CN112840028B - 包含变体RpoC编码序列的核酸分子 - Google Patents

包含变体RpoC编码序列的核酸分子 Download PDF

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CN112840028B
CN112840028B CN201980050327.0A CN201980050327A CN112840028B CN 112840028 B CN112840028 B CN 112840028B CN 201980050327 A CN201980050327 A CN 201980050327A CN 112840028 B CN112840028 B CN 112840028B
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李知宣
张东银
金素影
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Abstract

公开了包含变体rpoC编码序列的核酸分子。变体rpoC编码序列编码调控质粒的拷贝数的变体RpoC。还公开了包含该核酸分子的重组微生物、用于调控重组微生物中的对象载体的拷贝数的方法以及通过使用该重组微生物制备目标产物的方法。

Description

包含变体RpoC编码序列的核酸分子
技术领域
本发明涉及包含变体rpoC RNA聚合酶β'亚基蛋白编码序列(也称为“变体rpoC编码序列”)的核酸分子，其中变体rpoC编码序列编码变体RpoC RNA聚合酶β'亚基蛋白(也称为“变体RpoC”)，并且变体RpoC调控质粒的拷贝数。
背景技术
质粒在生物技术中具有重要作用，提供用于引入、修饰和从微生物去除目标基因以及用于生产由目标基因编码的相应蛋白质的手段。质粒是在细菌、古细菌和真核生物域的多种微生物中天然存在的、与其所在的微生物的染色体物理上分开并且独立于染色体复制的核酸分子。质粒通常是双链环状DNA分子，但也可以是线性DNA分子和/或RNA分子。质粒以约1kb至大于2Mb的尺寸范围存在。例如，根据Shintani et al.，Frontiers inMicrobiology 6:242(2015)的最新综述论文，在GenBank数据库中发现的4602种质粒中观察到宽泛变异的尺寸，其中质粒的尺寸范围为744bp至2.58Mb，并且平均尺寸为80kb。质粒还以一定范围的拷贝数/细胞存在。例如，质粒总体上被表征为低拷贝(例如1-20个拷贝/细胞)、中等拷贝(例如20-100个拷贝/细胞)或高拷贝(例如500-700或更多个拷贝/细胞)。质粒可以被修饰以包括目标基因。
在生物技术中使用质粒相关的挑战是，生物技术应用总体上需要在微生物中稳定并入目标基因，并在培养期间仔细控制目标基因及其相应蛋白质产物的产量，但是完成一者的工作可冲撞完成另一者的工作。在培养包括具有目标基因的质粒的微生物的过程中，质粒随着微生物细胞的生长和分裂而稳定地分离，使得在整个培养过程中高百分比的微生物细胞将包括目标基因，是总体上有利的。目标基因保持结构稳定，维持恒定的核苷酸序列以确保仅产生预期产物，也是总体上有利的。使目标基因以足以实现期望结果(例如相应蛋白质产物的足量和以活性形式产生)的高水平表达，也是总体上有利的。不幸的是，用于复制质粒和由质粒表达目标基因(特别是以高水平)的技术对细胞造成代谢负担。这可以导致质粒从细胞丢失和/或突变改变目标基因的表达水平或同一性。这还可以导致相应蛋白质产物的聚集和失活。因此，在生物技术应用中在质粒使用期间平衡稳定并入与产量控制总体上是经验过程，涉及试错(trial and error)。
质粒拷贝数是关于同时稳定并入和产量控制的重要考虑因素。质粒的拷贝数总体上由三个因素决定：质粒的复制起点、质粒的尺寸(包括其中所包含的目标基因)、和培养条件。关于复制起点，质粒可以基于其复制特征，特别是其复制起点，而归类为不相容性组。具体地，质粒总体上包括复制子(replicon)，相应于从单个复制起点起复制的质粒区域。质粒还总体上包括编码识别质粒复制起点并在此启动复制的蛋白质的基因。质粒蛋白与复制起点之间的相互作用决定了复制的特异性和复制子(和因此质粒)的拷贝数。基于关于分离稳定性方面质粒彼此不相容，具有相同复制起点的质粒被归类在同一不相容性组中。具有不同复制起点的质粒可以被归类在不同的不相容性组中——如果质粒彼此相容。关于质粒的尺寸，增加尺寸总体上导致质粒复制和由质粒表达目标基因相关的代谢负担增加，并因此导致质粒的拷贝数减少。关于培养条件，其也影响代谢负担，并且根据具体条件，可以导致拷贝数增加或减少。
在这三个因素中，复制起点总体上是针对具体应用选择质粒的主要考虑因素，因为复制起点建立拷贝数的基线。改变其它两个因素，即质粒的尺寸和培养条件，并非总是一个选择。质粒的尺寸可以由目标基因的尺寸来决定和/或限制。培养条件也可以由获得足量和足够活性的相应产物的需求来决定和/或限制。这也将是经验过程。
突变RNA聚合酶的使用是改变质粒的拷贝数的一种可能的方法。RNA聚合酶在转录中发挥作用。RNA聚合酶还在染色体和质粒的复制中发挥作用。多种细菌中的RNA聚合酶序列已被确定，为鉴定RNA聚合酶内的保守区域提供了基础。例如，Lee et al.，Antimicrobial Agents and Chemotherapy 57:56-65(2013)提供了来自21个菌株的RNA聚合酶β'亚基的C端结构域的比对。细菌RNA聚合酶的结构也已被确定。例如，Mukhopadhyayet al.，Cell 135:295-307(2008)报道了结构揭示，即RNA聚合酶的尺寸为～150埃x～100埃x～100埃，并且形状联想为蟹爪(crab claw)。RNA聚合酶β'亚基构成螯(pincer)(称为“钳”)和活动中心裂口(active center cleft)的部分。
大肠埃希氏菌(Escherichia coli)的编码RNA聚合酶β'亚基的rpoC基因中的两个突变已被报道导致ColE1型质粒的拷贝数减少。具体地，Ederth et al.，MolecularGenetics and Genomics 267:587-592(2002)鉴定了单个氨基酸取代(G1161R)和一个41氨基酸缺失。两者均位于rpoC基因的3'端区域附近。这两个突变分别导致ColE1质粒的拷贝数超过10倍和20倍减少(假设分别相应于超过90％和95％减少)。Ederth等提出自编码DNA聚合酶I的预引物和该预引物的反义抑制子的RNA II和RNA I的启动子的表达改变可引起所述减少。
rpoC中的突变也已被报道引起质粒pBR322的拷贝数增加。具体地，Petersen etal.，Journal of Bacteriology 173:5200-5206(1991)鉴定单个氨基酸取代(G1033D)——其也位于rpoC基因的3'端区域附近——在39℃的半许可生长温度下导致pBR322的拷贝数增加。Petersen指出突变还导致染色体拷贝数增加。
不幸的是，可预见地修饰RNA聚合酶β'亚基以获得改变质粒拷贝数的进一步突变体的通用方法是不存在的。确定具体突变是否以及在何种程度上可以改变质粒的拷贝数也将是经验过程。而且，用于可预见地修饰RNA聚合酶β'亚基以获得不会引起染色体拷贝数相应变化的这种突变体的通用方法是不存在的。
发明内容
技术问题
因此，对于被修饰以改变质粒拷贝数并且理想地不引起染色体拷贝数的相应变化的RNA聚合酶β'亚基突变体的需求是存在的。
问题解决方案
根据本公开的一个方面，公开了包含变体rpoC RNA聚合酶β'亚基蛋白编码序列(也称为“变体rpoC编码序列”)的核酸分子。变体rpoC编码序列编码变体RpoC RNA聚合酶β'亚基蛋白(也称为“变体RpoC”)。变体RpoC包含R47C取代，其中R47C取代的编号基于大肠埃希氏菌的野生型RpoC RNA聚合酶β'亚基蛋白(也称为“野生型RpoC”)来限定。
在一些实例中，相对于包含SEQ ID NO:26的野生型RpoC的表达，变体RpoC的表达使质粒的拷贝数减少。
而且，在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:28的N端结构域、(2)包含SEQ ID NO:29的中央结构域、和(3)包含SEQ ID NO:30的C端结构域。R47C取代存在于N端结构域内。
而且，在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:31的N端结构域、(2)包含SEQ ID NO:32的中央结构域、和(3)包含SEQ ID NO:33的C端结构域。R47C取代存在于N端结构域内。
而且，在一些实例中，变体RpoC包含与SEQ ID NO:27具有至少90％同一性的氨基酸序列。
而且，在一些实例中，变体RpoC包含SEQ ID NO:27。
根据本公开的另一方面，公开了包含该核酸分子的载体。
根据本公开的另一方面，公开了包含该核酸分子的重组微生物。
根据本公开的另一方面，公开了用于调控重组微生物中的对象载体的拷贝数的方法。该方法包括在足以进行对象载体复制的条件下在培养基中培养重组微生物，从而调控对象载体的拷贝数。
根据本公开的另一方面，公开了通过使用重组微生物制备目标产物的方法。该重组微生物包括目标基因载体。该目标基因载体包含用于制备目标产物的目标基因。方法包括以下步骤：(1)在重组微生物表达目标基因的条件下在培养基中培养重组微生物，从而制备目标产物，和(2)从重组微生物和/或培养基回收目标产物。
根据本公开的另一方面，公开了包含变体rpoC编码序列和基因置换序列的基因置换序列。变体rpoC编码序列编码包含SEQ ID NO:28的变体RpoC N端结构域。基因置换序列编码用于用变体rpoC编码序列置换微生物染色体中的内源rpoC编码序列的蛋白质。
发明的有利效果
本文公开的包含变体rpoC编码序列的核酸分子可用于调控载体(如质粒)的拷贝数，因此可用于通过使用该载体而提高目标产物的商业生产。
附图说明
图1显示了通过CLUSTAL O(1.2.4)进行的、下列的N端结构域的多序列比对：嗜热栖热菌(Thermus thermophilus)RpoC(Q8RQE8)(SEQ ID NO:34)、巴氏醋杆菌(Acetobacterpasteurianus)RpoC(BAH99075.1)(SEQ ID NO:35)、淋病奈瑟氏菌(Neisseriagonorrhoeae)RpoC(Q5F5R6)(SEQ ID NO:36)、嗜肺性军团杆菌(Legionella pneumophila)RpoC(Q5X865)(SEQ ID NO:37)、铜绿假单胞菌(Pseudomonas aeruginosa)RpoC(Q9HWC9)(SEQ ID NO:38)、霍乱弧菌(Vibrio cholerae)RpoC(Q9KV29)(SEQ ID NO:39)、大肠埃希氏菌(P0A8T7)(SEQ ID NO:26)、血清型鼠伤寒肠道沙门氏菌(Salmonella enterica serovarTyphimurium)RpoC(P0A2R4)(SEQ ID NO:40)、溶齿放线菌(Actinomyces odontolyticus)RpoC(EDN79927.1)(SEQ ID NO:41)、天蓝色链霉菌(Streptomyces coelicolor)RpoC(Q8CJT1)(SEQ ID NO:42)、白喉棒状杆菌(Corynebacterium diphtheriae)RpoC(Q6NJF6)(SEQ ID NO:43)、结核分支杆菌(Mycobacterium tuberculosis)RpoC(A5U053)(SEQ IDNO:44)、马红球菌(Rhodococcus equi)RpoC(CBH49656.1)(SEQ ID NO:45)、沙眼衣原体(Chlamydia trachomatis)RpoC(O84316)(SEQ ID NO:46)、肉毒梭状芽孢杆菌(Clostridium botulinum)RpoC(A7FZ76)(SEQ ID NO:47)、枯草芽孢杆菌(Bacillussubtilis)RpoC(P37871)(SEQ ID NO:48)、肺炎链球菌(Streptococcus pneumoniae)RpoC(Q97NQ8)(SEQ ID NO:49)、屎肠球菌(Enterococcus faecalis)RpoC(Q82Z41)(SEQ ID NO:50)和短乳杆菌(Lactobacillus brevis)RpoC(Q03PV0)(SEQ ID NO:51)。
图2显示了通过CLUSTAL O(1.2.4)进行的、图1所示RpoC蛋白(分别为SEQ ID NOS:34-39、26和40-51)的中央结构域的多序列比对。
图3A-B显示了通过CLUSTAL O(1.2.4)进行的、图1所示RpoC蛋白(分别为SEQ IDNOS:34-39、26和40-51)的C端结构域的多序列比对。
图4显示了大肠埃希氏菌的RpoC蛋白(SEQ ID NO:26)和变体RpoC(SEQ ID NO:27)的N端结构域的序列比对。
图5显示了大肠埃希氏菌的RpoC蛋白(SEQ ID NO:26)和变体RpoC(SEQ ID NO:27)的比对N端结构域的差异，其中大肠埃希氏菌的RpoC的N端结构域序列被完全提供并且变体RpoC的序列被提供显示差异。
图6A-B示例了构建用于置换染色体上的rpoC序列的重组质粒(称为pJSL47)的过程。
图7A-B示例了构建包括野生型RepFIC复制子的重组质粒(称为pJSL48)的过程。
图8A-B示例了构建包括修饰的RepFIC复制子的重组质粒(称为pJSL49)的过程。
具体实施方式
公开了包含变体rpoC编码序列的核酸分子。变体rpoC编码序列编码变体RpoC。变体RpoC包含R47C取代，其中R47C取代的编号基于大肠埃希氏菌的野生型RpoC RNA聚合酶β'亚基蛋白(也称为“野生型RpoC”)来限定。
惊人地，已确定包含变体rpoC编码序列的核酸分子(其中变体rpoC编码序列编码变体RpoC，并且变体RpoC包含R47C取代)可以用于导致包含该核苷酸序列的重组微生物中的质粒拷贝数显著减少，而不引起染色体拷贝数相应减少。这是惊人的，除其他原因外是因为本文公开的R47C取代发生在RpoC的N端结构域序列中，而如Ederth等和Petersen等所述的包括单一取代的大肠埃希氏菌RpoC突变体仅在rpoC基因的3'端区域附近并且因此在RpoC的C端结构域序列中包括突变。这之所以惊人还因为本文公开的R47C取代发生在九氨基酸残基N末端结构域序列中，该N末端结构域序列不然在各种细菌的RpoC中是严格保守的，而如Ederth等和Petersen等所述的大肠埃希氏菌RpoC突变体中的单一取代发生在RpoC序列内未被保守残基包围的位置并且因此其不发生在保守序列内。
