

Trp His Glu Arg Ala Lys Arg Leu Ala Pro Lys Gly Thr Gly Ala Val

340 345 350

Leu Ser Phe Glu Leu Ala Gly Gly Ile Glu Ala Gly Lys Ala Phe Val

355 360 365

Asn Ala Leu Lys Leu His Ser His Val Ala Asn Ile Gly Asp Val Arg

370 375 380

Ser Leu Val Ile His Pro Ala Ser Thr Thr His Ala Gln Leu Ser Pro

385 390 395 400

Ala Glu Gln Leu Ala Thr Gly Val Ser Pro Gly Leu Val Arg Leu Ala

405 410 415

Val Gly Ile Glu Gly Ile Asp Asp Ile Leu Ala Asp Leu Glu Leu Gly

420 425 430

Phe Ala Ala Ala Arg Arg Phe Ser Ala Asp Pro Gln Ser Val Ala Ala

435 440 445

Phe

<210>5

<211>1284

<212>DNA

<213>丙酮丁醇梭菌(Clostridium acetobutylicum)

<220>

<221>CDS

<222>(1)..(1284)

<223>

<400>5

atg agt gaa gaa aga aaa ttt ggt ttt gaa aca tta cag gtt cat gca 48

Met Ser Glu Glu Arg Lys Phe Gly Phe Glu Thr Leu Gln Val His Ala

1 5 10 15

gga caa gtt gct gat cca act aca gga tca aga gct gta cct att tat 96

Gly Gln Val Ala Asp Pro Thr Thr Gly Ser Arg Ala Val Pro Ile Tyr

20 25 30

caa aca aca tca tat gta ttt aaa aat gct gat cat gca gca aat tta 144

Gln Thr Thr Ser Tyr Val Phe Lys Asn Ala Asp His Ala Ala Asn Leu

35 40 45

ttt caa ttg aaa gaa cct gga aat gta tat aca agg ata atg aat cca 192

Phe Gln Leu Lys Glu Pro Gly Asn Val Tyr Thr Arg Ile Met Asn Pro

50 55 60

aca act gat gta ttt gaa caa aga gta gca gct ctt gag ggc gga gtt 240

Thr Thr Asp Val Phe Glu Gln Arg Val Ala Ala Leu Glu Gly Gly Val

65 70 75 80

gct gga ctt gca aca gca tca gga ctt gca gca att acc tat gct att 288

Ala Gly Leu Ala Thr Ala Ser Gly Leu Ala Ala Ile Thr Tyr Ala Ile

85 90 95

tta aat gtg gca agt gct ggg gat gaa att gtt gca gca agt acc tta 336

Leu Asn Val Ala Ser Ala Gly Asp Glu Ile Val Ala Ala Ser Thr Leu

100 105 110

tat ggt gga aca tat gaa tta ttt ggg gtt act ctt aag aag ctt gga 384

Tyr Gly Gly Thr Tyr Glu Leu Phe Gly Val Thr Leu Lys Lys Leu Gly

115 120 125

ata aag gtt gtt ttt gta gat cca gat aat cct gaa aat ata aga aaa 432

Ile Lys Val Val Phe Val Asp Pro Asp Asn Pro Glu Asn Ile Arg Lys

130 135 140

gca ata aat gat agg aca aaa gct gta tat ggg gaa act att gga aat 480

Ala Ile Asn Asp Arg Thr Lys Ala Val Tyr Gly Glu Thr Ile Gly Asn

145 150 155 160

cca aga ata aat gtt ttg gat ata gag gca gta gct aaa att gcc cat 528

Pro Arg Ile Asn Val Leu Asp Ile Glu Ala Val Ala Lys Ile Ala His

165 170 175

gaa aat aaa ata cca ctt ata atc gat aat aca ttt ggt aca ccg tat 576

Glu Asn Lys Ile Pro Leu Ile Ile Asp Asn Thr Phe Gly Thr Pro Tyr

180 185 190

ctt ata aga cct ata gaa ttt gga gca gat ata gtt gta cat tca gca 624

Leu Ile Arg Pro Ile Glu Phe Gly Ala Asp Ile Val Val His Ser Ala

195 200 205

aca aag ttt ata gga gga cat gga act act ata ggt gga att ata gtt 672

Thr Lys Phe Ile Gly Gly His Gly Thr Thr Ile Gly Gly Ile Ile Val

210 215 220

gat ggt gga aaa ttt gat tgg aga gct agt gga aag ttt cct gat ttt 720

Asp Gly Gly Lys Phe Asp Trp Arg Ala Ser Gly Lys Phe Pro Asp Phe

225 230 235 240

aca aca ccg gat aag agc tat aat gga ctt ata tat gct gat cta ggt 768

Thr Thr Pro Asp Lys Ser Tyr Asn Gly Leu Ile Tyr Ala Asp Leu Gly

245 250 255

gca cct gct ttt gct tta aaa gca aga gtt caa ctt tta aga aat aca 816

Ala Pro Ala Phe Ala Leu Lys Ala Arg Val Gln Leu Leu Arg Asn Thr

260 265 270

ggt gca acg ctt agt cca caa agt gct ttt tat ttc cta caa ggg ttg 864

Gly Ala Thr Leu Ser Pro Gln Ser Ala Phe Tyr Phe Leu Gln Gly Leu

275 280 285

gaa tca ctt tca ctt agg gtt caa aaa cat gtt gat aat aca aga aag 912

Glu Ser Leu Ser Leu Arg Val Gln Lys His Val Asp Asn Thr Arg Lys

290 295 300

gta gtt gaa ttc ttg aag aac cat cca aaa gtt tca tgg ata aat tat 960

Val Val Glu Phe Leu Lys Asn His Pro Lys Val Ser Trp Ile Asn Tyr

305 310 315 320

cct gaa ctt gag gaa agt cct tat aaa gag tta gca aat aaa tat ctt 1008

Pro Glu Leu Glu Glu Ser Pro Tyr Lys Glu Leu Ala Asn Lys Tyr Leu

325 330 335

cca aag ggt gca ggc tca ata ttt aca ttt gga ata aag gga gga ctt 1056

Pro Lys Gly Ala Gly Ser Ile Phe Thr Phe Gly Ile Lys Gly Gly Leu

340 345 350

gaa gct ggt aaa aga ttt ata aat agt gtt aaa cta ttc tct ctt ttg 1104

Glu Ala Gly Lys Arg Phe Ile Asn Ser Val Lys Leu Phe Ser Leu Leu

355 360 365

gca aat gtt gca gat gca aaa tca ctt gtt ata cat cct tca agt aca 1152

Ala Asn Val Ala Asp Ala Lys Ser Leu Val Ile His Pro Ser Ser Thr

370 375 380

act cat gct gaa ctt aat gaa gaa gaa caa aaa gca gct ggt gtt act 1200

Thr His Ala Glu Leu Asn Glu Glu Glu Gln Lys Ala Ala Gly Val Thr

385 390 395 400

cca gat atg ata aga ctt tca ata gga gta gag gat gca gag gat tta 1248

Pro Asp Met Ile Arg Leu Ser Ile Gly Val Glu Asp Ala Glu Asp Leu

405 410 415

ata tgg gac tta aat caa gct ctc gaa caa gct taa 1284

Ile Trp Asp Leu Asn Gln Ala Leu Glu Gln Ala

420 425

<210>6

<211>427

<212>PRT

<213>丙酮丁醇梭菌

<400>6

Met Ser Glu Glu Arg Lys Phe Gly Phe Glu Thr Leu Gln Val His Ala

1 5 10 15

Gly Gln Val Ala Asp Pro Thr Thr Gly Ser Arg Ala Val Pro Ile Tyr

20 25 30

Gln Thr Thr Ser Tyr Val Phe Lys Asn Ala Asp His Ala Ala Asn Leu

35 40 45

Phe Gln Leu Lys Glu Pro Gly Asn Val Tyr Thr Arg Ile Met Asn Pro

50 55 60

Thr Thr Asp Val Phe Glu Gln Arg Val Ala Ala Leu Glu Gly Gly Val

65 70 75 80

Ala Gly Leu Ala Thr Ala Ser Gly Leu Ala Ala Ile Thr Tyr Ala Ile

85 90 95

Leu Asn Val Ala Ser Ala Gly Asp Glu Ile Val Ala Ala Ser Thr Leu

100 105 110

Tyr Gly Gly Thr Tyr Glu Leu Phe Gly Val Thr Leu Lys Lys Leu Gly

115 120 125

Ile Lys Val Val Phe Val Asp Pro Asp Asn Pro Glu Asn Ile Arg Lys

130 135 140

Ala Ile Asn Asp Arg Thr Lys Ala Val Tyr Gly Glu Thr Ile Gly Asn

145 150 155 160

Pro Arg Ile Asn Val Leu Asp Ile Glu Ala Val Ala Lys Ile Ala His

165 170 175

Glu Asn Lys Ile Pro Leu Ile Ile Asp Asn Thr Phe Gly Thr Pro Tyr

180 185 190

Leu Ile Arg Pro Ile Glu Phe Gly Ala Asp Ile Val Val His Ser Ala

195 200 205

Thr Lys Phe Ile Gly Gly His Gly Thr Thr Ile Gly Gly Ile Ile Val

210 215 220

Asp Gly Gly Lys Phe Asp Trp Arg Ala Ser Gly Lys Phe Pro Asp Phe

225 230 235 240

Thr Thr Pro Asp Lys Ser Tyr Asn Gly Leu Ile Tyr Ala Asp Leu Gly

245 250 255

Ala Pro Ala Phe Ala Leu Lys Ala Arg Val Gin Leu Leu Arg Asn Thr

260 265 270

Gly Ala Thr Leu Ser Pro Gln Ser Ala Phe Tyr Phe Leu Gln Gly Leu

275 280 285

Glu Ser Leu Ser Leu Arg Val Gln Lys His Val Asp Asn Thr Arg Lys

290 295 300

Val Val Glu Phe Leu Lys Asn His Pro Lys Val Ser Trp Ile Asn Tyr

305 310 315 320

Pro Glu Leu Glu Glu Ser Pro Tyr Lys Glu Leu Ala Asn Lys Tyr Leu

325 330 335

Pro Lys Gly Ala Gly Ser Ile Phe Thr Phe Gly Ile Lys Gly Gly Leu

340 345 350

Glu Ala Gly Lys Arg Phe Ile Asn Ser Val Lys Leu Phe Ser Leu Leu

355 360 365

Ala Asn Val Ala Asp Ala Lys Ser Leu Val Ile His Pro Ser Ser Thr

370 375 380

Thr His Ala Glu Leu Asn Glu Glu Glu Gln Lys Ala Ala Gly Val Thr

385 390 395 400

Pro Asp Met Ile Arg Leu Ser Ile Gly Val Glu Asp Ala Glu Asp Leu

405 410 415

Ile Trp Asp Leu Asn Gln Ala Leu Glu Gln Ala

420 425

<210>7

<211>1293

<212>DNA

<213>嗜碱芽孢杆菌(Bacillus halodurans)

<220>

<221>CDS

<222>(1)..(1293)

<223>

<400>7

atg aat cat gaa aac caa tgg cag tta gaa aca aag gcc gtt cat tca 48

Met Asn His Glu Asn Gln Trp Gln Leu Glu Thr Lys Ala Val His Ser

1 5 10 15

gga cag gag atc gat ccg aca acg ttg tcg cga gcc gtc cca ttg tac 96

Gly Gln Glu Ile Asp Pro Thr Thr Leu Ser Arg Ala Val Pro Leu Tyr

20 25 30

caa acg acg tcc tac gga ttt aaa gat aca gac cat gcg gcg aat tta 144

Gln Thr Thr Ser Tyr Gly Phe Lys Asp Thr Asp His Ala Ala Asn Leu

35 40 45

ttt tca cta agt gaa ttt ggc aat atc tat acc cga ttg atg aac cca 192

Phe Ser Leu Ser Glu Phe Gly Asn Ile Tyr Thr Arg Leu Met Asn Pro

50 55 60

acg aca gat gtg ttt gaa aaa cgt gtg gct gcg tta gaa gga gga gcg 240

Thr Thr Asp Val Phe Glu Lys Arg Val Ala Ala Leu Glu Gly Gly Ala

65 70 75 80

gca gct tta gcg acg gcc tca ggg cag gcg gcc att acg tat tcg att 288

Ala Ala Leu Ala Thr Ala Ser Gly Gln Ala Ala Ile Thr Tyr Ser Ile

85 90 95

tta aat att gcg gag gct gga gat gaa atc gtg tcc gct agt agc ctt 336

Leu Asn Ile Ala Glu Ala Gly Asp Glu Ile Val Ser Ala Ser Ser Leu

100 105 110

tac ggc gga acg tat aat tta ttt tcg att acg ttg cca aag cta ggg 384

Tyr Gly Gly Thr Tyr Asn Leu Phe Ser Ile Thr Leu Pro Lys Leu Gly

115 120 125

gta aac gtc cgt ttc gtt gat cca tcg gac cca gaa aac ttc aaa gca 432

Val Asn Val Arg Phe Val Asp Pro Ser Asp Pro Glu Asn Phe Lys Ala

130 135 140

gcg att act gaa aag acg aaa gcc att ttc gct gag tcg att gga aac 480

Ala Ile Thr Glu Lys Thr Lys Ala Ile Phe Ala Glu Ser Ile Gly Asn

145 150 155 160

cct aag gga gac gtg tta gat att gaa gcg gtg gcg aaa gtt gca cac 528

Pro Lys Gly Asp Val Leu Asp Ile Glu Ala Val Ala Lys Val Ala His

165 170 175

gat cat cac ctt ccc ctc att gtc gat aac acg ttt cca agc cca tat 576

Asp His His Leu Pro Leu Ile Val Asp Asn Thr Phe Pro Ser Pro Tyr

180 185 190

ttg ctt caa ccg ata aag cac ggc gca gac att gtt gtg cat tca gca 624

Leu Leu Gln Pro Ile Lys His Gly Ala Asp Ile Val Val His Ser Ala

195 200 205

aca aaa ttt atc ggt ggt cat ggg acg tcg ata gga ggg atc att gtc 672

Thr Lys Phe Ile Gly Gly His Gly Thr Ser Ile Gly Gly Ile Ile Val

210 215 220

gat gga ggg acg ttt gat tgg gcg aaa acg gat cga tat cca ggg cta 720

Asp Gly Gly Thr Phe Asp Trp Ala Lys Thr Asp Arg Tyr Pro Gly Leu

225 230 235 240

aca aca cct gat ccg agt tac cac ggt gtt gta tat aca gat gcg gtc 768

Thr Thr Pro Asp Pro Ser Tyr His Gly Val Val Tyr Thr Asp Ala Val

245 250 255

ggt cca att gct tat att att aaa gcg cgt gtt cag cta ttg cgt gac 816

Gly Pro Ile Ala Tyr Ile Ile Lys Ala Arg Val Gln Leu Leu Arg Asp

260 265 270

atg ggg gca gcc ata tcg cca ttt aac tcg ttt tta ctg ttg caa ggg 864

Met Gly Ala Ala Ile Ser Pro Phe Asn Ser Phe Leu Leu Leu Gln Gly

275 280 285

ttg gaa acg ttg cat tta cgg atg gag aga cat agt gaa aat gcc tac 912

Leu Glu Thr Leu His Leu Arg Met Glu Arg His Ser Glu Asn Ala Tyr

290 295 300

aaa gta gca gag ttc ctt gag caa cat caa gcg gtc gaa tcg gtg agc 960

Lys Val Ala Glu Phe Leu Glu Gln His Gln Ala Val Glu Ser Val Ser

305 310 315 320

tac tct gga ctg cca tcc cat cca tcc tac cca tta gcg aaa aaa tac 1008

Tyr Ser Gly Leu Pro Ser His Pro Ser Tyr Pro Leu Ala Lys Lys Tyr

325 330 335

tta cct aaa ggc caa ggg gct atc tta acg ttc gag gta aag ggc ggc 1056

Leu Pro Lys Gly Gln Gly Ala Ile Leu Thr Phe Glu Val Lys Gly Gly

340 345 350

gtt gaa gca gga aag aaa ctc att cat tcg gtc cag cta ttc tcc cac 1104

Val Glu Ala Gly Lys Lys Leu Ile His Ser Val Gln Leu Phe Ser His

355 360 365

ctt gcc aac gta ggt gat tca aaa tcg ttg atc atc cat cct gca agc 1152

Leu Ala Asn Val Gly Asp Ser Lys Ser Leu Ile Ile His Pro Ala Ser

370 375 380

acg acc cac caa cag ctc tcg gaa gca gaa cag cga gac gca gga gtg 1200

Thr Thr His Gln Gln Leu Ser Glu Ala Glu Gln Arg Asp Ala Gly Val

385 390 395 400

aca cct ggg atg atc aga ctt tcg gta gga acc gaa tcg att cat gat 1248

Thr Pro Gly Met Ile Arg Leu Ser Val Gly Thr Glu Ser Ile His Asp

405 410 415

att atc acc gat ctc aaa cag gcg att gag gcg agt caa gcg taa 1293

Ile Ile Thr Asp Leu Lys Gln Ala Ile Glu Ala Ser Gln Ala

420 425 430

<210>8

<211>430

<212>PRT

<213>嗜碱芽孢杆菌

<400>8

Met Asn His Glu Asn Gln Trp Gln Leu Glu Thr Lys Ala Val His Ser

1 5 10 15

Gly Gln Glu Ile Asp Pro Thr Thr Leu Ser Arg Ala Val Pro Leu Tyr

20 25 30

Gln Thr Thr Ser Tyr Gly Phe Lys Asp Thr Asp His Ala Ala Asn Leu

35 40 45

Phe Ser Leu Ser Glu Phe Gly Asn Ile Tyr Thr Arg Leu Met Asn Pro

50 55 60

Thr Thr Asp Val Phe Glu Lys Arg Val Ala Ala Leu Glu Gly Gly Ala

65 70 75 80

Ala Ala Leu Ala Thr Ala Ser Gly Gln Ala Ala Ile Thr Tyr Ser Ile

85 90 95

Leu Asn Ile Ala Glu Ala Gly Asp Glu Ile Val Ser Ala Ser Ser Leu

100 105 110

Tyr Gly Gly Thr Tyr Asn Leu Phe Ser Ile Thr Leu Pro Lys Leu Gly

115 120 125

Val Asn Val Arg Phe Val Asp Pro Ser Asp Pro Glu Asn Phe Lys Ala

130 135 140

Ala Ile Thr Glu Lys Thr Lys Ala Ile Phe Ala Glu Ser Ile Gly Asn

145 150 155 160

Pro Lys Gly Asp Val Leu Asp Ile Glu Ala Val Ala Lys Val Ala His

165 170 175

Asp His His Leu Pro Leu Ile Val Asp Asn Thr Phe Pro Ser Pro Tyr

180 185 190

Leu Leu Gln Pro Ile Lys His Gly Ala Asp Ile Val Val His Ser Ala

195 200 205

Thr Lys Phe Ile Gly Gly His Gly Thr Ser Ile Gly Gly Ile Ile Val

210 215 220

Asp Gly Gly Thr Phe Asp Trp Ala Lys Thr Asp Arg Tyr Pro Gly Leu

225 230 235 240

Thr Thr Pro Asp Pro Ser Tyr His Gly Val Val Tyr Thr Asp Ala Val

245 250 255

Gly Pro Ile Ala Tyr Ile Ile Lys Ala Arg Val Gln Leu Leu Arg Asp

260 265 270

Met Gly Ala Ala Ile Ser Pro Phe Asn Ser Phe Leu Leu Leu Gln Gly

275 280 285

Leu Glu Thr Leu His Leu Arg Met Glu Arg His Ser Glu Asn Ala Tyr

290 295 300

Lys Val Ala Glu Phe Leu Glu Gln His Gln Ala Val Glu Ser Val Ser

305 310 315 320

Tyr Ser Gly Leu Pro Ser His Pro Ser Tyr Pro Leu Ala Lys Lys Tyr

325 330 335

Leu Pro Lys Gly Gln Gly Ala Ile Leu Thr Phe Glu Val Lys Gly Gly

340 345 350

Val Glu Ala Gly Lys Lys Leu Ile His Ser Val Gln Leu Phe Ser His

355 360 365

Leu Ala Asn Val Gly Asp Ser Lys Ser Leu Ile Ile His Pro Ala Ser

370 375 380

Thr Thr His Gln Gln Leu Ser Glu Ala Glu Gln Arg Asp Ala Gly Val

385 390 395 400

Thr Pro Gly Met Ile Arg Leu Ser Val Gly Thr Glu Ser Ile His Asp

405 410 415

Ile Ile Thr Asp Leu Lys Gln Ala Ile Glu Ala Ser Gln Ala

420 425 430

<210>9

<211>1203

<212>DNA

<213>嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)

<220>

<221>CDS

<222>(1)..(1203)

<223>

<400>9

atg tcg tat gta ttc cgc gac agc gag cac gcg gcc aat ttg ttt ggt 48

Met Ser Tyr Val Phe Arg Asp Ser Glu His Ala Ala Asn Leu Phe Gly

1 5 10 15

ttg aaa gag gaa ggt ttt att tat acg cgc att atg aat cca acg aac 96

Leu Lys Glu Glu Gly Phe Ile Tyr Thr Arg Ile Met Asn Pro Thr Asn

20 25 30

gac gtg ttc gaa aag cgg atc gcg gcg ctt gaa ggc ggc att ggg gcg 144

Asp Val Phe Glu Lys Arg Ile Ala Ala Leu Glu Gly Gly Ile Gly Ala

35 40 45

ctc gcg ctg tca tcg ggg cag gcg gcg gtg ttt tat tcg atc atc aac 192

Leu Ala Leu Ser Ser Gly Gln Ala Ala Val Phe Tyr Ser Ile Ile Asn

50 55 60

atc gcc tcg gcg ggc gat gaa atc gtc tcg tct tcg tcc att tac ggc 240

Ile Ala Ser Ala Gly Asp Glu Ile Val Ser Ser Ser Ser Ile Tyr Gly

65 70 75 80

gga acg tac aac ttg ttc gcc cat acg ctg cgc aag ttc ggc att acg 288

Gly Thr Tyr Asn Leu Phe Ala His Thr Leu Arg Lys Phe Gly Ile Thr

85 90 95

gtg aag ttt gtc gat ccg tcc gac ccc gaa aac ttt gag cgg gcg atc 336

Val Lys Phe Val Asp Pro Ser Asp Pro Glu Asn Phe Glu Arg Ala Ile

100 105 110

acc gac aaa acg aaa gcc ttg ttt gcg gaa acg atc ggc aac ccg aaa 384

Thr Asp Lys Thr Lys Ala Leu Phe Ala Glu Thr Ile Gly Asn Pro Lys

115 120 125

aac gat gtg ttg gac att gaa gcg gtg gcc gac atc gcc cat cgc cat 432

Asn Asp Val Leu Asp Ile Glu Ala Val Ala Asp Ile Ala His Arg His

130 135 140

gcc att ccg ctc att gtc gac aac acg gtg gcc agt cca tac tta ttg 480

Ala Ile Pro Leu Ile Val Asp Asn Thr Val Ala Ser Pro Tyr Leu Leu

145 150 155 160

cgg ccg att gaa ttc ggc gcc gat atc gtc gtc cac tca gcg acg aag 528

Arg Pro Ile Glu Phe Gly Ala Asp Ile Val Val His Ser Ala Thr Lys

165 170 175

ttc atc ggc ggg cac ggc aat tcg atc ggc ggt gtg att gtg gac agc 576

Phe Ile Gly Gly His Gly Asn Ser Ile Gly Gly Val Ile Val Asp Ser

180 185 190

ggc aag ttt gac tgg aaa ggg agc ggc aag ttt ccg gag ttc acc gag 624

Gly Lys Phe Asp Trp Lys Gly Ser Gly Lys Phe Pro Glu Phe Thr Glu

195 200 205

cca gac cca agc tac cac ggt ttg gtg tat gtg gac gcc gtc ggc gaa 672

Pro Asp Pro Ser Tyr His Gly Leu Val Tyr Val Asp Ala Val Gly Glu

210 215 220

gcg gcg tac atc acg aaa gcg cgc arc cag ctc ttg cgc gat ttg gga 720

Ala Ala Tyr Ile Thr Lys Ala Arg Ile Gln Leu Leu Arg Asp Leu Gly

225 230 235 240

gcg gcg ttg tcg ccg ttt aat gcg ttt ttg ctt ttg caa ggg ttg gag 768

Ala Ala Leu Ser Pro Phe Asn Ala Phe Leu Leu Leu Gln Gly Leu Glu

245 250 255

acg ctc cat ttg cgg atg cag cgc cat agc gaa aac gcc ctt gcc gtc 816

Thr Leu His Leu Arg Met Gln Arg His Ser Glu Asn Ala Leu Ala Val

260 265 270

gcc aag ttt tta gaa gag gaa gaa gcg gtc gaa tcg gtc aat tac cca 864

Ala Lys Phe Leu Glu Glu Glu Glu Ala Val Glu Ser Val Asn Tyr Pro

275 280 285

ggg ctt ccg agc cat ccg tcg cat gaa ctg gcg aaa aaa tat ttg cca 912

Gly Leu Pro Ser His Pro Ser His Glu Leu Ala Lys Lys Tyr Leu Pro

290 295 300

aac ggg caa gga gcg atc gtc acg ttt gaa atc aaa ggc ggc gtc gaa 960

Asn Gly Gln Gly Ala Ile Val Thr Phe Glu Ile Lys Gly Gly Val Glu

305 310 315 320

gcc ggc aaa aaa ctg atc gac tcg gtc aaa ctg ttc tct cat ttg gcc 1008

Ala Gly Lys Lys Leu Ile Asp Ser Val Lys Leu Phe Ser His Leu Ala

325 330 335

aac atc ggc gat tcg aaa tcg ctc atc atc cac ccg gcc agc aca acg 1056

Asn Ile Gly Asp Ser Lys Ser Leu Ile Ile His Pro Ala Ser Thr Thr

340 345 350

cac gag cag ctg agc cca gat gaa cag ctg tcc gcc ggc gtc acc cca 1104

His Glu Gln Leu Ser Pro Asp Glu Gln Leu Ser Ala Gly Val Thr Pro

355 360 365

ggc ctt gtg cgt ctg tcc gtc ggc aca gaa gcg atc gac gac att ttg 1152

Gly Leu Val Arg Leu Ser Val Gly Thr Glu Ala Ile Asp Asp Ile Leu

370 375 380

gac gac ttg cgc caa gcc att cgc caa agc cag acg gtg ggg gtg aag 1200

Asp Asp Leu Arg Gln Ala Ile Arg Gln Ser Gln Thr Val Gly Val Lys

385 390 395 400

tag 1203

<210>10

<21l>400

<212>PRT

<213>嗜热脂肪芽孢杆菌

<400>10

Met Ser Tyr Val Phe Arg Asp Ser Glu His Ala Ala Asn Leu Phe Gly

1 5 10 15

Leu Lys Glu Glu Gly Phe Ile Tyr Thr Arg Ile Met Asn Pro Thr Asn

20 25 30

Asp Val Phe Glu Lys Arg Ile Ala Ala Leu Glu Gly Gly Ile Gly Ala

35 40 45

Leu Ala Leu Ser Ser Gly Gln Ala Ala Val Phe Tyr Ser Ile Ile Asn

50 55 60

Ile Ala Ser Ala Gly Asp Glu Ile Val Ser Ser Ser Ser Ile Tyr Gly

65 70 75 80

Gly Thr Tyr Asn Leu Phe Ala His Thr Leu Arg Lys Phe Gly Ile Thr

85 90 95

Val Lys Phe Val Asp Pro Ser Asp Pro Glu Asn Phe Glu Arg Ala Ile

100 105 110

Thr Asp Lys Thr Lys Ala Leu Phe Ala Glu Thr Ile Gly Asn Pro Lys

115 120 125

Asn Asp Val Leu Asp Ile Glu Ala Val Ala Asp Ile Ala His Arg His

130 135 140

Ala Ile Pro Leu Ile Val Asp Asn Thr Val Ala Ser Pro Tyr Leu Leu

145 150 155 160

Arg Pro Ile Glu Phe Gly Ala Asp Ile Val Val His Ser Ala Thr Lys

165 170 175

Phe Ile Gly Gly His Gly Asn Ser Ile Gly Gly Val Ile Val Asp Ser

180 185 190

Gly Lys Phe Asp Trp Lys Gly Ser Gly Lys Phe Pro Glu Phe Thr Glu

195 200 205

Pro Asp Pro Ser Tyr His Gly Leu Val Tyr Val Asp Ala Val Gly Glu

210 215 220

Ala Ala Tyr Ile Thr Lys Ala Arg Ile Gln Leu Leu Arg Asp Leu Gly

225 230 235 240

Ala Ala Leu Ser Pro Phe Asn Ala Phe Leu Leu Leu Gln Gly Leu Glu

245 250 255

Thr Leu His Leu Arg Met Gln Arg His Ser Glu Asn Ala Leu Ala Val

260 265 270

Ala Lys Phe Leu Glu Glu Glu Glu Ala Val Glu Ser Val Asn Tyr Pro

275 280 285

Gly Leu Pro Ser His Pro Ser His Glu Leu Ala Lys Lys Tyr Leu Pro

290 295 300

Asn Gly Gln Gly Ala Ile Val Thr Phe Glu Ile Lys Gly Gly Val Glu

305 310 315 320

Ala Gly Lys Lys Leu Ile Asp Ser Val Lys Leu Phe Ser His Leu Ala

325 330 335

Asn Ile Gly Asp Ser Lys Ser Leu Ile Ile His Pro Ala Ser Thr Thr

340 345 350

His Glu Gln Leu Ser Pro Asp Glu Gln Leu Ser Ala Gly Val Thr Pro

355 360 365

Gly Leu Val Arg Leu Ser Val Gly Thr Glu Ala Ile Asp Asp Ile Leu

370 375 380

Asp Asp Leu Arg Gln Ala Ile Arg Gln Ser Gln Thr Val Gly Val Lys

385 390 395 400

<210>11

<211>1290

<212>DNA

<213>微温绿菌(Chlorobium tepidum)

<220>

<221>CDS

<222>(1)..(1290)

<223>

<400>11

atg agt gag gat aac acc ttc cgg ttc gag acc ttg cag gtt cac gcc 48

Met Ser Glu Asp Asn Thr Phe Arg Phe Glu Thr Leu Gln Val His Ala

1 5 10 15

ggg cag gag cct gat ccg gtg acc gga tcg cgc gcc gtg ccc att tac 96

Gly Gln Glu Pro Asp Pro Val Thr Gly Ser Arg Ala Val Pro Ile Tyr

20 25 30

cag acc acc tcc tac gtg ttc gag aac gcc gag cac ggc gct gac ctg 144

Gln Thr Thr Ser Tyr Val Phe Glu Asn Ala Glu His Gly Ala Asp Leu

35 40 45

ttc gcg ctt cgc aag gcg ggc aat atc tac acg cgc ctg atg aac ccg 192

Phe Ala Leu Arg Lys Ala Gly Asn Ile Tyr Thr Arg Leu Met Asn Pro

50 55 60

acc acc gac gtg ctc gaa aag cgc atg gcg gcg ctc gaa ggg ggc aag 240

Thr Thr Asp Val Leu Glu Lys Arg Met Ala Ala Leu Glu Gly Gly Lys

65 70 75 80

gcg gcc ctc ggc gtg gcg agc ggc cac tcg gcg cag ttc atc gct att 288

Ala Ala Leu Gly Val Ala Ser Gly His Ser Ala Gln Phe Ile Ala Ile

85 90 95

gcc acc atc tgc cag gct gga gac aac att gtg tca tcg agc tat ctc 336

Ala Thr Ile Cys Gln Ala Gly Asp Asn Ile Val Ser Ser Ser Tyr Leu

100 105 110

tac ggc ggc acc tac aac cag ttc aag gtc gcc ttc aag cgc ctc ggc 384

Tyr Gly Gly Thr Tyr Asn Gln Phe Lys Val Ala Phe Lys Arg Leu Gly

115 120 125

atc gag gtg agg ttc gtg gat ggc aac gat cag gag gcg ttc cgc aag 432

Ile Glu Val Arg Phe Val Asp Gly Asn Asp Gln Glu Ala Phe Arg Lys

130 135 140

gct atc gac gag aac acg aaa gcg ctc tac atg gag tcc agc ggc aat 480

Ala Ile Asp Glu Asn Thr Lys Ala Leu Tyr Met Glu Ser Ser Gly Asn

145 150 155 160

ccg gcg ttc cat gtg ccc gat ttc gac gct atc gcg aag att gcc cgt 528

Pro Ala Phe His Val Pro Asp Phe Asp Ala Ile Ala Lys Ile Ala Arg

165 170 175

gag aac ggc att ccg ctg atc gtc gat aac acc ttt ggc tgc gcg ggc 576

Glu Asn Gly Ile Pro Leu Ile Val Asp Asn Thr Phe Gly Cys Ala Gly

180 185 190

tat ctc tgc cgt ccc att gat cac ggc gcg tcg atc gtg gtc gag tcg 624

Tyr Leu Cys Arg Pro Ile Asp His Gly Ala Ser Ile Val Val Glu Ser

195 200 205

gcc acc aag tgg atc ggc ggg cac ggc acc tcg atg ggc ggc atc atc 672

Ala Thr Lys Trp Ile Gly Gly His Gly Thr Ser Met Gly Gly Ile Ile

210 215 220

gtc gat gcc gga acg ttc gac tgg ggc aac ggc aag ttt ccg ctc ttt 720

Val Asp Ala Gly Thr Phe Asp Trp Gly Asn Gly Lys Phe Pro Leu Phe

225 230 235 240

acc gag cca tcg gaa ggc tat cac ggc ctg aaa ttc tac gaa gcg gtc 768

Thr Glu Pro Ser Glu Gly Tyr His Gly Leu Lys Phe Tyr Glu Ala Val

245 250 255

ggc gag ctg gcc ttt atc atc cgg gcg cgg gtc gag gga ctg cgg gat 816

Gly Glu Leu Ala Phe Ile Ile Arg Ala Arg Val Glu Gly Leu Arg Asp

260 265 270

ttc ggc ccg gcg atc agc ccg ttc aac tcc ttc atg ctg ttg cag gga 864

Phe Gly Pro Ala Ile Ser Pro Phe Asn Ser Phe Met Leu Leu Gln Gly

275 280 285

ctt gaa acg ctc tcg ctt cgc gtg cag cgc cac ctc gac aac acg ctt 912

Leu Glu Thr Leu Ser Leu Arg Val Gln Arg His Leu Asp Asn Thr Leu

290 295 300

gaa ctg gcc cgc tgg ctc gaa agg cac gat gcg gtt gcg tgg gtg aac 960

Glu Leu Ala Arg Trp Leu Glu Arg His Asp Ala Val Ala Trp Val Asn

305 310 315 320

tat cca ggc ctc gaa agc cat ccg aca cac gcc ctg gca aaa aaa tat 1008

Tyr Pro Gly Leu Glu Ser His Pro Thr His Ala Leu Ala Lys Lys Tyr

325 330 335

ctc acg cat ggc ttc ggc tgc gtg ctg act ttc ggc gtg aag ggt ggt 1056

Leu Thr His Gly Phe Gly Cys Val Leu Thr Phe Gly Val Lys Gly Gly

340 345 350

tat gaa aac gcg gtg aag ttc atc gac agc gtg aag ctg gcg agc cac 1104

Tyr Glu Asn Ala Val Lys Phe Ile Asp Ser Val Lys Leu Ala Ser His

355 360 365

ctg gcc aac gtg ggt gat gca aaa acg ctc gtc att cat ccg gca tcg 1152

Leu Ala Asn Val Gly Asp Ala Lys Thr Leu Val Ile His Pro Ala Ser

370 375 380

acg acg cac cag cag ctc agc gcc gag gaa cag gta tcg gcg ggc gtc 1200

Thr Thr His Gln Gln Leu Ser Ala Glu Glu Gln Val Ser Ala Gly Val

385 390 395 400

acc gcc gat atg gtg cgc gtg tcg gtt ggt atc gag cat atc gat gac 1248

Thr Ala Asp Met Val Arg Val Ser Val Gly Ile Glu His Ile Asp Asp

405 410 415

atc aag gct gat ttc agc cag gct ttc gag aat tta gca tga 1290

Ile Lys Ala Asp Phe Ser Gln Ala Phe Glu Asn Leu Ala

420 425

<210>12

<211>429

<212>PRT

<213>微温绿菌

<400>12

Met Ser Glu Asp Asn Thr Phe Arg Phe Glu Thr Leu Gln Val His Ala

1 5 10 15

Gly Gln Glu Pro Asp Pro Val Thr Gly Ser Arg Ala Val Pro Ile Tyr

20 25 30

Gln Thr Thr Ser Tyr Val Phe Glu Asn Ala Glu His Gly Ala Asp Leu

35 40 45

Phe Ala Leu Arg Lys Ala Gly Asn Ile Tyr Thr Arg Leu Met Asn Pro

50 55 60

Thr Thr Asp Val Leu Glu Lys Arg Met Ala Ala Leu Glu Gly Gly Lys

65 70 75 80

Ala Ala Leu Gly Val Ala Ser Gly His Ser Ala Gln Phe Ile Ala Ile

85 90 95

Ala Thr Ile Cys Gln Ala Gly Asp Asn Ile Val Ser Ser Ser Tyr Leu

100 105 110

Tyr Gly Gly Thr Tyr Asn Gln Phe Lys Val Ala Phe Lys Arg Leu Gly

115 120 125

Ile Glu Val Arg Phe Val Asp Gly Asn Asp Gln Glu Ala Phe Arg Lys

130 135 140

Ala Ile Asp Glu Asn Thr Lys Ala Leu Tyr Met Glu Ser Ser Gly Asn

145 150 155 160

Pro Ala Phe His Val Pro Asp Phe Asp Ala Ile Ala Lys Ile Ala Arg

165 170 175

Glu Asn Gly Ile Pro Leu Ile Val Asp Asn Thr Phe Gly Cys Ala Gly

180 185 190

Tyr Leu Cys Arg Pro Ile Asp His Gly Ala Ser Ile Val Val Glu Ser

195 200 205

Ala Thr Lys Trp Ile Gly Gly His Gly Thr Ser Met Gly Gly Ile Ile

210 215 220

Val Asp Ala Gly Thr Phe Asp Trp Gly Asn Gly Lys Phe Pro Leu Phe

225 230 235 240

Thr Glu Pro Ser Glu Gly Tyr His Gly Leu Lys Phe Tyr Glu Ala Val

245 250 255

Gly Glu Leu Ala Phe Ile Ile Arg Ala Arg Val Glu Gly Leu Arg Asp

260 265 270

Phe Gly Pro Ala Ile Ser Pro Phe Asn Ser Phe Met Leu Leu Gln Gly

275 280 285

Leu Glu Thr Leu Ser Leu Arg Val Gln Arg His Leu Asp Asn Thr Leu

290 295 300

Glu Leu Ala Arg Trp Leu Glu Arg His Asp Ala Val Ala Trp Val Asn

305 310 315 320

Tyr Pro Gly Leu Glu Ser His Pro Thr His Ala Leu Ala Lys Lys Tyr

325 330 335

Leu Thr His Gly Phe Gly Cys Val Leu Thr Phe Gly Val Lys Gly Gly

340 345 350

Tyr Glu Asn Ala Val Lys Phe Ile Asp Ser Val Lys Leu Ala Ser His

355 360 365

Leu Ala Asn Val Gly Asp Ala Lys Thr Leu Val Ile His Pro Ala Ser

370 375 380

Thr Thr His Gln Gln Leu Ser Ala Glu Glu Gln Val Ser Ala Gly Val

385 390 395 400

Thr Ala Asp Met Val Arg Val Ser Val Gly Ile Glu His Ile Asp Asp

405 410 415

Ile Lys Ala Asp Phe Ser Gln Ala Phe Glu Asn Leu Ala

420 425

<210>13

<211>1281

<212>DNA

<213>乳酸乳球菌(Lactococcus lactis)

<220>

<221>CDS

<222>(1)..(1281)

<223>

<400>13

atg act aat cac aat tat aaa ttc gac act ttg caa gtc cat gca gga 48

Met Thr Asn His Asn Tyr Lys Phe Asp Thr Leu Gln Val His Ala Gly

1 5 10 15

caa gtc cct gat cct gtc acg ggt tca cgc gcc gtt ccg ctc tat caa 96

Gln Val Pro Asp Pro Val Thr Gly Ser Arg Ala Val Pro Leu Tyr Gln

20 25 30

aca act tct ttc gtt ttt aac aat tca gac cat gcc gaa gct cgt ttt 144

Thr Thr Ser Phe Val Phe Asn Asn Ser Asp His Ala Glu Ala Arg Phe

35 40 45

gct tta caa gat cct gga gct att tat tca cgt tta gga aat cca acc 192

Ala Leu Gln Asp Pro Gly Ala Ile Tyr Ser Arg Leu Gly Asn Pro Thr

50 55 60

aac gat gtt ttt gaa gca cgc atc gca gct ctt gaa ggt gga agt gca 240

Asn Asp Val Phe Glu Ala Arg Ile Ala Ala Leu Glu Gly Gly Ser Ala

65 70 75 80

gcc ctt ggt gtt ggt tct ggc tca gcc gct att acc tat gcc atc ttg 288

Ala Leu Gly Val Gly Ser Gly Ser Ala Ala Ile Thr Tyr Ala Ile Leu

85 90 95

aat atc gct aca gtc ggt gat aat att gtt tcc gca agt acc ctt tat 336

Asn Ile Ala Thr Val Gly Asp Asn Ile Val Ser Ala Ser Thr Leu Tyr

100 105 110

ggt gga acc tat cac ctt ttt tct ggg act tta cca aaa tat gga att 384

Gly Gly Thr Tyr His Leu Phe Ser Gly Thr Leu Pro Lys Tyr Gly Ile

115 120 125

aca act aaa ttt gtc aat cca gat gac ccg aag aat ttt gaa gag gcg 432

Thr Thr Lys Phe Val Asn Pro Asp Asp Pro Lys Asn Phe Glu Glu Ala

130 135 140

att gat gaa aaa acc aaa gct att tat tat gaa act ttg ggc aat ccg 480

Ile Asp Glu Lys Thr Lys Ala Ile Tyr Tyr Glu Thr Leu Gly Asn Pro

145 150 155 160

gga aat aat gtg att gat tat gat gcc att ggt caa att gct aaa aaa 528

Gly Asn Asn Val Ile Asp Tyr Asp Ala Ile Gly Gln Ile Ala Lys Lys

165 170 175

cat gga att ccc gtt att gtt gat gca acg ttt act acc cct gtg acc 576

His Gly Ile Pro Val Ile Val Asp Ala Thr Phe Thr Thr Pro Val Thr

180 185 190

ttt aaa cca ttt gaa cat ggt gct aat gta att gtt cat tca gca acg 624

Phe Lys Pro Phe Glu His Gly Ala Asn Val Ile Val His Ser Ala Thr

195 200 205

aaa ttc att ggc ggt cat ggt act tct att ggt gga gtc atc gtt gat 672

Lys Phe Ile Gly Gly His Gly Thr Ser Ile Gly Gly Val Ile Val Asp

210 215 220

ggc gga aac ttt gat tgg gca aat ggt aat ttt cct gat ttt aca caa 720

Gly Gly Asn Phe Asp Trp Ala Asn Gly Asn Phe Pro Asp Phe Thr Gln

225 230 235 240

gct gat gaa agc tac aat ggg att aaa ttt gcc gaa ttg ggt gaa att 768

Ala Asp Glu Ser Tyr Asn Gly Ile Lys Phe Ala Glu Leu Gly Glu Ile

245 250 255

gct ttt gtg act cgg gtt aga gct att tta tta cgt gat acg ggt gcg 816

Ala Phe Val Thr Arg Val Arg Ala Ile Leu Leu Arg Asp Thr Gly Ala

260 265 270

gct tta tca cct ttt cat tct tgg ctt ttc tta cag ggg cta gaa aca 864

Ala Leu Ser Pro Phe His Ser Trp Leu Phe Leu Gln Gly Leu Glu Thr

275 280 285

ctc tca ctc cgg gta gaa cgt cac atc tcc aat act aaa aag att gta 912

Leu Ser Leu Arg Val Glu Arg His Ile Ser Asn Thr Lys Lys Ile Val

290 295 300

gaa ttt tta gac aat cat cct aag gtg gaa ctt gtt aac cat cct ctg 960

Glu Phe Leu Asp Asn His Pro Lys Val Glu Leu Val Asn His Pro Leu

305 310 315 320

ctt gaa agt aat tcc tat cat gcg ctc tat cag aaa tat tat cca aaa 1008

Leu Glu Ser Asn Ser Tyr His Ala Leu Tyr Gln Lys Tyr Tyr Pro Lys

325 330 335

gat gct gga tct atc ttt acc ttt gaa ctc aaa gac aaa gat gag aaa 1056

Asp Ala Gly Ser Ile Phe Thr Phe Glu Leu Lys Asp Lys Asp Glu Lys

340 345 350

aaa gcg cgt gat ttg att gat cat ctt gaa att ttc tca ctt cta gcc 1104

Lys Ala Arg Asp Leu Ile Asp His Leu Glu Ile Phe Ser Leu Leu Ala

355 360 365

aac gtt gga gat acc aaa tca ttg gcc att cat cct gct tcg acc act 1152

Asn Val Gly Asp Thr Lys Ser Leu Ala Ile His Pro Ala Ser Thr Thr

370 375 380

cac cag cag ctg aat gcc gaa gaa ctt gct agt gca ggg att tcc aaa 1200

His Gln Gln Leu Asn Ala Glu Glu Leu Ala Ser Ala Gly Ile Ser Lys

385 390 395 400

gga acc att cga tta tcg gtt ggt att gaa gat gta act gac ttg att 1248

Gly Thr Ile Arg Leu Ser Val Gly Ile Glu Asp Val Thr Asp Leu Ile

405 410 415

gct gat tta gag caa gca tta gaa aaa ata taa 1281

Ala Asp Leu Glu Gln Ala Leu Glu Lys Ile

420 425

<210>14

<211>426

<212>PRT

<213>乳酸乳球菌

<400>14

Met Thr Asn His Asn Tyr Lys Phe Asp Thr Leu Gln Val His Ala Gly

1 5 10 15

Gln Val Pro Asp Pro Val Thr Gly Ser Arg Ala Val Pro Leu Tyr Gln

20 25 30

Thr Thr Ser Phe Val Phe Asn Asn Ser Asp His Ala Glu Ala Arg Phe

35 40 45

Ala Leu Gln Asp Pro Gly Ala Ile Tyr Ser Arg Leu Gly Asn Pro Thr

50 55 60

Asn Asp Val Phe Glu Ala Arg Ile Ala Ala Leu Glu Gly Gly Ser Ala

65 70 75 80

Ala Leu Gly Val Gly Ser Gly Ser Ala Ala Ile Thr Tyr Ala Ile Leu

85 90 95

Asn Ile Ala Thr Val Gly Asp Asn Ile Val Ser Ala Ser Thr Leu Tyr

100 105 110

Gly Gly Thr Tyr His Leu Phe Ser Gly Thr Leu Pro Lys Tyr Gly Ile

115 120 125

Thr Thr Lys Phe Val Asn Pro Asp Asp Pro Lys Asn Phe Glu Glu Ala

130 135 140

Ile Asp Glu Lys Thr Lys Ala Ile Tyr Tyr Glu Thr Leu Gly Asn Pro

145 150 155 160

Gly Asn Asn Val Ile Asp Tyr Asp Ala Ile Gly Gln Ile Ala Lys Lys

165 170 175

His Gly Ile Pro Val Ile Val Asp Ala Thr Phe Thr Thr Pro Val Thr

180 185 190

Phe Lys Pro Phe Glu His Gly Ala Asn Val Ile Val His Ser Ala Thr

195 200 205

Lys Phe Ile Gly Gly His Gly Thr Ser Ile Gly Gly Val Ile Val Asp

210 215 220

Gly Gly Asn Phe Asp Trp Ala Asn Gly Asn Phe Pro Asp Phe Thr Gln

225 230 235 240

Ala Asp Glu Ser Tyr Asn Gly Ile Lys Phe Ala Glu Leu Gly Glu Ile

245 250 255

Ala Phe Val Thr Arg Val Arg Ala Ile Leu Leu Arg Asp Thr Gly Ala

260 265 270

Ala Leu Ser Pro Phe His Ser Trp Leu Phe Leu Gln Gly Leu Glu Thr

275 280 285

Leu Ser Leu Arg Val Glu Arg His Ile Ser Asn Thr Lys Lys Ile Val

290 295 300

Glu Phe Leu Asp Asn His Pro Lys Val Glu Leu Val Asn His Pro Leu

305 310 315 320

Leu Glu Ser Asn Ser Tyr His Ala Leu Tyr Gln Lys Tyr Tyr Pro Lys

325 330 335

Asp Ala Gly Ser Ile Phe Thr Phe Glu Leu Lys Asp Lys Asp Glu Lys

340 345 350

Lys Ala Arg Asp Leu Ile Asp His Leu Glu Ile Phe Ser Leu Leu Ala

355 360 365

Asn Val Gly Asp Thr Lys Ser Leu Ala Ile His Pro Ala Ser Thr Thr

370 375 380

His Gln Gln Leu Asn Ala Glu Glu Leu Ala Ser Ala Gly Ile Ser Lys

385 390 395 400

Gly Thr Ile Arg Leu Ser Val Gly Ile Glu Asp Val Thr Asp Leu Ile

405 410 415

Ala Asp Leu Glu Gln Ala Leu Glu Lys Ile

420 425

<210>15

<211>1173

<212>DNA

<2l3>聚球藻属(Synechococcus)中的种

<220>

<221>CDS

<222>(1)..(1173)

<223>

<400>15

atg tct cag cgt ttc gaa acc ctc cag ctg cat gcc ggc cag tct cca 48

Met Ser Gln Arg Phe Glu Thr Leu Gln Leu His Ala Gly Gln Ser Pro

1 5 10 15

gac tcg gcc acc aat gcc aga gcg gtg ccg att tat cag acc agc tcc 96

Asp Ser Ala Thr Asn Ala Arg Ala Val Pro Ile Tyr Gln Thr Ser Ser

20 25 30

tac gtc ttc aac gac gcc gag cac ggc gcc aac ctg ttt gga ctg aag 144

Tyr Val Phe Asn Asp Ala Glu His Gly Ala Asn Leu Phe Gly Leu Lys

35 40 45

gaa ttc ggc aac atc tac acc cgt ctg atg aac ccg acg acg gat gtg 192

Glu Phe Gly Asn Ile Tyr Thr Arg Leu Met Asn Pro Thr Thr Asp Val

50 55 60

ttc gag aag cgg gtg gcg gcc ctg gaa ggg ggt gtg gcc gcg ctg gcc 240

Phe Glu Lys Arg Val Ala Ala Leu Glu Gly Gly Val Ala Ala Leu Ala

65 70 75 80

aca gcc tcc ggt cag tcg gct cag ttc ctg gcg atc acg aat tgc atg 288

Thr Ala Ser Gly Gln Ser Ala Gln Phe Leu Ala Ile Thr Asn Cys Met

85 90 95

cag gca ggg gat aac ttt gtg tcc acg tcg ttc ctt tac ggc ggc acc 336

Gln Ala Gly Asp Asn Phe Val Ser Thr Ser Phe Leu Tyr Gly Gly Thr

100 105 110

tac aac cag ttc aaa gtg caa ttc ccc cgg ctg ggc atc gac gtg cgc 384

Tyr Asn Gln Phe Lys Val Gln Phe Pro Arg Leu Gly Ile Asp Val Arg

115 120 125

ttc gct gat ggc gac gac gtg gag agc ttt gct gcg cag atc gac gac 432

Phe Ala Asp Gly Asp Asp Val Glu Ser Phe Ala Ala Gln Ile Asp Asp

130 135 140

aaa acc aaa ggc ctc tac gtc gaa gcg atg ggc aat cca cgc ttc aac 480

Lys Thr Lys Gly Leu Tyr Val Glu Ala Met Gly Asn Pro Arg Phe Asn

145 150 155 160

atc ccc gat ttc gag ggc ctc tca gcc ctg gct aaa gag cgc ggc atc 528

Ile Pro Asp Phe Glu Gly Leu Ser Ala Leu Ala Lys Glu Arg Gly Ile

165 170 175

cca ttg atc gtg gac aac acc ttg gga gct tgc ggt gcc ctg atg cgt 576

Pro Leu Ile Val Asp Asn Thr Leu Gly Ala Cys Gly Ala Leu Met Arg

180 185 190

ccg atc gat cat ggc gcg gat gtg gtg gtg gaa agc gcc acc aag tgg 624

Pro Ile Asp His Gly Ala Asp Val Val Val Glu Ser Ala Thr Lys Trp

195 200 205

att ggc ggc cat ggc acc agc ctc ggt ggc gtg atc gtt gat gcc ggc 672

Ile Gly Gly His Gly Thr Ser Leu Gly Gly Val Ile Val Asp Ala Gly

210 215 220

aca ttt aac tgg ggc aat ggc aaa ttc ccg ctg ctg agc caa ccc agt 720

Thr Phe Asn Trp Gly Asn Gly Lys Phe Pro Leu Leu Ser Gln Pro Ser

225 230 235 240

gcg gct tat cac ggc ctt gtg cac tgg gat gcc ttc ggc ttc ggc agc 768

Ala Ala Tyr His Gly Leu Val His Trp Asp Ala Phe Gly Phe Gly Ser

245 250 255

gac gtc tgc aag atg ctg gga gtg ccg gac aac cgc aac gtc gcc ttt 816

Asp Val Cys Lys Met Leu Gly Val Pro Asp Asn Arg Asn Val Ala Phe

260 265 270

gcc ctg cga gcc cgg gtc gag ggt cta cgg gac tgg ggt ccg gcg gtt 864

Ala Leu Arg Ala Arg Val Glu Gly Leu Arg Asp Trp Gly Pro Ala Val

275 280 285

agt ccc ttc aat agc ttc ctg ctg ctg caa ggt cta gaa acc ctc agc 912

Ser Pro Phe Asn Ser Phe Leu Leu Leu Gln Gly Leu Glu Thr Leu Ser

290 295 300

ctg cgg gtg gag cgc cac acg gag aac gcc atg gcg ctg gcc acc tgg 960

Leu Arg Val Glu Arg His Thr Glu Asn Ala Met Ala Leu Ala Thr Trp

305 310 315 320

cta gca acg cac ccc aat gtg gag cat gtg agc tac cca ggc ctg agc 1008

Leu Ala Thr His Pro Asn Val Glu His Val Ser Tyr Pro Gly Leu Ser

325 330 335

agc gat ccg tat cac gca gct gcc aag aaa tac ctg acg ggc cgg ggc 1056

Ser Asp Pro Tyr His Ala Ala Ala Lys Lys Tyr Leu Thr Gly Arg Gly

340 345 350

atg gga tgc atg ctg atg ttc tcg ctc aag ggc ggt tac gac gat gca 1104

Met Gly Cys Met Leu Met Phe Ser Leu Lys Gly Gly Tyr Asp Asp Ala

355 360 365

gtc cgt ttc atc aac agc ctt caa ctg gcc agt cac ctc gcc aat gtg 1152

Val Arg Phe Ile Asn Ser Leu Gln Leu Ala Ser His Leu Ala Asn Val

370 375 380

ggg gat gcc aaa acc tgg tga 1173

Gly Asp Ala Lys Thr Trp

385 390

<210>16

<211>390

<212>PRT

<213>聚球藻属中的种

<400>16

Met Ser Gln Arg Phe Glu Thr Leu Gln Leu His Ala Gly Gln Ser Pro

1 5 10 15

Asp Ser Ala Thr Asn Ala Arg Ala Val Pro Ile Tyr Gln Thr Ser Ser

20 25 30

Tyr Val Phe Asn Asp Ala Glu His Gly Ala Asn Leu Phe Gly Leu Lys

35 40 45

Glu Phe Gly Asn Ile Tyr Thr Arg Leu Met Asn Pro Thr Thr Asp Val

50 55 60

Phe Glu Lys Arg Val Ala Ala Leu Glu Gly Gly Val Ala Ala Leu Ala

65 70 75 80

Thr Ala Ser Gly Gln Ser Ala Gln Phe Leu Ala Ile Thr Asn Cys Met

85 90 95

Gln Ala Gly Asp Asn Phe Val Ser Thr Ser Phe Leu Tyr Gly Gly Thr

100 105 110

Tyr Asn Gln Phe Lys Val Gln Phe Pro Arg Leu Gly Ile Asp Val Arg

115 120 125

Phe Ala Asp Gly Asp Asp Val Glu Ser Phe Ala Ala Gln Ile Asp Asp

130 135 140

Lys Thr Lys Gly Leu Tyr Val Glu Ala Met Gly Asn Pro Arg Phe Asn

145 150 155 160

Ile Pro Asp Phe Glu Gly Leu Ser Ala Leu Ala Lys Glu Arg Gly Ile

165 170 175

Pro Leu Ile Val Asp Asn Thr Leu Gly Ala Cys Gly Ala Leu Met Arg

180 185 190

Pro Ile Asp His Gly Ala Asp Val Val Val Glu Ser Ala Thr Lys Trp

195 200 205

Ile Gly Gly His Gly Thr Ser Leu Gly Gly Val Ile Val Asp Ala Gly

210 215 220

Thr Phe Asn Trp Gly Asn Gly Lys Phe Pro Leu Leu Ser Gln Pro Ser

225 230 235 240

Ala Ala Tyr His Gly Leu Val His Trp Asp Ala Phe Gly Phe Gly Ser

245 250 255

Asp Val Cys Lys Met Leu Gly Val Pro Asp Asn Arg Asn Val Ala Phe

260 265 270

Ala Leu Arg Ala Arg Val Glu Gly Leu Arg Asp Trp Gly Pro Ala Val

275 280 285

Ser Pro Phe Asn Ser Phe Leu Leu Leu Gln Gly Leu Glu Thr Leu Ser

290 295 300

Leu Arg Val Glu Arg His Thr Glu Asn Ala Met Ala Leu Ala Thr Trp

305 310 315 320

Leu Ala Thr His Pro Asn Val Glu His Val Ser Tyr Pro Gly Leu Ser

325 330 335

Ser Asp Pro Tyr His Ala Ala Ala Lys Lys Tyr Leu Thr Gly Arg Gly

340 345 350

Met Gly Cys Met Leu Met Phe Ser Leu Lys Gly Gly Tyr Asp Asp Ala

355 360 365

Val Arg Phe Ile Asn Ser Leu Gln Leu Ala Ser His Leu Ala Asn Val

370 375 380

Gly Asp Ala Lys Thr Trp

385 390

<210>17

<211>1314

<212>DNA

<213>构巢裸孢壳(Emericella nidulans)

<220>

<221>CDS

<222>(1)..(1314)

<223>

<400>17

atg tcc gac cct tca ccg aaa cgt ttc gag acc ctc cag ctc cat gcg 48

Met Ser Asp Pro Ser Pro Lys Arg Phe Glu Thr Leu Gln Leu His Ala

1 5 10 15

ggc cag gag cct gac cct gca act aat tcc cgg gct gtc cca atc tat 96

Gly Gln Glu Pro Asp Pro Ala Thr Ash Ser Arg Ala Val Pro Ile Tyr

20 25 30

gcg aca acg tcc tac acc ttc aat gac tcc gca cac ggc gcc agg ctt 144

Ala Thr Thr Ser Tyr Thr Phe Asn Asp Ser Ala His Gly Ala Arg Leu

35 40 45

ttt ggc ctc aaa gag ttt ggc aat att tac agc cga att atg aat ccc 192

Phe Gly Leu Lys Glu Phe Gly Asn Ile Tyr Ser Arg Ile Met Asn Pro

50 55 60

aca gtc gat gtc ttc gaa aaa cgt att gct gca ctc gag gga ggt gtc 240

Thr Val Asp Val Phe Glu Lys Arg Ile Ala Ala Leu Glu Gly Gly Val

65 70 75 80

gct gcg gtg gct gcc tca tct ggc cag gca gcc cag ttc atg gcc atc 288

Ala Ala Val Ala Ala Ser Ser Gly Gln Ala Ala Gln Phe Met Ala Ile

85 90 95

tct gct cta gcc cat gct ggt gac aat atc gtt tcc aca agt aat ttg 336

Ser Ala Leu Ala His Ala Gly Asp Asn Ile Val Ser Thr Ser Asn Leu

100 105 110

tat ggt ggt aca tac aat cag ttt aag gtc ctt ttc cca cga ctg gga 384

Tyr Gly Gly Thr Tyr Asn Gln Phe Lys Val Leu Phe Pro Arg Leu Gly

115 120 125

att acc aca aaa ttc gtg cag gga gac aaa gca gag gac att gcc gcc 432

Ile Thr Thr Lys Phe Val Gln Gly Asp Lys Ala Glu Asp Ile Ala Ala

130 135 140

gct atc gat gac cgt acc aag gcc gtc tac gtc gag aca ata gga aac 480

Ala Ile Asp Asp Arg Thr Lys Ala Val Tyr Val Glu Thr Ile Gly Asn

145 150 155 160

cct cgc tac aat gtg ccc gac ttt gag gtc att gca aaa gta gcc cat 528

Pro Arg Tyr Asn Val Pro Asp Phe Glu Val Ile Ala Lys Val Ala His

165 170 175

gag aag gga att ccc ctt gtg gtt gac aac acc ttc ggt gcc gga ggc 576

Glu Lys Gly Ile Pro Leu Val Val Asp Asn Thr Phe Gly Ala Gly Gly

180 185 190

tac ttt gtt cga ccc att gaa cat ggc gcc gac att gtc gtg cac agt 624

Tyr Phe Val Arg Pro Ile Glu His Gly Ala Asp Ile Val Val His Ser

195 200 205

gca act aaa tgg att gga ggt cat ggc aca acc atc gga ggc gtt gtc 672

Ala Thr Lys Trp Ile Gly Gly His Gly Thr Thr Ile Gly Gly Val Val

210 215 220

gtg gac agc ggc aaa ttc gac tgg ggc aag aac gcc gcg cgg ttt cct 720

Val Asp Ser Gly Lys Phe Asp Trp Gly Lys Asn Ala Ala Arg Phe Pro

225 230 235 240

cag ttc acg cag cct tct gaa ggt tac cac ggg ttg aac ttc tgg gag 768

Gln Phe Thr Gln Pro Ser Glu Gly Tyr His Gly Leu Asn Phe Trp Glu

245 250 255

acc ttc ggc ccc att gcc ttc gcg att cgt gtc cgg gtc gaa atc ctg 816

Thr Phe Gly Pro Ile Ala Phe Ala Ile Arg Val Arg Val Glu Ile Leu

260 265 270

cgc gac ctc ggg tcc gcg ctg aac cct ttc gcc gcg cag cag ctc atc 864

Arg Asp Leu Gly Ser Ala Leu Asn Pro Phe Ala Ala Gln Gln Leu Ile

275 280 285

ctg ggt ctg gaa acc cta agc ttg cgc gct gag cgt cat gct tcc aac 912

Leu Gly Leu Glu Thr Leu Ser Leu Arg Ala Glu Arg His Ala Ser Asn

290 295 300

gct ctg gcc ctc gcc aac tgg cta aag aag aat gat cac gtc agc tgg 960

Ala Leu Ala Leu Ala Asn Trp Leu Lys Lys Asn Asp His Val Ser Trp

305 310 315 320

gtt tct tac gtg ggc cta gaa gag cac tcc agc cac gaa gtt gca aag 1008

Val Ser Tyr Val Gly Leu Glu Glu His Ser Ser His Glu Val Ala Lys

325 330 335

aag tac ctc aag cgt ggg ttc ggc ggt gtc cta tcc ttt ggt gtc aag 1056

Lys Tyr Leu Lys Arg Gly Phe Gly Gly Val Leu Ser Phe Gly Val Lys

340 345 350

ggt gag gca gcc gtc ggt agc cag gtt gtc gac aac ttt aag ctc atc 1104

Gly Glu Ala Ala Val Gly Ser Gln Val Val Asp Asn Phe Lys Leu Ile

355 360 365

tcc aat cta gca aat gtt gga gac tcc aag acc ctc gcg att cac ccc 1152

Ser Asn Leu Ala Asn Val Gly Asp Ser Lys Thr Leu Ala Ile His Pro

370 375 380

tgg agc acc act cac gag cag ttg acc gac cag gag cga atc gat tct 1200

Trp Ser Thr Thr His Glu Gln Leu Thr Asp Gln Glu Arg Ile Asp Ser

385 390 395 400

ggt gtt acg gaa gat gcc atc cgc atc tct gtc ggc act gag cac atc 1248

Gly Val Thr Glu Asp Ala Ile Arg Ile Ser Val Gly Thr Glu His Ile

405 410 415

gac gac atc atc gcc gac ttt gaa cag tca ttt gca gcg acc ttc aaa 1296

Asp Asp Ile Ile Ala Asp Phe Glu Gln Ser Phe Ala Ala Thr Phe Lys

420 425 430

gtt gtc cgg agt gct tag 1314

Val Val Arg Ser Ala

435

<210>18

<211>437

<212>PRT

<213>构巢裸孢壳

<400>18

Met Ser Asp Pro Ser Pro Lys Arg Phe Glu Thr Leu Gln Leu His Ala

1 5 10 15

Gly Gln Glu Pro Asp Pro Ala Thr Asn Ser Arg Ala Val Pro Ile Tyr

20 25 30

Ala Thr Thr Ser Tyr Thr Phe Asn Asp Ser Ala His Gly Ala Arg Leu

35 40 45

Phe Gly Leu Lys Glu Phe Gly Asn Ile Tyr Ser Arg Ile Met Asn Pro

50 55 60

Thr Val Asp Val Phe Glu Lys Arg Ile Ala Ala Leu Glu Gly Gly Val

65 70 75 80

Ala Ala Val Ala Ala Ser Ser Gly Gln Ala Ala Gln Phe Met Ala Ile

85 90 95

Ser Ala Leu Ala His Ala Gly Asp Asn Ile Val Ser Thr Ser Asn Leu

100 105 110

Tyr Gly Gly Thr Tyr Asn Gln Phe Lys Val Leu Phe Pro Arg Leu Gly

115 120 125

Ile Thr Thr Lys Phe Val Gln Gly Asp Lys Ala Glu Asp Ile Ala Ala

130 135 140

Ala Ile Asp Asp Arg Thr Lys Ala Val Tyr Val Glu Thr Ile Gly Asn

145 150 155 160

Pro Arg Tyr Asn Val Pro Asp Phe Glu Val Ile Ala Lys Val Ala His

165 170 175

Glu Lys Gly Ile Pro Leu Val Val Asp Asn Thr Phe Gly Ala Gly Gly

180 185 190

Tyr Phe Val Arg Pro Ile Glu His Gly Ala Asp Ile Val Val His Ser

195 200 205

Ala Thr Lys Trp Ile Gly Gly His Gly Thr Thr Ile Gly Gly Val Val

210 215 220

Val Asp Ser Gly Lys Phe Asp Trp Gly Lys Asn Ala Ala Arg Phe Pro

225 230 235 240

Gln Phe Thr Gln Pro Ser Glu Gly Tyr His Gly Leu Asn Phe Trp Glu

245 250 255

Thr Phe Gly Pro Ile Ala Phe Ala Ile Arg Val Arg Val Glu Ile Leu

260 265 270

Arg Asp Leu Gly Ser Ala Leu Asn Pro Phe Ala Ala Gln Gln Leu Ile

275 280 285

Leu Gly Leu Glu Thr Leu Ser Leu Arg Ala Glu Arg His Ala Ser Asn

290 295 300

Ala Leu Ala Leu Ala Asn Trp Leu Lys Lys Asn Asp His Val Ser Trp

305 310 315 320

Val Ser Tyr Val Gly Leu Glu Glu His Ser Ser His Glu Val Ala Lys

325 330 335

Lys Tyr Leu Lys Arg Gly Phe Gly Gly Val Leu Ser Phe Gly Val Lys

340 345 350

Gly Glu Ala Ala Val Gly Ser Gln Val Val Asp Asn Phe Lys Leu Ile

355 360 365

Ser Asn Leu Ala Asn Val Gly Asp Ser Lys Thr Leu Ala Ile His Pro

370 375 380

Trp Ser Thr Thr His Glu Gln Leu Thr Asp Gln Glu Arg Ile Asp Ser

385 390 395 400

Gly Val Thr Glu Asp Ala Ile Arg Ile Ser Val Gly Thr Glu His Ile

405 410 415

Asp Asp Ile Ile Ala Asp Phe Glu Gln Ser Phe Ala Ala Thr Phe Lys

420 425 430

Val Val Arg Ser Ala

435

<210>19

<211>1287

<212>DNA

<213>脆弱拟杆菌(Bacteroides fragilis)

<220>

<221>CDS

<222>(1)..(1287)

<223>

<400>19

atg gaa acg aaa aaa tta cat ttt gag act tta caa ctc cat gtt gga 48

Met Glu Thr Lys Lys Leu His Phe Glu Thr Leu Gln Leu His Val Gly

1 5 10 15

cag gag act ccc gac ccg gca acc gat gcg cgt gcc gta cct att tat 96

Gln Glu Thr Pro Asp Pro Ala Thr Asp Ala Arg Ala Val Pro Ile Tyr

20 25 30

cag aca act tcc tat gtg ttc cgg gat tcg gcc cat gcc gcc gca cga 144

Gln Thr Thr Ser Tyr Val Phe Arg Asp Ser Ala His Ala Ala Ala Arg

35 40 45

ttt gga ttg caa gac cct ggg aat att tat gga cga ctg acc aat tcc 192

Phe Gly Leu Gln Asp Pro Gly Asn Ile Tyr Gly Arg Leu Thr Asn Ser

50 55 60

act cag gga gta ttg gag gaa cgc atc gca gca ctt gaa ggg gga gta 240

Thr Gln Gly Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Gly Gly Val

65 70 75 80

ggt ggg ctt gcc gtg gct tcc gga gct gct gcc gtg acc tat gct atc 288

Gly Gly Leu Ala Val Ala Ser Gly Ala Ala Ala Val Thr Tyr Ala Ile

85 90 95

gag aat atc acc cgt tcc ggt gat cat att gtg gct gcc aag acc att 336

Glu Asn Ile Thr Arg Ser Gly Asp His Ile Val Ala Ala Lys Thr Ile

100 105 110

tat ggg ggc aca tat aac ttg ctg gcg cat act ctg cct gct tat gga 384

Tyr Gly Gly Thr Tyr Asn Leu Leu Ala His Thr Leu Pro Ala Tyr Gly

115 120 125

gta acg acc act ttt gta gat ccg tcc gat ctt ttt aat ttc gaa cgg 432

Val Thr Thr Thr Phe Val Asp Pro Ser Asp Leu Phe Asn Phe Glu Arg

130 135 140

gcg att cgt gaa aat aca aag gcg ata ttc att gaa act ctg gga aac 480

Ala Ile Arg Glu Asn Thr Lys Ala Ile Phe Ile Glu Thr Leu Gly Asn

145 150 155 160

ccc aat tcc aat att atc gat atg gat gcc gta gct gcc att gcc cat 528

Pro Asn Ser Asn Ile Ile Asp Met Asp Ala Val Ala Ala Ile Ala His

165 170 175

aaa tat cgg att ccg ctg att gtg gat aat act ttc ggt acg cct tac 576

Lys Tyr Arg Ile Pro Leu Ile Val Asp Asn Thr Phe Gly Thr Pro Tyr

180 185 190

ctt atc cgt ccc att gag cac ggg gca gac att gtg gta cat tct gcc 624

Leu Ile Arg Pro Ile Glu His Gly Ala Asp Ile Val Val His Ser Ala

195 200 205

aca aaa ttc att ggc gga cac ggc agt tcg ttg gga gga gtt att gtc 672

Thr Lys Phe Ile Gly Gly His Gly Ser Ser Leu Gly Gly Val Ile Val

210 215 220

gat tcc ggt aaa ttt gac tgg gtt gct tcc ggt aaa ttc ccg caa ctg 720

Asp Ser Gly Lys Phe Asp Trp Val Ala Ser Gly Lys Phe Pro Gln Leu

225 230 235 240

acc gag ccg gat gca agt tat cat ggg gta cgg ttt gtc gat gct gcc 768

Thr Glu Pro Asp Ala Ser Tyr His Gly Val Arg Phe Val Asp Ala Ala

245 250 255

ggg gct gct gcc tac att gtc cgt ata cgt gcc gtg ttg ctg cgc gat 816

Gly Ala Ala Ala Tyr Ile Val Arg Ile Arg Ala Val Leu Leu Arg Asp

260 265 270

acg ggt gct gcc atc agc ccg ttc aat gct ttt atc ttg ctg caa ggg 864

Thr Gly Ala Ala Ile Ser Pro Phe Asn Ala Phe Ile Leu Leu Gln Gly

275 280 285

ttg gag act ttg tct ttg cgt gta gaa cgg cat gtg gcc aat gct ttg 912

Leu Glu Thr Leu Ser Leu Arg Val Glu Arg His Val Ala Asn Ala Leu

290 295 300

aag gtt att gat ttt ctg gtg aac cat ccg aag gta gcg gct gtt aat 960

Lys Val Ile Asp Phe Leu Val Asn His Pro Lys Val Ala Ala Val Asn

305 310 315 320

cat cca tca ttg ccc ggt cat ccg gat cat gcc atc tat caa cgt tat 1008

His Pro Ser Leu Pro Gly His Pro Asp His Ala Ile Tyr Gln Arg Tyr

325 330 335

ttt cct ggc ggg gca ggt tct atc ttc act ttc gag gta aag gga gga 1056

Phe Pro Gly Gly Ala Gly Ser Ile Phe Thr Phe Glu Val Lys Gly Gly

340 345 350

acg gag gaa gcg cag aag ttt atc gat agt ctg cag ata ttc tct ttg 1104

Thr Glu Glu Ala Gln Lys Phe Ile Asp Ser Leu Gln Ile Phe Ser Leu

355 360 365

ctg gcc aat gtg gcc gat gtg aag tcg ctg gtg att cat ccg ggc act 1152

Leu Ala Asn Val Ala Asp Val Lys Ser Leu Val Ile His Pro Gly Thr

370 375 380

acc aca cac tcg cag ttg aat gcg cag gag ctg gag gaa cag ggg att 1200

Thr Thr His Ser Gln Leu Asn Ala Gln Glu Leu Glu Glu Gln Gly Ile

385 390 395 400

aaa ccc gga acg gtc aga ctt tcg ata ggt acg gag cat att gag gac 1248

Lys Pro Gly Thr Val Arg Leu Ser Ile Gly Thr Glu His Ile Glu Asp

405 410 415

att att gat gac tta cgt cag gca tta gag aaa att taa 1287

Ile Ile Asp Asp Leu Arg Gln Ala Leu Glu Lys Ile

420 425

<210>20

<211>428

<212>PRT

<213>脆弱拟杆菌

<400>20

Met Glu Thr Lys Lys Leu His Phe Glu Thr Leu Gln Leu His Val Gly

1 5 10 15

Gln Glu Thr Pro Asp Pro Ala Thr Asp Ala Arg Ala Val Pro Ile Tyr

20 25 30

Gln Thr Thr Ser Tyr Val Phe Arg Asp Ser Ala His Ala Ala Ala Arg

35 40 45

Phe Gly Leu Gln Asp Pro Gly Asn Ile Tyr Gly Arg Leu Thr Asn Ser

50 55 60

Thr Gln Gly Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Gly Gly Val

65 70 75 80

Gly Gly Leu Ala Val Ala Ser Gly Ala Ala Ala Val Thr Tyr Ala Ile

85 90 95

Glu Asn Ile Thr Arg Ser Gly Asp His Ile Val Ala Ala Lys Thr Ile

100 105 110

Tyr Gly Gly Thr Tyr Asn Leu Leu Ala His Thr Leu Pro Ala Tyr Gly

115 120 125

Val Thr Thr Thr Phe Val Asp Pro Ser Asp Leu Phe Asn Phe Glu Arg

130 135 140

Ala Ile Arg Glu Asn Thr Lys Ala Ile Phe Ile Glu Thr Leu Gly Asn

145 150 155 160

Pro Asn Ser Asn Ile Ile Asp Met Asp Ala Val Ala Ala Ile Ala His

165 170 175

Lys Tyr Arg Ile Pro Leu Ile Val Asp Asn Thr Phe Gly Thr Pro Tyr

180 185 190

Leu Ile Arg Pro Ile Glu His Gly Ala Asp Ile Val Val His Ser Ala

195 200 205

Thr Lys Phe Ile Gly Gly His Gly Ser Ser Leu Gly Gly Val Ile Val

210 215 220

Asp Ser Gly Lys Phe Asp Trp Val Ala Ser Gly Lys Phe Pro Gln Leu

225 230 235 240

Thr Glu Pro Asp Ala Ser Tyr His Gly Val Arg Phe Val Asp Ala Ala

245 250 255

Gly Ala Ala Ala Tyr Ile Val Arg Ile Arg Ala Val Leu Leu Arg Asp

260 265 270

Thr Gly Ala Ala Ile Ser Pro Phe Asn Ala Phe Ile Leu Leu Gln Gly

275 280 285

Leu Glu Thr Leu Ser Leu Arg Val Glu Arg His Val Ala Asn Ala Leu

290 295 300

Lys Val Ile Asp Phe Leu Val Asn His Pro Lys Val Ala Ala Val Asn

305 310 315 320

His Pro Ser Leu Pro Gly His Pro Asp His Ala Ile Tyr Gln Arg Tyr

325 330 335

Phe Pro Gly Gly Ala Gly Ser Ile Phe Thr Phe Glu Val Lys Gly Gly

340 345 350

Thr Glu Glu Ala Gln Lys Phe Ile Asp Ser Leu Gln Ile Phe Ser Leu

355 360 365

Leu Ala Asn Val Ala Asp Val Lys Ser Leu Val Ile His Pro Gly Thr

370 375 380

Thr Thr His Ser Gln Leu Asn Ala Gln Glu Leu Glu Glu Gln Gly Ile

385 390 395 400

Lys Pro Gly Thr Val Arg Leu Ser Ile Gly Thr Glu His Ile Glu Asp

405 410 415

Ile Ile Asp Asp Leu Arg Gln Ala Leu Glu Lys Ile

420 425

<210>21

<211>1278

<212>DNA

<213>铜绿假单胞菌(Pseudomonas aeruginosa)