在不希望受理论束缚的情况下认为，来自各种细菌的野生型RpoC包括相应于SEQID NO:26的残基40-48的、在野生型RpoC中严格保守的N端结构域序列。如图1所示，此序列在代表不同种系发生、代谢和环境的如下19种不同细菌中是严格保守的：嗜热栖热菌、巴氏醋杆菌、淋病奈瑟氏菌、嗜肺性军团杆菌、铜绿假单胞菌、霍乱弧菌、大肠埃希氏菌、血清型鼠伤寒肠道沙门氏菌、溶齿放线菌、天蓝色链霉菌、白喉棒状杆菌、结核分支杆菌、马红球菌、沙眼衣原体、肉毒梭状芽孢杆菌、枯草芽孢杆菌、肺炎链球菌、屎肠球菌和短乳杆菌。作为参考，这些序列相应于可从如Lee et al提供的21个菌株的RpoC的C端结构域比对获得的全长RpoC序列。还认为，来自各种细菌的野生型RpoC还包括相应于SEQ ID NO:29的中央结构域序列，其在野生型RpoC中也是严格保守的。如图2所示，此序列在19种不同细菌中也是严格保守的。还认为，来自各种细菌的野生型RpoC还包括相应于SEQ ID NO:30的C端结构域序列，其在野生型RpoC中也是严格保守的。如图3A-B所示，此序列在19种不同细菌中也是严格保守的。
进一步认为，相应于SEQ ID NO:26的残基33-57的较长N端结构域序列、相应于SEQID NO:32的较长中央结构域序列和相应于SEQ ID NO:33的较长C端结构域序列——其分别包括严格保守的N端、中央和C端结构域序列——包括在19种不同细菌的RpoC中总体上保守的多个残基。如图1、图2和图3A-B所示，大肠埃希氏菌RpoC包括这些较长序列。而且，来自其它细菌的RpoC包括与这些较长序列高度相似的序列。
由于来自不同细菌的野生型RpoC包括严格保守的N端、中央和C端结构域序列，认为这些序列在其它细菌的野生型RpoC中也是严格保守的。对于上下文，如表1所示，大肠埃希氏菌的RpoC相比于其它18种不同细菌的RpoC的配对序列比对结果表明相对高度的序列同一性和相似性，即使是细菌的RpoC，如从种系发生、代谢和环境方面远距大肠埃希氏菌的极端嗜热菌嗜热栖热菌。这与RpoC在转录和复制中所起的基本作用是一致的。[表1]大肠埃希氏菌的RpoC相比于其它细菌的RpoC的配对序列比对结果。*
*配对序列比对利用EMBOSS Needle Pairwise Sequence Alignment(PROTEIN)工具采用默认设置(矩阵：BLOSUM62；空隙开放(gap open)：10；空隙延伸：0.5；输出格式：对(pair)；末端空隙罚分(end gap penalty)：错误；末端空隙开放：10；末端空隙延伸：0.5)进行(网址：ebi.ac.uk/Tools/psa/emboss_needle/)。
而且，由于来自不同细菌的野生型RpoC包括与该较长N端、中央和C端结构域序列高度相似的序列，认为其它细菌的野生型RpoC也包括与这些序列高度相似的序列。另外，基于严格保守或总体上保守的各种序列，认为相应的N端、中央和C端结构域序列在结构上和/或功能上在RpoC在转录以及染色体和质粒复制中所起的作用中做出重要贡献。
同样，在不希望受理论束缚的情况下认为，在RpoC中，N端结构域尤其在决定(determining)质粒拷贝数方面发挥重要作用。如图4和图5所示，变体rpoC编码序列包括包含SEQ ID NO:28的N端结构域，其与野生型N端结构域的严格保守序列的区别在于氨基酸第47位处的单个取代，即R(即精氨酸)至C(即半胱氨酸)(也称为“R47C”)，其中编号基于大肠埃希氏菌的野生型RpoC来限定。包括此R47C取代并且在其它方面与大肠埃希氏菌的野生型RpoC相同的变体RpoC呈现质粒拷贝数减少，例如约25％至75％减少。由于N端结构域中的R47C取代是变体RpoC和大肠埃希氏菌野生型RpoC之间的唯一区别，并且由于R47C取代位于较长N端结构域序列内——该较长N端结构域序列包括在19种不同细菌中总体上保守的多个残基，并且由于这19种不同细菌的野生型RpoC中相应于SEQ ID NO:26的残基40-48的严格保守的N端结构域序列中不存在其它取代，N端结构域呈现在决定质粒拷贝数方面是非常重要的。
如本文所用，术语核酸分子意为DNA和/或RNA的分子，包括例如双链DNA分子、单链DNA分子、双链RNA分子、单链RNA分子或DNA/RNA混合分子，其中核酸分子的结构取决于核酸分子是包括DNA序列、RNA序列还是两者。
如本文所用，术语RNA聚合酶β'亚基蛋白意为RNA聚合酶的RNA聚合酶β'亚基。如上所述，RNA聚合酶在转录中发挥作用。RNA聚合酶还在染色体和质粒的复制中发挥作用。RNA聚合酶β'亚基蛋白可以基于与已知RNA聚合酶β'亚基蛋白的结构和/或功能相似性来鉴定，例如基于Lee等所示的序列比对，和/或Mukhopadhyay等讨论的结构特征。RNA聚合酶活性可以例如如Chamberlin et al.，The Journal of Biological Chemistry 254(20):10061-10069(1979)所述来测量。
如本文所用，术语rpoC RNA聚合酶β'亚基蛋白编码序列意为编码RNA聚合酶β'亚基蛋白的序列的DNA分子链或DNA分子链的部分。
如本文所用，术语野生型RNA聚合酶β'亚基蛋白意为在天然条件下在某个物种的个体中存在的RNA聚合酶β'亚基蛋白。
如本文所用，术语N端结构域意为存在于蛋白质的N端处或附近，例如蛋白质的氨基酸序列的前三分之一内，的蛋白质部分。
如本文所用，术语中央结构域意为存在于蛋白质的中央处或附近，例如蛋白质的氨基酸序列的中间三分之一内，的蛋白质部分。
如本文所用，术语C端结构域意为央蛋白质的C端处或附近，例如蛋白质的氨基酸序列的后三分之一内，的蛋白质部分。
如本文所用，术语复制子意为DNA分子的从单个复制起点起复制的区域。
如本文所用，术语载体意为可存在于微生物中(天然地或通过引入微生物)的核酸分子，如质粒、病毒载体、粘粒或人工染色体。
如本文所用，术语质粒意为可存在于微生物中(天然地或通过引入微生物)、与微生物的染色体(一个或多个)物理上分开并且独立于染色体(一个或多个)复制的核酸分子。如上所述，质粒通常是双链环状DNA分子，但也可以是线性DNA分子和/或RNA分子。质粒以约1kb至大于2Mb的尺寸范围存在。质粒还以从低拷贝数至高拷贝数范围的拷贝数/细胞存在。质粒可以被修饰以包括目标基因。
如本文所用，术语质粒拷贝数意为质粒在微生物细胞中的拷贝数。质粒在微生物中的质粒拷贝数可以被测量，例如通过利用实时PCR将以单个拷贝存在于质粒上的基因的拷贝数相对于以单个拷贝存在于微生物染色体上的基因进行比较和其它方法。
如本文所用，术语质粒拷贝数的调控因子意为诸如RNA分子或蛋白质的引起微生物中质粒拷贝数变化的因子，例如当微生物中存在该因子时比当微生物中不存在该因子时增加或减少。通过调控质粒拷贝数，可以稳定表达质粒，从而能够实现包括该质粒的微生物的稳定生长。
如述，公开了包含变体rpoC编码序列的核酸分子。该核酸分子可以是例如双链DNA分子，如其中已引入变体rpoC编码序列的染色体DNA，或其中已克隆变体rpoC编码序列的质粒。
还如述，变体rpoC编码序列编码变体RpoC。变体RpoC是基于包含R47C取代的变体，其中R47C取代的编号基于大肠埃希氏菌的野生型RpoC来限定。变体RpoC是一种RpoC，因此在转录、染色体复制以及质粒复制中发挥作用。正如所述19种不同细菌的野生型RpoC例如在非保守氨基酸位置处彼此相异，变体RpoC也可以根据变体rpoC编码序列的来源而变化。因此，例如，变体RpoC可以包括R47C取代，并且除此之外与大肠埃希氏菌的野生型RpoC具有至少90％的同一性。又例如，变体RpoC可以包括R47C取代，并且除此之外与其它18种不同细菌(即嗜热栖热菌、巴氏醋杆菌、淋病奈瑟氏菌、嗜肺性军团杆菌、铜绿假单胞菌、霍乱弧菌、血清型鼠伤寒肠道沙门氏菌、溶齿放线菌、天蓝色链霉菌、白喉棒状杆菌、结核分支杆菌、马红球菌、沙眼衣原体、肉毒梭状芽孢杆菌、枯草芽孢杆菌、肺炎链球菌、屎肠球菌、或短乳杆菌)中的任一种的野生型RpoC具有至少90％的同一性。又例如，变体RpoC可以包括R47C取代以及大肠埃希氏菌或其它18种不同细菌的野生型RpoC中的一种或多种的一个或多个部分。
在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:28的N端结构域、(2)包含SEQID NO:29的中央结构域、和(3)包含SEQ ID NO:30的C端结构域，其中R47C取代存在于N端结构域内。在这些实例中，变体RpoC包含相应于SEQ ID NO:28的N端结构域序列，其包括R47C取代。变体RpoC还包括相应于SEQ ID NO:29的严格保守的中央结构域序列和相应于SEQ IDNO:30的严格保守的C端结构域序列，与RpoC在转录以及染色体和质粒复制中所起的作用一致。
在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:31的N端结构域、(2)包含SEQID NO:32的中央结构域和(3)包含SEQ ID NO:33的C端结构域，其中R47C取代存在于N端结构域内。在这些实例中，变体RpoC包含相应于SEQ ID NO:31的N端结构域序列，其包括R47C取代。变体RpoC还包括相应于SEQ ID NO:32的总体上保守的较长中央结构域序列和相应于SEQ ID NO:33的总体上保守的较长C端结构域序列，也与RpoC在转录以及染色体和质粒复制中所起的作用一致。
在一些实例中，变体RpoC包含与SEQ ID NO:27具有至少90％同一性的氨基酸序列。作为参考，SEQ ID NO:27相应于包括R47C取代并且除此之外与大肠埃希氏菌的野生型RpoC相同的变体RpoC。同样作为参考，变体RpoC的氨基酸序列与SEQ ID NO:27之间的序列同一性百分比可以通过进行配对序列比对来确定。这可以使用EMBOSS Needle PairwiseSequence Alignment(PROTEIN)工具利用默认设置(矩阵：BLOSUM62；空隙开放：10；空隙延伸：0.5；输出格式：对；末端空隙罚分：错误；末端空隙开放：10；末端空隙延伸：0.5)来完成(网址：ebi.ac.uk/Tools/psa/emboss_needle/)。这也可以使用其它类似的配对序列比对工具来完成。
变体RpoC的氨基酸序列可以不同于SEQ ID NO:27，例如主要或完全基于大肠埃希氏菌的野生型RpoC与其它18种不同细菌的RpoC之间的非保守氨基酸残基的取代。参考表1，尽管大肠埃希氏菌的RpoC与其它18种不同细菌的RpoC相比的配对序列比对结果指示了相对高度的序列同一性和相似性，但该结果还表明与大肠埃希氏菌的野生型RpoC相比，所述不同细菌中17种的RpoC具有36.1％至82.4％范围内的序列同一性，因此显著低于90％。相似蛋白质之间的非保守氨基酸残基的取代总体上更可能被耐受，例如与保守氨基酸残基的取代相比不破坏结构和/或功能。表1的结果表明RpoC包括多个非保守的并且因此可适于取代的氨基酸残基。
变体RpoC的氨基酸序列也可以不同于SEQ ID NO:27——例如基于包括一些或多个保守取代，意为相对于SEQ ID NO:27，氨基酸残基被另一结构相似的氨基酸残基取代。保守取代通常包括以下分组中的取代：(1)甘氨酸和丙氨酸、(2)缬氨酸、异亮氨酸和亮氨酸、(3)天冬氨酸和谷氨酸、(4)天冬酰胺和谷氨酰胺、(5)丝氨酸和苏氨酸、(6)赖氨酸和精氨酸、和(7)苯丙氨酸和酪氨酸。与非保守取代相比，保守取代总体上更可能被耐受。
因此，在这些实例中，变体RpoC包括R47C取代。变体RpoC还包含与SEQ ID NO:27具有至少90％同一性的氨基酸序列。例如，变体RpoC可以包含与SEQ ID NO:27具有至少91％、92％、93％、94％、95％、96％、97％、98％、99％或100％同一性的氨基酸序列。
在一些实例中，变体RpoC包含SEQ ID NO:27。在一些实例中，变体RpoC由SEQ IDNO:27组成。
在一些实例中，相对于包含SEQ ID NO:26的野生型RpoC的表达，变体RpoC的表达使质粒的拷贝数减少。作为参考，SEQ ID NO:26相应于大肠埃希氏菌的野生型RpoC。如上所述，变体RpoC是一种RpoC，并且因此在转录、染色体复制以及质粒复制中发挥作用。还如述，包含R47C取代并且除此之外与大肠埃希氏菌的野生型RpoC相同的变体RpoC呈现质粒拷贝数减少例如约25％至75％。在一些实例中，相对于包含SEQ ID NO:26的野生型RpoC的表达，变体RpoC的表达使质粒的拷贝数减少10％至80％，例如25％至75％、30％至70％、35％至65％、40％至60％、45％至55％、10％至40％、20％至50％、30％至60％、40％至70％、或50％至80％。
还公开了包含核酸分子的载体。该核酸分子可以如上所述。在一些实例中，载体可以相应于质粒、病毒载体、粘粒或人工染色体中的一种或多种。
还公开了包含核酸分子的重组微生物。该核酸分子可以如上所述。因此，在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:28的N端结构域、(2)包含SEQ ID NO:29的中央结构域、和(3)包含SEQ ID NO:30的C端结构域，其中R47C取代存在于N端结构域内，如上所述。在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:31的N端结构域、(2)包含SEQ ID NO:32的中央结构域、和(3)包含SEQ ID NO:33的C端结构域，其中R47C取代存在于N端结构域内，如上所述。在一些实例中，变体RpoC包含与SEQ ID NO:27具有至少90％同一性的氨基酸序列，如上所述。在一些实例中，变体RpoC包含SEQ ID NO:27，如上所述。
在一些实例中，相对于对照微生物中包含SEQ ID NO:26的野生型RpoC的表达，重组微生物中变体RpoC的表达使质粒的拷贝数减少。对照微生物可以例如源自与重组微生物相同的属、种和/或菌株，并且可以包括相似或相同的质粒，因此除了关于重组微生物的变体rpoC编码序列与对照微生物的相应rpoC序列之间的差异，可以是在种系发生上相似的、紧密相关的和/或在遗传上相同的。如上所述类似地，所述减少可以是10％至80％，例如25％至75％、30％至70％、35％至65％、40％至60％、45％至55％、10％至40％、20％至50％、30％至60％、40％至70％或50％至80％。
例如，该包含核酸分子的重组微生物可以例如通过以下获得：通过转化、接合(conjugation)或转导将完整变体rpoC编码序列(例如克隆在载体中)引入前体微生物以获得重组微生物，然后在重组微生物中维持完整变体rpoC编码序列，例如通过载体选择。这可以通过标准分子生物学技术来完成。作为参考，载体可以相应于质粒、病毒载体、粘粒或人工染色体中的一种或多种。因此，在一些实例中，重组微生物可以通过转化、接合或转导中的一种或多种而将包含核酸序列的变体rpoC编码序列载体(例如质粒)引入前体微生物来制备。