<220>

<22l>CDS

<222>(1)..(1278)

<223>

<400>21

atg aaa ctg gaa acc ctg gcc gtc cac gcc ggc tac agc cct gac ccg 48

Met Lys Leu Glu Thr Leu Ala Val His Ala Gly Tyr Ser Pro Asp Pro

1 5 10 15

acc acc cgc gcg gtg gcg gtg ccg atc tac cag acc acc tcc tac gcc 96

Thr Thr Arg Ala Val Ala Val Pro Ile Tyr Gln Thr Thr Ser Tyr Ala

20 25 30

ttc gac gac acc cag cat ggc gcc gac ctg ttc gac ctg aag gta ccg 144

Phe Asp Asp Thr Gln His Gly Ala Asp Leu Phe Asp Leu Lys Val Pro

35 40 45

ggc aac atc tac aca cgg atc atg aac ccc acc aac gac gta ctg gaa 192

Gly Asn Ile Tyr Thr Arg Ile Met Asn Pro Thr Asn Asp Val Leu Glu

50 55 60

cag cgc gtc gcg gcg ctg gaa ggc ggg gtc ggg gcg ctg gcg gtg gcc 240

Gln Arg Val Ala Ala Leu Glu Gly Gly Val Gly Ala Leu Ala Val Ala

65 70 75 80

tcg ggg atg gcg gcc atc acc tac gcg atc cag acc gtc gcc gag gcc 288

Ser Gly Met Ala Ala Ile Thr Tyr Ala Ile Gln Thr Val Ala Glu Ala

85 90 95

ggc gac aac atc gtc tcg gtg gcc aag ctc tac ggc ggc acc tac aac 336

Gly Asp Asn Ile Val Ser Val Ala Lys Leu Tyr Gly Gly Thr Tyr Asn

100 105 110

ctg ctg gcc cac acc ctg cca cgc atc ggc atc cag gcg cgc ttc gcc 384

Leu Leu Ala His Thr Leu Pro Arg Ile Gly Ile Gln Ala Arg Phe Ala

115 120 125

gcc cac gac gac gtc gcc gcc ctg gaa gcg ctg atc gac gag cgg acc 432

Ala His Asp Asp Val Ala Ala Leu Glu Ala Leu Ile Asp Glu Arg Thr

130 135 140

aag gcc gtg ttc tgc gaa acc atc ggc aac ccg gcg ggc aac atc atc 480

Lys Ala Val Phe Cys Glu Thr Ile Gly Asn Pro Ala Gly Asn Ile Ile

145 150 155 160

gac ctg cag gca ctg gcc gac gcc gct cac cgc cac ggc gtg cca ctg 528

Asp Leu Gln Ala Leu Ala Asp Ala Ala His Arg His Gly Val Pro Leu

165 170 175

atc gtc gac aac acg gta gcc acc ccg gtg ctc tgc cgg ccg ttc gag 576

Ile Val Asp Asn Thr Val Ala Thr Pro Val Leu Cys Arg Pro Phe Glu

180 185 190

cac ggc gcc gac atc gtc gtg cac tcg ctg acc aag tac atg ggc ggc 624

His Gly Ala Asp Ile Val Val His Ser Leu Thr Lys Tyr Met Gly Gly

195 200 205

cac ggc acc agc atc ggc ggg atc gtg gtc gac tcc ggc aaa ttc gac 672

His Gly Thr Ser Ile Gly Gly Ile Val Val Asp Ser Gly Lys Phe Asp

210 215 220

tgg gcg gcg aac aag tcg cgc ttc ccg ctg ctg aac acg ccc gat ccg 720

Trp Ala Ala Asn Lys Ser Arg Phe Pro Leu Leu Asn Thr Pro Asp Pro

225 230 235 240

tcc tac cac ggc gtc acc tac acc gag gcc ttc gga ccc gcc gcc ttc 768

Ser Tyr His Gly Val Thr Tyr Thr Glu Ala Phe Gly Pro Ala Ala Phe

245 250 255

atc ggc cgc tgc cgg gtg gta ccg ctg cgc aac atg ggc gcg gcg ctc 816

Ile Gly Arg Cys Arg Val Val Pro Leu Arg Asn Met Gly Ala Ala Leu

260 265 270

tcg ccg ttc aac gcc ttc ctc atc ctc caa ggc ctg gag acc ctg gcg 864

Ser Pro Phe Asn Ala Phe Leu Ile Leu Gln Gly Leu Glu Thr Leu Ala

275 280 285

ctg cgc atg gag cgc cac tgc gac aac gcc ctc gcc gtg gcc cgc tac 912

Leu Arg Met Glu Arg His Cys Asp Asn Ala Leu Ala Val Ala Arg Tyr

290 295 300

ctg cag cag cat ccg cag gtg gcc tgg gtg aaa tac gcc ggc ctc gcc 960

Leu Gln Gln His Pro Gln Val Ala Trp Val Lys Tyr Ala Gly Leu Ala

305 310 315 320

gac aac ccc gag cac gcc ctg gcc cgg cgc tac ctg ggg ggc cgc ccg 1008

Asp Asn Pro Glu His Ala Leu Ala Arg Arg Tyr Leu Gly Gly Arg Pro

325 330 335

gcg gcg atc ctg tct ttc ggc atc cag ggc ggc agc gcc gcc ggc gcg 1056

Ala Ala Ile Leu Ser Phe Gly Ile Gln Gly Gly Ser Ala Ala Gly Ala

340 345 350

cgc ttc atc gac gcc ttg aag ctg gtg gtg cgg ctg gtc aac atc ggc 1104

Arg Phe Ile Asp Ala Leu Lys Leu Val Val Arg Leu Val Asn Ile Gly

355 360 365

gac gcc aag tcc ctg gcc tgc cac ccg gcg agc acc acc cac cgc cag 1152

Asp Ala Lys Ser Leu Ala Cys His Pro Ala Ser Thr Thr His Arg Gln

370 375 380

ttg aac gcg gag gaa ctg gcc cgc gcc gga gtc tcc gac gac atg gtg 1200

Leu Asn Ala Glu Glu Leu Ala Arg Ala Gly Val Ser Asp Asp Met Val

385 390 395 400

cgg ctg tcg atc ggc atc gag cac atc gac gac atc ctc gcc gac ctc 1248

Arg Leu Ser Ile Gly Ile Glu His Ile Asp Asp Ile Leu Ala Asp Leu

405 410 415

gac cag gcc ctg gcc gcc gcc gca cgc tga 1278

Asp Gln Ala Leu Ala Ala Ala Ala Arg

420 425

<210>22

<211>425

<212>PRT

<213>铜绿假单胞菌

<400>22

Met Lys Leu Glu Thr Leu Ala Val His Ala Gly Tyr Ser Pro Asp Pro

1 5 10 15

Thr Thr Arg Ala Val Ala Val Pro Ile Tyr Gln Thr Thr Ser Tyr Ala

20 25 30

Phe Asp Asp Thr Gln His Gly Ala Asp Leu Phe Asp Leu Lys Val Pro

35 40 45

Gly Asn Ile Tyr Thr Arg Ile Met Asn Pro Thr Asn Asp Val Leu Glu

50 55 60

Gln Arg Val Ala Ala Leu Glu Gly Gly Val Gly Ala Leu Ala Val Ala

65 70 75 80

Ser Gly Met Ala Ala Ile Thr Tyr Ala Ile Gln Thr Val Ala Glu Ala

85 90 95

Gly Asp Asn Ile Val Ser Val Ala Lys Leu Tyr Gly Gly Thr Tyr Asn

100 105 110

Leu Leu Ala His Thr Leu Pro Arg Ile Gly Ile Gln Ala Arg Phe Ala

115 120 125

Ala His Asp Asp Val Ala Ala Leu Glu Ala Leu Ile Asp Glu Arg Thr

130 135 140

Lys Ala Val Phe Cys Glu Thr Ile Gly Asn Pro Ala Gly Asn Ile Ile

145 150 155 160

Asp Leu Gln Ala Leu Ala Asp Ala Ala His Arg His Gly Val Pro Leu

165 170 175

Ile Val Asp Asn Thr Val Ala Thr Pro Val Leu Cys Arg Pro Phe Glu

180 185 190

His Gly Ala Asp Ile Val Val His Ser Leu Thr Lys Tyr Met Gly Gly

195 200 205

His Gly Thr Ser Ile Gly Gly Ile Val Val Asp Ser Gly Lys Phe Asp

210 215 220

Trp Ala Ala Asn Lys Ser Arg Phe Pro Leu Leu Asn Thr Pro Asp Pro

225 230 235 240

Ser Tyr His Gly Val Thr Tyr Thr Glu Ala Phe Gly Pro Ala Ala Phe

245 250 255

Ile Gly Arg Cys Arg Val Val Pro Leu Arg Asn Met Gly Ala Ala Leu

260 265 270

Ser Pro Phe Asn Ala Phe Leu Ile Leu Gln Gly Leu Glu Thr Leu Ala

275 280 285

Leu Arg Met Glu Arg His Cys Asp Asn Ala Leu Ala Val Ala Arg Tyr

290 295 300

Leu Gln Gln His Pro Gln Val Ala Trp Val Lys Tyr Ala Gly Leu Ala

305 310 315 320

Asp Asn Pro Glu His Ala Leu Ala Arg Arg Tyr Leu Gly Gly Arg Pro

325 330 335

Ala Ala Ile Leu Ser Phe Gly Ile Gln Gly Gly Ser Ala Ala Gly Ala

340 345 350

Arg Phe Ile Asp Ala Leu Lys Leu Val Val Arg Leu Val Asn Ile Gly

355 360 365

Asp Ala Lys Ser Leu Ala Cys His Pro Ala Ser Thr Thr His Arg Gln

370 375 380

Leu Asn Ala Glu Glu Leu Ala Arg Ala Gly Val Ser Asp Asp Met Val

385 390 395 400

Arg Leu Ser Ile Gly Ile Glu His Ile Asp Asp Ile Leu Ala Asp Leu

405 410 415

Asp Gln Ala Leu Ala Ala Ala Ala Arg

420 425

<210>23

<211>1296

<212>DNA

<213>支气管炎博德特氏菌(Bordetella broncniseptica)

<220>

<221>CDS

<222>(1)..(1296)

<223>

<400>23

atg agc gaa ccg aac caa ccc atc tgg cgg ctg gag acc atc gcc gta 48

Met Ser Glu Pro Asn Gln Pro Ile Trp Arg Leu Glu Thr Ile Ala Val

1 5 10 15

cat ggg ggc tac cgg ccc gac ccg acc acg cgc gcg gtg gcg gtg ccg 96

His Gly Gly Tyr Arg Pro Asp Pro Thr Thr Arg Ala Val Ala Val Pro

20 25 30

atc tac cag acc gtg gcc tat gcg ttc gac gac acc cag cat ggc gcg 144

Ile Tyr Gln Thr Val Ala Tyr Ala Phe Asp Asp Thr Gln His Gly Ala

35 40 45

gac ctg ttc gac ctg aag gtg ccg ggc aat atc tac acc cgc atc atg 192

Asp Leu Phe Asp Leu Lys Val Pro Gly Asn Ile Tyr Thr Arg Ile Met

50 55 60

aac ccc acc acc gac gtg ctg gag cag cgc gtg gcg gcg ctg gaa tgc 240

Asn Pro Thr Thr Asp Val Leu Glu Gln Arg Val Ala Ala Leu Glu Cys

65 70 75 80

ggc gtg gcc gcg ctg gcg ctg gcc tcc ggc cag gcg gcg gtg acc tat 288

Gly Val Ala Ala Leu Ala Leu Ala Ser Gly Gln Ala Ala Val Thr Tyr

85 90 95

gcg atc ctg acc atc gcc gag gcg ggc gac aac atc gtg tcg tcc agc 336

Ala Ile Leu Thr Ile Ala Glu Ala Gly Asp Asn Ile Val Ser Ser Ser

100 105 110

acg ctg tat ggc ggc acg tac aac ctg ttc gcc cac acg ctg ccg cag 384

Thr Leu Tyr Gly Gly Thr Tyr Asn Leu Phe Ala His Thr Leu Pro Gln

115 120 125

tac ggc atc acg acc cgc ttc gcc gat ccg cgc aac ctg gct tcg ttc 432

Tyr Gly Ile Thr Thr Arg Phe Ala Asp Pro Arg Asn Leu Ala Ser Phe

130 135 140

gag gcg ctg atc gac gag cgc acc aag gcc att ttc gcc gag tcg gtg 480

Glu Ala Leu Ile Asp Glu Arg Thr Lys Ala Ile Phe Ala Glu Ser Val

145 150 155 160

ggc aat ccg ctg ggc aac gtc acc gac atc gcc gcg ctg gcc gag atc 528

Gly Asn Pro Leu Gly Asn Val Thr Asp Ile Ala Ala Leu Ala Glu Ile

165 170 175

gcg cac cgc cat ggc gtg ccg ctg atc gtc gac aac acg gtg ccg tcg 576

Ala His Arg His Gly Val Pro Leu Ile Val Asp Asn Thr Val Pro Ser

180 185 190

ccc tac ctg ctg cgc ccc atc gag cac ggc gcc gac atc gtg gtg cag 624

Pro Tyr Leu Leu Arg Pro Ile Glu His Gly Ala Asp Ile Val Val Gln

195 200 205

tcg ctc acc aag tac ctg ggc ggg cac ggc acc agc ctg ggc ggg gcc 672

Ser Leu Thr Lys Tyr Leu Gly Gly His Gly Thr Ser Leu Gly Gly Ala

210 215 220

atc atc gat tcg ggc aag ttt ccc tgg gcc gag cac aag gcg cgc ttc 720

Ile Ile Asp Ser Gly Lys Phe Pro Trp Ala Glu His Lys Ala Arg Phe

225 230 235 240

aag cgc ctg aac gag ccc gac gtg agc tac cac ggc gtg gtc tac acc 768

Lys Arg Leu Asn Glu Pro Asp Val Ser Tyr His Gly Val Val Tyr Thr

245 250 255

gag gcg ttc ggc gcg gcg gcc tat atc ggc cgc gcc cgc gtg gtg ccg 816

Glu Ala Phe Gly Ala Ala Ala Tyr Ile Gly Arg Ala Arg Val Val Pro

260 265 270

ctg cgc aat acc ggc gcg gcc att tcg ccg ttc aac gcc ttc cag atc 864

Leu Arg Asn Thr Gly Ala Ala Ile Ser Pro Phe Asn Ala Phe Gln Ile

275 280 285

ctg cag ggc atc gag acg ctg gcg ctg cgc gtg gac cgc atc gtc gag 912

Leu Gln Gly Ile Glu Thr Leu Ala Leu Arg Val Asp Arg Ile Val Glu

290 295 300

aac tcg gtc aag gtg gcc ggg ttc ctg cgc gac cat ccc aag gtc gaa 960

Asn Ser Val Lys Val Ala Gly Phe Leu Arg Asp His Pro Lys Val Glu

305 310 315 320

tgg gtc aac tat gcc ggc ctg ccc gac cat gcc gac cat gcg ctg gtg 1008

Trp Val Asn Tyr Ala Gly Leu Pro Asp His Ala Asp His Ala Leu Val

325 330 335

cgc aag tac atg ggc ggc aag gcc ccc ggc ctg ttc act ttc ggc gtg 1056

Arg Lys Tyr Met Gly Gly Lys Ala Pro Gly Leu Phe Thr Phe Gly Val

340 345 350

aag ggc ggc cgc gag gcc ggc gcg cgc ttc cag gac gcc ttg cag ctg 1104

Lys Gly Gly Arg Glu Ala Gly Ala Arg Phe Gln Asp Ala Leu Gln Leu

355 360 365

ttc acc cgc ctg gtg aac atc ggc gac gcc aag tcg ctg gcc acg cac 1152

Phe Thr Arg Leu Val Asn Ile Gly Asp Ala Lys Ser Leu Ala Thr His

370 375 380

ccg gct tcc acc acg cac cgc cag ctc aac ccc gaa gag ctc gaa aag 1200

Pro Ala Ser Thr Thr His Arg Gln Leu Asn Pro Glu Glu Leu Glu Lys

385 390 395 400

gcc ggc gtg cgc gag gaa acg gtg cgc ctg tcg atc ggg atc gag cat 1248

Ala Gly Val Arg Glu Glu Thr Val Arg Leu Ser Ile Gly Ile Glu His

405 410 415

atc gac gac ctg atc gcc gac ctg gaa cag gcg ctg gcg caa gtc tga 1296

Ile Asp Asp Leu Ile Ala Asp Leu Glu Gln Ala Leu Ala Gln Val

420 425 430

<210>24

<211>431

<212>PRT

<213>支气管炎博德特氏菌

<400>24

Met Ser Glu Pro Asn Gln Pro Ile Trp Arg Leu Glu Thr Ile Ala Val

1 5 10 15

His Gly Gly Tyr Arg Pro Asp Pro Thr Thr Arg Ala Val Ala Val Pro

20 25 30

Ile Tyr Gln Thr Val Ala Tyr Ala Phe Asp Asp Thr Gln His Gly Ala

35 40 45

Asp Leu Phe Asp Leu Lys Val Pro Gly Asn Ile Tyr Thr Arg Ile Met

50 55 60

Asn Pro Thr Thr Asp Val Leu Glu Gln Arg Val Ala Ala Leu Glu Cys

65 70 75 80

Gly Val Ala Ala Leu Ala Leu Ala Ser Gly Gln Ala Ala Val Thr Tyr

85 90 95

Ala Ile Leu Thr Ile Ala Glu Ala Gly Asp Asn Ile Val Ser Ser Ser

100 105 110

Thr Leu Tyr Gly Gly Thr Tyr Asn Leu Phe Ala His Thr Leu Pro Gln

115 120 125

Tyr Gly Ile Thr Thr Arg Phe Ala Asp Pro Arg Asn Leu Ala Ser Phe

130 135 140

Glu Ala Leu Ile Asp Glu Arg Thr Lys Ala Ile Phc Ala Glu Ser Val

145 150 155 160

Gly Asn Pro Leu Gly Asn Val Thr Asp Ile Ala Ala Leu Ala Glu Ile

165 170 175

Ala His Arg His Gly Val Pro Leu Ile Val Asp Asn Thr Val Pro Ser

180 185 190

Pro Tyr Leu Leu Arg Pro Ile Glu His Gly Ala Asp Ile Val Val Gln

195 200 205

Ser Leu Thr Lys Tyr Leu Gly Gly His Gly Thr Ser Leu Gly Gly Ala

210 215 220

Ile Ile Asp Ser Gly Lys Phe Pro Trp Ala Glu His Lys Ala Arg Phe

225 230 235 240

Lys Arg Leu Asn Glu Pro Asp Val Ser Tyr His Gly Val Val Tyr Thr

245 250 255

Glu Ala Phe Gly Ala Ala Ala Tyr Ile Gly Arg Ala Arg Val Val Pro

260 265 270

Leu Arg Asn Thr Gly Ala Ala Ile Ser Pro Phe Asn Ala Phe Gln Ile

275 280 285

Leu Gln Gly Ile Glu Thr Leu Ala Leu Arg Val Asp Arg Ile Val Glu

290 295 300

Asn Ser Val Lys Val Ala Gly Phe Leu Arg Asp His Pro Lys Val Glu

305 310 315 320

Trp Val Asn Tyr Ala Gly Leu Pro Asp His Ala Asp His Ala Leu Val

325 330 335

Arg Lys Tyr Met Gly Gly Lys Ala Pro Gly Leu Phe Thr Phe Gly Val

340 345 350

Lys Gly Gly Arg Glu Ala Gly Ala Arg Phe Gln Asp Ala Leu Gln Leu

355 360 365

Phc Thr Arg Leu Val Asn Ile Gly Asp Ala Lys Ser Leu Ala Thr His

370 375 380

Pro Ala Ser Thr Thr His Arg Gln Leu Asn Pro Glu Glu Leu Glu Lys

385 390 395 400

Ala Gly Val Arg Glu Glu Thr Val Arg Leu Ser Ile Gly Ile Glu His

405 410 415

Ile Asp Asp Leu Ile Ala Asp Leu Glu Gln Ala Leu Ala Gln Val

420 425 430

<210>25

<211>1269

<212>DNA

<213>欧洲亚硝化单胞菌(Nitrosomonas europaea)

<220>

<221>CDS

<222>(1)..(1269)

<223>

<400>25

atg aaa cgg gaa aca ctc gcc att cat ggc ggt ttt gcc ggc gat ccg 48

Met Lys Arg Glu Thr Leu Ala Ile His Gly Gly Phe Ala Gly Asp Pro

1 5 10 15

cag act cat gca gtc gcg gtc ccc att tac cag acc acc agc tac tat 96

Gln Thr His Ala Val Ala Val Pro Ile Tyr Gln Thr Thr Ser Tyr Tyr

20 25 30

ttt gat gat act cag cac ggg gct gat ttg ttt gat ctg aag gtg cag 144

Phe Asp Asp Thr Gln His Gly Ala Asp Leu Phe Asp Leu Lys Val Gln

35 40 45

ggt aac atc tac aca cgc atc atg aac ccg act act gct gtc ctg gaa 192

Gly Asn Ile Tyr Thr Arg Ile Met Asn Pro Thr Thr Ala Val Leu Glu

50 55 60

gaa aga gtg gcg tta ctg gaa gga gga gtg gga gcg ctg gcc atg gct 240

Glu Arg Val Ala Leu Leu Glu Gly Gly Val Gly Ala Leu Ala Met Ala

65 70 75 80

tcc ggc atg gcc gcc att aca gcc tgt gtg cag act ctg gcc agg gcg 288

Ser Gly Met Ala Ala Ile Thr Ala Cys Val Gln Thr Leu Ala Arg Ala

85 90 95

ggc gac aac att atc tcc acc agc cag gtt tac ggt ggc acc tat aat 336

Gly Asp Asn Ile Ile Ser Thr Ser Gln Val Tyr Gly Gly Thr Tyr Asn

100 105 110

ttc ttt tgc cat acg ttg ccc aat ctg ggt att gaa gtt cgc atg gtg 384

Phe Phe Cys His Thr Leu Pro Asn Leu Gly Ile Glu Val Arg Met Val

115 120 125

gat ggt cgt aat ccg gcc gct ttt gcc gat gcc atc gat gac aat acc 432

Asp Gly Arg Asn Pro Ala Ala Phe Ala Asp Ala Ile Asp Asp Asn Thr

130 135 140

aga atg att tat tgc gag tcg atc gga aat ccg gcc ggt aat gtg gtg 480

Arg Met Ile Tyr Cys Glu Ser Ile Gly Asn Pro Ala Gly Asn Val Val

145 150 155 160

gat atc gcc gca ctg gct gaa gtg gcg cat gca gcg ggc gtg ccg ctg 528

Asp Ile Ala Ala Leu Ala Glu Val Ala His Ala Ala Gly Val Pro Leu

165 170 175

gta gtg gac aat acc gta cca acc ccg gtg ctt tgt cgt cct ttc gaa 576

Val Val Asp Asn Thr Val Pro Thr Pro Val Leu Cys Arg Pro Phe Glu

180 185 190

cat ggt gcc gat atc gtc gtc cat gcg ctg acc aaa tac atg ggt ggt 624

His Gly Ala Asp Ile Val Val His Ala Leu Thr Lys Tyr Met Gly Gly

195 200 205

cac ggc acc agc atc ggc gga atc atc gtg gat tcc ggc aag ttc ccc 672

His Gly Thr Ser Ile Gly Gly Ile Ile Val Asp Ser Gly Lys Phe Pro

210 215 220

tgg gaa ggc aac tcg cgt ttt cca caa ttc aac caa cct gat ccc agc 720

Trp Glu Gly Asn Ser Arg Phe Pro Gln Phe Asn Gln Pro Asp Pro Ser

225 230 235 240

tat cac ggt gtg gtt tat gtg gat gca ttt ggt ccg gct gcg ttt atc 768

Tyr His Gly Val Val Tyr Val Asp Ala Phe Gly Pro Ala Ala Phe Ile

245 250 255

ggc cgt gcg cgt gtg gta ccg ttg cgc aac atg gga gcg gca att tca 816

Gly Arg Ala Arg Val Val Pro Leu Arg Asn Met Gly Ala Ala Ile Ser

260 265 270

cct ttc aat tct ttt ctg att ctg caa ggt atc gaa acc ctg ccg ttg 864

Pro Phe Asn Ser Phe Leu Ile Leu Gln Gly Ile Glu Thr Leu Pro Leu

275 280 285

agg atg gaa cgg cat tgc acc aat gcg ctg gcg att gca cgt tat ctg 912

Arg Met Glu Arg His Cys Thr Asn Ala Leu Ala Ile Ala Arg Tyr Leu

290 295 300

caa agg cat ccc aaa gtc agc tgg gtc aat ttt gcc ggc ctt gaa gat 960

Gln Arg His Pro Lys Val Ser Trp Val Asn Phe Ala Gly Leu Glu Asp

305 310 315 320

aac cgt gat tac gca ctg gtg cag aaa tac atg gat ggc ggt att ccc 1008

Asn Arg Asp Tyr Ala Leu Val Gln Lys Tyr Met Asp Gly Gly Ile Pro

325 330 335

tca tcg att ctg agt ttt ggc atc aag ggc ggg cgc gag gct tgt gct 1056

Ser Ser Ile Leu Ser Phe Gly Ile Lys Gly Gly Arg Glu Ala Cys Ala

340 345 350

cgc ttt atg gac aga ctg atg ctg atc aaa cgg ctg gtc aac atc ggg 1104

Arg Phe Met Asp Arg Leu Met Leu Ile Lys Arg Leu Val Asn Ile Gly

355 360 365

gat gcc aaa acg ctg gcc tgc cac ccg gcg acg acc acc cac cgt cag 1152

Asp Ala Lys Thr Leu Ala Cys His Pro Ala Thr Thr Thr His Arg Gln

370 375 380

ctc aat gat gaa gaa ctg gca aaa gcc ggt gtc agt gct gat ctg gtg 1200

Leu Asn Asp Glu Glu Leu Ala Lys Ala Gly Val Ser Ala Asp Leu Val

385 390 395 400

cgt tta tgt gtc ggc atc gag cat att gac gat ctg att gcc gat gta 1248

Arg Leu Cys Val Gly Ile Glu His Ile Asp Asp Leu Ile Ala Asp Val

405 410 415

gag cag gct ttc cag gat tag 1269

Glu Gln Ala Phe Gln Asp

420

<210>26

<211>422

<212>PRT

<213>欧洲亚硝化单胞菌

<400>26

Met Lys Arg Glu Thr Leu Ala Ile His Gly Gly Phe Ala Gly Asp Pro

1 5 10 15

Gln Thr His Ala Val Ala Val Pro Ile Tyr Gln Thr Thr Ser Tyr Tyr

20 25 30

Phe Asp Asp Thr Gln His Gly Ala Asp Leu Phe Asp Leu Lys Val Gln

35 40 45

Gly Asn Ile Tyr Thr Arg Ile Met Asn Pro Thr Thr Ala Val Leu Glu

50 55 60

Glu Arg Val Ala Leu Leu Glu Gly Gly Val Gly Ala Leu Ala Met Ala

65 70 75 80

Ser Gly Met Ala Ala Ile Thr Ala Cys Val Gln Thr Leu Ala Arg Ala

85 90 95

Gly Asp Asn Ile Ile Ser Thr Ser Gln Val Tyr Gly Gly Thr Tyr Asn

100 105 110

Phe Phe Cys His Thr Leu Pro Asn Leu Gly Ile Glu Val Arg Met Val

115 120 125

Asp Gly Arg Asn Pro Ala Ala Phe Ala Asp Ala Ile Asp Asp Asn Thr

130 135 140

Arg Met Ile Tyr Cys Glu Ser Ile Gly Asn Pro Ala Gly Asn Val Val

145 150 155 160

Asp Ile Ala Ala Leu Ala Glu Val Ala His Ala Ala Gly Val Pro Leu

165 170 175

Val Val Asp Asn Thr Val Pro Thr Pro Val Leu Cys Arg Pro Phe Glu

180 185 190

His Gly Ala Asp Ile Val Val His Ala Leu Thr Lys Tyr Met Gly Gly

195 200 205

His Gly Thr Ser Ile Gly Gly Ile Ile Val Asp Ser Gly Lys Phe Pro

210 215 220

Trp Glu Gly Asn Ser Arg Phe Pro Gln Phe Asn Gln Pro Asp Pro Ser

225 230 235 240

Tyr His Gly Val Val Tyr Val Asp Als Phe Gly Pro Ala Ala Phe Ile

245 250 255

Gly Arg Ala Arg Val Val Pro Leu Arg Asn Met Gly Ala Ala Ile Ser

260 265 270

Pro Phe Asn Ser Phe Leu Ile Leu Gln Gly Ile Glu Thr Leu Pro Leu

275 280 285

Arg Met Glu Arg His Cys Thr Asn Ala Leu Ala Ile Ala Arg Tyr Leu

290 295 300

Gln Arg His Pro Lys Val Ser Trp Val Asn Phe Ala Gly Leu Glu Asp

305 310 315 320

Asn Arg Asp Tyr Ala Leu Val Gln Lys Tyr Met Asp Gly Gly Ile Pro

325 330 335

Ser Ser Ile Leu Ser Phe Gly Ile Lys Gly Gly Arg Glu Ala Cys Ala

340 345 350

Arg Phe Met Asp Arg Leu Met Leu Ile Lys Arg Leu Val Asn Ile Gly

355 360 365

Asp Ala Lys Thr Leu Ala Cys His Pro Ala Thr Thr Thr His Arg Gln

370 375 380

Leu Asn Asp Glu Glu Leu Ala Lys Ala Gly Val Ser Ala Asp Leu Val

385 390 395 400

Arg Leu Cys Val Gly Ile Glu His Ile Asp Asp Leu Ile Ala Asp Val

405 410 415

Glu Gln Ala Phe Gln Asp

420

<210>27

<211>1281

<212>DNA

<213>苜蓿中华根瘤菌(Sinorhizobium meliloti)

<220>

<221>CDS

<222>(1)..(1281)

<223>

<400>27

atg aaa gcc gga ccc gga ttc agc acg ctt gca att cac gcc ggg gcc 48

Met Lys Ala Gly Pro Gly Phe Ser Thr Leu Ala Ile His Ala Gly Ala

1 5 10 15

cag ccc gat ccg acg acc ggt gcg cgg gcg acg ccg atc tat cag acg 96

Gln Pro Asp Pro Thr Thr Gly Ala Arg Ala Thr Pro Ile Tyr Gln Thr

20 25 30

acc agc ttc gtc ttc aac gac acg gat cat gcg gcc gca ctc ttc ggc 144

Thr Ser Phe Val Phe Asn Asp Thr Asp His Ala Ala Ala Leu Phe Gly

35 40 45

ctc cag caa ttc ggc aat atc tat acc cgc atc atg aat ccg acg cag 192

Leu Gln Gln Phe Gly Asn Ile Tyr Thr Arg Ile Met Asn Pro Thr Gln

50 55 60

gcg gtg ctg gag gag cgg atc gcg gcg ctc gaa ggc ggg acc gcc ggg 240

Ala Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Gly Gly Thr Ala Gly

65 70 75 80

ctg gcc gtt tcc tcg ggg cat gcg gcc cag ctg ctg gtt ttc cat acg 288

Leu Ala Val Ser Ser Gly His Ala Ala Gln Leu Leu Val Phe His Thr

85 90 95

atc atg agg ccg ggt gac aat ttc gtt tcc gcc aga cag ctt tac ggc 336

Ile Met Arg Pro Gly Asp Asn Phe Val Ser Ala Arg Gln Leu Tyr Gly

100 105 110

ggg tcg gcc aat cag ttc ggc cat gcc ttc aag gcc ttc gac tgg cag 384

Gly Ser Ala Asn Gln Phe Gly His Ala Phe Lys Ala Phe Asp Trp Gln

115 120 125

gtc cgc tgg gcc gat tcg gcg gag ccc gaa agc ttc gat gcg cag atc 432

Val Arg Trp Ala Asp Ser Ala Glu Pro Glu Ser Phe Asp Ala Gln Ile

130 135 140

gac gaa cgc acc aag gcg atc ttc atc gaa agc ctc gcc aat ccg ggc 480

Asp Glu Arg Thr Lys Ala Ile Phe Ile Glu Ser Leu Ala Asn Pro Gly

145 150 155 160

ggc acc ttc gtc gac ata gcc gca atc gct gac gtt gcg cgg cga cac 528

Gly Thr Phe Val Asp Ile Ala Ala Ile Ala Asp Val Ala Arg Arg His

165 170 175

gga ctg ccg ctc atc gtc gac aat acg atg gcg acg ccc tat ctg atg 576

Gly Leu Pro Leu Ile Val Asp Asn Thr Met Ala Thr Pro Tyr Leu Met

180 185 190

cgg ccg ctc gaa cac ggc gcc gat atc gtc gtc cat tcg ctc acc aag 624

Arg Pro Leu Glu His Gly Ala Asp Ile Val Val His Ser Leu Thr Lys

195 200 205

ttc atc ggc ggt cac ggc aat tcg atg ggc ggc atc atc gtc gac ggc 672

Phe Ile Gly Gly His Gly Asn Ser Met Gly Gly Ile Ile Val Asp Gly

210 215 220

ggt acg ttc gac tgg tcg aaa tcc ggc aag tat ccg ctg ctg tcg gag 720

Gly Thr Phe Asp Trp Ser Lys Ser Gly Lys Tyr Pro Leu Leu Ser Glu

225 230 235 240

ccg agg ccc gaa tat ggc ggc gtc gtc ctg cac cag gcc ttc ggc aac 768

Pro Arg Pro Glu Tyr Gly Gly Val Val Leu His Gln Ala Phe Gly Asn

245 250 255

ttc gcc ttc gcc atc gcc gca cgg gta ttg ggt ctg agg gac ttc ggt 816

Phe Ala Phe Ala Ile Ala Ala Arg Val Leu Gly Leu Arg Asp Phe Gly

260 265 270

ccg gcc att tcg ccc ttc aac gcc ttc ctg atc cag acc ggc gtc gag 864

Pro Ala Ile Ser Pro Phe Asn Ala Phe Leu Ile Gln Thr Gly Val Glu

275 280 285

acg ctg ccg ctg agg atg cag cgc cat tgc gac aac gcg ctg gag gtc 912

Thr Leu Pro Leu Arg Met Gln Arg His Cys Asp Asn Ala Leu Glu Val

290 295 300

gcc aaa tgg ctg aag gga cat gaa aag gtc tcc tgg gtc cgc tat tcc 960

Ala Lys Trp Leu Lys Gly His Glu Lys Val Ser Trp Val Arg Tyr Ser

305 310 315 320

ggg ctc gaa gac gat ccg aac cac gca ctg cag aaa cgc tac tcg ccg 1008

Gly Leu Glu Asp Asp Pro Asn His Ala Leu Gln Lys Arg Tyr Ser Pro

325 330 335

aag ggg gcg gga gcc gtt ttc acc ttc ggg ctc gcg ggc gga tac gag 1056

Lys Gly Ala Gly Ala Val Phe Thr Phe Gly Leu Ala Gly Gly Tyr Glu

340 345 350

gcg gga aag cgc ttt gtc gag gca ctg gaa atg ttc tcc cat ctt gcc 1104

Ala Gly Lys Arg Phe Val Glu Ala Leu Glu Met Phe Ser His Leu Ala

355 360 365

aat atc ggc gac acg cgt tcg ctc gtc atc cac ccc gca tcg acc acg 1152

Asn Ile Gly Asp Thr Arg Ser Leu Val Ile His Pro Ala Ser Thr Thr

370 375 380

cac cgg cag ctc acg ccg gag cag cag gtc gcc gca ggc gcc gga ccc 1200

His Arg Gln Leu Thr Pro Glu Gln Gln Val Ala Ala Gly Ala Gly Pro

385 390 395 400

gac gtc atc cgg ttg tcg gtc ggc atc gag gat gtg gcc gac atc att 1248

Asp Val Ile Arg Leu Ser Val Gly Ile Glu Asp Val Ala Asp Ile Ile

405 410 415

gcc gat ctc gaa cag gcg ctg ggc aag gcc tga 1281

Ala Asp Leu Glu Gln Ala Leu Gly Lys Ala

420 425

<210>28

<211>426

<212>PRT

<213>苜蓿中华根瘤菌

<400>28

Met Lys Ala Gly Pro Gly Phe Ser Thr Leu Ala Ile His Ala Gly Ala

1 5 10 15

Gln Pro Asp Pro Thr Thr Gly Ala Arg Ala Thr Pro Ile Tyr Gln Thr

20 25 30

Thr Ser Phe Val Phe Asn Asp Thr Asp His Ala Ala Ala Leu Phe Gly

35 40 45

Leu Gln Gln Phe Gly Asn Ile Tyr Thr Arg Ile Met Asn Pro Thr Gln

50 55 60

Ala Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Gly Gly Thr Ala Gly

65 70 75 80

Leu Ala Val Ser Ser Gly His Ala Ala Gln Leu Leu Val Phe His Thr

85 90 95

Ile Met Arg Pro Gly Asp Asn Phe Val Ser Ala Arg Gln Leu Tyr Gly

100 105 110

Gly Ser Ala Asn Gln Phe Gly His Ala Phe Lys Ala Phe Asp Trp Gln

115 120 125

Val Arg Trp Ala Asp Ser Ala Glu Pro Glu Ser Phe Asp Ala Gln Ile

130 135 140

Asp Glu Arg Thr Lys Ala Ile Phe Ile Glu Ser Leu Ala Asn Pro Gly

145 150 155 160

Gly Thr Phe Val Asp Ile Ala Ala Ile Ala Asp Val Ala Arg Arg His

165 170 175

Gly Leu Pro Leu Ile Val Asp Asn Thr Met Ala Thr Pro Tyr Leu Met

180 185 190

Arg Pro Leu Glu His Gly Ala Asp Ile Val Val His Ser Leu Thr Lys

195 200 205

Phe Ile Gly Gly His Gly Asn Ser Met Gly Gly Ile Ile Val Asp Gly

210 215 220

Gly Thr Phe Asp Trp Ser Lys Ser Gly Lys Tyr Pro Leu Leu Ser Glu

225 230 235 240

Pro Arg Pro Glu Tyr Gly Gly Val Val Leu His Gln Ala Phe Gly Asn

245 250 255

Phe Ala Phe Ala Ile Ala Ala Arg Val Leu Gly Leu Arg Asp Phe Gly

260 265 270

Pro Ala Ile Ser Pro Phe Asn Ala Phe Leu Ile Gln Thr Gly Val Glu

275 280 285

Thr Leu Pro Leu Arg Met Gln Arg His Cys Asp Asn Ala Leu Glu Val

290 295 300

Ala Lys Trp Leu Lys Gly His Glu Lys Val Ser Trp Val Arg Tyr Ser

305 310 315 320

Gly Leu Glu Asp Asp Pro Asn His Ala Leu Gln Lys Arg Tyr Ser Pro

325 330 335

Lys Gly Ala Gly Ala Val Phe Thr Phe Gly Leu Ala Gly Gly Tyr Glu

340 345 350

Ala Gly Lys Arg Phe Val Glu Ala Leu Glu Met Phe Ser His Leu Ala

355 360 365

Asn Ile Gly Asp Thr Arg Ser Leu Val Ile His Pro Ala Ser Thr Thr

370 375 380

His Arg Gln Leu Thr Pro Glu Gln Gln Val Ala Ala Gly Ala Gly Pro

385 390 395 400

Asp Val Ile Arg Leu Ser Val Gly Ile Glu Asp Val Ala Asp Ile Ile

405 410 415

Ala Asp Leu Glu Gln Ala Leu Gly Lys Ala

420 425

<210>29

<211>1293

<212>DNA

<213>海栖热袍菌(Thermotoga maritima)

<220>

<221>CDS

<222>(1)..(1293)

<223>

<400>29

atg gac tgg aag aaa tac ggt tac aac aca agg gct ctt cac gca ggt 48

Met Asp Trp Lys Lys Tyr Gly Tyr Asn Thr Arg Ala Leu His Ala Gly

1 5 10 15

tat gaa cca ccc gag cag gcc aca gga tcg aga gcg gtc cct ata tat 96

Tyr Glu Pro Pro Glu Gln Ala Thr Gly Ser Arg Ala Val Pro Ile Tyr

20 25 30

caa acg act tct tac gtt ttc aga gac tct gat cac gcg gcg aga ctc 144

Gln Thr Thr Ser Tyr Val Phe Arg Asp Ser Asp His Ala Ala Arg Leu

35 40 45

ttc gca ctg gaa gaa cct ggg ttc atc tat aca agg att gga aat cct 192

Phe Ala Leu Glu Glu Pro Gly Phe Ile Tyr Thr Arg Ile Gly Asn Pro

50 55 60

acc gtc tca gtt ctt gaa gaa aga ata gcc gcc ctg gaa gaa ggg gtg 240

Thr Val Ser Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Glu Gly Val

65 70 75 80

gga gcc tta gcg gtt gcc agt gga caa gcc gct ata act tac gcc att 288

Gly Ala Leu Ala Val Ala Ser Gly Gln Ala Ala Ile Thr Tyr Ala Ile

85 90 95

ttg aac atc gcg ggc cca gga gat gag atc gtc agc ggg agc gcg ctg 336

Leu Asn Ile Ala Gly Pro Gly Asp Glu Ile Val Ser Gly Ser Ala Leu

100 105 110

tat ggg gga acg tac aat ctg ttc aga cac act ctc tat aaa aaa tcc 384

Tyr Gly Gly Thr Tyr Asn Leu Phe Arg His Thr Leu Tyr Lys Lys Ser

115 120 125

ggc atc atc gtg aag ttt gtg gat gag aca gat cca aag aac ata gaa 432

Gly Ile Ile Val Lys Phe Val Asp Glu Thr Asp Pro Lys Asn Ile Glu

130 135 140

gag gcc atc acc gag aaa aca aag gcg gtg tac ctt gaa act atc ggg 480

Glu Ala Ile Thr Glu Lys Thr Lys Ala Val Tyr Leu Glu Thr Ile Gly

145 150 155 160

aat ccc ggt ctc aca gtg ccg gac ttt gaa gcg ata gcg gag atc gct 528

Asn Pro Gly Leu Thr Val Pro Asp Phe Glu Ala Ile Ala Glu Ile Ala

165 170 175

cac aga cac ggt gtt cct ttg ata gtg gac aat acg gta gct ccg tac 576

His Arg His Gly Val Pro Leu Ile Val Asp Asn Thr Val Ala Pro Tyr

180 185 190

ata ttc agg ccc ttc gaa cac ggt gcc gac atc gtt gtt tat tcg gcc 624

Ile Phe Arg Pro Phe Glu His Gly Ala Asp Ile Val Val Tyr Ser Ala

195 200 205

acg aaa ttc atc gga gga cac gga aca tcg ata ggc ggt ctc atc gta 672

Thr Lys Phe Ile Gly Gly His Gly Thr Ser Ile Gly Gly Leu Ile Val

210 215 220

gac agc gga aaa ttc gac tgg acg aac gga aag ttt cca gaa ctc gtg 720

Asp Ser Gly Lys Phe Asp Trp Thr Asn Gly Lys Phe Pro Glu Leu Val

225 230 235 240

gaa cca gat ccc agc tac cac ggt gtg agt tat gtg gag acg ttc aaa 768

Glu Pro Asp Pro Ser Tyr His Gly Val Ser Tyr Val Glu Thr Phe Lys

245 250 255

gaa gca gcc tac ata gca aaa tgt aga acc cag ctt ttg agg gac ctg 816

Glu Als Ala Tyr Ile Ala Lys Cys Arg Thr Gln Leu Leu Arg Asp Leu

260 265 270

gga agc tgt atg agc ccg ttc aac gcg ttt ctg ttc atc ctc gga ctt 864

Gly Ser Cys Met Ser Pro Phe Asn Ala Phe Leu Phe Ile Leu Gly Leu

275 280 285

gaa acc ctc agc ttg agg atg aag aaa cac tgt gaa aac gca ctg aag 912

Glu Thr Leu Ser Leu Arg Met Lys Lys His Cys Glu Asn Ala Leu Lys

290 295 300

atc gtt gaa ttt ctg aaa tcg cat ccc gcc gtg agc tgg gtc aac tat 960

Ile Val Glu Phe Leu Lys Ser His Pro Ala Val Ser Trp Val Asn Tyr

305 310 315 320

ccg ata gct gaa ggc aat aaa acc aga gaa aat gcg ctg aaa tac ctc 1008

Pro Ile Ala Glu Gly Asn Lys Thr Arg Glu Asn Ala Leu Lys Tyr Leu

325 330 335

aaa gaa gga tac ggt gcg att gta acg ttc ggt gtg aaa ggc gga aaa 1056

Lys Glu Gly Tyr Gly Ala Ile Val Thr Phe Gly Val Lys Gly Gly Lys

340 345 350

gag gcg gga aag aag ttc ata gac agt ctc aca ctc att tcc cac ctc 1104

Glu Ala Gly Lys Lys Phe Ile Asp Ser Leu Thr Leu Ile Ser His Leu

355 360 365

gcc aac att ggt gat gca aga act ctg gct att cat ccc gct tcg aca 1152

Ala Asn Ile Gly Asp Ala Arg Thr Leu Ala Ile His Pro Ala Ser Thr

370 375 380

acc cat cag cag ctc acg gaa gaa gag cag ttg aaa acg ggt gtt act 1200

Thr His Gln Gln Leu Thr Glu Glu Glu Gln Leu Lys Thr Gly Val Thr

385 390 395 400

ccg gat atg ata aga ttg tct gtt gga ata gaa gat gtg gaa gat atc 1248

Pro Asp Met Ile Arg Leu Ser Val Gly Ile Glu Asp Val Glu Asp Ile

405 410 415

ata gcc gat ctg gat cag gct ctc aga aaa tct cag gag gga tga 1293

Ile Ala Asp Leu Asp Gln Ala Leu Arg Lys Ser Gln Glu Gly

420 425 430

<210>30

<211>430

<212>PRT

<213>海栖热袍菌

<400>30

Met Asp Trp Lys Lys Tyr Gly Tyr Asn Thr Arg Ala Leu His Ala Gly

1 5 10 15

Tyr Glu Pro Pro Glu Gln Ala Thr Gly Ser Arg Ala Val Pro Ile Tyr

20 25 30

Gln Thr Thr Ser Tyr Val Phe Arg Asp Ser Asp His Ala Ala Arg Leu

35 40 45

Phe Ala Leu Glu Glu Pro Gly Phe Ile Tyr Thr Arg Ile Gly Asn Pro

50 55 60

Thr Val Ser Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Glu Gly Val

65 70 75 80

Gly Ala Leu Ala Val Ala Ser Gly Gln Ala Ala Ile Thr Tyr Ala Ile

85 90 95

Leu Asn Ile Ala Gly Pro Gly Asp Glu Ile Val Ser Gly Ser Ala Leu

100 105 110

Tyr Gly Gly Thr Tyr Asn Leu Phe Arg His Thr Leu Tyr Lys Lys Ser

115 120 125

Gly Ile Ile Val Lys Phe Val Asp Glu Thr Asp Pro Lys Asn Ile Glu

130 135 140

Glu Ala Ile Thr Glu Lys Thr Lys Ala Val Tyr Leu Glu Thr Ile Gly

145 150 155 160

Asn Pro Gly Leu Thr Val Pro Asp Phe Glu Ala Ile Ala Glu Ile Ala

165 170 175

His Arg His Gly Val Pro Leu Ile Val Asp Asn Thr Val Ala Pro Tyr

180 185 190

Ile Phe Arg Pro Phe Glu His Gly Ala Asp Ile Val Val Tyr Ser Ala

195 200 205

Thr Lys Phe Ile Gly Gly His Gly Thr Ser Ile Gly Gly Leu Ile Val

210 215 220

Asp Ser Gly Lys Phe Asp Trp Thr Asn Gly Lys Phe Pro Glu Leu Val

225 230 235 240

Glu Pro Asp Pro Ser Tyr His Gly Val Ser Tyr Val Glu Thr Phe Lys

245 250 255

Glu Ala Ala Tyr Ile Ala Lys Cys Arg Thr Gln Leu Leu Arg Asp Leu

260 265 270

Gly Ser Cys Met Ser Pro Phe Asn Ala Phe Leu Phe Ile Leu Gly Leu

275 280 285

Glu Thr Leu Ser Leu Arg Met Lys Lys His Cys Glu Asn Ala Leu Lys

290 295 300

Ile Val Glu Phe Leu Lys Ser His Pro Ala Val Ser Trp Val Asn Tyr

305 310 315 320

Pro Ile Ala Glu Gly Asn Lys Thr Arg Glu Asn Ala Leu Lys Tyr Leu

325 330 335

Lys Glu Gly Tyr Gly Ala Ile Val Thr Phe Gly Val Lys Gly Gly Lys

340 345 350

Glu Ala Gly Lys Lys Phe Ile Asp Ser Leu Thr Leu Ile Ser His Leu

355 360 365

Ala Asn Ile Gly Asp Ala Arg Thr Leu Ala Ile His Pro Ala Ser Thr

370 375 380

Thr His Gln Gln Leu Thr Glu Glu Glu Gln Leu Lys Thr Gly Val Thr

385 390 395 400

Pro Asp Met Ile Arg Leu Ser Val Gly Ile Glu Asp Val Glu Asp Ile

405 410 415

Ile Ala Asp Leu Asp Gln Ala Leu Arg Lys Ser Gln Glu Gly

420 425 430

<210>31

<211>1314

<212>DNA

<213>变异链球菌(Streptococcus mutans)

<220>

<221>CDS

<222>(1)..(1314)

<223>

<400>31

atg gag cta att aat aat aaa agg aga gct tcc atg act cga gaa ttt 48

Met Glu Leu Ile Asn Asn Lys Arg Arg Ala Ser Met Thr Arg Glu Phe

1 5 10 15

tct ttt gaa act tta caa tta cat gcg gga caa agt gtt gat cct aca 96

Ser Phe Glu Thr Leu Gln Leu His Ala Gly Gln Ser Val Asp Pro Thr

20 25 30

aca aaa tcg cgt gca gta cca atc tat cag acg act tcc tat gtg ttt 144

Thr Lys Ser Arg Ala Val Pro Ile Tyr Gln Thr Thr Ser Tyr Val Phe

35 40 45

aat gat gca caa gat gct gaa gat tct ttt gca ctt cgt aca ccc ggc 192

Asn Asp Ala Gln Asp Ala Glu Asp Ser Phe Ala Leu Arg Thr Pro Gly

50 55 60

aat att tat acg cgg atc act aat ccg act aca gcc gtt ttt gaa gaa 240

Asn Ile Tyr Thr Arg Ile Thr Asn Pro Thr Thr Ala Val Phe Glu Glu

65 70 75 80

cgg atg gcc gct ctt gaa ggt ggt gtc ggt gca ctg gca aca gct tct 288

Arg Met Ala Ala Leu Glu Gly Gly Val Gly Ala Leu Ala Thr Ala Ser

85 90 95

ggt atg gca gca gta act tat att gcc ttg gct ctt gct cat gca ggt 336

Gly Met Ala Ala Val Thr Tyr Ile Ala Leu Ala Leu Ala His Ala Gly

100 105 110

gat cat att gtg tca gca gcg aca gtt tac ggt ggc act ttt aat ctt 384

Asp His Ile Val Ser Ala Ala Thr Val Tyr Gly Gly Thr Phe Asn Leu

115 120 125

ctt aag gaa act tta cct cgc tat ggc att act aca agt ttt gtt gat 432

Leu Lys Glu Thr Leu Pro Arg Tyr Gly Ile Thr Thr Ser Phe Val Asp

130 135 140

gtt gct aat ttc gct gaa att gaa gcg gct att aca gac aag act aag 480

Val Ala Asn Phe Ala Glu Ile Glu Ala Ala Ile Thr Asp Lys Thr Lys

145 150 155 160

ttt att atc gct gaa acg tta gga aat cct ctt gga aat atc gct gat 528

Phe Ile Ile Ala Glu Thr Leu Gly Asn Pro Leu Gly Asn Ile Ala Asp

165 170 175

ctt gaa aaa tta gct gag att gcc cat cga cat gct att ccc ttg gtt 576

Leu Glu Lys Leu Ala Glu Ile Ala His Arg His Ala Ile Pro Leu Val

180 185 190

att gat aat acc ttt ggt act cct tat ttg ctt aat gtc ttc tct tac 624

Ile Asp Asn Thr Phe Gly Thr Pro Tyr Leu Leu Asn Val Phe Ser Tyr

195 200 205

ggt gtt gat att gct gtt cat tct gcc act aaa ttt atc ggt gga cat 672

Gly Val Asp Ile Ala Val His Ser Ala Thr Lys Phe Ile Gly Gly His

210 215 220

ggg aca tct att ggc ggt gtc att gtt gat tct gga aac ttt gat tgg 720

Gly Thr Ser Ile Gly Gly Val Ile Val Asp Ser Gly Asn Phe Asp Trp

225 230 235 240

gaa aaa tct gga aaa ttc cca caa ttt gta gaa cca gat cct tcc tat 768

Glu Lys Ser Gly Lys Phe Pro Gln Phe Val Glu Pro Asp Pro Ser Tyr

245 250 255

cat gac att agt tat aca cgt gat att gga aaa gca gct ttt gta act 816

His Asp Ile Ser Tyr Thr Arg Asp Ile Gly Lys Ala Ala Phe Val Thr

260 265 270

gcg gtg cgt acg caa ctg ctg cgt gat aca ggc gcc tgc ctt tca cct 864

Ala Val Arg Thr Gln Leu Leu Arg Asp Thr Gly Ala Cys Leu Ser Pro

275 280 285

ttc aat gcc ttt ctt ttg cta caa ggt cta gaa acc tta tca ctt cgt 912

Phe Asn Ala Phe Leu Leu Leu Gln Gly Leu Glu Thr Leu Ser Leu Arg

290 295 300

gtt gag cgt cat gtg gaa aat gct aag aaa att gcg tac tat ctg gaa 960

Val Glu Arg His Val Glu Asn Ala Lys Lys Ile Ala Tyr Tyr Leu Glu

305 310 315 320

aat cat cct aaa gtc aca aaa gtt aat tat gct agt ttg cca tca agt 1008

Asn His Pro Lys Val Thr Lys Val Asn Tyr Ala Ser Leu Pro Ser Ser

325 330 335

cct tat tat gac ttg gct caa aaa tac ttg cca aaa gga gct agt tct 1056

Pro Tyr Tyr Asp Leu Ala Gln Lys Tyr Leu Pro Lys Gly Ala Ser Ser

340 345 350

atc ttt act ttt aat gtt gca ggc agt gcg aaa gcc gct cgc gag gtc 1104

Ile Phe Thr Phe Asn Val Ala Gly Ser Ala Lys Ala Ala Arg Glu Val

355 360 365

att gac agt ctt gaa atc ttt tct gat ttg gcg aat gtt gct gat gcc 1152

Ile Asp Ser Leu Glu Ile Phe Ser Asp Leu Ala Asn Val Ala Asp Ala

370 375 380

aaa tca cta gtt gtt cat ccg gca aca acc act cat ggt caa atg act 1200

Lys Ser Leu Val Val His Pro Ala Thr Thr Thr His Gly Gln Met Thr

385 390 395 400

gaa gaa gat cta cga gct tgc ggt att gaa cct gag caa atc cgt gtt 1248

Glu Glu Asp Leu Arg Ala Cys Gly Ile Glu Pro Glu Gln Ile Arg Val

405 410 415

tct att ggt ttg gaa aat gct gat gac tta atc gaa gat ttg cgc cta 1296

Ser Ile Gly Leu Glu Asn Ala Asp Asp Leu Ile Glu Asp Leu Arg Leu

420 425 430

gca ctt gaa aaa ata taa 1314

Ala Leu Glu Lys Ile

435

<210>32

<211>437

<212>PRT

<213>变异链球菌

<400>32

Met Glu Leu Ile Asn Asn Lys Arg Arg Ala Ser Met Thr Arg Glu Phe

1 5 10 15

Ser Phe Glu Thr Leu Gln Leu His Ala Gly Gln Ser Val Asp Pro Thr

20 25 30

Thr Lys Ser Arg Ala Val Pro Ile Tyr Gln Thr Thr Ser Tyr Val Phe

35 40 45

Asn Asp Ala Gln Asp Ala Glu Asp Ser Phe Ala Leu Arg Thr Pro Gly

50 55 60

Asn Ile Tyr Thr Arg Ile Thr Asn Pro Thr Thr Ala Val Phe Glu Glu

65 70 75 80

Arg Met Ala Ala Leu Glu Gly Gly Val Gly Ala Leu Ala Thr Ala Ser

85 90 95

Gly Met Ala Ala Val Thr Tyr Ile Ala Leu Ala Leu Ala His Ala Gly

100 105 110

Asp His Ile Val Ser Ala Ala Thr Val Tyr Gly Gly Thr Phe Asn Leu

115 120 125

Leu Lys Glu Thr Leu Pro Arg Tyr Gly Ile Thr Thr Ser Phe Val Asp

130 135 140

Val Ala Asn Phe Ala Glu Ile Glu Ala Ala Ile Thr Asp Lys Thr Lys

145 150 155 160

Phe Ile Ile Ala Glu Thr Leu Gly Asn Pro Leu Gly Asn Ile Ala Asp

165 170 175

Leu Glu Lys Leu Ala Glu Ile Ala His Arg His Ala Ile Pro Leu Val

180 185 190

Ile Asp Asn Thr Phe Gly Thr Pro Tyr Leu Leu Asn Val Phe Ser Tyr

195 200 205

Gly Val Asp Ile Ala Val His Ser Ala Thr Lys Phe Ile Gly Gly His

210 215 220

Gly Thr Ser Ile Gly Gly Val Ile Val Asp Ser Gly Asn Phe Asp Trp

225 230 235 240

Glu Lys Ser Gly Lys Phe Pro Gln Phe Val Glu Pro Asp Pro Ser Tyr

245 250 255

His Asp Ile Ser Tyr Thr Arg Asp Ile Gly Lys Ala Ala Phe Val Thr

260 265 270

Ala Val Arg Thr Gln Leu Leu Arg Asp Thr Gly Ala Cys Leu Ser Pro

275 280 285

Phe Asn Ala Phe Leu Leu Leu Gln Gly Leu Glu Thr Leu Ser Leu Arg

290 295 300

Val Glu Arg His Val Glu Asn Ala Lys Lys Ile Ala Tyr Tyr Leu Glu

305 310 315 320

Asn His Pro Lys Val Thr Lys Val Asn Tyr Ala Ser Leu Pro Ser Ser

325 330 335

Pro Tyr Tyr Asp Leu Ala Gln Lys Tyr Leu Pro Lys Gly Ala Ser Ser

340 345 350

Ile Phe Thr Phe Asn Val Ala Gly Ser Ala Lys Ala Ala Arg Glu Val

355 360 365

Ile Asp Ser Leu Glu Ile Phe Ser Asp Leu Ala Asn Val Ala Asp Ala

370 375 380

Lys Ser Leu Val Val His Pro Ala Thr Thr Thr His Gly Gln Met Thr

385 390 395 400

Glu Glu Asp Leu Arg Ala Cys Gly Ile Glu Pro Glu Gln Ile Arg Val

405 410 415

Ser Ile Gly Leu Glu Asn Ala Asp Asp Leu Ile Glu Asp Leu Arg Leu

420 425 430

Ala Leu Glu Lys Ile

435

<210>33

<211>1431

<212>DNA

<213>洋葱伯克霍尔德氏菌(Burkholderia cepacia)

<220>

<221>CDS

<222>(1)..(1431)

<223>

<400>33

ttg aag cgc cgc acg ccg gtg ata gga tgg ccg cca ctt tca cct ttc 48

Leu Lys Arg Arg Thr Pro Val Ile Gly Trp Pro Pro Leu Ser Pro Phe

1 5 10 15

gcg agg ccg tcc gtg gcc ccg ccg ccc agc atg tcc gcg aac cgt ttc 96

Ala Arg Pro Ser Val Ala Pro Pro Pro Ser Met Ser Ala Asn Arg Phe

20 25 30

gac acg ctt gcg ctg cac gcc ggc gct gct ccc gac ccg acc acc ggc 144

Asp Thr Leu Ala Leu His Ala Gly Ala Ala Pro Asp Pro Thr Thr Gly

35 40 45

gcg cgc gcc acg ccg att tac cag act acc tcg ttt tcg ttc cgc gat 192

Ala Arg Ala Thr Pro Ile Tyr Gln Thr Thr Ser Phe Ser Phe Arg Asp

50 55 60

tcc gac cac gcc gcg gcg ctc ttc aat atg gag cgc gcc ggt cat gtt 240

Ser Asp His Ala Ala Ala Leu Phe Asn Met Glu Arg Ala Gly His Val

65 70 75 80

tat tcg cgc att tcg aac ccg acc gtg gcc gtg ttc gag gaa cgc gtg 288

Tyr Ser Arg Ile Ser Asn Pro Thr Val Ala Val Phe Glu Glu Arg Val

85 90 95

gcc gcg ctg gaa aac ggc gcg ggc gcg atc ggc acg gca agc ggc cag 336

Ala Ala Leu Glu Asn Gly Ala Gly Ala Ile Gly Thr Ala Ser Gly Gln

100 105 110

gcg gcc ctg cat ctg gcc att gcc acg ctg atg ggc gcg ggt tcg cat 384

Ala Ala Leu His Leu Ala Ile Ala Thr Leu Met Gly Ala Gly Ser His

115 120 125

arc gtc gcc tcc agc gcg ctg tac ggc ggc tcg cac aat ctg ctg cac 432

Ile Val Ala Ser Ser Ala Leu Tyr Gly Gly Ser His Asn Leu Leu His

130 135 140

tac acg ttg cgg cgc ttc ggc atc gag acg act ttc gtc aaa ccc ggc 480

Tyr Thr Leu Arg Arg Phe Gly Ile Glu Thr Thr Phe Val Lys Pro Gly

145 150 155 160

gac ctg gac gcg tgg cgc gcc gcg ctg cgc cca aac acg cgg ctg ctg 528

Asp Leu Asp Ala Trp Arg Ala Ala Leu Arg Pro Asn Thr Arg Leu Leu

165 170 175

ttc ggc gag acg ctc ggc aat ccg ggg ctc gac gtg ctc gat atc gcc 576

Phe Gly Glu Thr Leu Gly Asn Pro Gly Leu Asp Val Leu Asp Ile Ala

180 185 190

gcc gtc gcg cag atc gcg cat gag cac cgc gtg ccg ctg ctg gtc gac 624

Ala Val Ala Gln Ile Ala His Glu His Arg Val Pro Leu Leu Val Asp

195 200 205

tcg acc ttc acc aca cct tac ctg ctc aaa ccg ttc gaa cat ggc gcg 672

Ser Thr Phe Thr Thr Pro Tyr Leu Leu Lys Pro Phe Glu His Gly Ala

210 215 220

gac ttc gtc tat cac tcg gcc acc aaa ttc ctc ggc ggc cac ggc acg 720

Asp Phe Val Tyr His Ser Ala Thr Lys Phe Leu Gly Gly His Gly Thr

225 230 235 240

acg atc ggc ggc gtg ctg gtg gac ggc ggc acg ttc gac ttc gac gcc 768

Thr Ile Gly Gly Val Leu Val Asp Gly Gly Thr Phe Asp Phe Asp Ala

245 250 255

tcg ggg cgc ttc ccc gaa ttc acc gaa cct tac gac ggc ttt cac ggc 816

Ser Gly Arg Phe Pro Glu Phe Thr Glu Pro Tyr Asp Gly Phe His Gly

260 265 270

atg gtg ttc gcc gag gag agc acc gtc gcg ccg ttt ctg ctg cga gca 864

Met Val Phe Ala Glu Glu Ser Thr Val Ala Pro Phe Leu Leu Arg Ala

275 280 285

cgc cgc gag ggg ctg cgc gac ttc ggc gca tgc ctg cat ccg caa gcc 912

Arg Arg Glu Gly Leu Arg Asp Phe Gly Ala Cys Leu His Pro Gln Ala

290 295 300

gca tgg caa ctg ctg caa ggc atc gag acg ctg ccg ttg cga atg gaa 960

Ala Trp Gln Leu Leu Gln Gly Ile Glu Thr Leu Pro Leu Arg Met Glu

305 310 315 320

cgg cac gtt gcc aac acg cgc cgg gtg gtc gag ttc ctc gcc ggt cac 1008

Arg His Val Ala Asn Thr Arg Arg Val Val Glu Phe Leu Ala Gly His

325 330 335

gcc gcg gtc ggg gcc gtc gcc tat ccg gaa ctg ccc acg cac ccc gac 1056

Ala Ala Val Gly Ala Val Ala Tyr Pro Glu Leu Pro Thr His Pro Asp

340 345 350

cac gcg ctc gcg aag cgg ctg ctg ccg cgc ggc gcc ggt gcc gtg ttc 1104

His Ala Leu Ala Lys Arg Leu Leu Pro Arg Gly Ala Gly Ala Val Phe

355 360 365

agc ttc gat ctg cgc ggc gac cgc gcc gcc gga cgc agc ttt atc gaa 1152

Ser Phe Asp Leu Arg Gly Asp Arg Ala Ala Gly Arg Ser Phe Ile Glu

370 375 380

gcg ctc tcg ctg ttc tcg cat ctc gcg aac gtg ggc gac gcg cgc tcg 1200

Ala Leu Ser Leu Phe Ser His Leu Ala Asn Val Gly Asp Ala Arg Ser

385 390 395 400

ctc gtg atc cat ccc gcc tcg acc acc cac ttt cgc atg gac gcc gct 1248

Leu Val Ile His Pro Ala Ser Thr Thr His Phe Arg Met Asp Ala Ala

405 410 415

gcc ctt gcc gcg gcc ggt atc gcc gaa ggc acg atc cgc ctc tcg atc 1296

Ala Leu Ala Ala Ala Gly Ile Ala Glu Gly Thr Ile Arg Leu Ser Ile

420 425 430

ggc ctc gaa gat ccc gac gat ctg atc gac gat ctc aag cgc gcg cta 1344

Gly Leu Glu Asp Pro Asp Asp Leu Ile Asp Asp Leu Lys Arg Ala Leu

435 440 445

aag gcc gca cag aaa gcg ggc agt tcg agc gca gcg cac ggc ggc gca 1392

Lys Ala Ala Gln Lys Ala Gly Ser Ser Ser Ala Ala His Gly Gly Ala

450 455 460

tcc ggc agt gcc gcc caa ccc cgc ccg gag tcc gca tga 1431

Ser Gly Ser Ala Ala Gln Pro Arg Pro Glu Ser Ala

465 470 475

<210>34

<211>476

<212>PRT

<213>洋葱伯克霍尔德氏菌

<400>34

Leu Lys Arg Arg Thr Pro Val Ile Gly Trp Pro Pro Leu Ser Pro Phe

1 5 10 15

Ala Arg Pro Ser Val Ala Pro Pro Pro Ser Met Ser Ala Asn Arg Phe

20 25 30

Asp Thr Leu Ala Leu His Ala Gly Ala Ala Pro Asp Pro Thr Thr Gly

35 40 45

Ala Arg Ala Thr Pro Ile Tyr Gln Thr Thr Ser Phe Ser Phe Arg Asp

50 55 60

Ser Asp His Ala Ala Ala Leu Phe Asn Met Glu Arg Ala Gly His Val

65 70 75 80

Tyr Ser Arg Ile Ser Asn Pro Thr Val Ala Val Phe Glu Glu Arg Val

85 90 95

Ala Ala Leu Glu Asn Gly Ala Gly Ala Ile Gly Thr Ala Ser Gly Gln

100 105 110

Ala Ala Leu His Leu Ala Ile Ala Thr Leu Met Gly Ala Gly Ser His

115 120 125

Ile Val Ala Ser Ser Ala Leu Tyr Gly Gly Ser His Asn Leu Leu His

130 135 140

Tyr Thr Leu Arg Arg Phe Gly Ile Glu Thr Thr Phe Val Lys Pro Gly

145 150 155 160

Asp Leu Asp Ala Trp Arg Ala Ala Leu Arg Pro Asn Thr Arg Leu Leu

165 170 175

Phe Gly Glu Thr Leu Gly Asn Pro Gly Leu Asp Val Leu Asp Ile Ala

180 185 190

Ala Val Ala Gln Ile Ala His Glu His Arg Val Pro Leu Leu Val Asp

195 200 205

Ser Thr Phe Thr Thr Pro Tyr Leu Leu Lys Pro Phe Glu His Gly Ala

210 215 220

Asp Phe Val Tyr His Ser Ala Thr Lys Phe Leu Gly Gly His Gly Thr

225 230 235 240

Thr Ile Gly Gly Val Leu Val Asp Gly Gly Thr Phe Asp Phe Asp Ala

245 250 255

Ser Gly Arg Phe Pro Glu Phe Thr Glu Pro Tyr Asp Gly Phe His Gly

260 265 270

Met Val Phe Ala Glu Glu Ser Thr Val Ala Pro Phe Leu Leu Arg Ala

275 280 285

Arg Arg Glu Gly Leu Arg Asp Phe Gly Ala Cys Leu His Pro Gln Ala

290 295 300

Ala Trp Gln Leu Leu Gln Gly Ile Glu Thr Leu Pro Leu Arg Met Glu

305 310 315 320

Arg His Val Ala Asn Thr Arg Arg Val Val Glu Phe Leu Ala Gly His

325 330 335

Ala Ala Val Gly Ala Val Ala Tyr Pro Glu Leu Pro Thr His Pro Asp

340 345 350

His Ala Leu Ala Lys Arg Leu Leu Pro Arg Gly Ala Gly Ala Val Phe

355 360 365

Ser Phe Asp Leu Arg Gly Asp Arg Ala Ala Gly Arg Ser Phe Ile Glu

370 375 380

Ala Leu Ser Leu Phe Ser His Leu Ala Asn Val Gly Asp Ala Arg Ser

385 390 395 400

Leu Val Ile His Pro Ala Ser Thr Thr His Phe Arg Met Asp Ala Ala

405 410 415

Ala Leu Ala Ala Ala Gly Ile Ala Glu Gly Thr Ile Arg Leu Ser Ile

420 425 430

Gly Leu Glu Asp Pro Asp Asp Leu Ile Asp Asp Leu Lys Arg Ala Leu

435 440 445

Lys Ala Ala Gln Lys Ala Gly Ser Ser Ser Ala Ala His Gly Gly Ala

450 455 460

Ser Gly Ser Ala Ala Gln Pro Arg Pro Glu Ser Ala

465 470 475

<210>35

<211>1722

<212>DNA

<213>耐辐射奇异球菌(Deinococcus radiodurans)

<220>

<221>CDS

<222>(1)..(1722)