包含核酸分子的重组微生物还可以例如通过以下获得：引入变体rpoC编码序列的一部分(例如克隆在载体中)，并使用该部分置换内源染色体野生型rpoC编码序列的相应部分，例如通过同源重组进行基因置换，例如通过使用sacB载体。这也可以通过标准分子生物学技术来完成。因此，在一些实例中，重组微生物包含染色体，并且基于内源rpoC编码序列被变体rpoC编码序列置换，变体rpoC编码序列存在于染色体中。
重组微生物可以由各种细菌中的某个细菌来制备。如上所述，基于各种N端、中央和C端结构域序列在所述19种不同细菌中是严格或总体上保守的，认为相应的序列在结构上和/或功能上在RpoC在各种细菌的转录以及染色体和质粒复制中所起的作用中做出重要贡献。而且，认为在RpoC中，N端结构域尤其在决定质粒的拷贝数方面发挥重要作用。因此，在一些实例中，重组微生物可以由以下细菌中的一种或多种制备：栖热菌属(Thermus)(例如，嗜热栖热菌)、醋菌属(Acetobacter)(例如，巴氏醋杆菌)、奈瑟氏菌属(Neisseria)(例如，淋病奈瑟氏菌)、军团杆菌属(Legionella)(例如，嗜肺性军团杆菌)、假单胞菌属(Pseudomonas)(例如，铜绿假单胞菌)、弧菌属(Vibrio)(例如，霍乱弧菌)、埃希氏菌属(Escherichia)(例如，大肠埃希氏菌)、沙门氏菌属(Salmonella)(例如，血清型鼠伤寒肠道沙门氏菌)、放线菌属(Actinomyces)(例如，溶齿放线菌)、链霉菌属(Streptomyces)(例如，天蓝色链霉菌)、棒状杆菌属(Corynebacterium)(例如，白喉棒状杆菌)、分枝杆菌属(Mycobacterium)(例如，结核分支杆菌)、红球菌属(Rhodococcus)(例如，马红球菌)、衣原体属(Chlamydia)(例如，沙眼衣原体)、梭菌属(Clostridium)(例如，肉毒梭状芽孢杆菌)、杆菌属(Bacillus)，(例如，枯草芽孢杆菌)、链球菌属(Streptococcus)(例如，肺炎链球菌)、肠道球菌属(Enterococcus)(例如，屎肠球菌)、或乳杆菌属(Lactobacillus)(例如，短乳杆菌)。
如上所述，包括R47C取代并且除此之外与大肠埃希氏菌的野生型RpoC相同的变体RpoC呈现质粒拷贝数减少例如约25％至75％。如下所述，这已经在各种大肠埃希氏菌菌株中实现。因此，包含变体rpoC编码链的核酸分子可以调控质粒拷贝数，具体地在埃希氏菌属的细菌中，特别是大肠埃希氏菌种的细菌中。因此，在一些实例中，重组微生物可以由埃希氏菌属细菌或大肠埃希氏菌种细菌中的一种或多种制备。
还公开了用于调控重组微生物中的对象载体的拷贝数的方法。还公开了变体rpoC编码序列对于调控重组微生物中的对象载体的拷贝数的应用。重组微生物可以如上所述。因此，在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:28的N端结构域、(2)包含SEQ IDNO:29的中央结构域、和(3)包含SEQ ID NO:30的C端结构域，其中R47C取代存在于N端结构域内，如上所述。在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:31的N端结构域、(2)包含SEQ ID NO:32的中央结构域、和(3)包含SEQ ID NO:33的C端结构域，其中R47C取代存在于N端结构域内，如上所述。在一些实例中，变体RpoC包含与SEQ ID NO:27具有至少90％同一性的氨基酸序列，如上所述。在一些实例中，变体RpoC包含SEQ ID NO:27，如上所述。在一些实例中，重组微生物可以由埃希氏菌属细菌或大肠埃希氏菌种细菌中的一种或多种制备。在一些实例中，相对于对照微生物中包含SEQ ID NO:26的野生型RpoC的表达，重组微生物中变体RpoC的表达使质粒的拷贝数减少例如10％至80％，例如25％至75％、30％至70％、35％至65％、40％至60％、45％至55％、10％至40％、20％至50％、30％至60％、40％至70％或50％至80％。
该方法包括在足以进行对象载体复制的条件下在培养基中培养重组微生物，从而调控对象载体的拷贝数。如上所述，载体可以相应于质粒、病毒载体、粘粒或人工染色体中的一种或多种。而且如述，RpoC在质粒的复制中发挥作用。因此，在一些实例中，对象载体包括质粒。
培养可以通过标准微生物学技术进行，例如在培养管、烧瓶和/或生物反应器中，其细节对于本领域普通技术人员而言将是显而易见的。培养可以在合适的培养基中进行，例如富营养物的培养基或最低限度培养基，其细节对于本领域普通技术人员而言也将是显而易见的。培养可以在合适的培育温度下进行，例如，处于或大约25-38℃、28-37℃或37℃，其细节对于本领域普通技术人员而言也将是显而易见的。随着重组微生物在培养过程中生长和分裂，载体将复制。因此，例如，关于由大肠埃希氏菌制备的重组微生物，可以通过发酵技术在生物反应器中分批或连续培养重组微生物。培养可以在最低限度培养基中进行，例如包括限定量的盐(如M9最低限度盐培养基)以及一种或多种碳源(例如葡萄糖、蔗糖、或木质纤维素材料)等的培养基。培养可以在约37℃下进行。这种条件支持大肠埃希氏菌的生长和分裂，因此将支持载体的复制。其他适于培养由大肠埃希氏菌以及其它微生物制备的重组微生物的条件是本领域普通技术人员已知的和将显而易见的。
还公开了通过使用重组微生物来制备目标产物的方法。还公开了重组微生物对于制备目标产物的应用。同样，重组微生物可以如上所述。因此，在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:28的N端结构域、(2)包含SEQ ID NO:29的中央结构域和(3)包含SEQID NO:30的C端结构域，其中R47C取代存在于N端结构域内，如上所述。在一些实例中，变体RpoC包含：(1)包含SEQ ID NO:31的N端结构域、(2)包含SEQ ID NO:32的中央结构域和(3)包含SEQ ID NO:33的C端结构域，其中R47C取代存在于N端结构域内，如上所述。在一些实例中，变体RpoC包含与SEQ ID NO:27具有至少90％同一性的氨基酸序列，如上所述。在一些实例中，变体RpoC包含SEQ ID NO:27，如上所述。在一些实例中，重组微生物可以由埃希氏菌属细菌或大肠埃希氏菌种细菌中的一种或多种制备。在一些实例中，相对于对照微生物中包含SEQ ID NO:26的野生型RpoC的表达，重组微生物中变体RpoC的表达使质粒的拷贝数减少例如10％至80％，例如25％至75％、30％至70％、35％至65％、40％至60％、45％至55％、10％至40％、20％至50％、30％至60％、40％至70％或50％至80％。
根据此方法，重组微生物包括目标基因载体，并且该目标基因载体包含用于制备目标产物的目标基因。载体可以是例如重组质粒，该重组质粒包括来自生物体(如细菌、古细菌或真核生物域的微生物、动物、和/或植物和其它生物体)的目标基因。具体地关于细菌域的微生物，目标基因可以来自例如埃希氏菌属的细菌(如大肠埃希氏菌)、棒状杆菌属的细菌(如谷氨酸棒状杆菌)、或杆菌属的细菌(如枯草芽孢杆菌)等。多种用于基因工程的技术已被开发，允许重组表达各种生物体(包括细菌、古细菌和真核生物域的微生物以及动物和植物和其它生物体)的基因。对于本领域普通技术人员将显而易见，根据本公开，这种技术可以用于在重组微生物中克隆和表达来自各种生物体的目标基因。
在一些实例中，目标产物包含以下一种或多种：(i)目标RNA、(ii)目标蛋白、(iii)目标生物材料、(iv)目标聚合物、其前体和/或用于其生产的酶、(v)目标增甜剂、其前体和/或用于其生产的酶、(vi)目标油、其前体和/或用于其生产的酶、(vii)目标脂肪、其前体和/或用于其生产的酶、(viii)目标多糖、其前体和/或用于其生产的酶、(ix)目标氨基酸、其前体和/或用于其生产的酶、(x)目标核苷酸、其前体和/或用于其生产的酶、(xi)目标疫苗、其前体和/或用于其生产的酶、或(xii)目标药物产品、其前体和/或用于其生产的酶。因此，在一些实例中，目标基因编码目标RNA，并且目标产物相应于目标RNA。而且，在一些实例中，目标基因编码目标蛋白，并且目标产物相应于目标蛋白。而且，在一些实例中，目标基因编码目标RNA和/或目标蛋白，并且目标RNA和/或目标蛋白进而在生产目标生物材料、目标聚合物、其前体和/或用于其生产的酶、目标增甜剂、其前体和/或用于其生产的酶、目标油、其前体和/或用于其生产的酶、目标脂肪、其前体和/或用于其生产的酶、目标多糖、其前体和/或用于其生产的酶、目标氨基酸、其前体和/或用于其生产的酶、目标核苷酸、其前体和/或用于其生产的酶、目标疫苗、其前体和/或用于其生产的酶、或目标药物产品、其前体和/或用于其生产的酶中发挥作用。例如，关于目标聚合物、目标增甜剂、目标油、目标脂肪、目标多糖、目标氨基酸和/或目标核苷酸，目标基因可以编码目标蛋白，该目标蛋白相应于，直接地或通过前体，生产目标聚合物、目标增甜剂、目标油、目标脂肪、目标多糖、目标氨基酸和/或目标核苷酸的酶。又例如，关于疫苗，目标基因可以编码目标蛋白，该目标蛋白相应于可以用作抗病原性微生物疫苗组分的抗原或抗原片段，例如蛋白亚基、受体或病原性微生物的其它蛋白质或其片段。又例如，关于目标药物产品，目标基因可以编码目标蛋白，该目标蛋白可以用作药物产品的组分，如抗体、受体或激素。
在一些实例中，载体包含多个目标基因，例如来自具体生物体的多个目标基因和/或来自多个生物体中的每一个的一个或多个目标基因。而且，在一些实例中，目标基因用于制备多种目标产物。
在一些实例中，目标基因载体，例如质粒，具有3至120kb的尺寸。重组载体通常以3至120kb的尺寸存在，因为这些是其中已克隆目标基因的载体的一般尺寸。
方法包括以下步骤：(1)在重组微生物表达目标基因的条件下在培养基中培养重组微生物，从而制备目标产物。同样，培养可以通过标准微生物学技术进行，例如在培养管、烧瓶和/或生物反应器中，在合适的培养基(例如富营养物的培养基或最低限度培养基)中，在合适的培育温度下，例如处于或大约25-38℃、28-37℃、或37℃，其细节对于本领域普通技术人员而言将是显而易见的。
方法还包括以下步骤：(2)从重组微生物和/或培养基回收目标产物。适于回收目标产物的方法可以基于目标产物的细节而建立，例如相应于目标蛋白的目标产物的标准蛋白质纯化技术、或相应于目标聚合物的目标产物的标准聚合物提取和沉淀技术和其它方式，其细节对于本领域普通技术人员而言将是显而易见的——根据例如目标产物的类型(例如RNA、蛋白质、聚合物等)、目标产物的具体细节(例如化学结构、分子量、亲和标签等)和期望的纯度(例如低至高)。例如，关于相应于目标RNA的目标产物，在培养重组微生物后，可以通过利用如Stead et al.,Nucleic Acids Research 40(20),e156:1-9(2012)所述的RNAsnap(TM)方法，或通过商业可得的方法，如TRIzol(R)Max(TM)细菌RNA分离试剂盒(ThermoFisher Scientific)、RNeasy(R)Protect细菌分离试剂盒(Qiagen)或RiboPure(TM)细菌RNA分离(ThermoFisher Scientific)和其它本领域已知的方法从重组微生物回收目标RNA。对于相应于目标蛋白的目标产物，可以按照本领域公知的程序，通过提取、离子交换色谱法、亲和色谱法和/或通过沉淀浓缩，从重组微生物回收目标蛋白。对于相应于目标聚合物的目标产物，同样按照本领域公知的程序，可以通过提取、洗涤和浓缩来回收目标聚合物，其中用于洗涤的组合物和浓缩的沉淀剂基于化学结构和分子量来选择。对于相应于目标增甜剂、目标油、目标脂肪、目标多糖、目标氨基酸或目标核苷酸的目标产物，如果目标产物在重组微生物中积累，则也可以使用类似的方法，而如果目标产物在细胞外积累，则可以例如通过从培养基沉淀(析出，precipitation)而回收目标产物，同样是按照本领域公知的程序。对于相应于目标疫苗或目标药物产品的目标产物，目标产物可以例如如上文关于目标蛋白所述来回收，例如对于相应于抗原或抗原片段的目标疫苗而言基于离子交换色谱法，或对于相应于单克隆抗体的药物产品而言基于亲和色谱法，以及其它方法，同样按照本领域公知的程序。
在一些实例中，可以通过转化、接合或转导中的一种或多种而将目标基因载体引入重组微生物，来制备重组微生物，如上所述。
更详细地考虑目标产物，在一些实例中，目标产物包含以下中的一种或多种：(i)目标聚合物、其前体和/或用于其生产的酶，或(ii)目标生物聚合物、其前体和/或用于其生产的酶。基于对重组微生物中多个目标基因的协调引入和表达的需求，特别是对于基于具有复杂化学结构的单体的聚合物和/或包括两种或更多种单体的共聚物的需求，包括生物聚合物在内的聚合物的生物学生产可以是挑战性的。类似的考虑关于上述其它目标产物，特别是目标增甜剂、目标脂肪、目标多糖、目标氨基酸、目标核苷酸、目标疫苗和目标药物产品也适用。
该方法可以有效用于快速确定包含多个目标基因(例如用于生产目标聚合物、目标生物聚合物、目标增甜剂、目标脂肪、目标多糖、目标氨基酸、目标核苷酸、目标疫苗或目标药物产品)的载体的适当拷贝数，以平衡载体在重组微生物中的稳定并入和目标基因产物的产量控制。可以制备一组载体。载体可以包括一个或多个目标基因。载体可以相对于其基线拷贝数变化。可以将该载体引入第一细菌菌株，例如包含野生型rpoC编码序列因此表达野生型RpoC的大肠埃希氏菌菌株，以获得包括该载体并表达野生型RpoC的第一组大肠埃希氏菌菌株。还可以将载体引入相应第二重组细菌菌株，例如包含编码包含R47C取代的变体RpoC的变体rpoC编码序列因此表达变体RpoC的重组大肠埃希氏菌菌株，该第二菌株除此之外与该第一菌株相同，从而获得包括该载体并表达变体RpoC的第二组相应大肠埃希氏菌菌株。该方法可以通过以下进行：在菌株表达所述一个或多个目标基因的条件下在培养基中培养第一组和第二组菌株，从而制备目标产物，和回收目标产物。可以在培养过程中确定各载体的拷贝数。可以在回收过程中确定目标产物的产量或其它相关特性。此方法可以显著降低对于第一组菌株所能实现的拷贝数下限，这可有利于在目标基因的表达有害的情况下(例如编码对细胞有毒的目标RNA和/或目标蛋白的目标基因，当在细胞中被表达超过某些水平时)维持菌株细胞的活力。此方法还可以有效地翻倍样品尺寸——关于测试载体拷贝数对目标产物产量的效果。此方法可以具体地用于相应于质粒(包括尺寸为3至120kb的和/或包括多个目标基因的质粒)的载体。
还公开了包含变体rpoC编码序列和基因置换序列的基因置换载体。变体rpoC编码序列编码包含SEQ ID NO:28的变体RpoC N端结构域。在这种情况下，变体rpoC编码序列无需是全长变体rpoC编码序列，并且优选地仅包括变体rpoC编码序列的足以与基因置换载体所要引入的微生物的染色体中的内源rpoC编码序列进行同源重组的(长度)。变体rpoC编码序列可以包括例如全长变体rpoC编码序列的0.2至5kb、0.5至3kb、0.7至2.5kb、0.8至2kb、0.9至1.5kb或约1kb。