<223>

<400>35

gtg gcc ttc ccg tgc ggt cag gcg ggg aac aag ata aca agg ccg ggc 48

Val Ala Phe Pro Cys Gly Gln Ala Gly Asn Lys Ile Thr Arg Pro Gly

1 5 10 15

caa tgt gtc aac ggg ggc agg gca cgc tca gcc ccg tct aag ttt cgc 96

Gln Cys Val Asn Gly Gly Arg Ala Arg Ser Ala Pro Ser Lys Phe Arg

20 25 30

ctt gac ccc tta ccc gcc tcc gcg cta ctt ttt gag gag ctc ccg cag 144

Leu Asp Pro Leu Pro Ala Ser Ala Leu Leu Phe Glu Glu Leu Pro Gln

35 40 45

cag gag cca ccc act tca gag cgc ccg aga gac ctg gct cga cga cgg 192

Gln Glu Pro Pro Thr Ser Glu Arg Pro Arg Asp Leu Ala Arg Arg Arg

50 55 60

cgc ggc aac cgg acc cca tca cgt cac ggt gcc aag gcc agc ccc ctg 240

Arg Gly Asn Arg Thr Pro Ser Arg His Gly Ala Lys Ala Ser Pro Leu

65 70 75 80

ggc gtg tca acg atg agc cgc cgg gcg gga cca agc ggg aag gcc acg 288

Gly Val Ser Thr Met Ser Arg Arg Ala Gly Pro Ser Gly Lys Ala Thr

85 90 95

cgg atg acg ata ttc aag tgt ccc ttc tcg att cac agc agg cag ggg 336

Arg Met Thr Ile Phe Lys Cys Pro Phe Ser Ile His Ser Arg Gln Gly

100 105 110

gag tgc cgt gac tgg cgc ccc cga acc tgc ttc ccc cga gga gcc gcc 384

Glu Cys Arg Asp Trp Arg Pro Arg Thr Cys Phe Pro Arg Gly Ala Ala

115 120 125

acc atg acc gat acc aaa cag ccg cag cct ctg cac ttc gag acc ttg 432

Thr Met Thr Asp Thr Lys Gln Pro Gln Pro Leu His Phe Glu Thr Leu

130 135 140

cag gtg cac gcc gga caa cgc ccc gac ccc gtg acc gga gcg cag caa 480

Gln Val His Ala gly Gln Arg Pro Asp Pro Val Thr Gly Ala Gln Gln

145 150 155 160

acg ccc atc tac gcc acc aac tcc tac gtg ttc gag tcg ccc gag cac 528

Thr Pro Ile Tyr Ala Thr Asn Ser Tyr Val Phe Glu Ser Pro Glu His

165 170 175

gcc gcc gac ctc ttc ggg ctg cgg caa ttc ggc aac atc tac agc cgc 576

Ala Ala Asp Leu Phe Gly Leu Arg Gln Phe Gly Asn Ile Tyr Ser Arg

180 185 190

atc atg aac ccc acc aac gac gtg ttc gag cag cgg gtg gcc gcc ctc 624

Ile Met Asn Pro Thr Asn Asp Val Phe Glu Gln Arg Val Ala Ala Leu

195 200 205

gaa ggg ggc gtg ggg gcg ctg tcg gtg tcg agc ggg cac gcg ggg cag 672

Glu Gly Gly Val Gly Ala Leu Ser Val Ser Ser Gly His Ala Gly Gln

210 215 220

ctc gtg aca ttg ctc acg ctg gcg cag gcg gga gac aac atc gtc tcg 720

Leu Val Thr Leu Leu Thr Leu Ala Gln Ala Gly Asp Asn Ile Val Ser

225 230 235 240

tcg ccc aac ctg tac ggc ggc acc gtc aac cag ttc cgc gtc acg ctc 768

Ser Pro Asn Leu Tyr Gly Gly Thr Val Asn Gln Phe Arg Val Thr Leu

245 250 255

aag cgg ctc ggc atc gag gtg cgg ttt acc agc aaa gac gag cgc ccc 816

Lys Arg Leu Gly Ile Glu Val Arg Phe Thr Ser Lys Asp Glu Arg Pro

260 265 270

gag gaa ttc gcc gcg ctg atc gac gag cgc acg cgg gcc gta tat ctg 864

Glu Glu Phe Ala Ala Leu Ile Asp Glu Arg Thr Arg Ala Val Tyr Leu

275 280 285

gaa acc atc ggc aac ccg gcg ctg aac att ccc gat ttc gag ggc gtg 912

Glu Thr Ile Gly Asn Pro Ala Leu Asn Ile Pro Asp Phe Glu Gly Val

290 295 300

gcg aaa gtc gcg cac gag cac ggc gtc gcg gtg gtc gtg gac aac acc 960

Ala Lys Val Ala His Glu His Gly Val Ala Val Val Val Asp Asn Thr

305 310 315 320

ttc ggg gcc ggc gga tac tac tgc cag ccg ctg cgg cac ggc gcc aac 1008

Phe Gly Ala Gly Gly Tyr Tyr Cys Gln Pro Leu Arg His Gly Ala Asn

325 330 335

atc gtg ctg cac tcg gcg agc aag tgg atc ggc ggg cac ggc aac ggc 1056

Ile Val Leu His Ser Ala Ser Lys Trp Ile Gly Gly His Gly Asn Gly

340 345 350

atc ggc ggg gtc atc gtg gac ggc ggg aac ttc gac tgg ggc agc ggg 1104

Ile Gly Gly Val Ile Val Asp Gly Gly Asn Phe Asp Trp Gly Ser Gly

355 360 365

cgg tat ccg ctg atg acc gag ccc tcg ccg agt tat cac ggg ctg aag 1152

Arg Tyr Pro Leu Met Thr Glu Pro Ser Pro Ser Tyr His Gly Leu Lys

370 375 380

ttc tgg gag acg ttc ggg gaa ggc aac ggg ctg ggg ctg ccg aac atc 1200

Phe Trp Glu Thr Phe Gly Glu Gly Asn Gly Leu Gly Leu Pro Asn Ile

385 390 395 400

gcc ttc atc acc cgc gcc cgc acc gag ggg ctg cgc gac ctg gga acg 1248

Ala Phe Ile Thr Arg Ala Arg Thr Glu Gly Leu Arg Asp Leu Gly Thr

405 410 415

acc ctg gcg ccg cag cag gcg tgg cag ttt ctg caa ggc ctt gaa acc 1296

Thr Leu Ala Pro Gln Gln Ala Trp Gln Phe Leu Gln Gly Leu Glu Thr

420 425 430

ctg agc ctg cgc gcc gag cgc cac gcc gag aac acc ctg gcg ctg gcg 1344

Leu Ser Leu Arg Ala Glu Arg His Ala Glu Asn Thr Leu Ala Leu Ala

435 440 445

cac tgg ctc atc agc cac ccg gac gtg aag cag gtc act tac ccc ggc 1392

His Tru Leu Ile Ser His Pro Asp Val Lys Gln Val Thr Tyr Pro Gly

450 455 460

ctg agc aac cac ccc cac tac gac cgg gcg cag acc tac ttg ccg cgc 1440

Leu Ser Asn His Pro His Tyr Asp Arg Ala Gln Thr Tyr Leu Pro Arg

465 470 475 480

ggg gcg ggc gcg gtg ctc acc ttc gag ctg cgc ggg ggc cgg gcg gcg 1488

Gly Ala Gly Ala Val Leu Thr Phe Glu Leu Arg Gly Gly Arg Ala Ala

485 490 495

ggc gaa gcg ttt att cgc tcg gtc aag ctc gcg cag cac gtc gcc aac 1536

Gly Glu Ala Phe Ile Arg Ser Val Lys Leu Ala Gln His Val Ala Asn

500 505 510

gtg ggc gac acc cgc acg ctg gtc att cat ccg gcg agc acc acc cac 1584

Val Gly Asp Thr Arg Thr Leu Val Ile His Pro Ala Ser Thr Thr His

515 520 525

agc cag ctc gac gag gtg acg cag acg aac gcc ggg gtc acg ccg ggc 1632

Ser Gln Leu Asp Glu Val Thr Gln Thr Asn Ala Gly Val Thr Pro Gly

530 535 540

ctc atc cgg gtg tcg gtg ggc atc gag cac gta gac gac atc cgc gag 1680

Leu Ile Arg Val Ser Val Gly Ile Glu His Val Asp Asp Ile Arg Glu

545 550 555 560

gac ttc gcg cag gcc ctg gcg agc gct ggg gag cgg gcg tga 1722

Asp Phe Ala Gln Ala Leu Ala Ser Ala Gly Glu Arg Ala

565 570

<210>36

<211>573

<212>PRT

<213>耐辐射奇异球菌

<400>36

Val Ala Phe Pro Cys Gly Gln Ala Gly Asn Lys Ile Thr Arg Pro Gly

1 5 10 15

Gln Cys Val Asn Gly Gly Arg Ala Arg Ser Ala Pro Ser Lys Phe Arg

20 25 30

Leu Asp Pro Leu Pro Ala Ser Ala Leu Leu Phe Glu Glu Leu Pro Gln

35 40 45

Gln Glu Pro Pro Thr Ser Glu Arg Pro Arg Asp Leu Ala Arg Arg Arg

50 55 60

Arg Gly Asn Arg Thr Pro Ser Arg His Gly Ala Lys Ala Ser Pro Leu

65 70 75 80

Gly Val Ser Thr Met Ser Arg Arg Ala Gly Pro Ser Gly Lys Ala Thr

85 90 95

Arg Met Thr Ile Phe Lys Cys Pro Phe Ser Ile His Ser Arg Gln Gly

100 105 110

Glu Cys Arg Asp Trp Arg Pro Arg Thr Cys Phe Pro Arg Gly Ala Ala

115 120 125

Thr Met Thr Asp Thr Lys Gln Pro Gln Pro Leu His Phe Glu Thr Leu

130 135 140

Gln Val His Ala Gly Gln Arg Pro Asp Pro Val Thr Gly Ala Gln Gln

145 150 155 160

Thr Pro Ile Tyr Ala Thr Asn Ser Tyr Val Phe Glu Ser Pro Glu His

165 170 175

Ala Ala Asp Leu Phe Gly Leu Arg Gln Phe Gly Asn Ile Tyr Ser Arg

180 185 190

Ile Met Asn Pro Thr Asn Asp Val Phe Glu Gln Arg Val Ala Ala Leu

195 200 205

Glu Gly Gly Val Gly Ala Leu Ser Val Ser Ser Gly His Ala Gly Gln

210 215 220

Leu Val Thr Leu Leu Thr Leu Ala Gln Ala Gly Asp Asn Ile Val Ser

225 230 235 240

Ser Pro Asn Leu Tyr Gly Gly Thr Val Asn Gln Phe Arg Val Thr Leu

245 250 255

Lys Arg Leu Gly Ile Glu Val Arg Phe Thr Ser Lys Asp Glu Arg Pro

260 265 270

Glu Glu Phe Ala Ala Leu Ile Asp Glu Arg Thr Arg Ala Val Tyr Leu

275 280 285

Glu Thr Ile Gly Asn Pro Ala Leu Asn Ile Pro Asp Phe Glu Gly Val

290 295 300

Ala Lys Val Ala His Glu His Gly Val Ala Val Val Val Asp Asn Thr

305 310 315 320

Phe Gly Ala Gly Gly Tyr Tyr Cys Gln Pro Leu Arg His Gly Ala Asn

325 330 335

Ile Val Leu His Ser Ala Ser Lys Trp Ile Gly Gly His Gly Asn Gly

340 345 350

Ile Gly Gly Val Ile Val Asp Gly Gly Asn Phe Asp Trp Gly Ser Gly

355 360 365

Arg Tyr Pro Leu Met Thr Glu Pro Ser Pro Ser Tyr His Gly Leu Lys

370 375 380

Phe Trp Glu Thr Phe Gly Glu Gly Asn Gly Leu Gly Leu Pro Asn Ile

385 390 395 400

Ala Phe Ile Thr Arg Ala Arg Thr Glu Gly Leu Arg Asp Leu Gly Thr

405 410 415

Thr Leu Ala Pro Gln Gln Ala Trp Gln Phe Leu Gln Gly Leu Glu Thr

420 425 430

Leu Ser Leu Arg Ala Glu Arg His Ala Glu Asn Thr Leu Ala Leu Ala

435 440 445

His Trp Leu Ile Ser His Pro Asp Val Lys Gln Val Thr Tyr Pro Gly

450 455 460

Leu Ser Asn His Pro His Tyr Asp Arg Ala Gln Thr Tyr Leu Pro Arg

465 470 475 480

Gly Ala Gly Ala Val Leu Thr Phe Glu Leu Arg Gly Gly Arg Ala Ala

485 490 495

Gly Glu Ala Phe Ile Arg Ser Val Lys Leu Ala Gln His Val Ala Asn

500 505 510

Val Gly Asp Thr Arg Thr Leu Val Ile His Pro Ala Ser Thr Thr His

515 520 525

Ser Gln Leu Asp Glu Val Thr Gln Thr Asn Ala Gly Val Thr Pro Gly

530 535 540

Leu Ile Arg Val Ser Val Gly Ile Glu His Val Asp Asp Ile Arg Glu

545 550 555 560

Asp Phe Ala Gln Ala Leu Ala Ser Ala Gly Glu Arg Ala

565 570

<210>37

<211>1284

<212>DNA

<213>荚膜红细菌(Rhodobacter capsulatus)

<220>

<221>CDS

<222>(1)..(1284)

<223>

<400>37

atg acc gac cag gcc ttt gac acg ctg caa att cac gcg ggc gcc gaa 48

Met Thr Asp Gln Ala Phe Asp Thr Leu Gln Ile His Ala Gly Ala Glu

1 5 10 15

ccc gat ccc gcg acg ggc gcg cgg cag gtg ccg att tac cag acc acc 96

Pro Asp Pro Ala Thr Gly Ala Arg Gln Val Pro Ile Tyr Gln Thr Thr

20 25 30

tcc tat gtc ttc aag gac gcc gac cat gcc gcg cgc ctg ttc ggg ctg 144

Ser Tyr Val Phe Lys Asp Ala Asp His Ala Ala Arg Leu Phe Gly Leu

30 40 45

cag gag gtg ggc tat atc tat tcc cgc ctg acc aac ccg acc gtt tcg 192

Gln Glu Val Gly Tyr Ile Tyr Ser Arg Leu Thr Asn Pro Thr Val Ser

50 55 60

gca ctg gcc gcc cgc gtt gcg gcg ctt gaa ggc ggc gtg ggc gcg gtc 240

Ala Leu Ala Ala Arg Val Ala Ala Leu Glu Gly Gly Val Gly Ala Val

65 70 75 80

tgc tgc tcg tcc ggc cat gcg gcg cag atc atg gcg ctg ttt ccg ctg 288

Cys Cys Ser Ser Gly His Ala Ala Gln Ile Met Ala Leu Phe Pro Leu

85 90 95

atg ggg ccg ggg ctg aac atc gtc gcc tcg acc cgg ctt tac ggc ggc 336

Met Gly Pro Gly Leu Asn Ile Val Ala Ser Thr Arg Leu Tyr Gly Gly

100 105 110

acg atc acc cag ttc agc cag acc atc aaa cgc ttc ggc tgg tcc tgc 384

Thr Ile Thr Gln Phe Ser Gln Thr Ile Lys Arg Phe Gly Trp Ser Cys

115 120 125

acc ttt gtc gat ttc gac gat ctg gcg gcg ctc gag gcc gcg gtg gat 432

Thr Phe Val Asp Phe Asp Asp Leu Ala Ala Leu Glu Ala Ala Val Asp

130 135 140

gac aac acc cgg gcg atc ttt tgc gaa tcg atc tcg aac ccg ggc ggc 480

Asp Asn Thr Arg Ala Ile Phe Cys Glu Ser Ile Ser Asn Pro Gly Gly

145 150 155 160

tac atc acc gac ctg ccc gcc gtc gcg gcg gtg gcg aac aag gtc ggc 528

Tyr Ile Thr Asp Leu Pro Ala Val Ala Ala Val Ala Asn Lys Val Gly

165 170 175

ctg ccg ctc att gtc gac aac acg ctg gcc tcg cct tat ctc tgc cgc 576

Leu Pro Leu Ile Val Asp Asn Thr Leu Ala Ser Pro Tyr Leu Cys Arg

180 185 190

ccg atc gag cat ggc gcg acg ctg gtt gtc cat tcc gcc acg aaa tac 624

Pro Ile Glu His Gly Ala Thr Leu Val Val His Ser Ala Thr Lys Tyr

195 200 205

ctg acc ggc aac ggc acg gtg acg ggc ggg gtg atc gtc gat tcg ggc 672

Leu Thr Gly Asn Gly Thr Val Thr Gly Gly Val Ile Val Asp Ser Gly

210 215 220

aag ttc gac tgg tcg gcc tcg ggc aag ttc ccc agc ctt tcg gcg ccc 720

Lys Phe Asp Trp Ser Ala Ser Gly Lys Phe Pro Ser Leu Ser Ala Pro

225 230 235 240

gaa ccc gcc tat cac ggg ctg aag ttc cac gag gca ctc ggc ccg atg 768

Glu Pro Ala Tyr His Gly Leu Lys Phe His Glu Ala Leu Gly Pro Met

245 250 255

gcc ttc acc ttc cat tcg atc gcc gtc ggg ctg cgc gat ctg ggc atg 816

Ala Phe Thr Phe His Ser Ile Ala Val Gly Leu Arg Asp Leu Gly Met

260 265 270

acg atg aac ccg cag ggc gcg cat tac acg ctg atg ggg atc gag acg 864

Thr Met Asn Pro Gln Gly Ala His Tyr Thr Leu Met Gly Ile Glu Thr

275 280 285

ctc agc ctg cgc atg gac aag cac gtc gcc aat gcg aag gcg gtg gcg 912

Leu Ser Leu Arg Met Asp Lys His Val Ala Asn Ala Lys Ala Val Ala

290 295 300

gaa tgg ctg gcc aaa gac ccg cgc atc gac ttc gtc acc tgg gcc ggg 960

Glu Trp Leu Ala Lys Asp Pro Arg Ile Asp Phe Val Thr Trp Ala Gly

305 310 315 320

ctg ccc tcc tcg ccc tgg cac gaa cgc gcc gag cgg ctt tgc ccg aag 1008

Leu Pro Ser Ser Pro Trp His Glu Arg Ala Glu Arg Leu Cys Pro Lys

325 330 335

ggg gcg ggg gcg ctt ttc acc gtc gcg gtc aag ggc ggc tat gag gcc 1056

Gly Ala Gly Ala Leu Phe Thr Val Ala Val Lys Gly Gly Tyr Glu Ala

340 345 350

tgc gtg aaa ttg gtc aac aat ctc aag ctg ttc agc cat gtg gca aac 1104

Cys Val Lys Leu Val Asn Asn Leu Lys Leu Phe Ser His Val Ala Asn

355 360 365

ctg ggc gac gcg cgc tcg ctg atc atc cat tcg gcc tcg acc acg cac 1152

Leu Gly Asp Ala Arg Ser Leu Ile Ile His Ser Ala Ser Thr Thr His

370 375 380

cgt cag ctg acc gag gaa cag cag atc aag gcg ggg gcg gcg ccg aat 1200

Arg Gln Leu Thr Glu Glu Gln Gln Ile Lys Ala Gly Ala Ala Pro Asn

385 390 395 400

gtg gtg cgg ctc tcg atc ggg atc gag aat gcc gcc gat ctg atc gcc 1248

Val Val Arg Leu Ser Ile Gly Ile Glu Asn Ala Ala Asp Leu Ile Ala

405 410 415

gat ctg gat cag gct ctg gcc gcc gcc acc gcc tga 1284

Asp Leu Asp Gln Ala Leu Ala Ala Ala Thr Ala

420 425

<210>38

<211>427

<212>PRT

<213>荚膜红细菌

<400>38

Met Thr Asp Gln Ala Phe Asp Thr Leu Gln Ile His Ala Gly Ala Glu

1 5 10 15

Pro Asp Pro Ala Thr Gly Ala Arg Gln Val Pro Ile Tyr Gln Thr Thr

20 25 30

Ser Tyr Val Phe Lys Asp Ala Asp His Ala Ala Arg Leu Phe Gly Leu

35 40 45

Gln Glu Val Gly Tyr Ile Tyr Ser Arg Leu Thr Asn Pro Thr Val Ser

50 55 60

Ala Leu Ala Ala Arg Val Ala Ala Leu Glu Gly Gly Val Gly Ala Val

65 70 75 80

Cys Cys Ser Ser Gly His Ala Ala Gln Ile Met Ala Leu Phe Pro Leu

85 90 95

Met Gly Pro Gly Leu Asn Ile Val Ala Ser Thr Arg Leu Tyr Gly Gly

100 105 110

Thr Ile Thr Gln Phe Ser Gln Thr Ile Lys Arg Phe Gly Trp Ser Cys

115 120 125

Thr Phe Val Asp Phe Asp Asp Leu Ala Ala Leu Glu Ala Ala Val Asp

130 135 140

Asp Asn Thr Arg Ala Ile Phe Cys Glu Ser Ile Ser Asn Pro Gly Gly

145 150 155 160

Tyr Ile Thr Asp Leu Pro Ala Val Ala Ala Val Ala Asn Lys Val Gly

165 170 175

Leu Pro Leu Ile Val Asp Asn Thr Leu Ala Ser Pro Tyr Leu Cys Arg

180 185 190

Pro Ile Glu His Gly Ala Thr Leu Val Val His Ser Ala Thr Lys Tyr

195 200 205

Leu Thr Gly Asn Gly Thr Val Thr Gly Gly Val Ile Val Asp Ser Gly

210 215 220

Lys Phe Asp Trp Ser Ala Ser Gly Lys Phe Pro Ser Leu Ser Ala Pro

225 230 235 240

Glu Pro Ala Tyr His Gly Leu Lys Phe His Glu Ala Leu Gly Pro Met

245 250 255

Ala Phe Thr Phe His Ser Ile Ala Val Gly Leu Arg Asp Leu Gly Met

260 265 270

Thr Met Asn Pro Gln Gly Ala His Tyr Thr Leu Met Gly Ile Glu Thr

275 280 285

Leu Ser Leu Arg Met Asp Lys His Val Ala Asn Ala Lys Ala Val Ala

290 295 300

Glu Trp Leu Ala Lys Asp Pro Arg Ile Asp Phe Val Thr Trp Ala Gly

305 310 315 320

Leu Pro Ser Ser Pro Trp His Glu Arg Ala Glu Arg Leu Cys Pro Lys

325 330 335

Gly Ala Gly Ala Leu Phe Thr Val Ala Val Lys Gly Gly Tyr Glu Ala

340 345 350

Cys Val Lys Leu Val Asn Asn Leu Lys Leu Phe Ser His Val Ala Asn

355 360 365

Leu Gly Asp Ala Arg Ser Leu Ile Ile His Ser Ala Ser Thr Thr His

370 375 380

Arg Gln Leu Thr Glu Glu Gln Gln Ile Lys Ala Gly Ala Ala Pro Asn

385 390 395 400

Val Val Arg Leu Ser Ile Gly Ile Glu Asn Ala Ala Asp Leu Ile Ala

405 410 415

Asp Leu Asp Gln Ala Leu Ala Ala Ala Thr Ala

420 425

<210>39

<211>1269

<212>DNA

<213>多杀巴斯德氏菌(Pasteurella multocida)

<220>

<221>CDS

<222>(1)..(1269)

<223>

<400>39

atg gaa ttt gca aca aaa tgt cta cat gcc ggt tat aca ccg aaa aat 48

Met Glu Phe Ala Thr Lys Cys Leu His Ala Gly Tyr Thr Pro Lys Asn

1 5 10 15

ggt gag cct cgt gtt caa ccg atc gta caa agt acc act ttt acc tac 96

Gly Glu Pro Arg Val Gln Pro Ile Val Gln Ser Thr Thr Phe Thr Tyr

20 25 30

gat tcc gcc gaa gaa att ggt aag tta ttt gat tta caa gcg gct ggc 144

Asp Ser Ala Glu Glu Ile Gly Lys Leu Phe Asp Leu Gln Ala Ala Gly

35 40 45

tat ttt tac acc cgc ctt tca aat cct act acc aat gcg gca gaa gaa 192

Tyr Phe Tyr Thr Arg Leu Ser Asn Pro Thr Thr Asn Ala Ala Glu Glu

50 55 60

aaa att acc gca ctt gaa ggc ggt gta gca acc atg tgt acc gca tca 240

Lys Ile Thr Ala Leu Glu Gly Gly Val Ala Thr Met Cys Thr Ala Ser

65 70 75 80

ggg caa gcc gcc gtg ttt tac gcg atg ctc aat att tta caa gcc ggt 288

Gly Gln Ala Ala Val Phe Tyr Ala Met Leu Asn Ile Leu Gln Ala Gly

85 90 95

gat cac ttt att tct tca tcg tat gtt tac ggt ggt agc tac aac tta 336

Asp His Phe Ile Ser Ser Ser Tyr Val Tyr Gly Gly Ser Tyr Asn Leu

100 105 110

ttt gca cat acc ttc aaa aaa atg gga att gag gtc act ttt gtg gat 384

Phe Ala His Thr Phe Lys Lys Met Gly Ile Glu Val Thr Phe Val Asp

115 120 125

caa gat tta cct ctt gag gaa tta aaa aaa gct att cgc cca aat acg 432

Gln Asp Leu Pro Leu Glu Glu Leu Lys Lys Ala Ile Arg Pro Asn Thr

130 135 140

aaa gcc att ttt gcc gaa act att gcc aat ccc gca tta cgc gtg ttg 480

Lys Ala Ile Phe Ala Glu Thr Ile Ala Asn Pro Ala Leu Arg Val Leu

145 150 155 160

gat att gaa aag ttt gtt gca ctt gcg aag gca gca caa gcc cct tta 528

Asp Ile Glu Lys Phe Val Ala Leu Ala Lys Ala Ala Gln Ala Pro Leu

165 170 175

tta gtt gac aat act ttt gca acc ccg tat ttt tgt cgc cct atc gaa 576

Leu Val Asp Asn Thr Phe Ala Thr Pro Tyr Phe Cys Arg Pro Ile Glu

180 185 190

ttt ggt gct aac gtg gta att cat agt acg tca aaa tat tta gat ggg 624

Phe Gly Ala Asn Val Val Ile His Ser Thr Ser Lys Tyr Leu Asp Gly

195 200 205

cat gcg att gcg ttg gga ggt tcg atc aca gat ggc ggg aat ttt gat 672

His Ala Ile Ala Leu Gly Gly Ser Ile Thr Asp Gly Gly Asn Phe Asp

210 215 220

tgg aat aat ggt aaa ttc cca caa tta agc aca cct gat caa act tat 720

Trp Asn Asn Gly Lys Phe Pro Gln Leu Ser Thr Pro Asp Gln Thr Tyr

225 230 235 240

cac ggt tta gtt tat acc gaa acc ttt gtt cca gcc gct tat att gtc 768

His Gly Leu Val Tyr Thr Glu Thr Phe Val Pro Ala Ala Tyr Ile Val

245 250 255

aaa gcc cgt gtg caa tta atg cgt gat tta ggt gcc aca cca gca cca 816

Lys Ala Arg Val Gln Leu Met Arg Asp Leu Gly Ala Thr Pro Ala Pro

260 265 270

caa aat agt ttc ttg ctc aat gtg ggc atg gaa act ctt gca ctg cgt 864

Gln Asn Ser Phe Leu Leu Asn Val Gly Met Glu Thr Leu Ala Leu Arg

275 280 285

atg caa cgt cat tat gaa aat gca caa gcg gtc gcc gaa ttt tta gaa 912

Met Gln Arg His Tyr Glu Asn Ala Gln Ala Val Ala Glu Phe Leu Glu

290 295 300

aat cat cca caa gtg gca aaa gtg agt tat ccg ggc ttg gca agt tca 960

Asn His Pro Gln Val Ala Lys Val Ser Tyr Pro Gly Leu Ala Ser Ser

305 310 315 320

cct gat cat gca cta aaa caa aaa tat tta cca aac ggt tta tgt ggt 1008

Pro Asp His Ala Leu Lys Gln Lys Tyr Leu Pro Asn Gly Leu Cys Gly

325 330 335

gtg att tcc ttt gaa att aga ggg gga aga gaa act gca gca aaa tgg 1056

Val Ile Ser Phe Glu Ile Arg Gly Gly Arg Glu Thr Ala Ala Lys Trp

340 345 350

ctg aat gcg cta caa ctg gct tct cgt gaa gtc cat gta gcg gat att 1104

Leu Asn Ala Leu Gln Leu Ala Ser Arg Glu Val His Val Ala Asp Ile

355 360 365

cgc act tgt gct tta cat ccg gcg acg tca aca cac cgt caa tta agt 1152

Arg Thr Cys Ala Leu His Pro Ala Thr Ser Thr His Arg Gln Leu Ser

370 375 380

gag gct gaa tta gaa aaa gtg ggg att tct gcg ggt tta att cgt ctt 1200

Glu Ala Glu Leu Glu Lys Val Gly Ile Ser Ala Gly Leu Ile Arg Leu

385 390 395 400

tct tgc ggt att gaa agt atc caa gat att ttg gct gac tta gaa caa 1248

Ser Cys Gly Ile Glu Ser Ile Gln Asp Ile Leu Ala Asp Leu Glu Gln

405 410 415

gca ttc cac gcg gca aaa taa 1269

Ala Phe His Ala Ala Lys

420

<210>40

<211>422

<212>PRT

<213>多杀巴斯德氏菌

<400>40

Met Glu Phe Ala Thr Lys Cys Leu His Ala Gly Tyr Thr Pro Lys Asn

1 5 10 15

Gly Glu Pro Arg Val Gln Pro Ile Val Gln Ser Thr Thr Phe Thr Tyr

20 25 30

Asp Ser Ala Glu Glu Ile Gly Lys Leu Phe Asp Leu Gln Ala Ala Gly

35 40 45

Tyr Phe Tyr Thr Arg Leu Ser Asn Pro Thr Thr Asn Ala Ala Glu Glu

50 55 60

Lys Ile Thr Ala Leu Glu Gly Gly Val Ala Thr Met Cys Thr Ala Ser

65 70 75 80

Gly Gln Ala Ala Val Phe Tyr Ala Met Leu Asn Ile Leu Gln Ala Gly

85 90 95

Asp His Phe Ile Ser Ser Ser Tyr Val Tyr Gly Gly Ser Tyr Asn Leu

100 105 110

Phe Ala His Thr Phe Lys Lys Met Gly Ile Glu Val Thr Phe Val Asp

115 120 125

Gln Asp Leu Pro Leu Glu Glu Leu Lys Lys Ala Ile Arg Pro Asn Thr

130 135 140

Lys Ala Ile Phe Ala Glu Thr Ile Ala Asn Pro Ala Leu Arg Val Leu

145 150 155 160

Asp Ile Glu Lys Phe Val Ala Leu Ala Lys Ala Ala Gln Ala Pro Leu

165 170 175

Leu Val Asp Asn Thr Phe Ala Thr Pro Tyr Phe Cys Arg Pro Ile Glu

180 185 190

Phe Gly Ala Asn Val Val Ile His Ser Thr Ser Lys Tyr Leu Asp Gly

195 200 205

His Ala Ile Ala Leu Gly Gly Ser Ile Thr Asp Gly Gly Asn Phe Asp

210 215 220

Trp Asn Asn Gly Lys Phe Pro Gln Leu Ser Thr Pro Asp Gln Thr Tyr

225 230 235 240

His Gly Leu Val Tyr Thr Glu Thr Phe Val Pro Ala Ala Tyr Ile Val

245 250 255

Lys Ala Arg Val Gln Leu Met Arg Asp Leu Gly Ala Thr Pro Ala Pro

260 265 270

Gln Asn Ser Phe Leu Leu Asn Val Gly Met Glu Thr Leu Ala Leu Arg

275 280 285

Met Gln Arg His Tyr Glu Asn Ala Gln Ala Val Ala Glu Phe Leu Glu

290 295 300

Asn His Pro Gln Val Ala Lys Val Ser Tyr Pro Gly Leu Ala Ser Ser

305 310 315 320

Pro Asp His Ala Leu Lys Gln Lys Tyr Leu Pro Asn Gly Leu Cys Gly

325 330 335

Val Ile Ser Phe Glu Ile Arg Gly Gly Arg Glu Thr Ala Ala Lys Trp

340 345 350

Leu Asn Ala Leu Gln Leu Ala Ser Arg Glu Val His Val Ala Asp Ile

355 360 365

Arg Thr Cys Ala Leu His Pro Ala Thr Ser Thr His Arg Gln Leu Ser

370 375 380

Glu Ala Glu Leu Glu Lys Val Gly Ile Ser Ala Gly Leu Ile Arg Leu

385 390 395 400

Ser Cys Gly Ile Glu Ser Ile Gln Asp Ile Leu Ala Asp Leu Glu Gln

405 410 415

Ala Phe His Ala Ala Lys

420

<210>41

<211>1266

<212>DNA

<213>艰难梭菌(Clostridium difficile)

<220>

<221>CDS

<222>(1)..(1266)

<223>

<400>41

atg tat aat aaa gaa aca ata tgt gtg caa gga aat tat aaa cca ggt 48

Met Tyr Asn Lys Glu Thr Ile Cys Val Gln Gly Asn Tyr Lys Pro Gly

1 5 10 15

aat gga gaa cca aga gta cta cct tta tat caa agt aca act ttt aaa 96

Asn Gly Glu Pro Arg Val Leu Pro Leu Tyr Gln SerThr Thr Phe Lys

20 25 30

tat agc agt ata gac caa ctt gct gaa tta ttt gat tta aaa gtt gat 144

Tyr Ser Ser Ile Asp Gln Leu Ala Glu Leu Phe Asp Leu Lys Val Asp

35 40 45

gga cat ata tat tca aga ata agc aat cct act att caa gct ttt gaa 192

Gly His Ile Tyr Ser Arg Ile Ser Asn Pro Thr Ile Gln Ala Phe Glu

50 55 60

gaa aaa ata agt tta cta gag ggt gga gta tct tct gta gct gta tca 240

Glu Lys Ile Ser Leu Leu Glu Gly Gly Val Ser Ser Val Ala Val Ser

65 70 75 80

tca ggg caa tct gca aat atg ttg gca gtt tta aat ata tgt aaa tca 288

Ser Gly Gln Ser Ala Asn Met Leu Ala Val Leu Asn Ile Cys Lys Ser

85 90 95

gga gat agt ata ctt tgt tct tca aaa gta tat gga gga aca ttc aat 336

Gly Asp Ser Ile Leu Cys Ser Ser Lys Val Tyr Gly Gly Thr Phe Asn

100 105 110

tta cta gga cct agt ctt aaa aaa ttt ggt ata gat tta ata tcg ttt 384

Leu Leu Gly Pro Ser Leu Lys Lys Phe Gly Ile Asp Leu Ile Ser Phe

115 120 125

gac tta gat tca agt gaa gat gag ata gta gaa ctt gca aag gaa aat 432

Asp Leu Asp Ser Ser Glu Asp Glu Ile Val Glu Leu Ala Lys Glu Asn

130 135 140

act aag gtt gtg ttt gca gaa aca ctt gca aat cca act ctt gaa gtc 480

Thr Lys Val Val Phe Ala Glu Thr Leu Ala Asn Pro Thr Leu Glu Val

145 150 155 160

ata gat ttt gaa aaa ata gca aat gta gct aag aga att aat gtt cca 528

Ile Asp Phe Glu Lys Ile Ala Asn Val Ala Lys Arg Ile Asn Val Pro

165 170 175

ttt att gtt gat aat tca tta gca tct cca gtg ctt tgt aac cct tta 576

Phe Ile Val Asp Asn Ser Leu Ala Ser Pro Val Leu Cys Asn Pro Leu

180 185 190

aag tat gga gca aat ata gtt act cat tct acc aca aaa tat tta gat 624

Lys Tyr Gly Ala Asn Ile Val Thr His Ser Thr Thr Lys Tyr Leu Asp

195 200 205

ggg cat gct tca agt gtt gga gga att ata gtg gat ggt gga aac ttt 672

Gly His Ala Ser Ser Val Gly Gly Ile Ile Val Asp Gly Gly Asn Phe

210 215 220

aac tgg gat aat gga aaa ttt cca gaa tta gtt gag cca gac cca aca 720

Asn Trp Asp Asn Gly Lys Phe Pro Glu Leu Val Glu Pro Asp Pro Thr

225 230 235 240

tat cat ggt ata agc tat act caa aaa ttt gga aat gcc gca tat gca 768

Tyr His Gly Ile Ser Tyr Thr Gln Lys Phe Gly Asn Ala Ala Tyr Ala

245 250 255

act aaa gca aga gtt cag ttg ctt aga gac tat gga aat tgt tta agc 816

Thr Lys Ala Arg Val Gln Leu Leu Arg Asp Tyr Gly Asn Cys Leu Ser

260 265 270

cca ttc aat gcg tat ctt act aat tta aat gtt gaa aca cta cat ctt 864

Pro Phe Asn Ala Tyr Leu Thr Asn Leu Asn Val Glu Thr Leu His Leu

275 280 285

aga atg gag aga cat agt gaa aat gca ctt aaa ata gct aga ttt tta 912

Arg Met Glu Arg His Ser Glu Asn Ala Leu Lys Ile Ala Arg Phe Leu

290 295 300

gaa aaa cat gaa aat gta gat tgg att aat tac cca gga ctt gaa gat 960

Glu Lys His Glu Asn Val Asp Trp Ile Asn Tyr Pro Gly Leu Glu Asp

305 310 315 320

aac aag tat tat gag aat gcc aaa aag tat tta tca aga gga tgt agt 1008

Asn Lys Tyr Tyr Glu Asn Ala Lys Lys Tyr Leu Ser Arg Gly Cys Ser

325 330 335

ggt gtt tta tca ttt gga gta aga ggt ggg tta gaa aat gcc aaa aaa 1056

Gly Val Leu Ser Phe Gly Val Arg Gly Gly Leu Glu Asn Ala Lys Lys

340 345 350

ttt gtg gaa aaa tta cag ata gca tct ttg gtt aca cat gtt tca gat 1104

Phe Val Glu Lys Leu Gln Ile Ala Ser Leu Val Thr His Val Ser Asp

355 360 365

gta aga act tgt gtt ata cat cca gct tca act act cat aga caa tta 1152

Val Arg Thr Cys Val Ile His Pro Ala Ser Thr Thr His Arg Gln Leu

370 375 380

aca gaa gaa caa tta att gca tct gga gta ttg cct tca cta ata aga 1200

Thr Glu Glu Gln Leu Ile Ala Ser Gly Val Leu Pro Ser Leu Ile Arg

385 390 395 400

tta tct gtt gga ata gaa aat gta gag gat tta ata gct gat tta aat 1248

Leu Ser Val Gly Ile Glu Asn Val Glu Asp Leu Ile Ala Asp Leu Asn

405 410 415

caa gct tta aat ttc taa 1266

Gln Ala Leu Asn Phe

420

<210>42

<211>421

<212>PRT

<213>艰难梭菌

<400>42

Met Tyr Asn Lys Glu Thr Ile Cys Val Gln Gly Asn Tyr Lys Pro Gly

1 5 10 15

Asn Gly Glu Pro Arg Val Leu Pro Leu Tyr Gln Ser Thr Thr Phe Lys

20 25 30

Tyr Ser Ser Ile Asp Gln Leu Ala Glu Leu Phe Asp Leu Lys Val Asp

35 40 45

Gly His Ile Tyr Ser Arg Ile Ser Asn Pro Thr Ile Gln Ala Phe Glu

50 55 60

Glu Lys Ile Ser Leu Leu Glu Gly Gly Val Ser Ser Val Ala Val Ser

65 70 75 80

Ser Gly Gln Ser Ala Asn Met Leu Ala Val Leu Asn Ile Cys Lys Ser

85 90 95

Gly Asp Ser Ile Leu Cys Ser Ser Lys Val Tyr Gly Gly Thr Phe Asn

100 105 110

Leu Leu Gly Pro Ser Leu Lys Lys Phe Gly Ile Asp Leu Ile Ser Phe

115 120 125

Asp Leu Asp Ser Ser Glu Asp Glu Ile Val Glu Leu Ala Lys Glu Asn

130 135 140

Thr Lys Val Val Phe Ala Glu Thr Leu Ala Asn Pro Thr Leu Glu Val

145 150 155 160

Ile Asp Phe Glu Lys Ile Ala Asn Val Ala Lys Arg Ile Asn Val Pro

165 170 175

Phe Ile Val Asp Asn Ser Leu Ala Ser Pro Val Leu Cys Asn Pro Leu

180 185 190

Lys Tyr Gly Ala Asn Ile Val Thr His Ser Thr Thr Lys Tyr Leu Asp

195 200 205

Gly His Ala Ser Ser Val Gly Gly Ile Ile Val Asp Gly Gly Asn Phe

210 215 220

Asn Trp Asp Asn Gly Lys Phe Pro Glu Leu Val Glu Pro Asp Pro Thr

225 230 235 240

Tyr His Gly Ile Ser Tyr Thr Gln Lys Phe Gly Asn Ala Ala Tyr Ala

245 250 255

Thr Lys Ala Arg Val Gln Leu Leu Arg Asp Tyr Gly Asn Cys Leu Ser

260 265 270

Pro Phe Asn Ala Tyr Leu Thr Asn Leu Asn Val Glu Thr Leu His Leu

275 280 285

Arg Met Glu Arg His Ser Glu Asn Ala Leu Lys Ile Ala Arg Phe Leu

290 295 300

Glu Lys His Glu Asn Val Asp Trp Ile Asn Tyr Pro Gly Leu Glu Asp

305 310 315 320

Asn Lys Tyr Tyr Glu Asn Ala Lys Lys Tyr Leu Ser Arg Gly Cys Ser

325 330 335

Gly Val Leu Ser Phe Gly Val Arg Gly Gly Leu Glu Asn Ala Lys Lys

340 345 350

Phe Val Glu Lys Leu Gln Ile Ala Ser Leu Val Thr His Val Ser Asp

355 360 365

Val Arg Thr Cys Val Ile His Pro Ala Ser Thr Thr His Arg Gln Leu

370 375 380

Thr Glu Glu Gln Leu Ile Ala Ser Gly Val Leu Pro Ser Leu Ile Arg

385 390 395 400

Leu Ser Val Gly Ile Glu Asn Val Glu Asp Leu Ile Ala Asp Leu Asn

405 410 415

Gln Ala Leu Asn Phe

420

<210>43

<211>1272

<212>DNA

<213>空肠弯曲杆菌(campylobacter jejuni)