基因置换序列编码用于用变体rpoC编码序列置换微生物的染色体中的内源rpoC编码序列的蛋白质。在一些实例中，基因置换序列包含sacB基因，并且蛋白质包含SacB。示例性sacB载体包括如Link et al.，Journal of Bacteriology 179:6228-6237(1997)和以下网址：arep.med.harvard.edu/labgc/pko3.html中所述的pKO3和pKOV。
基因置换载体可以通过将变体rpoC编码序列，例如全长变体rpoC编码序列的0.2至5kb、0.5至3kb、0.7至2.5kb、0.8至2kb、0.9至1.5kb或约1kb，克隆到前体载体中来制备。前体载体可以包括基因置换序列，例如编码蛋白质(例如SacB)的sacB基因，用于用变体rpoC编码序列置换微生物的染色体中的内源rpoC编码序列。
通过标准分子生物学技术，基因置换载体可以用于用包含SEQ ID NO:28的变体rpoC编码序列置换微生物的染色体中的内源rpoC编码序列。sacB载体对于基因置换的应用也被Link等和网址：arep.med.harvard.edu/labgc/pko3.html描述。
发明方式
实施例
实施例1：构建用于置换染色体上的rpoC序列的质粒
(1)制备rpoC片段和sacB载体。
为了扩增含有具有修饰核苷酸序列(SEQ ID NO:1)的部分变体rpoC序列的两个0.5kb DNA片段，利用QIAGEN Genomic-tip系统提取从Coli Genetic Stock Center(CGSC)获得的大肠埃希氏菌菌株LS5218(菌株6966)的基因组DNA(gDNA)，并利用该gDNA作为模板以及PfuUltra II Fusion HS DNA聚合酶(由Agilent制造)进行聚合酶链反应(PCR)。相应的修饰的RpoC蛋白(由核苷酸序列推导)是SEQ ID NO:27。修饰的RpoC蛋白序列考虑，rpoC核苷酸序列包括替代性起始密码子GTG代替ATG。尽管GTG总体上编码缬氨酸，但GTG作为替代性起始密码子编码甲硫氨酸。利用SEQ ID NO:3和SEQ ID NO:4的引物如下进行PCR：30个循环的在95℃下变性30秒，在56℃下退火30秒，和在72℃下延伸30秒。SEQ ID NO:5和SEQID NO:6的引物用于在72℃下延伸30秒，进行另一PCR。将混合物用QIAGEN纯化试剂盒纯化，然后洗脱以获得两种不同的0.5kb DNA片段。
为了制备含有sacB基因和R6K起点的基因置换载体(图6A-B)，用限制酶EcoRV消化pSKH130。将PCR混合物和EcoRV消化的反应混合物用QIAGEN纯化试剂盒纯化，然后洗脱以获得第一0.5kb DNA片段、第二0.5kb DNA片段和4.7kb载体DNA片段(也称为“sacB载体切割段”)。
(2)构建用于置换rpoC序列的质粒。
将实施例1-(1)中描述的第一0.5kb DNA片段、第二0.5kb DNA片段和sacB载体切割段用于构建pJSL47。利用NEBuilder HiFi DNA Assembly Master Mix(由NEB制造)和BW25113(其是Coli Genetic Stock Center(CGSC)的菌株7636)构建pJSL47质粒。
(3)制备重组大肠埃希氏菌CC06-9642。
为了用变体rpoC序列(SEQ ID NO:1)取代大肠埃希氏菌LS5218的染色体上的rpoC(SEQ ID NO:7)，将pJSL47质粒通过电穿孔引入大肠埃希氏菌菌株LS5218中，然后选择在含有50mg/L卡那霉素的Luria-Bertani(LB)琼脂板上生长的单个菌落。利用SEQ ID NO:8和SEQ ID NO:9的引物，通过PCR确认pJSL47插入到所选菌落的染色体中。使选择的菌株在缺少NaCl但含有10％蔗糖的LB琼脂板上生长，以“弹出(pop out)”sacB基因和R6K起点。通过PCR和序列确认，验证转化体的LS5218 rpoC(SEQ ID NO:7)被变体rpoC序列(SEQ ID NO:1)置换。具有正确基因型的所得菌株被命名为大肠埃希氏菌CC06-9642。
实施例2：测量质粒拷贝数
野生型菌株LS5218和菌株CC06-9642均含有F样质粒(67,502bp)。创建CC06-9642后，通过PCR方法确认F样质粒的存在。所用引物是例如SEQ ID NO:10和11。当创建CC06-9642时，还创建了CC06-9637。其之间的区别是CC06-9642含有F样质粒，但CC06-9637不含有F样质粒。CC06-9637的RpoC也是变体rpoC。
利用实时PCR(也称为“qPCR”)方法测量这两个菌株的质粒拷贝数，该实时PCR方法利用SYBR(R)Green I染料通过结合PCR过程中形成的双链DNA来检测PCR产物。方案采用Fast SYBR(R)Green Cells-to-Ct(TM)试剂盒，在Applied Biosystems 7500Fast实时PCR系统上，以“一锅法(one-pot)”进行细胞裂解和PCR反应。
为了制备细胞裂解物，将在LB肉汤中各菌株的过夜培养物用冷(4℃)1xPBS缓冲剂稀释，然后添加裂解液、终止液(Fast SYBR(R)Green Cells-to-Ct试剂盒，目录号4402956)和RNA酶A(Life Technologies，目录号12091-021，20mg/ml)。通过将4μL细胞裂解物添加至16μL PCR混合物来制备qPCR反应混合物，其组成如表2所示。
[表2]
PCR混合物组成
LS5218和CC06-9642细胞样品中的F样质粒的拷贝数由质粒上的标记物DNA序列(具体地RepFIA和RepFIC)相对于编码β-半乳糖苷酶的单拷贝染色体lacZ基因的相对丰度来估测。用于RepFIA的引物是SEQ ID NO:24和SEQ ID NO:25。用于RepFIC的引物是SEQ IDNO:10和SEQ ID NO:11。用于lacZ的引物是SEQ ID NO:12和SEQ ID NO:13。
利用7500Fast实时PCR默认程序如下进行实时PCR反应：1个循环的在95℃下酶活化20秒，40个循环的在95℃下变性3秒，在60℃下退火和延伸30秒，以及解离曲线。
通过计算2DCt来测量质粒拷贝数，其中DCt通过从lacZ基因Ct值减去RepFIC Ct值来计算(DCt＝Ct_lacZ-Ct_repFIC.)。
[表3]
利用qPCR的拷贝数测量结果。
如表3所示，CC06-9642的F样质粒的拷贝数低于对照LS5218。因此，确认变体rpoC序列导致F样质粒的拷贝数减少。当质粒拷贝数过多时，可在微生物的细胞中导致代谢负担。以上结果表明变体rpoC可以具有调控质粒拷贝数的功能，并且因此从以上结果可知，该菌株可以稳定生长，并且质粒可以被稳定表达。
实施例3：构建含有RepFIC复制子的修饰DNA序列的质粒
(1)制备RepFIC片段和卡那霉素抗性基因片段。
为了扩增含有RepFIC复制子(SEQ ID NO:14)的5.2kb DNA片段，利用QIAGENGenomic-tip系统提取大肠埃希氏菌LS5218的基因组DNA(gDNA)，并利用该gDNA作为模板以及PfuUltra II融合HS DNA聚合酶(由Agilent制造)进行聚合酶链反应(PCR)。利用SEQ IDNO:15和SEQ ID NO:16的引物如下进行PCR：30个循环的在95℃下变性30秒，在56℃下退火30秒和在72℃下延伸5分钟。
为了扩增含有卡那霉素抗性基因的1.4kb DNA片段，利用质粒pKD4作为模板以及PfuUltra II Fusion HS DNA聚合酶进行PCR。利用SEQ ID NO:17和SEQ ID NO:18的引物如下进行PCR：30个循环的在94℃下变性30秒，在56℃下退火30秒和在72℃下延伸1分30秒。
在PCR反应完成并且然后混合后，将1.3mL的DpnI和5.7ml的来自Thermo FisherScientific的10X缓冲剂Tango(目录号ER1701)添加至每一个50μL PCR混合物，然后将其在37℃下培育1小时，以去除模板DNA。将混合物用QIAGEN纯化试剂盒纯化，然后洗脱以获得5.2kb DNA片段(也称为“RepFIC片段”)和1.4kb DNA片段(也称为“KanR片段”)。
(2)制备含有修饰序列的RepFIC片段。
已获得修饰的RepFIC复制子(SEQ ID NO:19)，其包括相对于大肠埃希氏菌LS5218的RepFIC复制子的单个核苷酸取代并导致质粒拷贝数增加。为了扩增含有修饰RepFIC复制子(SEQ ID NO:19)的5.2kb DNA片段，利用大肠埃希氏菌LS5218的gDNA作为模板以及PfuUltra II Fusion HS DNA聚合酶进行PCR。利用SEQ ID NO:15和SEQ ID NO:20的引物如下进行PCR：30个循环的在95℃下变性30秒，在56℃下退火30秒和在72℃下延伸3分钟。利用SEQ ID NO:21和SEQ ID NO:16的引物用于在72℃下延伸2分30秒，进行另一PCR。
在PCR反应完成后，将1.3mL的DpnI和5.7ml的来自Thermo Fisher Scientific的10X缓冲剂Tango(目录号ER1701)添加至50μL的PCR反应混合物中，然后将其在37℃培育1小时以去除模板DNA。将混合物用QIAGEN纯化试剂盒纯化，然后洗脱以获得2.8kb DNA片段和2.4kb DNA片段。
(3)构建含有RepFIC复制子的野生型或修饰序列的质粒。
将实施例3-(1)中描述的RepFIC片段和KanR片段用于构建质粒，称为pJSL48。利用NEBuilder HiFi DNA AssemblyMaster Mix(由NEB制造)构建pJSL48质粒，如图7A-B所示。
将实施例3-(2)中描述的2.8kb和2.4kb DNA片段和实施例3-(1)中描述的KanR片段用于构建另一质粒，称为pJSL49。通过NEBuilder HiFi DNA AssemblyMaster Mix进行三个片段的Gibson组装，如图8A-B所示。
实施例4：测量质粒拷贝数
为了测量质粒拷贝数，将质粒pJSL48和pJSL49引入大肠埃希氏菌LS5218中，产生大肠埃希氏菌菌株CC06-9665和CC06-9666。将质粒pJSL48和pJSL49也引入CC06-9642中，分别产生大肠埃希氏菌菌株CC06-9638和CC06-9639。
如实施例2中所述，利用实时PCR方法测量大肠埃希氏菌菌株CC06-9665、9666、9638和9639中的质粒拷贝数。
细胞样品中的质粒拷贝数由质粒RepFIC复制子上的标记物DNA序列相对于编码β-半乳糖苷酶的单拷贝染色体lacZ基因的相对丰度来估测。用于RepFIC的引物为SEQ ID NO:10和SEQ ID NO:11。用于lacZ的引物为SEQ ID NO:12和SEQ ID NO:13。用于卡那霉素抗性基因的引物为SEQ ID NO:22和SEQ ID NO:23。
[表4]
qPCR拷贝数测量结果
如表4所示，菌株CC06-9666和9639(含有引入质粒pJSL49中修饰RepFIC序列的菌株)的质粒拷贝数高于菌株CC06-9665和9638的质粒拷贝数。菌株CC06-9665的质粒拷贝数高于CC06-9638的质粒拷贝数，并且菌株CC06-9666的质粒拷贝数高于CC06-9639的质粒拷贝数。因此确认，置换的rpoC序列导致质粒拷贝数减少，而与使用哪个RepFIC复制子(即大肠埃希氏菌LS5218的RepFIC复制子或修饰的RepFIC)无关。
工业应用性
本文公开的包含变体rpoC编码序列的核酸分子可用于调控载体(如质粒)的拷贝数，因此可用于通过使用该载体来提高目标产物的商业生产。
关于生物保藏的信息
被转化以包括包含编码包括R47C取代的变体RpoC的变体rpoC编码序列的核酸分子的大肠埃希氏菌菌株如上所述被制备，被命名为大肠埃希氏菌(Escherichia coli)CC06-9637，并于2018年6月15日以登录号KCCM12276P被保藏在韩国微生物培养中心(Korean Culture Center of Microorganisms)(韩国，首尔)，其是布达佩斯条约规定的国际保藏机构。此菌株按照布达佩斯条约规定由国际保藏机构保藏。
关于用于专利程序的微生物保藏的国际承认的布达佩斯条约国际形式
CJ第一制糖株式会社由本页底部确定的国际保藏
CJ CHEILJEDANG CENTER，330，机构依照第7.1条发布
DONGHO-RO，JUNG-GU，
首尔100-400，大韩民国
1第6.4(d)条适用时，该日期是获得国际保藏机构状态的日期。
序列表
<110> CJ第一制糖株式会社
<120> 包含变体RpoC编码序列的核酸分子
<130> OPA18456-PCT
<150> US 62/715,530
<151> 2018-08-07
<160> 51
<170> PatentIn version 3.5
<210> 1
<211> 4224
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体修饰rpoC
<400> 1
gtgaaagatt tattaaagtt tctgaaagcg cagactaaaa ccgaagagtt tgatgcgatc 60
aaaattgctc tggcttcgcc agacatgatc cgttcatggt ctttcggtga agttaaaaag 120
ccggaaacca tcaactactg tacgttcaaa ccagaacgtg acggcctttt ctgcgcccgt 180
atctttgggc cggtaaaaga ttacgagtgc ctgtgcggta agtacaagcg cctgaaacac 240
cgtggcgtca tctgtgagaa gtgcggcgtt gaagtgaccc agactaaagt acgccgtgag 300
cgtatgggcc acatcgaact ggcttccccg actgcgcaca tctggttcct gaaatcgctg 360
ccgtcccgta tcggtctgct gctcgatatg ccgctgcgcg atatcgaacg cgtactgtac 420
tttgaatcct atgtggttat cgaaggcggt atgaccaacc tggaacgtca gcagatcctg 480
actgaagagc agtatctgga cgcgctggaa gagttcggtg acgaattcga cgcgaagatg 540
ggggcggaag caatccaggc tctgctgaag agcatggatc tggagcaaga gtgcgaacag 600
ctgcgtgaag agctgaacga aaccaactcc gaaaccaagc gtaaaaagct gaccaagcgt 660