<220>

<221>CDS

<222>(1)..(1272)

<223>

<400>43

atg aat ttc aat aaa gaa act tta gca tta cac gga gct tat aat ttt 48

Met Asn Phe Asn Lys Glu Thr Leu Ala Leu His Gly Ala Tyr Asn Phe

1 5 10 15

gat act caa aga agt att agt gtg cct ata tat caa aac act gcg tat 96

Asp Thr Gln Arg Ser Ile Ser Val Pro Ile Tyr Gln Asn Thr Ala Tyr

20 25 30

aat ttt gaa aat ttg gat caa gct gca gca agg ttt aat ctt caa gaa 144

Asn Phe Glu Asn Leu Asp Gln Ala Ala Ala Arg Phe Asn Leu Gln Glu

35 40 45

ctt ggc aat att tac tca aga ctt agc aat cct aca agc gat gtt tta 192

Leu Gly Asn Ile Tyr Ser Arg Leu Ser Asn Pro Thr Ser Asp Val Leu

50 55 60

gga caa aga ctt gct aat gtc gaa gga ggg gct ttt gga att cct gtt 240

Gly Gln Arg Leu Ala Asn Val Glu Gly Gly Ala Phe Gly Ile Pro Val

65 70 75 80

gct agc ggt atg gca gct tgt ttt tat gct ctt atc aat tta gca agt 288

Ala Ser Gly Met Ala Ala Cys Phe Tyr Ala Leu Ile Asn Leu Ala Ser

85 90 95

tcg gga gat aat gtc gcg tat tcg aac aaa att tat ggt ggg act caa 336

Ser Gly Asp Asn Val Ala Tyr Ser Asn Lys Ile Tyr Gly Gly Thr Gln

100 105 110

act tta att tct cac aca ctt aaa aat ttt ggc ata gaa gct agg gaa 384

Thr Leu Ile Ser His Thr Leu Lys Asn Phe Gly Ile Glu Ala Arg Glu

115 120 125

ttt gat atc gat gat tta gat agc ttg gaa aaa gtt ata gat caa aac 432

Phe Asp Ile Asp Asp Leu Asp Ser Leu Glu Lys Val Ile Asp Gln Asn

130 135 140

aca aaa gcg att ttt ttc gaa agt ctt tca aat cct caa att gcc ata 480

Thr Lys Ala Ile Phe Phe Glu Ser Leu Ser Asn Pro Gln Ile Ala Ile

145 150 155 160

gct gat ata gaa aaa ata aac caa ata gca aaa aaa cat aaa atc gtt 528

Ala Asp Ile Glu Lys Ile Asn Gln Ile Ala Lys Lys His Lys Ile Val

165 170 175

agc att tgt gat aat acc gtt gct act cct ttc tta ctc caa cct ttt 576

Ser Ile Cys Asp Asn Thr Val Ala Thr Pro Phe Leu Leu Gln Pro Phe

180 185 190

aaa cat ggc gtg gat gta atc gtg cat agt tta agt aaa tat gta agc 624

Lys His Gly Val Asp Val Ile Val His Ser Leu Ser Lys Tyr Val Ser

195 200 205

ggt caa ggc act gct ttg ggt gga gca ctt ata gaa aga aaa gat tta 672

Gly Gln Gly Thr Ala Leu Gly Gly Ala Leu Ile Glu Arg Lys Asp Leu

210 215 220

aac gac ttg ctt aaa aat aac gat aga tat aaa gct ttt aac act cct 720

Asn Asp Leu Leu Lys Asn Asn Asp Arg Tyr Lys Ala Phe Asn Thr Pro

225 230 235 240

gat cca agt tat cat gga ctg aat tta aat aca ctt gat ttg ccg att 768

Asp Pro Ser Tyr His Gly Leu Asn Leu Asn Thr Leu Asp Leu Pro Ile

245 250 255

ttt agt att aga gtc atc atc act tgg ctt aga gat cta gga gct agc 816

Phe Ser Ile Arg Val Ile Ile Thr Trp Leu Arg Asp Leu Gly Ala Ser

260 265 270

tta gca cct caa aat gct tgg tta ctt tta caa gga ctt gaa acc ttg 864

Leu Ala Pro Gln Asn Ala Trp Leu Leu Leu Gln Gly Leu Glu Thr Leu

275 280 285

gca gtg cgt ata gaa aaa cac agt caa aat gct gaa aaa gtt gcg aat 912

Ala Val Arg Ile Glu Lys His Ser Gln Asn Ala Glu Lys Val Ala Asn

290 295 300

ttt tta aat tct cat cct gat atc aag ggc gta aat tat cct act tta 960

Phe Leu Asn Ser His Pro Asp Ile Lys Gly Val Asn Tyr Pro Thr Leu

305 310 315 320

gca agt aat gct tat cat aat tta ttt aaa aaa tat ttt gat aaa aat 1008

Ala Ser Asn Ala Tyr His Asn Leu Phe Lys Lys Tyr Phe Asp Lys Asn

325 330 335

ttt gct agc ggg ctt tta agc ttt gaa gct aaa gat tat gag cat gct 1056

Phe Ala Ser Gly Leu Leu Ser Phe Glu Ala Lys Asp Tyr Glu His Ala

340 345 350

aga aga att tgt gat aaa act caa ctt ttc tta ctt gct gca aat ttg 1104

Arg Arg Ile Cys Asp Lys Thr Gln Leu Phe Leu Leu Ala Ala Asn Leu

355 360 365

ggt gat agc aag tct ttg atc atc cat cct gct tct act act cat tcg 1152

Gly Asp Ser Lys Ser Leu Ile Ile His Pro Ala Ser Thr Thr His Ser

370 375 380

caa cta agc gaa gaa gaa ctc caa aaa gca ggc att acg aaa gct act 1200

Gln Leu Ser Glu Glu Glu Leu Gln Lys Ala Gly Ile Thr Lys Ala Thr

385 390 395 400

ata cgc tta agc ata gga ctt gaa aat agc gat gat ttg ata gcg gat 1248

Ile Arg Leu Ser Ile Gly Leu Glu Asn Ser Asp Asp Leu Ile Ala Asp

405 410 415

tta aaa caa gct ata gaa agt taa 1272

Leu Lys Gln Ala Ile Glu Ser

420

<210>44

<211>423

<212>PRT

<213>空肠弯曲杆菌

<400>44

Met Asn Phe Asn Lys Glu Thr Leu Ala Leu His Gly Ala Tyr Asn Phe

1 5 10 15

Asp Thr Gln Arg Ser Ile Ser Val Pro Ile Tyr Gln Asn Thr Ala Tyr

20 25 30

Asn Phe Glu Asn Leu Asp Gln Ala Ala Ala Arg Phe Asn Leu Gln Glu

35 40 45

Leu Gly Asn Ile Tyr Ser Arg Leu Ser Asn Pro Thr Ser Asp Val Leu

50 55 60

Gly Gln Arg Leu Ala Asn Val Glu Gly Gly Ala Phe Gly Ile Pro Val

65 70 75 80

Ala Ser Gly Met Ala Ala Cys Phe Tyr Ala Leu Ile Asn Leu Ala Ser

85 90 95

Ser Gly Asp Asn Val Ala Tyr Ser Asn Lys Ile Tyr Gly Gly Thr Gln

100 105 110

Thr Leu Ile Ser His Thr Leu Lys Asn Phe Gly Ile Glu Ala Arg Glu

115 120 125

Phe Asp Ile Asp Asp Leu Asp Ser Leu Glu Lys Val Ile Asp Gln Asn

130 135 140

Thr Lys Ala Ile Phe Phe Glu Ser Leu Ser Asn Pro Gln Ile Ala Ile

145 150 155 160

Ala Asp Ile Glu Lys Ile Asn Gln Ile Ala Lys Lys His Lys Ile Val

165 170 175

Ser Ile Cys Asp Asn Thr Val Ala Thr Pro Phe Leu Leu Gln Pro Phe

180 185 190

Lys His Gly Val Asp Val Ile Val His Ser Leu Ser Lys Tyr Val Ser

195 200 205

Gly Gln Gly Thr Ala Leu Gly Gly Ala Leu Ile Glu Arg Lys Asp Leu

210 215 220

Asn Asp Leu Leu Lys Asn Asn Asp Arg Tyr Lys Ala Phe Asn Thr Pro

225 230 235 240

Asp Pro Ser Tyr His Gly Leu Asn Leu Asn Thr Leu Asp Leu Pro Ile

245 250 255

Phe Ser Ile Arg Val Ile Ile Thr Trp Leu Arg Asp Leu Gly Ala Ser

260 265 270

Leu Ala Pro Gln Asn Ala Trp Leu Leu Leu Gln Gly Leu Glu Thr Leu

275 280 285

Ala Val Arg Ile Glu Lys His Ser Gln Asn Ala Glu Lys Val Ala Asn

290 295 300

Phe Leu Asn Ser His Pro Asp Ile Lys Gly Val Asn Tyr Pro Thr Leu

305 310 315 320

Ala Ser Asn Ala Tyr His Asn Leu Phe Lys Lys Tyr Phe Asp Lys Asn

325 330 335

Phe Ala Ser Gly Leu Leu Ser Phe Glu Ala Lys Asp Tyr Glu His Ala

340 345 350

Arg Arg Ile Cys Asp Lys Thr Gln Leu Phe Leu Leu Ala Ala Asn Leu

355 360 365

Gly Asp Ser Lys Ser Leu Ile Ile His Pro Ala Ser Thr Thr His Ser

370 375 380

Gln Leu Ser Glu Glu Glu Leu Gln Lys Ala Gly Ile Thr Lys Ala Thr

385 390 395 400

Ile Arg Leu Ser Ile Gly Leu Glu Asn Ser Asp Asp Leu Ile Ala Asp

405 410 415

Leu Lys Gln Ala Ile Glu Ser

420

<210>45

<211>1041

<212>DNA

<213>肺炎链球菌(Streptococcus pneumoniae)

<220>

<221>CDS

<222>(1)..(1041)

<223>

<400>45

ttg agg aaa cca ggg aac att tat act cgt atc acc aat cct aca aca 48

Leu Arg Lys Pro Gly Asn Ile Tyr Thr Arg Ile Thr Asn Pro Thr Thr

1 5 10 15

gct gcc ctt gaa ggt ggt gtt gaa gcg cta gca aca gca tca ggt atg 96

Ala Ala Leu Glu Gly Gly Val Glu Ala Leu Ala Thr Ala Ser Gly Met

20 25 30

act gca gtg act tat acg att ttg gcg att gcc cat gct ggt gac cat 144

Thr Ala Val Thr Tyr Thr Ile Leu Ala Ile Ala His Ala Gly Asp His

35 40 45

gta gtg gct gct tcg act att tac ggt gga acc ttc aat ctt ttg aaa 192

Val Val Ala Ala Ser Thr Ile Tyr Gly Gly Thr Phe Asn Leu Leu Lys

50 55 60

gaa ccc ctt cct cgt tat ggt atc aca aca acc ttt ttc gat att gat 240

Glu Pro Leu Pro Arg Tyr Gly Ile Thr Thr Thr Phe Phe Asp Ile Asp

65 70 75 80

aat ttg gag gaa gta gaa gca gct atc aaa gac aat acc aag ctt gtc 288

Asn Leu Glu Glu Val Glu Ala Ala Ile Lys Asp Asn Thr Lys Leu Val

85 90 95

ttg att gaa acc ttg ggt aac ccc ttg att aat att cca gac ctg gaa 336

Leu Ile Glu Thr Leu Gly Asn Pro Leu Ile Asn Ile Pro Asp Leu Glu

100 105 110

aaa ctg gca gag att gct cat aaa cat caa atc cca ctt gtg tca gac 384

Lys Leu Ala Glu Ile Ala His Lys His Gln Ile Pro Leu Val Ser Asp

115 120 125

aat act ttt gca aca cct tat ttg att aac gtc ttc tct cat ggc gtt 432

Asn Thr Phe Ala Thr Pro Tyr Leu Ile Asn Val Phe Ser His Gly Val

130 135 140

gac att gcc att cac tct gtg act aag ttt atc ggt ggg cat ggt aca 480

Asp Ile Ala Ile His Ser Val Thr Lys Phe Ile Gly Gly His Gly Thr

145 150 155 160

act att gga gga ata att gtc gat agt ggt cgt ttt gac tgg acg gct 528

Thr Ile Gly Gly Ile Ile Val Asp Ser Gly Arg Phe Asp Trp Thr Ala

165 170 175

tca ggg aaa ttc cct caa ttt gtt gac gag ggt cca agc tgc cac aat 576

Ser Gly Lys Phe Pro Gln Phe Val Asp Glu Gly Pro Ser Cys His Asn

180 185 190

ttg agc tat act cgt gat gtg ggt gca gca gcc ttt att ata gct gtt 624

Leu Ser Tyr Thr Arg Asp Val Gly Ala Ala Ala Phe Ile Ile Ala Val

195 200 205

cga gtt caa ttg ctt cgt gat aca ggt gca gcc ttg tca cca ttc aat 672

Arg Val Gln Leu Leu Arg Asp Thr Gly Ala Ala Leu Ser Pro Phe Asn

210 215 220

gct ttc ctc ttg cta caa aga ctt gaa acc tct tca ctt cgt gtg gaa 720

Ala Phe Leu Leu Leu Gln Arg Leu Glu Thr Ser Ser Leu Arg Val Glu

225 230 235 240

cgc cat gta caa aat gct gag aca att gtt gat ttt ctt gtc aac cat 768

Arg His Val Gln Asn Ala Glu Thr Ile Val Asp Phe Leu Val Asn His

245 250 255

cct aag gta gaa aag gta aat tat cca aaa ctt gca gat agt cct tat 816

Pro Lys Val Glu Lys Val Asn Tyr Pro Lys Leu Ala Asp Ser Pro Tyr

260 265 270

cat gcc ttg gct gag aaa tac ttg cca aaa ggt gtc ggt tca atc ttt 864

His Ala Leu Ala Glu Lys Tyr Leu Pro Lys Gly Val Gly Ser Ile Phe

275 280 285

acc ttc cac gtc aaa ggt ggc gag gaa gaa gca cgc aag gtc att gat 912

Thr Phe His Val Lys Gly Gly Glu Glu Glu Ala Arg Lys Val Ile Asp

290 295 300

aat tta gaa atc ttt tct gac ctt gca aac gcg gca gat gct aaa tcg 960

Asn Leu Glu Ile Phe Ser Asp Leu Ala Asn Ala Ala Asp Ala Lys Ser

305 310 315 320

ctt gtt gtc cat cca gca aca acc act cac ggt caa ttg tca gaa aaa 1008

Leu Val Val His Pro Ala Thr Thr Thr His Gly Gln Leu Ser Glu Lys

325 330 335

gac cta gaa gca gca ggt gtc aca cca aac taa 1041

Asp Leu Glu Ala Ala Gly Val Thr Pro Asn

340 345

<210>46

<211>346

<212>PRT

<213>肺炎链球菌

<400>46

Leu Arg Lys Pro Gly Asn Ile Tyr Thr Arg Ile Thr Asn Pro Thr Thr

1 5 10 15

Ala Ala Leu Glu Gly Gly Val Glu Ala Leu Ala Thr Ala Ser Gly Met

20 25 30

Thr Ala Val Thr Tyr Thr Ile Leu Ala Ile Ala His Ala Gly Asp His

35 40 45

Val Val Ala Ala Ser Thr Ile Tyr Gly Gly Thr Phe Asn Leu Leu Lys

50 55 60

Glu Pro Leu Pro Arg Tyr Gly Ile Thr Thr Thr Phe Phe Asp Ile Asp

65 70 75 80

Asn Leu Glu Glu Val Glu Ala Ala Ile Lys Asp Asn Thr Lys Leu Val

85 90 95

Leu Ile Glu Thr Leu Gly Asn Pro Leu Ile Asn Ile Pro Asp Leu Glu

100 105 110

Lys Leu Ala Glu Ile Ala His Lys His Gln Ile Pro Leu Val Ser Asp

115 120 125

Asn Thr Phe Ala Thr Pro Tyr Leu Ile Asn Val Phe Ser His Gly Val

130 135 140

Asp Ile Ala Ile His Ser Val Thr Lys Phe Ile Gly Gly His Gly Thr

145 150 155 160

Thr Ile Gly Gly Ile Ile Val Asp Ser Gly Arg Phe Asp Trp Thr Ala

165 170 175

Ser Gly Lys Phe Pro Gln Phe Val Asp Glu Gly Pro Ser Cys His Asn

180 185 190

Leu Ser Tyr Thr Arg Asp Val Gly Ala Ala Ala Phe Ile Ile Ala Val

195 200 205

Arg Val Gln Leu Leu Arg Asp Thr Gly Ala Ala Leu Ser Pro Phe Asn

210 215 220

Ala Phe Leu Leu Leu Gln Arg Leu Glu Thr Ser Ser Leu Arg Val Glu

225 230 235 240

Arg His Val Gln Asn Ala Glu Thr Ile Val Asp Phe Leu Val Asn His

245 250 255

Pro Lys Val Glu Lys Val Asn Tyr Pro Lys Leu Ala Asp Ser Pro Tyr

260 265 270

His Ala Leu Ala Glu Lys Tyr Leu Pro Lys Gly Val Gly Ser Ile Phe

275 280 285

Thr Phe His Val Lys Gly Gly Glu Glu Glu Ala Arg Lys Val Ile Asp

290 295 300

Asn Leu Glu Ile Phe Ser Asp Leu Ala Asn Ala Ala Asp Ala Lys Ser

305 310 315 320

Leu Val Val His Pro Ala Thr Thr Thr His Gly Gln Leu Ser Glu Lys

325 330 335

Asp Leu Glu Ala Ala Gly Val Thr Pro Asn

340 345

<210>47

<211>1335

<212>DNA

<213>酿酒酵母(Saccharomyces cerevisiae)

<220>

<221>CDS

<222>(1)..(1335)

<223>

<400>47

atg cca tct cat ttc gat act gtt caa cta cac gcc ggc caa gag aac 48

Met Pro Ser His Phe Asp Thr Val Gln Leu His Ala Gly Gln Glu Asn

1 5 10 15

cct ggt gac aat gct cac aga tcc aga gct gta cca att tac gcc acc 96

Pro Gly Asp Asn Ala His Arg Ser Arg Ala Val Pro Ile Tyr Ala Thr

20 25 30

act tct tat gtt ttc gaa aac tct aag cat ggt tcg caa ttg ttt ggt 144

Thr Ser Tyr Val Phe Glu Asn Ser Lys His Gly Ser Gln Leu Phe Gly

35 40 45

cta gaa gtt cca ggt tac gtc tat tcc cgt ttc caa aac cca acc agt 192

Leu Glu Val Pro Gly Tyr Val Tyr Ser Arg Phe Gln Asn Pro Thr Ser

50 55 60

aat gtt ttg gaa gaa aga att gct gct tta gaa ggt ggt gct gct gct 240

Asn Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Gly Gly Ala Ala Ala

65 70 75 80

ttg gct gtt tcc tcc ggt caa gcc gct caa acc ctt gcc atc caa ggt 288

Leu Ala Val Ser Ser Gly Gln Ala Ala Gln Thr Leu Ala Ile Gln Gly

85 90 95

ttg gca cac act ggt gac aac atc gtt tcc act tct tac tta tac ggt 336

Leu Ala His Thr Gly Asp Asn Ile Val Ser Thr Ser Tyr Leu Tyr Gly

100 105 110

ggt act tat aac cag ttc aaa atc tcg ttc aaa aga ttt ggt atc gag 384

Gly Thr Tyr Asn Gln Phe Lys Ile Ser Phe Lys Arg Phe Gly Ile Glu

115 120 125

gct aga ttt gtt gaa ggt gac aat cca gaa gaa ttc gaa aag gtc ttt 432

Ala Arg Phe Val Glu Gly Asp Asn Pro Glu Glu Phe Glu Lys Val Phe

130 135 140

gat gaa aga acc aag gct gtt tat ttg gaa acc att ggt aat cca aag 480

Asp Glu Arg Thr Lys Ala Val Tyr Leu Glu Thr Ile Gly Asn Pro Lys

145 150 155 160

tac aat gtt ccg gat ttt gaa aaa att gtt gca att gct cac aaa cac 528

Tyr Asn Val Pro Asp Phe Glu Lys Ile Val Ala Ile Ala His Lys His

165 170 175

ggt att cca gtt gtc gtt gac aac aca ttt ggt gcc ggt ggt tac ttc 576

Gly Ile Pro Val Val Val Asp Asn Thr Phe Gly Ala Gly Gly Tyr Phe

180 185 190

tgt cag cca att aaa tac ggt gct gat att gta aca cat tct gct acc 624

Cys Gln Pro Ile Lys Tyr Gly Ala Asp Ile Val Thr His Ser Ala Thr

195 200 205

aaa tgg att ggt ggt cat ggt act act atc ggt ggt att att gtt gac 672

Lys Trp Ile Gly Gly His Gly Thr Thr Ile Gly Gly Ile Ile Val Asp

210 215 220

tct ggt aag ttc cca tgg aag gac tac cca gaa aag ttc cct caa ttc 720

Ser Gly Lys Phe Pro Trp Lys Asp Tyr Pro Glu Lys Phe Pro Gln Phe

225 230 235 240

tct caa cct gcc gaa gga tat cac ggt act atc tac aat gaa gcc tac 768

Ser Gln Pro Ala Glu Gly Tyr His Gly Thr Ile Tyr Asn Glu Ala Tyr

245 250 255

ggt aac ttg gca tac atc gtt cat gtt aga act gaa cta tta aga gat 816

Gly Asn Leu Ala Tyr Ile Val His Val Arg Thr Glu Leu Leu Arg Asp

260 265 270

ttg ggt cca ttg atg aac cca ttt gcc tct ttc ttg cta cta caa ggt 864

Leu Gly Pro Leu Met Asn Pro Phe Ala Ser Phe Leu Leu Leu Gln Gly

275 280 285

gtt gaa aca tta tct ttg aga gct gaa aga cac ggt gaa aat gca ttg 912

Val Glu Thr Leu Ser Leu Arg Ala Glu Arg His Gly Glu Asn Ala Leu

290 295 300

aag tta gcc aaa tgg tta gaa caa tcc cca tac gta tct tgg gtt tca 960

Lys Leu Ala Lys Trp Leu Glu Gln Ser Pro Tyr Val Ser Trp Val Ser

305 310 315 320

tac cct ggt tta gca tct cat tct cat cat gaa aat gct aag aag tat 1008

Tyr Pro Gly Leu Ala Ser His Ser His His Glu Asn Ala Lys Lys Tyr

325 330 335

cta tct aac ggt ttc ggt ggt gtc tta tct ttc ggt gta aaa gac tta 1056

Leu Ser Asn Gly Phe Gly Gly Val Leu Ser Phe Gly Val Lys Asp Leu

340 345 350

cca aat gcc gac aag gaa act gac cca ttc aaa ctt tct ggt gct caa 1104

Pro Asn Ala Asp Lys Glu Thr Asp Pro Phe Lys Leu Ser Gly Ala Gln

355 360 365

gtt gtt gac aat tta aag ctt gcc tct aac ttg gcc aat gtt ggt gat 1152

Val Val Asp Asn Leu Lys Leu Ala Ser Asn Leu Ala Asn Val Gly Asp

370 375 380

gcc aag acc tta gtc att gct cca tac ttc act acc cac aaa caa tta 1200

Ala Lys Thr Leu Val Ile Ala Pro Tyr Phe Thr Thr His Lys Gln Leu

385 390 395 400

aat gac aaa gaa aag ttg gca tct ggt gtt acc aag gac tta att cgt 1248

Asn Asp Lys Glu Lys Leu Ala Ser Gly Val Thr Lys Asp Leu Ile Arg

405 410 415

gtc tct gtt ggt atc gaa ttt att gat gac att att gca gac ttc cag 1296

Val Ser Val Gly Ile Glu Phe Ile Asp Asp Ile Ile Ala Asp Phe Gln

420 425 430

caa tct ttt gaa act gtt ttc gct ggc caa aaa cca tga 1335

Gln Ser Phe Glu Thr Val Phe Ala Gly Gln Lys Pro

435 440

<210>48

<211>444

<212>PRT

<213>酿酒酵母

<400>48

Met Pro Ser His Phe Asp Thr Val Gln Leu His Ala Gly Gln Glu Asn

1 5 10 15

Pro Gly Asp Asn Ala His Arg Ser Arg Ala Val Pro Ile Tyr Ala Thr

20 25 30

Thr Ser Tyr Val Phe Glu Asn Ser Lys His Gly Ser Gln Leu Phe Gly

35 40 45

Leu Glu Val Pro Gly Tyr Val Tyr Ser Arg Phe Gln Asn Pro Thr Ser

50 55 60

Asn Val Leu Glu Glu Arg Ile Ala Ala Leu Glu Gly Gly Ala Ala Ala

65 70 75 80

Leu Ala Val Ser Ser Gly Gln Ala Ala Gln Thr Leu Ala Ile Gln Gly

85 90 95

Leu Ala His Thr Gly Asp Asn Ile Val Ser Thr Ser Tyr Leu Tyr Gly

100 105 110

Gly Thr Tyr Asn Gln Phe Lys Ile Ser Phe Lys Arg Phe Gly Ile Glu

115 120 125

Ala Arg Phe Val Glu Gly Asp Asn Pro Glu Glu Phe Glu Lys Val Phe

130 135 140

Asp Glu Arg Thr Lys Ala Val Tyr Leu Glu Thr Ile Gly Asn Pro Lys

145 150 155 160

Tyr Asn Val Pro Asp Phe Glu Lys Ile Val Ala Ile Ala His Lys His

165 170 175

Gly Ile Pro Val Val Val Asp Asn Thr Phe Gly Ala Gly Gly Tyr Phe

180 185 190

Cys Gln Pro Ile Lys Tyr Gly Ala Asp Ile Val Thr His Ser Ala Thr

195 200 205

Lys Trp Ile Gly Gly His Gly Thr Thr Ile Gly Gly Ile Ile Val Asp

210 215 220

Ser Gly Lys Phe Pro Trp Lys Asp Tyr Pro Glu Lys Phe Pro Gln Phe

225 230 235 240

Ser Gln Pro Ala Glu Gly Tyr His Gly Thr Ile Tyr Asn Glu Ala Tyr

245 250 255

Gly Asn Leu Ala Tyr Ile Val His Val Arg Thr Glu Leu Leu Arg Asp

260 265 270

Leu Gly Pro Leu Met Asn Pro Phe Ala Ser Phe Leu Leu Leu Gln Gly

275 280 285

Val Glu Thr Leu Ser Leu Arg Ala Glu Arg His Gly Glu Asn Ala Leu

290 295 300

Lys Leu Ala Lys Trp Leu Glu Gln Ser Pro Tyr Val Ser Trp Val Ser

305 310 315 320

Tyr Pro Gly Leu Ala Ser His Ser His His Glu Asn Ala Lys Lys Tyr

325 330 335

Leu Ser Asn Gly Phe Gly Gly Val Leu Ser Phe Gly Val Lys Asp Leu

340 345 350

Pro Asn Ala Asp Lys Glu Thr Asp Pro Phe Lys Leu Ser Gly Ala Gln

355 360 365

Val Val Asp Asn Leu Lys Leu Ala Ser Asn Leu Ala Asn Val Gly Asp

370 375 380

Ala Lys Thr Leu Val Ile Ala Pro Tyr Phe Thr Thr His Lys Gln Leu

385 390 395 400

Asn Asp Lys Glu Lys Leu Ala Ser Gly Val Thr Lys Asp Leu Ile Arg

405 410 415

Val Ser Val Gly Ile Glu Phe Ile Asp Asp Ile Ile Ala Asp Phe Gln

420 425 430

Gln Ser Phe Glu Thr Val Phe Ala Gly Gln Lys Pro

435 440

<210>49

<211>1335

<212>DNA

<213>乳酸克鲁维酵母(Kluyveromyces lactis)

<220>

<221>CDS

<222>(1)..(1335)

<223>

<400>49

atg cca tct cac ttc gat act ttg caa ttg cac gct ggt caa gaa aag 48

Met Pro Ser His Phe Asp Thr Leu Gln Leu His Ala Gly Gln Glu Lys

1 5 10 15

act gct gat gct cat aac cca aga gcc gtc cca att tac gct acc act 96

Thr Ala Asp Ala His Asn Pro Arg Ala Val Pro Ile Tyr Ala Thr Thr

20 25 30

tct tac gtc ttc aac gac tct aag cat ggt gct caa ttg ttc ggt tta 144

Ser Tyr Val Phe Asn Asp Ser Lys His Gly Ala Gln Leu Phe Gly Leu

35 40 45

gaa act cca ggt tac att tac tct cgt att atg aac cct act cta gac 192

Glu Thr Pro Gly Tyr Ile Tyr Ser Arg Ile Met Asn Pro Thr Leu Asp

50 55 60

gtc ttg gaa aag aga ttg gca gcc tta gaa ggt ggt att gct gct ttg 240

Val Leu Glu Lys Arg Leu Ala Ala Leu Glu Gly Gly Ile Ala Ala Leu

65 70 75 80

gct act tct tct ggc caa gct gct caa acc ttg gct gtc act ggt ttg 288

Ala Thr Ser Ser Gly Gln Ala Ala Gln Thr Leu Ala Val Thr Gly Leu

85 90 95

gcc cac act ggt gac aat att gtc tct acc tct ttc tta tac ggt ggt 336

Ala His Thr Gly Asp Asn Ile Val Ser Thr Ser Phe Leu Tyr Gly Gly

100 105 110

act tat aac caa ttc aag gtt gcc ttc aag aga tta gga att gaa gct 384

Thr Tyr Asn Gln Phe Lys Val Ala Phe Lys Arg Leu Gly Ile Glu Ala

115 120 125

aga ttt gtc gat ggt gac aag cca gaa gac ttc gaa aag ttg ttc gat 432

Arg Phe Val Asp Gly Asp Lys Pro Glu Asp Phe Glu Lys Leu Phe Asp

130 135 140

gaa aag act aag gct ctc tat ctg gaa tct atc ggt aat cct aag tac 480

Glu Lys Thr Lys Ala Leu Tyr Leu Glu Ser Ile Gly Asn Pro Lys Tyr

145 150 155 160

aat gtc cca gac ttc gaa aag att gtt gct gtt gct cat aag cat ggt 528

Asn Val Pro Asp Phe Glu Lys Ile Val Ala Val Ala His Lys His Gly

165 170 175

atc cca gtt gtt gtt gac aac act ttc ggt gcc ggt ggt ttc ttc tgc 576

Ile Pro Val Val Val Asp Asn Thr Phe Gly Ala Gly Gly Phe Phe Cys

180 185 190

caa cct atc aaa tac ggt gct gat atc gtt act cac tct gct acc aag 624

Gln Pro Ile Lys Tyr Gly Ala Asp Ile Val Thr His Ser Ala Thr Lys

195 200 205

tgg atc ggt ggt cat ggt gtc acc gtt ggt ggt gtc atc att gac tct 672

Trp Ile Gly Gly His Gly Val Thr Val Gly Gly Val Ile Ile Asp Ser

210 215 220

ggt aag ttc cca tgg aag gat tac ccg gaa aag ttc cct caa ttc tct 720

Gly Lys Phe Pro Trp Lys Asp Tyr Pro Glu Lys Phe Pro Gln Phe Ser

225 230 235 240

cag cca tct gaa ggt tat cat ggt ttg atc ttc aat gat gcc ttt ggt 768

Gln Pro Ser Glu Gly Tyr His Gly Leu Ile Phe Asn Asp Ala Phe Gly

245 250 255

cca gct gct ttc att ggt cat gta aga acc gaa ttg cta aga gat tta 816

Pro Ala Ala Phe Ile Gly His Val Arg Thr Glu Leu Leu Arg Asp Leu

260 265 270

ggt cca gtg ttg agt cca ttc gct ggt ttc ttg ttg tta cag ggt ctt 864

Gly Pro Val Leu Ser Pro Phe Ala Gly Phe Leu Leu Leu Gln Gly Leu

275 280 285

gaa act ttg tct cta aga ggt gaa aga cac ggt tcc aac gct ttg aag 912

Glu Thr Leu Ser Leu Arg Gly Glu Arg His Gly Ser Asn Ala Leu Lys

290 295 300

ttg gct caa tac ttg gaa agt tct cca tac gtt tca tgg gtc tct tac 960

Leu Ala Gln Tyr Leu Glu Ser Ser Pro Tyr Val Ser Trp Val Ser Tyr

305 310 315 320

cca ggt ttg cca tct cac tct cac cac gaa aac gct aag aaa tac ttg 1008

Pro Gly Leu Pro Ser His Ser His His Glu Asn Ala Lys Lys Tyr Leu

325 330 335

gaa aat ggt ttc ggt ggt gtt tta tcc ttc ggt gtc aaa gat ttg cct 1056

Glu Asn Gly Phe Gly Gly Val Leu Ser Phe Gly Val Lys Asp Leu Pro

340 345 350

aac gct tcc gag gaa tct gat cca ttc aag gct tct ggt gcc caa gtt 1104

Asn Ala Ser Glu Glu Ser Asp Pro Phe Lys Ala Ser Gly Ala Gln Val

355 360 365

gtt gac aac ttg aag ctg gct tct aac ttg gca aac gtt ggt gac tcc 1152

Val Asp Asn Leu Lys Leu Ala Ser Asn Leu Ala Asn Val Gly Asp Ser

370 375 380

aag acc ttg gtc att gct cca tac ttc act aca cat caa caa ttg acc 1200

Lys Thr Leu Val Ile Ala Pro Tyr Phe Thr Thr His Gln Gln Leu Thr

385 390 395 400

gac gaa gaa aag tta gct tct ggt gtt acc aag gac ttg atc cgt gtt 1248

Asp Glu Glu Lys Leu Ala Ser Gly Val Thr Lys Asp Leu Ile Arg Val

405 410 415

tct gtt ggt act gaa ttc att gac gac att att gct gac ttt gaa gca 1296

Ser Val Gly Thr Glu Phe Ile Asp Asp Ile Ile Ala Asp Phe Glu Ala

420 425 430

tct ttc gct act gtc ttc aat ggc caa aaa cct gaa taa 1335

Ser Phe Ala Thr Val Phe Asn Gly Gln Lys Pro Glu

435 440

<210>50

<211>444

<212>PRT

<213>乳酸克鲁维酵母

<400>50

Met Pro Ser His Phe Asp Thr Leu Gln Leu His Ala Gly Gln Glu Lys

1 5 10 15

Thr Ala Asp Ala His Asn Pro Arg Ala Val Pro Ile Tyr Ala Thr Thr

20 25 30

Ser Tyr Val Phe Asn Asp Ser Lys His Gly Ala Gln Leu Phe Gly Leu

35 40 45

Glu Thr Pro Gly Tyr Ile Tyr Ser Arg Ile Met Asn Pro Thr Leu Asp

50 55 60

Val Leu Glu Lys Arg Leu Ala Ala Leu Glu Gly Gly Ile Ala Ala Leu

65 70 75 80

Ala Thr Ser Ser Gly Gln Ala Ala Gln Thr Leu Ala Val Thr Gly Leu

85 90 95

Ala His Thr Gly Asp Asn Ile Val Ser Thr Ser Phe Leu Tyr Gly Gly

100 105 110

Thr Tyr Asn Gln Phe Lys Val Ala Phe Lys Arg Leu Gly Ile Glu Ala

115 120 125

Arg Phe Val Asp Gly Asp Lys Pro Glu Asp Phe Glu Lys Leu Phe Asp

130 135 140

Glu Lys Thr Lys Ala Leu Tyr Leu Glu Ser Ile Gly Asn Pro Lys Tyr

145 150 155 160

Asn Val Pro Asp Phe Glu Lys Ile Val Ala Val Ala His Lys His Gly

165 170 175

Ile Pro Val Val Val Asp Asn Thr Phe Gly Ala Gly Gly Phe Phe Cys

180 185 190

Gln Pro Ile Lys Tyr Gly Ala Asp Ile Val Thr His Ser Ala Thr Lys

195 200 205

Trp Ile Gly Gly His Gly Val Thr Val Gly Gly Val Ile Ile Asp Ser

210 215 220

Gly Lys Phe Pro Trp Lys Asp Tyr Pro Glu Lys Phe Pro Gln Phe Ser

225 230 235 240

Gln Pro Ser Glu Gly Tyr His Gly Leu Ile Phe Asn Asp Ala Phe Gly

245 250 255

Pro Ala Ala Phe Ile Gly His Val Arg Thr Glu Leu Leu Arg Asp Leu

260 265 270

Gly Pro Val Leu Ser Pro Phe Ala Gly Phe Leu Leu Leu Gln Gly Leu

275 280 285

Glu Thr Leu Ser Leu Arg Gly Glu Arg His Gly Ser Asn Ala Leu Lys

290 295 300

Leu Ala Gln Tyr Leu Glu Ser Ser Pro Tyr Val Ser Trp Val Ser Tyr

305 310 315 320

Pro Gly Leu Pro Ser His Ser His His Glu Asn Ala Lys Lys Tyr Leu

325 330 335

Glu Asn Gly Phe Gly Gly Val Leu Ser Phe Gly Val Lys Asp Leu Pro

340 345 350

Asn Ala Ser Glu Glu Ser Asp Pro Phe Lys Ala Ser Gly Ala Gln Val

355 360 365

Val Asp Asn Leu Lys Leu Ala Ser Asn Leu Ala Asn Val Gly Asp Ser

370 375 380

Lys Thr Leu Val Ile Ala Pro Tyr Phe Thr Thr His Gln Gln Leu Thr

385 390 395 400

Asp Glu Glu Lys Leu Ala Ser Gly Val Thr Lys Asp Leu Ile Arg Val

405 410 415

Ser Val Gly Thr Glu Phe Ile Asp Asp Ile Ile Ala Asp Phe Glu Ala

420 425 430

Ser Phe Ala Thr Val Phe Asn Gly Gln Lys Pro Glu

435 440

<210>51

<211>1323

<212>DNA

<213>白假丝酵母(Candida albicans)