atcaaactgc tggaagcgtt cgttcagtct ggtaacaaac cagagtggat gatcctgacc 720
gttctgccgg tactgccgcc agatctgcgt ccgctggttc cgctggatgg tggtcgtttc 780
gcgacttctg acctgaacga tctgtatcgt cgcgtcatta accgtaacaa ccgtctgaaa 840
cgtctgctgg atctggctgc gccggacatc atcgtacgta acgaaaaacg tatgctgcag 900
gaagcggtag acgccctgct ggataacggt cgtcgcggtc gtgcgatcac cggttctaac 960
aagcgtcctc tgaaatcttt ggccgacatg atcaaaggta aacagggtcg tttccgtcag 1020
aacctgctcg gtaagcgtgt tgactactcc ggtcgttctg taatcaccgt aggtccatac 1080
ctgcgtctgc atcagtgcgg tctgccgaag aaaatggcac tggagctgtt caaaccgttc 1140
atctacggca agctggaact gcgtggtctt gctaccacca ttaaagctgc gaagaaaatg 1200
gttgagcgcg aagaagctgt cgtttgggat atcctggacg aagttatccg cgaacacccg 1260
gtactgctga accgtgcacc gactctgcac cgtctgggta tccaggcatt tgaaccggta 1320
ctgatcgaag gtaaagctat ccagctgcac ccgctggttt gtgcggcata taacgccgac 1380
ttcgatggtg accagatggc tgttcacgta ccgctgacgc tggaagccca gctggaagcg 1440
cgtgcgctga tgatgtctac caacaacatc ctgtccccgg cgaacggcga accaatcatc 1500
gttccgtctc aggacgttgt actgggtctg tactacatga cccgtgactg tgttaacgcc 1560
aaaggcgaag gcatggtgct gactggcccg aaagaagcag aacgtctgta tcgctctggt 1620
ctggcttctc tgcatgcgcg cgttaaagtg cgtatcaccg agtatgaaaa agatgctaac 1680
ggtgaattag tagcgaaaac cagcctgaaa gacacgactg ttggccgtgc cattctgtgg 1740
atgattgtac cgaaaggtct gccttactcc atcgtcaacc aggcgctggg taaaaaagca 1800
atctccaaaa tgctgaacac ctgctaccgc attctcggtc tgaaaccgac cgttattttt 1860
gcggaccaga tcatgtacac cggcttcgcc tatgcagcgc gttctggtgc atctgttggt 1920
atcgatgaca tggtcatccc ggagaagaaa cacgaaatca tctccgaggc agaagcagaa 1980
gttgctgaaa ttcaggagca gttccagtct ggtctggtaa ctgcgggcga acgctacaac 2040
aaagttatcg atatctgggc tgcggcgaac gatcgtgtat ccaaagcgat gatggataac 2100
ctgcaaactg aaaccgtgat taaccgtgac ggtcaggaag agaagcaggt ttccttcaac 2160
agcatctaca tgatggccga ctccggtgcg cgtggttctg cggcacagat tcgtcagctt 2220
gctggtatgc gtggtctgat ggcgaagccg gatggctcca tcatcgaaac gccaatcacc 2280
gcgaacttcc gtgaaggtct gaacgtactc cagtacttca tctccaccca cggtgctcgt 2340
aaaggtctgg cggataccgc actgaaaact gcgaactccg gttacctgac tcgtcgtctg 2400
gttgacgtgg cgcaggacct ggtggttacc gaagacgatt gtggtaccca tgaaggtatc 2460
atgatgactc cggttatcga gggtggtgac gttaaagagc cgctgcgcga tcgcgtactg 2520
ggtcgtgtaa ctgctgaaga cgttctgaag ccgggtactg ctgatatcct cgttccgcgc 2580
aacacgctgc tgcacgaaca gtggtgtgac ctgctggaag agaactctgt cgacgcggtt 2640
aaagtacgtt ctgttgtatc ttgtgacacc gactttggtg tatgtgcgca ctgctacggt 2700
cgtgacctgg cgcgtggcca catcatcaac aagggtgaag caatcggtgt tatcgcggca 2760
cagtccatcg gtgaaccggg tacacagctg accatgcgta cgttccacat cggtggtgcg 2820
gcatctcgtg cggctgctga atccagcatc caagtgaaaa acaaaggtag catcaagctc 2880
agcaacgtga agtcggttgt gaactccagc ggtaaactgg ttatcacttc ccgtaatact 2940
gaactgaaac tgatcgacga attcggtcgt actaaagaaa gctacaaagt accttacggt 3000
gcggtactgg cgaaaggcga tggcgaacag gttgctggcg gcgaaaccgt tgcaaactgg 3060
gacccgcaca ccatgccggt tatcaccgaa gtaagcggtt ttgtacgctt tactgacatg 3120
atcgacggcc agaccattac gcgtcagacc gacgaactga ccggtctgtc ttcgctggtg 3180
gttctggatt ccgcagaacg taccgcaggt ggtaaagatc tgcgtccggc actgaaaatc 3240
gttgatgctc agggtaacga cgttctgatc ccaggtaccg atatgccagc gcagtacttc 3300
ctgccgggta aagcgattgt tcagctggaa gatggcgtac agatcagctc tggtgacacc 3360
ctggcgcgta ttccgcagga atccggcggt accaaggaca tcaccggtgg tctgccgcgc 3420
gttgcggacc tgttcgaagc acgtcgtccg aaagagccgg caatcctggc tgaaatcagc 3480
ggtatcgttt ccttcggtaa agaaaccaaa ggtaaacgtc gtctggttat caccccggta 3540
gacggtagcg atccgtacga agagatgatt ccgaaatggc gtcagctcaa cgtgttcgaa 3600
ggtgaacgtg tagaacgtgg tgacgtaatt tccgacggtc cggaagcgcc gcacgacatt 3660
ctgcgtctgc gtggtgttca tgctgttact cgttacatcg ttaacgaagt acaggacgta 3720
taccgtctgc agggcgttaa gattaacgat aaacacatcg aagttatcgt tcgtcagatg 3780
ctgcgtaaag ctaccatcgt taacgcgggt agctccgact tcctggaagg cgaacaggtt 3840
gaatactctc gcgtcaagat cgcaaaccgc gaactggaag cgaacggcaa agtgggtgca 3900
acttactccc gcgatctgct gggtatcacc aaagcgtctc tggcaaccga gtccttcatc 3960
tccgcggcat cgttccagga gaccactcgc gtgctgaccg aagcagccgt tgcgggcaaa 4020
cgcgacgaac tgcgcggcct gaaagagaac gttatcgtgg gtcgtctgat cccggcaggt 4080
accggttacg cgtaccacca ggatcgtatg cgtcgccgtg ctgcgggtga agctccggct 4140
gcaccgcagg tgactgcaga agacgcatct gccagcctgg cagaactgct gaacgcaggt 4200
ctgggcggtt ctgataacga gtaa 4224
<210> 2
<211> 1407
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体修饰RpoC
<400> 2
Val Lys Asp Leu Leu Lys Phe Leu Lys Ala Gln Thr Lys Thr Glu Glu
1 5 10 15
Phe Asp Ala Ile Lys Ile Ala Leu Ala Ser Pro Asp Met Ile Arg Ser
20 25 30
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Cys Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Arg Ile Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Val Thr Gln Thr Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ser Pro Thr Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Leu
115 120 125
Asp Met Pro Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Ser Tyr
130 135 140
Val Val Ile Glu Gly Gly Met Thr Asn Leu Glu Arg Gln Gln Ile Leu
145 150 155 160
Thr Glu Glu Gln Tyr Leu Asp Ala Leu Glu Glu Phe Gly Asp Glu Phe
165 170 175
Asp Ala Lys Met Gly Ala Glu Ala Ile Gln Ala Leu Leu Lys Ser Met
180 185 190
Asp Leu Glu Gln Glu Cys Glu Gln Leu Arg Glu Glu Leu Asn Glu Thr
195 200 205
Asn Ser Glu Thr Lys Arg Lys Lys Leu Thr Lys Arg Ile Lys Leu Leu
210 215 220
Glu Ala Phe Val Gln Ser Gly Asn Lys Pro Glu Trp Met Ile Leu Thr
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Asp Leu Ala Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ser Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Thr Val Gly Pro Tyr Leu Arg Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Tyr Gly Lys
370 375 380
Leu Glu Leu Arg Gly Leu Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Glu Ala Val Val Trp Asp Ile Leu Asp Glu Val Ile
405 410 415
Arg Glu His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Ala Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Asn Gly
485 490 495
Glu Pro Ile Ile Val Pro Ser Gln Asp Val Val Leu Gly Leu Tyr Tyr
500 505 510
Met Thr Arg Asp Cys Val Asn Ala Lys Gly Glu Gly Met Val Leu Thr
515 520 525
Gly Pro Lys Glu Ala Glu Arg Leu Tyr Arg Ser Gly Leu Ala Ser Leu
530 535 540
His Ala Arg Val Lys Val Arg Ile Thr Glu Tyr Glu Lys Asp Ala Asn
545 550 555 560
Gly Glu Leu Val Ala Lys Thr Ser Leu Lys Asp Thr Thr Val Gly Arg
565 570 575
Ala Ile Leu Trp Met Ile Val Pro Lys Gly Leu Pro Tyr Ser Ile Val
580 585 590
Asn Gln Ala Leu Gly Lys Lys Ala Ile Ser Lys Met Leu Asn Thr Cys
595 600 605
Tyr Arg Ile Leu Gly Leu Lys Pro Thr Val Ile Phe Ala Asp Gln Ile
610 615 620