<220>

<221>CDS

<222>(L)..(1323)

<223>

<400>51

atg cct tct cac ttt gat aca ctt caa tta cat gct ggt caa cca gtt 48

Met Pro Ser His Phe Asp Thr Leu Gln Leu His Ala Gly Gln Pro Val

1 5 10 15

gaa aaa cca cac caa cca aga gcc cca cca att tat gca acc acc tcc 96

Glu Lys Pro His Gln Pro Arg Ala Pro Pro Ile Tyr Ala Thr Thr Ser

20 25 30

tat gtt ttc aat gac tct aaa cac ggt gct caa tta ttt ggt tta gaa 144

Tyr Val Phe Asn Asp Ser Lys His Gly Ala Gln Leu Phe Gly Leu Glu

35 40 45

acc cca gga tac att tac tcc aga att atg aat cca aca aac gat gtg 192

Thr Pro Gly Tyr Ile Tyr Ser Arg Ile Met Asn Pro Thr Asn Asp Val

50 55 60

ttt gaa caa aga att gct gcc ttg gaa ggt ggt att ggt gca ttg gcc 240

Phe Glu Gln Arg Ile Ala Ala Leu Glu Gly Gly Ile Gly Ala Leu Ala

65 70 75 80

act tct tct ggt caa tca gct caa ttc ttg gcc att gct ggg ttg gct 288

Thr Ser Ser Gly Gln Ser Ala Gln Phe Leu Ala Ile Ala Gly Leu Ala

85 90 95

cat gct ggt gat aac att atc agt aca tcc tac ttg tat ggt ggt act 336

His Ala Gly Asp Asn Ile Ile Ser Thr Ser Tyr Leu Tyr Gly Gly Thr

100 105 110

tat aat caa ttc aaa gtt gct ttc aaa cgt ttg ggc att gaa acc aaa 384

Tyr Asn Gln Phe Lys Val Ala Phe Lys Arg Leu Gly Ile Glu Thr Lys

115 120 125

ttc gtt aat ggt gac gcc gct gaa gat ttt gct aaa ttg att gac gac 432

Phe Val Asn Gly Asp Ala Ala Glu Asp Phe Ala Lys Leu Ile Asp Asp

130 135 140

aag aca aaa gct att tat att gaa acc att gga aac cct aaa tat aat 480

Lys Thr Lys Ala Ile Tyr Ile Glu Thr Ile Gly Asn Pro Lys Tyr Asn

145 150 155 160

gtt ccg gac ttt gaa aaa atc acc aaa ttg gcc cat gaa cac ggt att 528

Val Pro Asp Phe Glu Lys Ile Thr Lys Leu Ala His Glu His Gly Ile

165 170 175

cct gtt gtt gtc gac aac act ttt ggt gct ggt gga ttt tta gtt aac 576

Pro Val Val Val Asp Asn Thr Phe Gly Ala Gly Gly Phe Leu Val Asn

180 185 190

cca att gcc cac ggt gct gat att gtt gtt cat tct gct act aaa tgg 624

Pro Ile Ala His Gly Ala Asp Ile Val Val His Ser Ala Thr Lys Trp

195 200 205

att ggt ggt cac ggt act aca att gct ggt gtt att gtt gat tcc ggt 672

Ile Gly Gly His Gly Thr Thr Ile Ala Gly Val Ile Val Asp Ser Gly

210 215 220

aac ttc cca tgg acc gag tac cca gaa aaa tac cca caa ttc tct aaa 720

Asn Phe Pro Trp Thr Glu Tyr Pro Glu Lys Tyr Pro Gln Phe Ser Lys

225 230 235 240

cca tca gaa ggt tac cac ggg ttg atc ttg aat gat gct tta ggt aag 768

Pro Ser Glu Gly Tyr His Gly Leu Ile Leu Asn Asp Ala Leu Gly Lys

245 250 255

gcc gca tac att ggt cac ttg aga att gaa ttg ttg aga gac ttg ggt 816

Ala Ala Tyr Ile Gly His Leu Arg Ile Glu Leu Leu Arg Asp Leu Gly

260 265 270

cca gct ttg aat cca ttt gga agt ttt ttg ttg ttg caa ggt tta gaa 864

Pro Ala Leu Asn Pro Phe Gly Ser Phe Leu Leu Leu Gln Gly Leu Glu

275 280 285

act ttg tct ttg aga gtt gaa aga caa tct gaa aat gct ttg aaa ttg 912

Thr Leu Ser Leu Arg Val Glu Arg Gln Ser Glu Asn Ala Leu Lys Leu

290 295 300

gcc caa tgg ttg gaa aag aac cca aat gtt gag tct gtg tcc tat ttg 960

Ala Gln Trp Leu Glu Lys Asn Pro Asn Val Glu Ser Val Ser Tyr Leu

305 310 315 320

gga ttg cca tct cac gaa tcc cac gaa ttg agt aaa aaa tac ttg aac 1008

Gly Leu Pro Ser His Glu Ser His Glu Leu Ser Lys Lys Tyr Leu Asn

325 330 335

aat gac gct aag tac ttt ggt ggt gct tta gca ttt act gtc aag gac 1056

Asn Asp Ala Lys Tyr Phe Gly Gly Ala Leu Ala Phe Thr Val Lys Asp

340 345 350

atc acc aac acc tcc agc gac cca ttc aat gaa gcc tca cca aag ttg 1104

Ile Thr Asn Thr Ser Ser Asp Pro Phe Asn Glu Ala Ser Pro Lys Leu

355 360 365

gtt gac aat ttg gag att gct tca aac ttg gct aat gtg ggt gac tct 1152

Val Asp Asn Leu Glu Ile Ala Ser Asm Leu Ala Asn Val Gly Asp Ser

370 375 380

aag act ttg gtt att gct cca tgg ttt act aca cat caa caa ttg tct 1200

Lys Thr Leu Val Ile Ala Pro Trp Phe Thr Thr His Gln Gln Leu Ser

385 390 395 400

gat gaa gaa aag ttg gct tct ggt gtt acc aag ggc tta atc aga gtt 1248

Asp Glu Glu Lys Leu Ala Ser Gly Val Thr Lys Gly Leu Ile Arg Val

405 410 415

tct act ggt act gaa tat att gat gat att att aac gac ttt gaa caa 1296

Ser Thr Gly Thr Glu Tyr Ile Asp Asp Ile Ile Asn Asp Phe Glu Gln

420 425 430

gca ttc aag aag gtt tat aac aac taa 1323

Ala Phe Lys Lys Val Tyr Asn Asn

435 440

<210>52

<211>440

<212>PRT

<213>白假丝酵母

<400>52

Met Pro Ser His Phe Asp Thr Leu Gln Leu His Ala Gly Gln Pro Val

1 5 10 15

Glu Lys Pro His Gln Pro Arg Ala Pro Pro Ile Tyr Ala Thr Thr Ser

20 25 30

Tyr Val Phe Asn Asp Ser Lys His Gly Ala Gln Leu Phe Gly Leu Glu

35 40 45

Thr Pro Gly Tyr Ile Tyr Ser Arg Ile Met Asn Pro Thr Asn Asp Val

50 55 60

Phe Glu Gln Arg Ile Ala Ala Leu Glu Gly Gly Ile Gly Ala Leu Ala

65 70 75 80

Thr Ser Ser Gly Gln Ser Ala Gln Phe Leu Ala Ile Ala Gly Leu Ala

85 90 95

His Ala Gly Asp Asn Ile Ile Ser Thr Ser Tyr Leu Tyr Gly Gly Thr

100 105 110

Tyr Asn Gln Phe Lys Val Ala Phe Lys Arg Leu Gly Ile Glu Thr Lys

115 120 125

Phe Val Asn Gly Asp Ala Ala Glu Asp Phe Ala Lys Leu Ile Asp Asp

130 135 140

Lys Thr Lys Ala Ile Tyr Ile Glu Thr Ile Gly Asn Pro Lys Tyr Asn

145 150 155 160

Val Pro Asp Phe Glu Lys Ile Thr Lys Leu Ala His Glu His Gly Ile

165 170 175

Pro Val Val Val Asp Asn Thr Phe Gly Ala Gly Gly Phe Leu Val Asn

180 185 190

Pro Ile Ala His Gly Ala Asp Ile Val Val His Ser Ala Thr Lys Trp

195 200 205

Ile Gly Gly His Gly Thr Thr Ile Ala Gly Val Ile Val Asp Ser Gly

210 215 220

Asn Phe Pro Trp Thr Glu Tyr Pro Glu Lys Tyr Pro Gln Phe Ser Lys

225 230 235 240

Pro Ser Glu Gly Tyr His Gly Leu Ile Leu Asn Asp Ala Leu Gly Lys

245 250 255

Ala Ala Tyr Ile Gly His Leu Arg Ile Glu Leu Leu Arg Asp Leu Gly

260 265 270

Pro Ala Leu Asn Pro Phe Gly Ser Phe Leu Leu Leu Gln Gly Leu Glu

275 280 285

Thr Leu Ser Leu Arg Val Glu Arg Gln Ser Glu Asn Ala Leu Lys Leu

290 295 300

Ala Gln Trp Leu Glu Lys Asn Pro Asn Val Glu Ser Val Ser Tyr Leu

305 310 315 320

Gly Leu Pro Ser His Glu Ser His Glu Leu Ser Lys Lys Tyr Leu Asn

325 330 335

Asn Asp Ala Lys Tyr Phe Gly Gly Ala Leu Ala Phe Thr Val Lys Asp

340 345 350

Ile Thr Asn Thr Ser Ser Asp Pro Phe Asn Glu Ala Ser Pro Lys Leu

355 360 365

Val Asp Asn Leu Glu Ile Ala Ser Asn Leu Ala Asn Val Gly Asp Ser

370 375 380

Lys Thr Leu Val Ile Ala Pro Trp Phe Thr Thr His Gln Gln Leu Ser

385 390 395 400

Asp Glu Glu Lys Leu Ala Ser Gly Val Thr Lys Gly Leu Ile Arg Val

405 410 415

Ser Thr Gly Thr Glu Tyr Ile Asp Asp Ile Ile Asn Asp Phe Glu Gln

420 425 430

Ala Phe Lys Lys Val Tyr Asn Asn

435 440

<210>53

<211>1290

<212>DNA

<213>粟酒裂殖酵母(Schizosaccharomyces pombe)

<220>

<221>CDS

<222>(1)..(1290)

<223>

<400>53

atg cca gtc gag agt gaa cat ttc gaa act tta caa tta cat gct ggc 48

Met Pro Val Glu Ser Glu His Phe Glu Thr Leu Gln Leu His Ala Gly

1 5 10 15

caa gag cct gat gct gct acc agc tct cgt gcc gtt ccc atc tac gct 96

Gln Glu Pro Asp Ala Ala Thr Ser Ser Arg Ala Val Pro Ile Tyr Ala

20 25 30

act act tcc tat gtt ttc cgt gat tgc gac cat ggc ggc cgc ttg ttc 144

Thr Thr Ser Tyr Val Phe Arg Asp Cys Asp His Gly Gly Arg Leu Phe

35 40 45

gga tta cag gaa cca ggt tac atc tac tcg cgt atg atg aat ccc acc 192

Gly Leu Gln Glu Pro Gly Tyr Ile Tyr Ser Arg Met Met Asn Pro Thr

50 55 60

gcc gac gtt ttt gag aaa cgt att gcc gcc ttg gag cat ggc gct gct 240

Ala Asp Val Phe Glu Lys Arg Ile Ala Ala Leu Glu His Gly Ala Ala

65 70 75 80

gca atc gct act agt tcc ggt act tcc gct ctc ttc atg gct ttg acc 288

Ala Ile Ala Thr Ser Ser Gly Thr Ser Ala Leu Phe Met Ala Leu Thr

85 90 95

acg ttg gct aag gcc ggt gat aac att gtc tcc act tct tac ctt tat 336

Thr Leu Ala Lys Ala Gly Asp Asn Ile Val Ser Thr Ser Tyr Leu Tyr

100 105 110

ggt ggt act tac aac ctc ttc aag gtt acc ctg cct aga ttg gga att 384

Gly Gly Thr Tyr Asn Leu Phe Lys Val Thr Leu Pro Arg Leu Gly Ile

115 120 125

act acc aag ttt gtc aat ggt gat gat cct aat gat ctt gca gct cag 432

Thr Thr Lys Phe Val Asn Gly Asp Asp Pro Asn Asp Leu Ala Ala Gln

130 135 140

att gat gaa aac aca aag gct gtt tac gtt gag tcc atc ggc aat ccc 480

Ile Asp Glu Asn Thr Lys Ala Val Tyr Val Glu Ser Ile Gly Asn Pro

145 150 155 160

atg tac aac gtt ccc gat ttt gag cgt atc gct gag gtt gct cat gcc 528

Met Tyr Asn Val Pro Asp Phe Glu Arg Ile Ala Glu Val Ala His Ala

165 170 175

gct ggt gtg cct tta atg gtc gat aac act ttt ggc ggc ggt ggt tat 576

Ala Gly Val Pro Leu Met Val Asp Asn Thr Phe Gly Gly Gly Gly Tyr

180 185 190

ttg gtt cgt ccc att gac cac ggt gcc gat atc gtt acc cac tct gcc 624

Leu Val Arg Pro Ile Asp His Gly Ala Asp Ile Val Thr His Ser Ala

195 200 205

act aag tgg atc ggt ggt cat ggc act act att ggc ggt gtg att gtt 672

Thr Lys Trp Ile Gly Gly His Gly Thr Thr Ile Gly Gly Val Ile Val

210 215 220

gat agt ggt aag ttt gac tgg aag aag aac agc aag cgt ttc cct gaa 720

Asp Ser Gly Lys Phe Asp Trp Lys Lys Asn Ser Lys Arg Phe Pro Glu

225 230 235 240

ttc aac gag cct cat ccc ggt tac cat ggc atg gtc ttt act gaa act 768

Phe Asn Glu Pro His Pro Gly Tyr His Gly Met Val Phe Thr Glu Thr

245 250 255

ttt ggt aac ttg gca tat gct ttt gct tgc cgt act caa act ctc cgt 816

Phe Gly Asn Leu Ala Tyr Ala Phe Ala Cys Arg Thr Gln Thr Leu Arg

260 265 270

gat gtt ggt ggc aat gcc aat cca ttc ggt gtc ttt ttg ctt ctt caa 864

Asp Val Gly Gly Asn Ala Asn Pro Phe Gly Val Phe Leu Leu Leu Gln

275 280 285

ggt ctt gaa acg ctt tct ctt cgt atg gag cgt cac gtt caa aat gca 912

Gly Leu Glu Thr Leu Ser Leu Arg Met Glu Arg His Val Gln Asn Ala

290 295 300

ttt gct ctt gca aaa tat ttg gaa aag cac ccc aag gtt aac tgg gtt 960

Phe Ala Leu Ala Lys Tyr Leu Glu Lys His Pro Lys Val Asn Trp Val

305 310 315 320

tct tac cct ggt ctt gaa tct cac gtc tct cac aaa ctt gcc aag aag 1008

Ser Tyr Pro Gly Leu Glu Ser His Val Ser His Lys Leu Ala Lys Lys

325 330 335

tac ttg aaa aat ggt tac ggc gcc gtt ctc agc ttt ggc gct aaa ggt 1056

Tyr Leu Lys Asn Gly Tyr Gly Ala Val Leu Ser Phe Gly Ala Lys Gly

340 345 350

ggc cct gat caa agt cgt aag gta gtc aat gcc tta aag ctt gct agt 1104

Gly Pro Asp Gln Ser Arg Lys Val Val Asn Ala Leu Lys Leu Ala Ser

355 360 365

cag ttg gcc aat gtt ggt gat gcc aaa act ttg gtt atc gct cct gcc 1152

Gln Leu Ala Asn Val Gly Asp Ala Lys Thr Leu Val Ile Ala Pro Ala

370 375 380

tat acc act cat tta caa tta act gat gag gag caa att tct gcc ggt 1200

Tyr Thr Thr His Leu Gln Leu Thr Asp Glu Glu Gln Ile Ser Ala Gly

385 390 395 400

gtc act aag gat ctt att cgt gtg gcc gtc ggt att gag cac atc gat 1248

Val Thr Lys Asp Leu Ile Arg Val Ala Val Gly Ile Glu His Ile Asp

405 410 415

gat att atc gcc gac ttt gct caa gct ttg gaa gtt gcc taa 1290

Asp Ile Ile Ala Asp Phe Ala Gln Ala Leu Glu Val Ala

420 425

<210>54

<211>429

<212>PRT

<213>粟酒裂殖酵母

<400>54

Met Pro Val Glu Ser Glu His Phe Glu Thr Leu Gln Leu His Ala Gly

1 5 10 15

Gln Glu Pro Asp Ala Ala Thr Ser Ser Arg Ala Val Pro Ile Tyr Ala

20 25 30

Thr Thr Ser Tyr Val Phe Arg Asp Cys Asp His Gly Gly Arg Leu Phe

35 40 45

Gly Leu Gln Glu Pro Gly Tyr Ile Tyr Ser Arg Met Met Asn Pro Thr

50 55 60

Ala Asp Val Phe Glu Lys Arg Ile Ala Ala Leu Glu His Gly Ala Ala

65 70 75 80

Ala Ile Ala Thr Ser Ser Gly Thr Ser Ala Leu Phe Met Ala Leu Thr

85 90 95

Thr Leu Ala Lys Ala Gly Asp Asn Ile Val Ser Thr Ser Tyr Leu Tyr

100 105 110

Gly Gly Thr Tyr Asn Leu Phe Lys Val Thr Leu Pro Arg Leu Gly Ile

115 120 125

Thr Thr Lys Phe Val Asn Gly Asp Asp Pro Asn Asp Leu Ala Ala Gln

130 135 140

Ile Asp Glu Asn Thr Lys Ala Val Tyr Val Glu Ser Ile Gly Asn Pro

145 150 155 160

Met Tyr Asn Val Pro Asp Phe Glu Arg Ile Ala Glu Val Ala His Ala

165 170 175

Ala Gly Val Pro Leu Met Val Asp Asn Thr Phe Gly Gly Gly Gly Tyr

180 185 190

Leu Val Arg Pro Ile Asp His Gly Ala Asp Ile Val Thr His Ser Ala

195 200 205

Thr Lys Trp Ile Gly Gly His Gly Thr Thr Ile Gly Gly Val Ile Val

210 215 220

Asp Ser Gly Lys Phe Asp Trp Lys Lys Asn Ser Lys Arg Phe Pro Glu

225 230 235 240

Phe Asn Glu Pro His Pro Gly Tyr His Gly Met Val Phe Thr Glu Thr

245 250 255

Phe Gly Asn Leu Ala Tyr Ala Phe Ala Cys Arg Thr Gln Thr Leu Arg

260 265 270

Asp Val Gly Gly Asn Ala Asn Pro Phe Gly Val Phe Leu Leu Leu Gln

275 280 285

Gly Leu Glu Thr Leu Ser Leu Arg Met Glu Arg His Val Gln Asn Ala

290 295 300

Phe Ala Leu Ala Lys Tyr Leu Glu Lys His Pro Lys Val Asn Trp Val

305 310 315 320

Ser Tyr Pro Gly Leu Glu Ser His Val Ser His Lys Leu Ala Lys Lys

325 330 335

Tyr Leu Lys Asn Gly Tyr Gly Ala Val Leu Ser Phe Gly Ala Lys Gly

340 345 350

Gly Pro Asp Gln Ser Arg Lys Val Val Asn Ala Leu Lys Leu Ala Ser

355 360 365

Gln Leu Ala Asn Val Gly Asp Ala Lys Thr Leu Val Ile Ala Pro Ala

370 375 380

Tyr Thr Thr His Leu Gln Leu Thr Asp Glu Glu Gln Ile Ser Ala Gly

385 390 395 400

Val Thr Lys Asp Leu Ile Arg Val Ala Val Gly Ile Glu His Ile Asp

405 410 415

Asp Ile Ile Ala Asp Phe Ala Gln Ala Leu Glu Val Ala

420 425

<210>55

<211>52

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>55

cccgggatcc gctagcggcg cgccggccgg cccggtgtga aataccgcac ag 52

<210>56

<211>53

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>56

tctagactcg agcggccgcg gccggccttt aaattgaaga cgaaagggcc tcg 53

<210>57

<211>47

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>57

gagatctaga cccggggatc cgctagcggg ctgctaaagg aagcgga 47

<210>58

<211>38

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>58

gagaggcgcg ccgctagcgt gggcgaagaa ctccagca 38

<210>59

<211>34

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>59

gagagggcgg ccgcgcaaag tcccgcttcg tgaa 34

<210>60

<211>34

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>60

gagagggcgg ccgctcaagt cggtcaagcc acgc 34

<210>61

<211>140

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>61

tcgaatttaa atctcgagag gcctgacgtc gggcccggta ccacgcgtca tatgactagt 60

tcggacctag ggatatcgtc gacatcgatg ctcttctgcg ttaattaaca attgggatcc 120

tctagacccg ggatttaaat 140

<210>62

<211>140

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>62

gatcatttaa atcccgggtc tagaggatcc caattgttaa ttaacgcaga agagcatcga 60

tgtcgacgat atccctaggt ccgaactagt catatgacgc gtggtaccgg gcccgacgtc 120

aggcctctcg agatttaaat 140

<210>63

<211>33

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>63

gagagcggcc gccgatcctt ttteacccat cac 33

<210>64

<211>32

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>64

aggagcggcc gccatcggca ttttcttttg cg 32

<210>65

<211>5091

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：质粒

<400>65

gccgcgactg ccttcgcgaa gccttgcccc gcggaaattt cctccaccga gttcgtgcac 60

acccctatgc caagcttctt tcaccctaaa ttcgagagat tggattctta ccgtggaaat 120

tcttcgcaaa aatcgtcccc tgatcgccct tgcgacgttg gcgtcggtgc cgctggttgc 180

gcttggcttg accgacttga tcagcggccg ctcgatttaa atctcgagag gcctgacgtc 240

gggcccggta ccacgcgtca tatgactagt tcggacctag ggatatcgtc gacatcgatg 300

ctcttctgcg ttaattaaca attgggatcc tctagacccg ggatttaaat cgctagcggg 360

ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat 420

gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt 480

agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga 540

accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg 600

gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac 660

aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 720

ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 780

cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 840

cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 900

cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 960

gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 1020

catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 1080

ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 1140

tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 1200

caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 1260

gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 1320

ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 1380

cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 1440

cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 1500

gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 1560

aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 1620

ctcatgctgg agttcttcgc ccacgctagc ggcgcgccgg ccggcccggt gtgaaatacc 1680

gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga 1740

ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 1800

acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 1860

aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 1920

tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 1980

aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 2040

gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 2100

acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 2160

accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 2220

ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 2280

gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 2340

gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 2400

ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 2460

gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 2520

cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 2580

cttcacctag atccttttaa aggccggccg cggccgcgca aagtcccgct tcgtgaaaat 2640

tttcgtgccg cgtgattttc cgccaaaaac tttaacgaac gttcgttata atggtgtcat 2700

gaccttcacg acgaagtact aaaattggcc cgaatcatca gctatggatc tctctgatgt 2760

cgcgctggag tccgacgcgc tcgatgctgc cgtcgattta aaaacggtga tcggattttt 2820

ccgagctctc gatacgacgg acgcgccagc atcacgagac tgggccagtg ccgcgagcga 2880

cctagaaact ctcgtggcgg atcttgagga gctggctgac gagctgcgtg ctcggccagc 2940

gccaggagga cgcacagtag tggaggatgc aatcagttgc gcctactgcg gtggcctgat 3000

tcctccccgg cctgacccgc gaggacggcg cgcaaaatat tgctcagatg cgtgtcgtgc 3060

cgcagccagc cgcgagcgcg ccaacaaacg ccacgccgag gagctggagg cggctaggtc 3120

gcaaatggcg ctggaagtgc gtcccccgag cgaaattttg gccatggtcg tcacagagct 3180

ggaagcggca gcgagaatta tcgcgatcgt ggcggtgccc gcaggcatga caaacatcgt 3240

aaatgccgcg tttcgtgtgc cgtggccgcc caggacgtgt cagcgccgcc accacctgca 3300

ccgaatcggc agcagcgtcg cgcgtcgaaa aagcgcacag gcggcaagaa gcgataagct 3360

gcacgaatac ctgaaaaatg ttgaacgccc cgtgagcggt aactcacagg gcgtcggcta 3420

acccccagtc caaacctggg agaaagcgct caaaaatgac tctagcggat tcacgagaca 3480

ttgacacacc ggcctggaaa ttttccgctg atctgttcga cacccatccc gagctcgcgc 3540

tgcgatcacg tggctggacg agcgaagacc gccgcgaatt cctcgctcac ctgggcagag 3600

aaaatttcca gggcagcaag acccgcgact tcgccagcgc ttggatcaaa gacccggaca 3660

cggagaaaca cagccgaagt tataccgagt tggttcaaaa tcgcttgccc ggtgccagta 3720

tgttgctctg acgcacgcgc agcacgcagc cgtgcttgtc ctggacattg atgtgccgag 3780

ccaccaggcc ggcgggaaaa tcgagcacgt aaaccccgag gtctacgcga ttttggagcg 3840

ctgggcacgc ctggaaaaag cgccagcttg gatcggcgtg aatccactga gcgggaaatg 3900

ccagctcatc tggctcattg atccggtgta tgccgcagca ggcatgagca gcccgaatat 3960

gcgcctgctg gctgcaacga ccgaggaaat gacccgcgtt ttcggcgctg accaggcttt 4020

ttcacatagg ctgagccgtg gccactgcac tctccgacga tcccagccgt accgctggca 4080

tgcccagcac aatcgcgtgg atcgcctagc tgatcttatg gaggttgctc gcatgatctc 4140

aggcacagaa aaacctaaaa aacgctatga gcaggagttt tctagcggac gggcacgtat 4200

cgaagcggca agaaaagcca ctgcggaagc aaaagcactt gccacgcttg aagcaagcct 4260

gccgagcgcc gctgaagcgt ctggagagct gatcgacggc gtccgtgtcc tctggactgc 4320

tccagggcgt gccgcccgtg atgagacggc ttttcgccac gctttgactg tgggatacca 4380

gttaaaagcg gctggtgagc gcctaaaaga caccaagggt catcgagcct acgagcgtgc 4440

ctacaccgtc gctcaggcgg tcggaggagg ccgtgagcct gatctgccgc cggactgtga 4500

ccgccagacg gattggccgc gacgtgtgcg cggctacgtc gctaaaggcc agccagtcgt 4560

ccctgctcgt cagacagaga cgcagagcca gccgaggcga aaagctctgg ccactatggg 4620

aagacgtggc ggtaaaaagg ccgcagaacg ctggaaagac ccaaacagtg agtacgcccg 4680

agcacagcga gaaaaactag ctaagtccag tcaacgacaa gctaggaaag ctaaaggaaa 4740

tcgcttgacc attgcaggtt ggtttatgac tgttgaggga gagactggct cgtggccgac 4800

aatcaatgaa gctatgtctg aatttagcgt gtcacgtcag accgtgaata gagcacttaa 4860

ggtctgcggg cattgaactt ccacgaggac gccgaaagct tcccagtaaa tgtgccatct 4920

cgtaggcaga aaacggttcc cccgtagggt ctctctcttg gcctcctttc taggtcgggc 4980

tgattgctct tgaagctctc taggggggct cacaccatag gcagataacg ttccccaccg 5040

gctcgcctcg taagcgcaca aggactgctc ccaaagatct tcaaagccac t 5091

<210>66

<211>4323

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：质粒

<400>66

tctctcagcg tatggttgtc gcctgagctg tagttgcctt catcgatgaa ctgctgtaca 60

ttttgatacg tttttccgtc accgtcaaag attgatttat aatcctctac accgttgatg 120

ttcaaagagc tgtctgatgc tgatacgtta acttgtgcag ttgtcagtgt ttgtttgccg 180

taatgtttac cggagaaatc agtgtagaat aaacggattt ttccgtcaga tgtaaatgtg 240

gctgaacctg accattcttg tgtttggtct tttaggatag aatcatttgc atcgaatttg 300

tcgctgtctt taaagacgcg gccagcgttt ttccagctgt caatagaagt ttcgccgact 360

ttttgataga acatgtaaat cgatgtgtca tccgcatttt taggatctcc ggctaatgca 420

aagacgatgt ggtagccgtg atagtttgcg acagtgccgt cagcgttttg taatggccag 480

ctgtcccaaa cgtccaggcc ttttgcagaa gagatatttt taattgtgga cgaatcaaat 540

tcagaaactt gatatttttc atttttttgc tgttcaggga tttgcagcat atcatggcgt 600

gtaatatggg aaatgccgta tgtttcctta tatggctttt ggttcgtttc tttcgcaaac 660

gcttgagttg cgcctcctgc cagcagtgcg gtagtaaagg ttaatactgt tgcttgtttt 720

gcaaactttt tgatgttcat cgttcatgtc tcctttttta tgtactgtgt tagcggtctg 780

cttcttccag ccctcctgtt tgaagatggc aagttagtta cgcacaataa aaaaagacct 840

aaaatatgta aggggtgacg ccaaagtata cactttgccc tttacacatt ttaggtcttg 900

cctgctttat cagtaacaaa cccgcgcgat ttacttttcg acctcattct attagactct 960

cgtttggatt gcaactggtc tattttcctc ttttgtttga tagaaaatca taaaaggatt 1020

tgcagactac gggcctaaag aactaaaaaa tctatctgtt tcttttcatt ctctgtattt 1080

tttatagttt ctgttgcatg ggcataaagt tgccttttta atcacaattc agaaaatatc 1140

ataatatctc atttcactaa ataatagtga acggcaggta tatgtgatgg gttaaaaagg 1200

atcggcggcc gctcgattta aatctcgaga ggcctgacgt cgggcccggt accacgcgtc 1260

atatgactag ttcggaccta gggatatcgt cgacatcgat gctcttctgc gttaattaac 1320

aattgggatc ctctagaccc gggatttaaa tcgctagcgg gctgctaaag gaagcggaac 1380

acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct 1440

atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca 1500

tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg 1560

gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca 1620

aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc 1680

atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 1740

ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 1800

gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 1860

caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 1920

ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 1980

gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 2040

cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 2100

atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 2160

gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 2220

ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 2280

ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 2340

atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 2400

ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 2460

gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 2520

tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 2580

ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 2640

cccacgctag cggcgcgccg gccggcccgg tgtgaaatac cgcacagatg cgtaaggaga 2700

aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 2760

cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 2820

ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 2880

aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 2940

cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 3000

cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 3060

gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 3120

tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 3180

cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 3240

ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 3300

gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 3360

gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 3420

accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 3480

ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 3540

tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 3600

aaggccggcc gcggccgcca tcggcatttt cttttgcgtt tttatttgtt aactgttaat 3660

tgtccttgtt caaggatgct gtctttgaca acagatgttt tcttgccttt gatgttcagc 3720

aggaagctcg gcgcaaacgt tgattgtttg tctgcgtaga atcctctgtt tgtcatatag 3780

cttgtaatca cgacattgtt tcctttcgct tgaggtacag cgaagtgtga gtaagtaaag 3840

gttacatcgt taggatcaag atccattttt aacacaaggc cagttttgtt cagcggcttg 3900

tatgggccag ttaaagaatt agaaacataa ccaagcatgt aaatatcgtt agacgtaatg 3960

ccgtcaatcg tcatttttga tccgcgggag tcagtgaaca ggtaccattt gccgttcatt 4020

ttaaagacgt tcgcgcgttc aatttcatct gttactgtgt tagatgcaat cagcggtttc 4080

atcacttttt tcagtgtgta atcatcgttt agctcaatca taccgagagc gccgtttgct 4140

aactcagccg tgcgtttttt atcgctttgc agaagttttt gactttcttg acggaagaat 4200

gatgtgcttt tgccatagta tgctttgtta aataaagatt cttcgccttg gtagccatct 4260

tcagttccag tgtttgcttc aaatactaag tatttgtggc ctttatcttc tacgtagtga 4320

gga 4323

<210>67

<211>35

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>67

gagagagaga cgcgtcccag tggctgagac gcatc 35

<210>68

<211>34

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>68

ctctctctgt cgacgaattc aatcttacgg cctg 34

<210>69

<211>5860

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：质粒

<400>69

cccggtacca cgcgtcccag tggctgagac gcatccgcta aagccccagg aaccctgtgc 60

agaaagaaaa cactcctctg gctaggtaga cacagtttat aaaggtagag ttgagcgggt 120

aactgtcagc acgtagatcg aaaggtgcac aaaggtggcc ctggtcgtac agaaatatgg 180

cggttcctcg cttgagagtg cggaacgcat tagaaacgtc gctgaacgga tcgttgccac 240

caagaaggct ggaaatgatg tcgtggttgt ctgctccgca atgggagaca ccacggatga 300

acttctagaa cttgcagcgg cagtgaatcc cgttccgcca gctcgtgaaa tggatatgct 360

cctgactgct ggtgagcgta tttctaacgc tctcgtcgcc atggctattg agtcccttgg 420

cgcagaagcc caatctttca cgggctctca ggctggtgtg ctcaccaccg agcgccacgg 480

aaacgcacgc attgttgatg tcactccagg tcgtgtgcgt gaagcactcg atgagggcaa 540

gatctgcatt gttgctggtt tccagggtgt taataaagaa acccgcgatg tcaccacgtt 600

gggtcgtggt ggttctgaca ccactgcagt tgcgttggca gctgctttga acgctgatgt 660

gtgtgagatt tactcggacg ttgacggtgt gtataccgct gacccgcgca tcgttcctaa 720

tgcacagaag ctggaaaagc tcegcttcga agaaatgctg gaacttgctg ctgttggctc 780

caagattttg gtgctgcgca gtgttgaata cgctcgtgca ttcaatgtgc cacttcgcgt 840

acgctcgtct tatagtaatg atcccggcac tttgattgcc ggctctatgg aggatattcc 900

tgtggaagaa gcagtcctta ccggtgtcgc aaccgacaag tccgaagcca aagtaaccgt 960

tctgggtatt tccgataagc caggcgaggc tgcgaaggtt ttccgtgcgt tggctgatgc 1020

agaaatcaac attgacatgg ttctgcagaa cgtctcttct gtagaagacg gcaccaccga 1080

catcaccttc acctgccctc gttccgacgg ccgccgcgcg atggagatct tgaagaagct 1140

tcaggttcag ggcaactgga ccaatgtgct ttacgacgac caggtcggca aagtctccct 1200

cgtgggtgct ggcatgaagt ctcacccagg tgttaccgca gagttcatgg aagctctgcg 1260

cgatgtcaac gtgaacatcg aattgatttc cacctctgag attcgtattt ccgtgctgat 1320

ccgtgaagat gatctggatg ctgctgcacg tgcattgcat gagcagttcc agctgggcgg 1380

cgaagacgaa gccgtcgttt atgcaggcac cggacgctaa agttttaaag gagtagtttt 1440

acaatgacca ccatcgcagt tgttggtgca accggccagg tcggccaggt tatgcgcacc 1500

cttttggaag agcgcaattt cccagctgac actgttcgtt tctttgcttc cccacgttcc 1560

gcaggccgta agattgaatt cgtcgacatc gatgctcttc tgcgttaatt aacaattggg 1620

atcctctaga cccgggattt aaatcgctag cgggctgcta aaggaagcgg aacacgtaga 1680

aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga 1740

caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat 1800

agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct 1860

ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct 1920

gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg 1980

aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 2040

actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 2100

ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 2160

aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 2220

ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 2280

tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 2340

tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 2400

gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 2460

aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 2520

atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 2580

tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 2640

tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 2700

tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 2760

tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 2820

acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 2880

ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccacgc 2940

tagcggcgcg ccggccggcc cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 3000

gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 3060

ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 3120

acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 3180

cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 3240

caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 3300

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 3360

tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 3420

aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 3480

ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 3540

cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 3600

tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 3660

tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3720

ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3780

aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3840

aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaaggccg 3900

gccgcggccg ccatcggcat tttcttttgc gtttttattt gttaactgtt aattgtcctt 3960

gttcaaggat gctgtctttg acaacagatg ttttcttgcc tttgatgttc agcaggaagc 4020

tcggcgcaaa cgttgattgt ttgtctgcgt agaatcctct gtttgtcata tagcttgtaa 4080

tcacgacatt gtttcctttc gcttgaggta cagcgaagtg tgagtaagta aaggttacat 4140

cgttaggatc aagatccatt tttaacacaa ggccagtttt gttcagcggc ttgtatgggc 4200

cagttaaaga attagaaaca taaccaagca tgtaaatatc gttagacgta atgccgtcaa 4260

tcgtcatttt tgatccgcgg gagtcagtga acaggtacca tttgccgttc attttaaaga 4320

cgttcgcgcg ttcaatttca tctgttactg tgttagatgc aatcagcggt ttcatcactt 4380

ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt gctaactcag 4440

ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag aatgatgtgc 4500

ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca tcttcagttc 4560

cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag tgaggatctc 4620

tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc tgtacatttt 4680

gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg ttgatgttca 4740

aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt ttgccgtaat 4800

gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta aatgtggctg 4860

aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg aatttgtcgc 4920

tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg ccgacttttt 4980

gatagaacat gtaaatcgat gtgtcatccg catttttagg atctccggct aatgcaaaga 5040

cgatgtggta gccgtgatag tttgcgacag tgccgtcagc gttttgtaat ggccagctgt 5100

cccaaacgtc caggcctttt gcagaagaga tatttttaat tgtggacgaa tcaaattcag 5160

aaacttgata tttttcattt ttttgctgtt cagggatttg cagcatatca tggcgtgtaa 5220

tatgggaaat gccgtatgtt tccttatatg gcttttggtt cgtttctttc gcaaacgctt 5280

gagttgcgcc tcctgccagc agtgcggtag taaaggttaa tactgttgct tgttttgcaa 5340

actttttgat gttcatcgtt catgtctcct tttttatgta ctgtgttagc ggtctgcttc 5400

ttccagccct cctgtttgaa gatggcaagt tagttacgca caataaaaaa agacctaaaa 5460

tatgtaaggg gtgacgccaa agtatacact ttgcccttta cacattttag gtcttgcctg 5520

ctttatcagt aacaaacccg cgcgatttac ttttcgacct cattctatta gactctcgtt 5580

tggattgcaa ctggtctatt ttcctctttt gtttgataga aaatcataaa aggatttgca 5640

gactacgggc ctaaagaact aaaaaatcta tctgtttctt ttcattctct gtatttttta 5700

tagtttctgt tgcatgggca taaagttgcc tttttaatca caattcagaa aatatcataa 5760

tatctcattt cactaaataa tagtgaacgg caggtatatg tgatgggtta aaaaggatcg 5820

gcggccgctc gatttaaatc tcgagaggcc tgacgtcggg 5860

<210>70

<211>38

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>70

cggcaccacc gacatcatct tcacctgccc tcgttccg 38

<210>71

<211>38

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PcR引物

<400>71

cggaacgagg gcaggtgaag atgatgtcgg tggtgccg 38

<210>72

<211>1266

<212>DNA

<213>LysC突变体

<220>

<221>CDS

<222>(1)..(1266)

<223>

<400>72

gtg gcc ctg gtc gta cag aaa tat ggc ggt tcc tcg ctt gag agt gcg 48

Val Ala Leu Val Val Gln Lys Tyr Gly Gly Ser Ser Leu Glu Ser Ala

1 5 10 15

gaa cgc att aga aac gtc gct gaa cgg atc gtt gcc acc aag aag gct 96

Glu Arg Ile Arg Asn Val Ala Glu Arg Ile Val Ala Thr Lys Lys Ala

20 25 30

gga aat gat gtc gtg gtt gtc tgc tcc gca atg gga gac acc acg gat 144

Gly Asn Asp Val Val Val Val Cys Ser Ala Met Gly Asp Thr Thr Asp

35 40 45

gaa ctt cta gaa ctt gca gcg gca gtg aat ccc gtt ccg cca gct cgt 192

Glu Leu Leu Glu Leu Ala Ala Ala Val Asn Pro Val Pro Pro Ala Arg

50 55 60

gaa atg gat atg ctc ctg act gct ggt gag cgt att tct aac gct ctc 240

Glu Met Asp Met Leu Leu Thr Ala Gly Glu Arg Ile Ser Asn Ala Leu

65 70 75 80

gtc gcc atg gct att gag tcc ctt ggc gca gaa gcc caa tct ttc acg 288

Val Ala Met Ala Ile Glu Ser Leu Gly Ala Glu Ala Gln Ser Phe Thr

85 90 95

ggc tct cag gct ggt gtg ctc acc acc gag cgc cac gga aac gca cgc 336

Gly Ser Gln Ala Gly Val Leu Thr Thr Glu Arg His Gly Asn Ala Arg

100 105 110

att gtt gat gtc act cca ggt cgt gtg cgt gaa gca ctc gat gag ggc 384

Ile Val Asp Val Thr Pro Gly Arg Val Arg Glu Ala Leu Asp Glu Gly

115 120 125

aag atc tgc att gtt gct ggt ttc cag ggt gtt aat aaa gaa acc cgc 432

Lys Ile Cys Ile Val Ala Gly Phe Gln Gly Val Asn Lys Glu Thr Arg

130 135 140

gat gtc acc acg ttg ggt cgt ggt ggt tct gac acc act gca gtt gcg 480

Asp Val Thr Thr Leu Gly Arg Gly Gly Ser Asp Thr Thr Ala Val Ala

145 150 155 160

ttg gca gct gct ttg aac gct gat gtg tgt gag att tac tcg gac gtt 528

Leu Ala Ala Ala Leu Asn Ala Asp Val Cys Glu Ile Tyr Ser Asp Val

165 170 175

gac ggt gtg tat acc gct gac ccg cgc atc gtt cct aat gca cag aag 576

Asp Gly Val Tyr Thr Ala Asp Pro Arg Ile Val Pro Asn Ala Gln Lys

180 185 190

ctg gaa aag ctc agc ttc gaa gaa atg ctg gaa ctt gct gct gtt ggc 624

Leu Glu Lys Leu Ser Phe Glu Glu Met Leu Glu Leu Ala Ala Val Gly

195 200 205

tcc aag att ttg gtg ctg cgc agt gtt gaa tac gct cgt gca ttc aat 672

Ser Lys Ile Leu Val Leu Arg Ser Val Glu Tyr Ala Arg Ala Phe Asn

210 215 220

gtg cca ctt cgc gta cgc tcg tct tat agt aat gat ccc ggc act ttg 720

Val Pro Leu Arg Val Arg Ser Ser Tyr Ser Asn Asp Pro Gly Thr Leu

225 230 235 240

att gcc ggc tct atg gag gat att cct gtg gaa gaa gca gtc ctt acc 768

Ile Ala Gly Ser Met Glu Asp Ile Pro Val Glu Glu Ala Val Leu Thr

245 250 255

ggt gtc gca acc gac aag tcc gaa gcc aaa gta acc gtt ctg ggt att 816

Gly Val Ala Thr Asp Lys Ser Glu Ala Lys Val Thr Val Leu Gly Ile

260 265 270

tcc gat aag cca ggc gag gct gcg aag gtt ttc cgt gcg ttg gct gat 864

Ser Asp Lys Pro Gly Glu Ala Ala Lys Val Phe Arg Ala Leu Ala Asp

275 280 285

gca gaa atc aac att gac atg gtt ctg cag aac gtc tct tct gta gaa 912

Ala Glu Ile Asn Ile Asp Met Val Leu Gln Asn Val Ser Ser Val Glu

290 295 300

gac ggc acc acc gac atc atc ttc acc tgc cct cgt tcc gac ggc cgc 960

Asp Gly Thr Thr Asp Ile Ile Phe Thr Cys Pro Arg Ser Asp Gly Arg

305 310 315 320

cgc gcg atg gag atc ttg aag aag ctt cag gtt cag ggc aac tgg acc 1008

Arg Ala Met Glu Ile Leu Lys Lys Leu Gln Val Gln Gly Asn Trp Thr

325 330 335

aat gtg ctt tac gac gac cag gtc ggc aaa gtc tcc ctc gtg ggt gct 1056

Asn Val Leu Tyr Asp Asp Gln Val Gly Lys Val Ser Leu Val Gly Ala

340 345 350

ggc atg aag tct cac cca ggt gtt acc gca gag ttc atg gaa gct ctg 1104

Gly Met Lys Ser His Pro Gly Val Thr Ala Glu Phe Met Glu Ala Leu

355 360 365

cgc gat gtc aac gtg aac atc gaa ttg att tcc acc tct gag att cgt 1152

Arg Asp Val Asn Val Asn Ile Glu Leu Ile Ser Thr Ser Glu Ile Arg

370 375 380

att tcc gtg ctg atc cgt gaa gat gat ctg gat gct gct gca cgt gca 1200

Ile Ser Val Leu Ile Arg Glu Asp Asp Leu Asp Ala Ala Ala Arg Ala

385 390 395 400

ttg cat gag cag ttc cag ctg ggc ggc gaa gac gaa gcc gtc gtt tat 1248

Leu His Glu Gln Phe Gln Leu Gly Gly Glu Asp Glu Ala Val Val Tyr

405 410 415

gca ggc acc gga cgc taa 1266

Ala Gly Thr Gly Arg

420

<210>73

<211>421

<212>PRT

<213>LysC突变体

<400>73

Val Ala Leu Val Val Gln Lys Tyr Gly Gly Ser Ser Leu Glu Ser Ala

1 5 10 15

Glu Arg Ile Arg Asn Val Ala Glu Arg Ile Val Ala Thr Lys Lys Ala

20 25 30

Gly Asn Asp Val Val Val Val Cys Ser Ala Met Gly Asp Thr Thr Asp

35 40 45

Glu Leu Leu Glu Leu Ala Ala Ala Val Asn Pro Val Pro Pro Ala Arg

50 55 60

Glu Met Asp Met Leu Leu Thr Ala Gly Glu Arg Ile Ser Asn Ala Leu

65 70 75 80

Val Ala Met Ala Ile Glu Ser Leu Gly Ala Glu Ala Gln Ser Phe Thr

85 90 95

Gly Ser Gln Ala Gly Val Leu Thr Thr Glu Arg His Gly Asn Ala Arg

100 105 110

Ile Val Asp Val Thr Pro Gly Arg Val Arg Glu Ala Leu Asp Glu Gly

115 120 125

Lys Ile Cys Ile Val Ala Gly Phe Gln Gly Val Asn Lys Glu Thr Arg

130 135 140

Asp Val Thr Thr Leu Gly Arg Gly Gly Ser Asp Thr Thr Ala Val Ala

145 150 155 160

Leu Ala Ala Ala Leu Asn Ala Asp Val Cys Glu Ile Tyr Ser Asp Val

165 170 175

Asp Gly Val Tyr Thr Ala Asp Pro Arg Ile Val Pro Asn Ala Gln Lys

180 185 190

Leu Glu Lys Leu Ser Phe Glu Glu Met Leu Glu Leu Ala Ala Val Gly

195 200 205

Ser Lys Ile Leu Val Leu Arg Ser Val Glu Tyr Ala Arg Ala Phe Asn

210 215 220

Val Pro Leu Arg Val Arg Ser Ser Tyr Ser Asn Asp Pro Gly Thr Leu

225 230 235 240

Ile Ala Gly Ser Met Glu Asp Ile Pro Val Glu Glu Ala Val Leu Thr

245 250 255

Gly Val Ala Thr Asp Lys Ser Glu Ala Lys Val Thr Val Leu Gly Ile

260 265 270

Ser Asp Lys Pro Gly Glu Ala Ala Lys Val Phe Arg Ala Leu Ala Asp

275 280 285

Ala Glu Ile Asn Ile Asp Met Val Leu Gln Asn Val Ser Ser Val Glu

290 295 300

Asp Gly Thr Thr Asp Ile Ile Phe Thr Cys Pro Arg Ser Asp Gly Arg

305 310 315 320

Arg Ala Met Glu Ile Leu Lys Lys Leu Gln Val Gln Gly Asn Trp Thr

325 330 335

Asn Val Leu Tyr Asp Asp Gln Val Gly Lys Val Ser Leu Val Gly Ala

340 345 350

Gly Met Lys Ser His Pro Gly Val Thr Ala Glu Phe Met Glu Ala Leu

355 360 365

Arg Asp Val Asn Val Asn Ile Glu Leu Ile Ser Thr Ser Glu Ile Arg

370 375 380

Ile Ser Val Leu Ile Arg Glu Asp Asp Leu Asp Ala Ala Ala Arg Ala

385 390 395 400

Leu His Glu Gln Phe Gln Leu Gly Gly Glu Asp Glu Ala Val Val Tyr

405 410 415

Ala Gly Thr Gly Arg

420

<210>74

<211>5860

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：质粒

<400>74

cccggtacca cgcgtcccag tggctgagac gcatccgcta aagccccagg aaccctgtgc 60

agaaagaaaa cactcctctg gctaggtaga cacagtttat aaaggtagag ttgagcgggt 120

aactgtcagc acgtagatcg aaaggtgcac aaaggtggcc ctggtcgtac agaaatatgg 180

cggttcctcg cttgagagtg cggaacgcat tagaaacgtc gctgaacgga tcgttgccac 240

caagaaggct ggaaatgatg tcgtggttgt ctgctccgca atgggagaca ccacggatga 300

acttctagaa cttgcagcgg cagtgaatcc cgttccgcca gctcgtgaaa tggatatgct 360

cctgactgct ggtgagcgta tttctaacgc tctcgtcgcc atggctattg agtcccttgg 420

cgcagaagcc caatctttca cgggctctca ggctggtgtg ctcaccaccg agcgccacgg 480

aaacgcacgc attgttgatg tcactccagg tcgtgtgcgt gaagcactcg atgagggcaa 540

gatctgcatt gttgctggtt tccagggtgt taataaagaa acccgcgatg tcaccacgtt 600

gggtcgtggt ggttctgaca ccactgcagt tgcgttggca gctgctttga acgctgatgt 660

gtgtgagatt tactcggacg ttgacggtgt gtataccgct gacccgcgca tcgttcctaa 720

tgcacagaag ctggaaaagc tcagcttcga agaaatgctg gaacttgctg ctgttggctc 780

caagattttg gtgctgcgca gtgttgaata cgctcgtgca ttcaatgtgc cacttcgcgt 840

acgctcgtct tatagtaatg atcccggcac tttgattgcc ggctctatgg aggatattcc 900

tgtggaagaa gcagtcctta ccggtgtcgc aaccgacaag tccgaagcca aagtaaccgt 960

tctgggtatt tccgataagc caggcgaggc tgcgaaggtt ttccgtgcgt tggctgatgc 1020

agaaatcaac attgacatgg ttctgcagaa cgtctcttct gtagaagacg gcaccaccga 1080

catcatcttc acctgccctc gttccgacgg ccgccgcgcg atggagatct tgaagaagct 1140

tcaggttcag ggcaactgga ccaatgtgct ttacgacgac caggtcggca aagtctccct 1200

cgtgggtgct ggcatgaagt ctcacccagg tgttaccgca gagttcatgg aagctctgcg 1260

cgatgtcaac gtgaacatcg aattgatttc cacctctgag attcgtattt ccgtgctgat 1320

ccgtgaagat gatctggatg ctgctgcacg tgcattgcat gagcagttcc agctgggcgg 1380

cgaagacgaa gccgtcgttt atgcaggcac cggacgctaa agttttaaag gagtagtttt 1440

acaatgacca ccatcgcagt tgttggtgca accggccagg tcggccaggt tatgcgcacc 1500

cttttggaag agcgcaattt cccagctgac actgttcgtt tctttgcttc cccacgttcc 1560

gcaggccgta agattgaatt cgtcgacatc gatgctcttc tgcgttaatt aacaattggg 1620

atcctctaga cccgggattt aaatcgctag cgggctgcta aaggaagcgg aacacgtaga 1680

aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga 1740

caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat 1800

agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct 1860

ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct 1920

gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg 1980

aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 2040

actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 2100

ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 2160

aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 2220

ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 2280

tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 2340

tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 2400

gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 2460

aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 2520

atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 2580

tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 2640

tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 2700

tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 2760

tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 2820

acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 2880

ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccacgc 2940

tagcggcgcg ccggccggcc cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 3000

gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 3060

ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 3120

acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 3180

cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 3240

caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 3300

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 3360

tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 3420

aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 3480

ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 3540

cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 3600

tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 3660

tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3720

ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3780

aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3840

aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaaggccg 3900

gccgcggccg ccatcggcat tttcttttgc gtttttattt gttaactgtt aattgtcctt 3960

gttcaaggat gctgtctttg acaacagatg ttttcttgcc tttgatgttc agcaggaagc 4020

tcggcgcaaa cgttgattgt ttgtctgcgt agaatcctct gtttgtcata tagcttgtaa 4080

tcacgacatt gtttcctttc gcttgaggta cagcgaagtg tgagtaagta aaggttacat 4140

cgttaggatc aagatccatt tttaacacaa ggccagtttt gttcagcggc ttgtatgggc 4200

cagttaaaga attagaaaca taaccaagca tgtaaatatc gttagacgta atgccgtcaa 4260

tcgtcatttt tgatccgcgg gagtcagtga acaggtacca tttgccgttc attttaaaga 4320

cgttcgcgcg ttcaatttca tctgttactg tgttagatgc aatcagcggt ttcatcactt 4380

ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt gctaactcag 4440

ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag aatgatgtgc 4500

ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca tcttcagttc 4560

cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag tgaggatctc 4620

tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc tgtacatttt 4680

gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg ttgatgttca 4740

aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt ttgccgtaat 4800

gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta aatgtggctg 4860

aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg aatttgtcgc 4920

tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg ccgacttttt 4980

gatagaacat gtaaatcgat gtgtcatccg catttttagg atctccggct aatgcaaaga 5040

cgatgtggta gccgtgatag tttgcgacag tgccgtcagc gttttgtaat ggccagctgt 5100

cccaaacgtc caggcctttt gcagaagaga tatttttaat tgtggacgaa tcaaattcag 5160

aaacttgata tttttcattt ttttgctgtt cagggatttg cagcatatca tggcgtgtaa 5220

tatgggaaat gccgtatgtt tccttatatg gcttttggtt cgtttctttc gcaaacgctt 5280

gagttgcgcc tcctgccagc agtgcggtag taaaggttaa tactgttgct tgttttgcaa 5340

actttttgat gttcatcgtt catgtctcct tttttatgta ctgtgttagc ggtctgcttc 5400

ttccagccct cctgtttgaa gatggcaagt tagttacgca caataaaaaa agacctaaaa 5460

tatgtaaggg gtgacgccaa agtatacact ttgcccttta cacattttag gtcttgcctg 5520

ctttatcagt aacaaacccg cgcgatttac ttttcgacct cattctatta gactctcgtt 5580

tggattgcaa ctggtctatt ttcctctttt gtttgataga aaatcataaa aggatttgca 5640

gactacgggc ctaaagaact aaaaaatcta tctgtttctt ttcattctct gtatttttta 5700

tagtttctgt tgcatgggca taaagttgcc tttttaatca caattcagaa aatatcataa 5760

tatctcattt cactaaataa tagtgaacgg caggtatatg tgatgggtta aaaaggatcg 5820

gcggccgctc gatttaaatc tcgagaggcc tgacgtcggg 5860

<210>75

<211>31

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>75

gagaggatcc ggaaggtgaa tcgaatttcg g 31

<210>76

<211>40

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>76

ctattgctgt cggcgctcat gattctccaa aaataatcgc 40

<210>77

<211>20

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>77

atgagcgccg acagcaatag 20

<210>78

<211>29

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：PCR引物

<400>78

gaactctaga tcagaacgcc gccacggac 29

<210>79

<211>6591

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：质粒

<400>79

gatccggaag gtgaatcgaa tttcggggct ttaaagcaaa aatgaacagc ttggtctata 60

gtggctaggt accctttttg ttttggacac atgtagggtg gccgaaacaa agtaatagga 120

caacaacgct cgaccgcgat tatttttgga gaatcatgag cgccgacagc aatagcaccg 180

acgccgatcc gaccgcgcat tggtcgttcg aaaccaaaca gatacacgct ggtcagcacc 240

ctgatccgac caccaacgcc cgggctctgc cgatctatgc gaccacgtcg tacaccttcg 300

acgacaccgc gcacgccgcc gccctgttcg gactggaaat tccgggcaat atctacaccc 360

ggatcggcaa ccccaccacc gacgtcgtcg agcagcgcat cgccgcgctc gagggcggtg 420

tggccgcgct gttcctgtcg tcggggcagg ccgcggagac gttcgccatc ttgaacctgg 480

ccggcgcggg cgatcacatc gtgtccagcc cgcgcctgta cggcggcacc tacaacctgt 540

tccactattc gctggccaag ctcggcatcg aggtcagctt cgtcgacgat ccggacgatc 600

tggacacctg gcaggcggcg gtacggccca acaccaaggc gttcttcgcc gagaccatct 660

ccaacccgca gatcgacctg ctggacaccc cggcggtttc cgaggtcgcc catcgcaacg 720

gggtgccgtt gatcgtcgac aacaccatcg ccacgccata cctgatccaa ccgttggccc 780

agggcgccga catcgtcgtg cattcggcca ccaagtacct gggcgggcac ggtgccgcca 840

tcgcgggtgt gatcgtcgac ggcggcaact tcgattggac ccagggccgc ttccccggct 900

tcaccacccc cgaccccagc taccacggcg tggtgttcgc cgagctgggt ccaccggcgt 960

ttgcgctcaa agctcgagtg cagctgctcc gtgactacgg ctcggcggct tcgccgttca 1020

acgcgttctt ggtggcgcag ggtctggaaa cgctgagcct gcggatcgag cggcacgtcg 1080

ccaacgcgca gcgcgtcgcc gagttcctgg ccgcccgcga cgacgtgctt tcggtcaact 1140

atgcggggct gccctcctcg ccctggcatg agcgggccaa gaggctggcg cccaagggaa 1200

ccggggccgt gctgtccttc gagttggccg gcggcatcga ggccggcaag gcattcgtga 1260

acgcgttgaa gctgcacagc cacgtcgcca acatcggtga cgtgcgctcg ctggtgatcc 1320

acccggcatc gaccactcat gcccagctga gcccggccga gcagctggcg accggggtca 1380

gcccgggcct ggtgcgtttg gctgtgggca tcgaaggtat cgacgatatc ctggccgacc 1440

tggagcttgg ctttgccgcg gcccgcagat tcagcgccga cccgcagtcc gtggcggcgt 1500

tctgatctag acccgggatt taaatcgcta gcgggctgct aaaggaagcg gaacacgtag 1560

aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg 1620

acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga 1680

tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc 1740

tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc 1800

tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 1860

gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 1920

gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 1980

gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 2040

gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 2100

gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 2160

ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 2220

ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 2280

cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 2340

caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 2400

gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 2460

ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 2520

ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 2580

ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 2640

ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 2700

cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 2760

gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacg 2820

ctagcggcgc gccggccggc ccggtgtgaa ataccgcaca gatgcgtaag gagaaaatac 2880

cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 2940

cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3000

aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 3060

gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 3120

tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 3180

agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 3240

ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 3300

taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 3360

gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 3420

gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 3480

ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 3540

ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 3600

gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 3660

caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 3720

taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaaggcc 3780

ggccgcggcc gcgcaaagtc ccgcttcgtg aaaattttcg tgccgcgtga ttttccgcca 3840

aaaactttaa cgaacgttcg ttataatggt gtcatgacct tcacgacgaa gtactaaaat 3900

tggcccgaat catcagctat ggatctctct gatgtcgcgc tggagtccga cgcgctcgat 3960

gctgccgtcg atttaaaaac ggtgatcgga tttttccgag ctctcgatac gacggacgcg 4020

ccagcatcac gagactgggc cagtgccgcg agcgacctag aaactctcgt ggcggatctt 4080

gaggagctgg ctgacgagct gcgtgctcgg ccagcgccag gaggacgcac agtagtggag 4140

gatgcaatca gttgcgccta ctgcggtggc ctgattcctc cccggcctga cccgcgagga 4200

cggcgcgcaa aatattgctc agatgcgtgt cgtgccgcag ccagccgcga gcgcgccaac 4260

aaacgccacg ccgaggagct ggaggcggct aggtcgcaaa tggcgctgga agtgcgtccc 4320

ccgagcgaaa ttttggccat ggtcgtcaca gagctggaag cggcagcgag aattatcgcg 4380

atcgtggcgg tgcccgcagg catgacaaac atcgtaaatg ccgcgtttcg tgtgccgtgg 4440

ccgcccagga cgtgtcagcg ccgccaccac ctgcaccgaa tcggcagcag cgtcgcgcgt 4500

cgaaaaagcg cacaggcggc aagaagcgat aagctgcacg aatacctgaa aaatgttgaa 4560

cgccccgtga gcggtaactc acagggcgtc ggctaacccc cagtccaaac ctgggagaaa 4620

gcgctcaaaa atgactctag cggattcacg agacattgac acaccggcct ggaaattttc 4680

cgctgatctg ttcgacaccc atcccgagct cgcgctgcga tcacgtggct ggacgagcga 4740

agaccgccgc gaattcctcg ctcacctggg cagagaaaat ttccagggca gcaagacccg 4800

cgacttcgcc agcgcttgga tcaaagaccc ggacacggag aaacacagcc gaagttatac 4860

cgagttggtt caaaatcgct tgcccggtgc cagtatgttg ctctgacgca cgcgcagcac 4920

gcagccgtgc ttgtcctgga cattgatgtg ccgagccacc aggccggcgg gaaaatcgag 4980

cacgtaaacc ccgaggtcta cgcgattttg gagcgctggg cacgcctgga aaaagcgcca 5040

gcttggatcg gcgtgaatcc actgagcggg aaatgccagc tcatctggct cattgatccg 5100

gtgtatgccg cagcaggcat gagcagcccg aatatgcgcc tgctggctgc aacgaccgag 5160

gaaatgaccc gcgttttcgg cgctgaccag gctttttcac ataggctgag ccgtggccac 5220

tgcactctcc gacgatccca gccgtaccgc tggcatgccc agcacaatcg cgtggatcgc 5280

ctagctgatc ttatggaggt tgctcgcatg atctcaggca cagaaaaacc taaaaaacgc 5340

tatgagcagg agttttctag cggacgggca cgtatcgaag cggcaagaaa agccactgcg 5400

gaagcaaaag cacttgccac gcttgaagca agcctgccga gcgccgctga agcgtctgga 5460

gagctgatcg acggcgtccg tgtcctctgg actgctccag ggcgtgccgc ccgtgatgag 5520

acggcttttc gccacgcttt gactgtggga taccagttaa aagcggctgg tgagcgccta 5580

aaagacacca agggtcatcg agcctacgag cgtgcctaca ccgtcgctca ggcggtcgga 5640

ggaggccgtg agcctgatct gccgccggac tgtgaccgcc agacggattg gccgcgacgt 5700

gtgcgcggct acgtcgctaa aggccagcca gtcgtccctg ctcgtcagac agagacgcag 5760

agccagccga ggcgaaaagc tctggccact atgggaagac gtggcggtaa aaaggccgca 5820

gaacgctgga aagacccaaa cagtgagtac gcccgagcac agcgagaaaa actagctaag 5880

tccagtcaac gacaagctag gaaagctaaa ggaaatcgct tgaccattgc aggttggttt 5940

atgactgttg agggagagac tggctcgtgg ccgacaatca atgaagctat gtctgaattt 6000

agcgtgtcac gtcagaccgt gaatagagca cttaaggtct gcgggcattg aacttccacg 6060

aggacgccga aagcttccca gtaaatgtgc catctcgtag gcagaaaacg gttcccccgt 6120

agggtctctc tcttggcctc ctttctaggt cgggctgatt gctcttgaag ctctctaggg 6180

gggctcacac cataggcaga taacgttccc caccggctcg cctcgtaagc gcacaaggac 6240

tgctcccaaa gatcttcaaa gccactgccg cgactgcctt cgcgaagcct tgccccgcgg 6300

aaatttcctc caccgagttc gtgcacaccc ctatgccaag cttctttcac cctaaattcg 6360

agagattgga ttcttaccgt ggaaattctt cgcaaaaatc gtcccctgat cgcccttgcg 6420

acgttggcgt cggtgccgct ggttgcgctt ggcttgaccg acttgatcag cggccgctcg 6480

atttaaatct cgagaggcct gacgtcgggc ccggtaccac gcgtcatatg actagttcgg 6540

acctagggat atcgtcgaca tcgatgctct tctgcgttaa ttaacaattg g 6591

Claims

1.发酵产生至少一种含硫精细化学品的方法，该方法包括下面的步骤：

a)发酵产生目的含硫精细化学品的棒状细菌培养物，其中所述棒状细菌表达至少一种这样的异源核苷酸序列，该序列编码具有O-乙酰基高丝氨酸硫化氢解酶(metY)活性的蛋白质；

b)浓缩培养基或细菌细胞中的含硫精细化学品；和

c)分离含硫精细化学品。

2.权利要求1所述的方法，其中含硫精细化学品包含L-甲硫氨酸。

3.前述权利要求任一项中所述的方法，其中编码异源metY的核苷酸序列与来自谷氨酸棒杆菌(Corynebacterium glutamicum)ATCC 13032的metY编码序列的同源性小于100％。

4.权利要求3所述的方法，其中metY编码序列来自任一下列生物体：白喉棒杆菌(Corynebacterium diphteriae) ATCC 14779 结核分枝杆菌(Mycobacterium tuberculosis)CDC1551 ATCC 25584 丙酮丁醇梭菌(Clostridium acetobutylicum) ATCC 824 嗜碱芽孢杆菌(Bacillus halodurans) ATCC 21591 嗜热脂肪芽孢杆菌(Bacillus stearothermophilus) ATCC 12980 微温绿菌(Chlorobium tepidum) ATCC 49652 聚球藻属(Synechococcus)中的种 ATCC 27104 构巢裸孢壳(Emericella nidulans) ATCC 36104 脆弱拟杆菌(Bacteroides fragilis) ATCC 25285 乳酸乳球菌(Lactococcus lactis) ATCC 7962 支气管炎博德特氏菌(Bordetella bronchiseptica) ATCC 19395 铜绿假单胞菌(Pseudomonas aeruginosa) ATCC 17933 欧洲亚硝化单胞菌(Nitrosomonas europaea) ATCC 19718

苜蓿中华根瘤菌(Sinorhizobium meliloti) ATCC 4399 海栖热袍菌(Thermotoga maritima) ATCC 43589 变异链球菌(Streptococcus mutans) ATCC 25175 洋葱伯克霍尔德氏菌(Burkholderia cepacia) ATCC 25416 耐辐射奇异球菌(Deinococcus radiodurans) ATCC 13939 荚膜红细菌(Rhodobacter capsulatus) ATCC 11166 多杀巴斯德氏菌(Pasteurella multocida) ATCC 6530 艰难梭菌(Clostridium difficile) ATCC 9689 空肠弯曲杆菌(Campylobacter jejuni) ATCC 33560 肺炎链球菌(Streptococcus pneumoniae) ATCC 6308 酿酒酵母(Saccharomyces cerevisiae) ATCC 2704 乳酸克鲁维酵母(Kluyveromyces lactis) ATCC 8585 白假丝酵母(Candida albicans) ATCC 10231 粟酒裂殖酵母(Schizosaccharomyces pombe) ATCC 24969

5.前述权利要求任一项中所述的方法，其中metY编码序列包含根据SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、41、43、45、47、49、51和53的编码序列或者包含与它们同源的编码具有metY活性的蛋白质的核苷酸序列。

6.前述权利要求任一项中所述的方法，其中metY编码序列编码具有metY活性的蛋白质，所述蛋白质包含根据SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52和54的氨基酸序列或包含与它们同源的代表具有metY活性的蛋白质的氨基酸序列。

7.前述权利要求任一项中所述的方法，其中编码metY的序列为可以在棒状细菌中复制或者被稳定整合到染色体中的DNA或RNA。

8.权利要求7所述的方法，其中

a)使用经这样的质粒载体转化的细菌菌株，其中所述质粒载体携带处于调节序列控制下的至少一个拷贝的metY编码序列，或者

b)使用其中metY编码序列已经被整合入该细菌染色体中的菌株。

9.前述权利要求任一项中所述的方法，其中编码metY的序列被过量表达。

10.前述权利要求任一项中所述的方法，其中发酵这样的细菌，该细菌中目的含硫精细化学品生物合成途径的至少另一种基因额外地被扩增或突变从而其活性不受代谢的代谢物影响。

11.前述权利要求任一项中所述的方法，其中发酵这样的细菌，该细菌中降低目的含硫精细化学品产生的至少一条代谢途径至少被部分关闭。

12.前述权利要求任一项中所述的方法，其中发酵这样的棒状细菌，该细菌中同时对选自：

a)基因lysC，其编码天冬氨酸激酶，

b)甘油醛-3-磷酸脱氢酶编码基因gap，

c)3-磷酸甘油酸激酶编码基因pgk，

d)丙酮酸羧化酶编码基因pyc，

e)磷酸丙糖异构酶编码基因tpi，

f)高丝氨酸O-乙酰转移酶编码基因metA，

g)γ胱硫醚合酶编码基因metB，

h)γ胱硫醚裂合酶编码基因metC，

i)丝氨酸羟甲基转移酶编码基因glyA，

j)亚甲基四氢叶酸还原酶编码基因metF，

k)依赖维生素B12的甲硫氨酸合酶编码基因metH，

l)磷酸丝氨酸氨基转移酶编码基因serC，

m)磷酸丝氨酸磷酸酶编码基因serB，

n)丝氨酸乙酰转移酶编码基因cysE，和

o)高丝氨酸脱氢酶编码基因hom

的至少一种基因以某种方式过量表达或者突变，从而使得相应蛋白质的活性与未突变的蛋白质相比，受代谢的代谢物影响程度如果有的话较小。

13.前述权利要求任一项中所述的方法，其中发酵这样的棒状细菌，该细菌中同时对选自：

a)高丝氨酸激酶编码基因thrB，

b)苏氨酸脱水酶编码基因ilvA，

c)苏氨酸合酶编码基因thrC，

d)内消旋-二氨基庚二酸D-脱氢酶编码基因ddh，

e)磷酸烯醇丙酮酸羧激酶编码基因pck，

f)葡萄糖-6-磷酸6-异构酶编码基因pgi，

g)丙酮酸氧化酶编码基因poxB，

h)二氢吡啶二羧酸合酶编码基因dapA，

i)二氢吡啶二羧酸还原酶编码基因dapB；和

j)二氨基吡啶甲酸脱羧酶编码基因

的至少一种基因通过改变表达速率或者通过导入特定的突变而被弱化。

14.前述权利要求中一项或多项所述的方法，其中使用谷氨酸棒杆菌种的微生物。

15.从发酵液产生含L-甲硫氨酸的动物饲料添加剂的方法，该方法包括下面的步骤：

a)在发酵培养基中培养并发酵产生L-甲硫氨酸的微生物；

b)从含L-甲硫氨酸的发酵液除去水；

c)除去发酵过程中形成的生物量重量的0到100％；和

d)干燥根据b)和/或c)所得发酵液，以得到目的粉剂或粒剂形式的动物饲料添加剂。

16.权利要求15所述的方法，其中使用根据权利要求1到14任一项中所定义的微生物。