Met Tyr Thr Gly Phe Ala Tyr Ala Ala Arg Ser Gly Ala Ser Val Gly
625 630 635 640
Ile Asp Asp Met Val Ile Pro Glu Lys Lys His Glu Ile Ile Ser Glu
645 650 655
Ala Glu Ala Glu Val Ala Glu Ile Gln Glu Gln Phe Gln Ser Gly Leu
660 665 670
Val Thr Ala Gly Glu Arg Tyr Asn Lys Val Ile Asp Ile Trp Ala Ala
675 680 685
Ala Asn Asp Arg Val Ser Lys Ala Met Met Asp Asn Leu Gln Thr Glu
690 695 700
Thr Val Ile Asn Arg Asp Gly Gln Glu Glu Lys Gln Val Ser Phe Asn
705 710 715 720
Ser Ile Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln
725 730 735
Ile Arg Gln Leu Ala Gly Met Arg Gly Leu Met Ala Lys Pro Asp Gly
740 745 750
Ser Ile Ile Glu Thr Pro Ile Thr Ala Asn Phe Arg Glu Gly Leu Asn
755 760 765
Val Leu Gln Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Leu Val Val Thr Glu Asp Asp Cys Gly Thr
805 810 815
His Glu Gly Ile Met Met Thr Pro Val Ile Glu Gly Gly Asp Val Lys
820 825 830
Glu Pro Leu Arg Asp Arg Val Leu Gly Arg Val Thr Ala Glu Asp Val
835 840 845
Leu Lys Pro Gly Thr Ala Asp Ile Leu Val Pro Arg Asn Thr Leu Leu
850 855 860
His Glu Gln Trp Cys Asp Leu Leu Glu Glu Asn Ser Val Asp Ala Val
865 870 875 880
Lys Val Arg Ser Val Val Ser Cys Asp Thr Asp Phe Gly Val Cys Ala
885 890 895
His Cys Tyr Gly Arg Asp Leu Ala Arg Gly His Ile Ile Asn Lys Gly
900 905 910
Glu Ala Ile Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
915 920 925
Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser Arg Ala
930 935 940
Ala Ala Glu Ser Ser Ile Gln Val Lys Asn Lys Gly Ser Ile Lys Leu
945 950 955 960
Ser Asn Val Lys Ser Val Val Asn Ser Ser Gly Lys Leu Val Ile Thr
965 970 975
Ser Arg Asn Thr Glu Leu Lys Leu Ile Asp Glu Phe Gly Arg Thr Lys
980 985 990
Glu Ser Tyr Lys Val Pro Tyr Gly Ala Val Leu Ala Lys Gly Asp Gly
995 1000 1005
Glu Gln Val Ala Gly Gly Glu Thr Val Ala Asn Trp Asp Pro His Thr
1010 1015 1020
Met Pro Val Ile Thr Glu Val Ser Gly Phe Val Arg Phe Thr Asp Met
1025 1030 1035 1040
Ile Asp Gly Gln Thr Ile Thr Arg Gln Thr Asp Glu Leu Thr Gly Leu
1045 1050 1055
Ser Ser Leu Val Val Leu Asp Ser Ala Glu Arg Thr Ala Gly Gly Lys
1060 1065 1070
Asp Leu Arg Pro Ala Leu Lys Ile Val Asp Ala Gln Gly Asn Asp Val
1075 1080 1085
Leu Ile Pro Gly Thr Asp Met Pro Ala Gln Tyr Phe Leu Pro Gly Lys
1090 1095 1100
Ala Ile Val Gln Leu Glu Asp Gly Val Gln Ile Ser Ser Gly Asp Thr
1105 1110 1115 1120
Leu Ala Arg Ile Pro Gln Glu Ser Gly Gly Thr Lys Asp Ile Thr Gly
1125 1130 1135
Gly Leu Pro Arg Val Ala Asp Leu Phe Glu Ala Arg Arg Pro Lys Glu
1140 1145 1150
Pro Ala Ile Leu Ala Glu Ile Ser Gly Ile Val Ser Phe Gly Lys Glu
1155 1160 1165
Thr Lys Gly Lys Arg Arg Leu Val Ile Thr Pro Val Asp Gly Ser Asp
1170 1175 1180
Pro Tyr Glu Glu Met Ile Pro Lys Trp Arg Gln Leu Asn Val Phe Glu
1185 1190 1195 1200
Gly Glu Arg Val Glu Arg Gly Asp Val Ile Ser Asp Gly Pro Glu Ala
1205 1210 1215
Pro His Asp Ile Leu Arg Leu Arg Gly Val His Ala Val Thr Arg Tyr
1220 1225 1230
Ile Val Asn Glu Val Gln Asp Val Tyr Arg Leu Gln Gly Val Lys Ile
1235 1240 1245
Asn Asp Lys His Ile Glu Val Ile Val Arg Gln Met Leu Arg Lys Ala
1250 1255 1260
Thr Ile Val Asn Ala Gly Ser Ser Asp Phe Leu Glu Gly Glu Gln Val
1265 1270 1275 1280
Glu Tyr Ser Arg Val Lys Ile Ala Asn Arg Glu Leu Glu Ala Asn Gly
1285 1290 1295
Lys Val Gly Ala Thr Tyr Ser Arg Asp Leu Leu Gly Ile Thr Lys Ala
1300 1305 1310
Ser Leu Ala Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe Gln Glu Thr
1315 1320 1325
Thr Arg Val Leu Thr Glu Ala Ala Val Ala Gly Lys Arg Asp Glu Leu
1330 1335 1340
Arg Gly Leu Lys Glu Asn Val Ile Val Gly Arg Leu Ile Pro Ala Gly
1345 1350 1355 1360
Thr Gly Tyr Ala Tyr His Gln Asp Arg Met Arg Arg Arg Ala Ala Gly
1365 1370 1375
Glu Ala Pro Ala Ala Pro Gln Val Thr Ala Glu Asp Ala Ser Ala Ser
1380 1385 1390
Leu Ala Glu Leu Leu Asn Ala Gly Leu Gly Gly Ser Asp Asn Glu
1395 1400 1405
<210> 3
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体rpoC引物
<400> 3
ctgcaggaat tcgatcggtt cttacagcct ggttac 36
<210> 4
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体rpoC引物
<400> 4
gaacgtacag tagttgatgg 20
<210> 5
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体rpoC引物
<400> 5
ccatcaacta ctgtacgttc 20
<210> 6
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体rpoC引物
<400> 6
gtcgactagc gtgatcttgg tttcggagtt ggtttc 36
<210> 7
<211> 4224
<212> DNA
<213> 大肠埃希氏菌（Escherichia coli）
<400> 7
gtgaaagatt tattaaagtt tctgaaagcg cagactaaaa ccgaagagtt tgatgcgatc 60
aaaattgctc tggcttcgcc agacatgatc cgttcatggt ctttcggtga agttaaaaag 120
ccggaaacca tcaactaccg tacgttcaaa ccagaacgtg acggcctttt ctgcgcccgt 180
atctttgggc cggtaaaaga ttacgagtgc ctgtgcggta agtacaagcg cctgaaacac 240
cgtggcgtca tctgtgagaa gtgcggcgtt gaagtgaccc agactaaagt acgccgtgag 300
cgtatgggcc acatcgaact ggcttccccg actgcgcaca tctggttcct gaaatcgctg 360
ccgtcccgta tcggtctgct gctcgatatg ccgctgcgcg atatcgaacg cgtactgtac 420
tttgaatcct atgtggttat cgaaggcggt atgaccaacc tggaacgtca gcagatcctg 480
actgaagagc agtatctgga cgcgctggaa gagttcggtg acgaattcga cgcgaagatg 540
ggggcggaag caatccaggc tctgctgaag agcatggatc tggagcaaga gtgcgaacag 600
ctgcgtgaag agctgaacga aaccaactcc gaaaccaagc gtaaaaagct gaccaagcgt 660
atcaaactgc tggaagcgtt cgttcagtct ggtaacaaac cagagtggat gatcctgacc 720
gttctgccgg tactgccgcc agatctgcgt ccgctggttc cgctggatgg tggtcgtttc 780
gcgacttctg acctgaacga tctgtatcgt cgcgtcatta accgtaacaa ccgtctgaaa 840
cgtctgctgg atctggctgc gccggacatc atcgtacgta acgaaaaacg tatgctgcag 900
gaagcggtag acgccctgct ggataacggt cgtcgcggtc gtgcgatcac cggttctaac 960
aagcgtcctc tgaaatcttt ggccgacatg atcaaaggta aacagggtcg tttccgtcag 1020
aacctgctcg gtaagcgtgt tgactactcc ggtcgttctg taatcaccgt aggtccatac 1080
ctgcgtctgc atcagtgcgg tctgccgaag aaaatggcac tggagctgtt caaaccgttc 1140
atctacggca agctggaact gcgtggtctt gctaccacca ttaaagctgc gaagaaaatg 1200
gttgagcgcg aagaagctgt cgtttgggat atcctggacg aagttatccg cgaacacccg 1260
gtactgctga accgtgcacc gactctgcac cgtctgggta tccaggcatt tgaaccggta 1320
ctgatcgaag gtaaagctat ccagctgcac ccgctggttt gtgcggcata taacgccgac 1380
ttcgatggtg accagatggc tgttcacgta ccgctgacgc tggaagccca gctggaagcg 1440
cgtgcgctga tgatgtctac caacaacatc ctgtccccgg cgaacggcga accaatcatc 1500
gttccgtctc aggacgttgt actgggtctg tactacatga cccgtgactg tgttaacgcc 1560
aaaggcgaag gcatggtgct gactggcccg aaagaagcag aacgtctgta tcgctctggt 1620
ctggcttctc tgcatgcgcg cgttaaagtg cgtatcaccg agtatgaaaa agatgctaac 1680
ggtgaattag tagcgaaaac cagcctgaaa gacacgactg ttggccgtgc cattctgtgg 1740
atgattgtac cgaaaggtct gccttactcc atcgtcaacc aggcgctggg taaaaaagca 1800
atctccaaaa tgctgaacac ctgctaccgc attctcggtc tgaaaccgac cgttattttt 1860
gcggaccaga tcatgtacac cggcttcgcc tatgcagcgc gttctggtgc atctgttggt 1920
atcgatgaca tggtcatccc ggagaagaaa cacgaaatca tctccgaggc agaagcagaa 1980
gttgctgaaa ttcaggagca gttccagtct ggtctggtaa ctgcgggcga acgctacaac 2040
aaagttatcg atatctgggc tgcggcgaac gatcgtgtat ccaaagcgat gatggataac 2100
ctgcaaactg aaaccgtgat taaccgtgac ggtcaggaag agaagcaggt ttccttcaac 2160
agcatctaca tgatggccga ctccggtgcg cgtggttctg cggcacagat tcgtcagctt 2220
gctggtatgc gtggtctgat ggcgaagccg gatggctcca tcatcgaaac gccaatcacc 2280
gcgaacttcc gtgaaggtct gaacgtactc cagtacttca tctccaccca cggtgctcgt 2340
aaaggtctgg cggataccgc actgaaaact gcgaactccg gttacctgac tcgtcgtctg 2400
gttgacgtgg cgcaggacct ggtggttacc gaagacgatt gtggtaccca tgaaggtatc 2460
atgatgactc cggttatcga gggtggtgac gttaaagagc cgctgcgcga tcgcgtactg 2520
ggtcgtgtaa ctgctgaaga cgttctgaag ccgggtactg ctgatatcct cgttccgcgc 2580
aacacgctgc tgcacgaaca gtggtgtgac ctgctggaag agaactctgt cgacgcggtt 2640
aaagtacgtt ctgttgtatc ttgtgacacc gactttggtg tatgtgcgca ctgctacggt 2700
cgtgacctgg cgcgtggcca catcatcaac aagggtgaag caatcggtgt tatcgcggca 2760
cagtccatcg gtgaaccggg tacacagctg accatgcgta cgttccacat cggtggtgcg 2820
gcatctcgtg cggctgctga atccagcatc caagtgaaaa acaaaggtag catcaagctc 2880
agcaacgtga agtcggttgt gaactccagc ggtaaactgg ttatcacttc ccgtaatact 2940
gaactgaaac tgatcgacga attcggtcgt actaaagaaa gctacaaagt accttacggt 3000
gcggtactgg cgaaaggcga tggcgaacag gttgctggcg gcgaaaccgt tgcaaactgg 3060
gacccgcaca ccatgccggt tatcaccgaa gtaagcggtt ttgtacgctt tactgacatg 3120
atcgacggcc agaccattac gcgtcagacc gacgaactga ccggtctgtc ttcgctggtg 3180
gttctggatt ccgcagaacg taccgcaggt ggtaaagatc tgcgtccggc actgaaaatc 3240
gttgatgctc agggtaacga cgttctgatc ccaggtaccg atatgccagc gcagtacttc 3300
ctgccgggta aagcgattgt tcagctggaa gatggcgtac agatcagctc tggtgacacc 3360
ctggcgcgta ttccgcagga atccggcggt accaaggaca tcaccggtgg tctgccgcgc 3420
gttgcggacc tgttcgaagc acgtcgtccg aaagagccgg caatcctggc tgaaatcagc 3480
ggtatcgttt ccttcggtaa agaaaccaaa ggtaaacgtc gtctggttat caccccggta 3540
gacggtagcg atccgtacga agagatgatt ccgaaatggc gtcagctcaa cgtgttcgaa 3600
ggtgaacgtg tagaacgtgg tgacgtaatt tccgacggtc cggaagcgcc gcacgacatt 3660
ctgcgtctgc gtggtgttca tgctgttact cgttacatcg ttaacgaagt acaggacgta 3720
taccgtctgc agggcgttaa gattaacgat aaacacatcg aagttatcgt tcgtcagatg 3780
ctgcgtaaag ctaccatcgt taacgcgggt agctccgact tcctggaagg cgaacaggtt 3840
gaatactctc gcgtcaagat cgcaaaccgc gaactggaag cgaacggcaa agtgggtgca 3900
acttactccc gcgatctgct gggtatcacc aaagcgtctc tggcaaccga gtccttcatc 3960
tccgcggcat cgttccagga gaccactcgc gtgctgaccg aagcagccgt tgcgggcaaa 4020
cgcgacgaac tgcgcggcct gaaagagaac gttatcgtgg gtcgtctgat cccggcaggt 4080
accggttacg cgtaccacca ggatcgtatg cgtcgccgtg ctgcgggtga agctccggct 4140
gcaccgcagg tgactgcaga agacgcatct gccagcctgg cagaactgct gaacgcaggt 4200
ctgggcggtt ctgataacga gtaa 4224
<210> 8
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体pJSL47引物
<400> 8
gagcgtccgg taaccgttgg 20
<210> 9
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体pJSL47引物
<400> 9
gtcaggatca tccactctgg 20
<210> 10
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC引物
<400> 10
cgggtgggat tgaatcagat 20
<210> 11
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC引物
<400> 11
tacgtttgcc atgcgctatt a 21
<210> 12
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体lacZ引物
<400> 12
ccttactgcc gcctgttttg 20
<210> 13
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体lacZ引物
<400> 13
ccactggtgt gggccataat 20
<210> 14
<211> 2459
<212> DNA
<213> 大肠埃希氏菌
<400> 14
atgtcgcaga cagaaaatgc agtgacttcc tcatcaggta acaagcgtgc ataccggaaa 60
ggtaaccctg ttccggccag agagaggcaa agggcttctc tagctcgcag aagcaacact 120
cataaggctt ttcatgcggt tatccaggcc cggttaaaag acaggctgag tgaactggca 180
gatgaggaag gtattaccca ggcgcagatg cttgaaaaac tgattgaatc agagctgaaa 240
cgcagagcga ctttgtaaat attcacattc ttgcttatct caggcgtgag tgatagattg 300
ctgatcgttt aaggaatttt gtggctggcc acgccataag gtggcaggga actggttctg 360
atgtggattt acaggagcca gaaaagcaaa aaccccgata atcttcatct agttttgcga 420
cgaggagaag attaccggga tccacttaaa ccgtatagcc aacaattcag ctatgcgggg 480
agtatagtta tatgcccgga aaagttcaag acttctttct gtgctcactc cttctgtgca 540
ttgtaagtgc aggatggtgt ggctaatcat gaaacacatt cagtaatagc gggtgggatt 600
gaatcagatc ttcacattga ttccagcaag tatcctcacc cgttttgcag ccttctccag 660
aaaagggctc attttgactc cttcaagcat ctgatcttca tcagaggttt gcttgtaata 720
gcgcatggca aacgtaaaaa taaaatcagc gcgtcgatgg ttagttttta tgtttccctc 780
gtacaagtaa tgtgcgcaca ctacatccct gatacgaaca aagttaactt atctgttaaa 840
gagctttcag tttgtagtgg aggttcatac actcgtgttt ggagagccct caaaactctt 900
gataatgatc ttcatctcat cgcttttgat ggaggaacca tctggttcag accagatatg 960
ttcgaaactt tacgtgtcgg cccagacgag ctagttgccg cccgtaggag ggggaatagt 1020
gttggagggg gacatggctg atctccttca aaaatactat tcacaggtta aaaacccgaa 1080
tccggtgttc acaccccgtg aaggtgccgg aacgctgaag ttctgcgaaa aactgatgga 1140
aaaggcggtg ggcttcacct cccgttttga tttcgccatt catgtggcac atgctcgttc 1200
gaagggactg cgtcggcgca tgccaccggt actgcgtcgc cgggctattg atgcgctgct 1260
gcaggggctg tgttttcact atgacccgct ggccaaccgc gtccagtgct ccatcactac 1320
gctggccatt gagtgcggac tggcgacgga gtctgctgcc ggaaaactct ccatcacccg 1380
ggccacccga gccttgacgt tccttgcaga gctgggactg attacctacc agacggaata 1440
tgatccgctt atcgggtgct acattccgac cgatatcacg ttcacaccgg cgctatttgc 1500
cgcccttgat gtgtctgagg atgcagtggt tgctgcgcgc cgcagtcgtg ttgaatggga 1560
aaacagacag cgcaaaaagc agggactgga taccctgggt atggatgaac tgatagcgaa 1620
agcctggcgt tttgtgcgtg agcgtttccg cagttaccag acagagctta agtcccgtgg 1680
aataaagcgt gcccgtgcgc gtcgtgatgc gaacagggaa cgtcaggata tcgtcaccct 1740
ggtgaaacgg cagctgacgc gtgaaatctc ggaagggcgc ttcactgcca atcgtgaggc 1800
ggtaaaacgc gaagtggagc gtcgtgtgaa agatcgcatg attctgtcac gtaaccgtaa 1860
ttacagccgg ctggccacag cttccccctg aaagtgacct cctctgaata atccggcccg 1920
caccggaggc atctgcacgc ctgaagcctg tcggcgaaca aaaaaacagc accgcataca 1980
aaaaacaacc tcatcatcca ccttcaggtg catccggtcc ctcctgtttt tgatacaaaa 2040
cacgcctcac agacgaggaa ttttgcttat ccacatttaa ctgcaaggga cttccccata 2100
aggttacaac cgttcatgtc ataaagcgcc atccgccagc cttacagggt gcaatgtatc 2160
ttttaaacac ctgtttatat ctcctttaaa ctacttaact acattcattt aaaaagaaaa 2220
cctattcact gcctgtcctg tggacagaca ggtatgcacc tcccaccgca agcggcgggc 2280
cccgaccgga gccactttaa ttacaacact cagatacaac caccagaaaa accccggtcc 2340
cgcgcagaac tgaaaccaca aagccccccc tcataactga aaagcggccc cgccccggcc 2400
caaagggccg gaacagagtc gcttttaatt atgaatgttg taactacaca gcatcatcg 2459
<210> 15
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC引物
<400> 15
ttccggaagt gtgaggcggc cgcacttgtg tataa 35
<210> 16
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC引物
<400> 16
gtcgacaagc tttacgcggc cagatctgat caaga 35
<210> 17
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体卡那霉素引物
<400> 17
atcagatctg gccgcgtaaa gcttgtcgac gaatt 35
<210> 18
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体卡那霉素引物
<400> 18
cacaagtgcg gccgcctcac acttccggaa agcgg 35
<210> 19
<211> 98
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC序列
<400> 19
aagcaaaaac cccgataatc ttcatctagt tttgcgacaa ggagaagatt accgggatcc 60
acttaaaccg tatagccaac aattcagcta tgcgggga 98
<210> 20
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC引物
<400> 20
tcttctcctt gtcgcaaaac 20
<210> 21
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIC引物
<400> 21
gttttgcgac aaggagaaga 20
<210> 22
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体卡那霉素引物
<400> 22
gcagccgatt gtctgttgtg 20
<210> 23
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体卡那霉素引物
<400> 23
atggattgca cgcaggttct 20
<210> 24
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIA引物
<400> 24
atggctcagg catcgtctct 20
<210> 25
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体repFIA引物
<400> 25
agaggcgcat tggagttctg 20
<210> 26
<211> 1407
<212> PRT
<213> 大肠埃希氏菌
<400> 26
Met Lys Asp Leu Leu Lys Phe Leu Lys Ala Gln Thr Lys Thr Glu Glu
1 5 10 15
Phe Asp Ala Ile Lys Ile Ala Leu Ala Ser Pro Asp Met Ile Arg Ser
20 25 30
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Arg Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Arg Ile Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Val Thr Gln Thr Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ser Pro Thr Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Leu
115 120 125
Asp Met Pro Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Ser Tyr
130 135 140
Val Val Ile Glu Gly Gly Met Thr Asn Leu Glu Arg Gln Gln Ile Leu
145 150 155 160
Thr Glu Glu Gln Tyr Leu Asp Ala Leu Glu Glu Phe Gly Asp Glu Phe
165 170 175
Asp Ala Lys Met Gly Ala Glu Ala Ile Gln Ala Leu Leu Lys Ser Met
180 185 190
Asp Leu Glu Gln Glu Cys Glu Gln Leu Arg Glu Glu Leu Asn Glu Thr
195 200 205
Asn Ser Glu Thr Lys Arg Lys Lys Leu Thr Lys Arg Ile Lys Leu Leu
210 215 220
Glu Ala Phe Val Gln Ser Gly Asn Lys Pro Glu Trp Met Ile Leu Thr
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Asp Leu Ala Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ser Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Thr Val Gly Pro Tyr Leu Arg Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Tyr Gly Lys
370 375 380
Leu Glu Leu Arg Gly Leu Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Glu Ala Val Val Trp Asp Ile Leu Asp Glu Val Ile
405 410 415
Arg Glu His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Ala Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Asn Gly
485 490 495
Glu Pro Ile Ile Val Pro Ser Gln Asp Val Val Leu Gly Leu Tyr Tyr
500 505 510
Met Thr Arg Asp Cys Val Asn Ala Lys Gly Glu Gly Met Val Leu Thr
515 520 525
Gly Pro Lys Glu Ala Glu Arg Leu Tyr Arg Ser Gly Leu Ala Ser Leu
530 535 540
His Ala Arg Val Lys Val Arg Ile Thr Glu Tyr Glu Lys Asp Ala Asn
545 550 555 560
Gly Glu Leu Val Ala Lys Thr Ser Leu Lys Asp Thr Thr Val Gly Arg
565 570 575
Ala Ile Leu Trp Met Ile Val Pro Lys Gly Leu Pro Tyr Ser Ile Val
580 585 590
Asn Gln Ala Leu Gly Lys Lys Ala Ile Ser Lys Met Leu Asn Thr Cys
595 600 605
Tyr Arg Ile Leu Gly Leu Lys Pro Thr Val Ile Phe Ala Asp Gln Ile
610 615 620
Met Tyr Thr Gly Phe Ala Tyr Ala Ala Arg Ser Gly Ala Ser Val Gly
625 630 635 640
Ile Asp Asp Met Val Ile Pro Glu Lys Lys His Glu Ile Ile Ser Glu
645 650 655
Ala Glu Ala Glu Val Ala Glu Ile Gln Glu Gln Phe Gln Ser Gly Leu
660 665 670
Val Thr Ala Gly Glu Arg Tyr Asn Lys Val Ile Asp Ile Trp Ala Ala
675 680 685
Ala Asn Asp Arg Val Ser Lys Ala Met Met Asp Asn Leu Gln Thr Glu
690 695 700
Thr Val Ile Asn Arg Asp Gly Gln Glu Glu Lys Gln Val Ser Phe Asn
705 710 715 720
Ser Ile Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln
725 730 735
Ile Arg Gln Leu Ala Gly Met Arg Gly Leu Met Ala Lys Pro Asp Gly
740 745 750
Ser Ile Ile Glu Thr Pro Ile Thr Ala Asn Phe Arg Glu Gly Leu Asn
755 760 765
Val Leu Gln Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Leu Val Val Thr Glu Asp Asp Cys Gly Thr
805 810 815
His Glu Gly Ile Met Met Thr Pro Val Ile Glu Gly Gly Asp Val Lys
820 825 830
Glu Pro Leu Arg Asp Arg Val Leu Gly Arg Val Thr Ala Glu Asp Val
835 840 845
Leu Lys Pro Gly Thr Ala Asp Ile Leu Val Pro Arg Asn Thr Leu Leu
850 855 860
His Glu Gln Trp Cys Asp Leu Leu Glu Glu Asn Ser Val Asp Ala Val
865 870 875 880
Lys Val Arg Ser Val Val Ser Cys Asp Thr Asp Phe Gly Val Cys Ala
885 890 895
His Cys Tyr Gly Arg Asp Leu Ala Arg Gly His Ile Ile Asn Lys Gly
900 905 910
Glu Ala Ile Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
915 920 925
Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser Arg Ala
930 935 940
Ala Ala Glu Ser Ser Ile Gln Val Lys Asn Lys Gly Ser Ile Lys Leu
945 950 955 960
Ser Asn Val Lys Ser Val Val Asn Ser Ser Gly Lys Leu Val Ile Thr
965 970 975
Ser Arg Asn Thr Glu Leu Lys Leu Ile Asp Glu Phe Gly Arg Thr Lys
980 985 990
Glu Ser Tyr Lys Val Pro Tyr Gly Ala Val Leu Ala Lys Gly Asp Gly
995 1000 1005
Glu Gln Val Ala Gly Gly Glu Thr Val Ala Asn Trp Asp Pro His Thr
1010 1015 1020
Met Pro Val Ile Thr Glu Val Ser Gly Phe Val Arg Phe Thr Asp Met
1025 1030 1035 1040
Ile Asp Gly Gln Thr Ile Thr Arg Gln Thr Asp Glu Leu Thr Gly Leu
1045 1050 1055
Ser Ser Leu Val Val Leu Asp Ser Ala Glu Arg Thr Ala Gly Gly Lys
1060 1065 1070
Asp Leu Arg Pro Ala Leu Lys Ile Val Asp Ala Gln Gly Asn Asp Val
1075 1080 1085
Leu Ile Pro Gly Thr Asp Met Pro Ala Gln Tyr Phe Leu Pro Gly Lys
1090 1095 1100
Ala Ile Val Gln Leu Glu Asp Gly Val Gln Ile Ser Ser Gly Asp Thr
1105 1110 1115 1120
Leu Ala Arg Ile Pro Gln Glu Ser Gly Gly Thr Lys Asp Ile Thr Gly
1125 1130 1135
Gly Leu Pro Arg Val Ala Asp Leu Phe Glu Ala Arg Arg Pro Lys Glu
1140 1145 1150
Pro Ala Ile Leu Ala Glu Ile Ser Gly Ile Val Ser Phe Gly Lys Glu
1155 1160 1165
Thr Lys Gly Lys Arg Arg Leu Val Ile Thr Pro Val Asp Gly Ser Asp
1170 1175 1180
Pro Tyr Glu Glu Met Ile Pro Lys Trp Arg Gln Leu Asn Val Phe Glu
1185 1190 1195 1200
Gly Glu Arg Val Glu Arg Gly Asp Val Ile Ser Asp Gly Pro Glu Ala
1205 1210 1215
Pro His Asp Ile Leu Arg Leu Arg Gly Val His Ala Val Thr Arg Tyr
1220 1225 1230
Ile Val Asn Glu Val Gln Asp Val Tyr Arg Leu Gln Gly Val Lys Ile
1235 1240 1245
Asn Asp Lys His Ile Glu Val Ile Val Arg Gln Met Leu Arg Lys Ala
1250 1255 1260
Thr Ile Val Asn Ala Gly Ser Ser Asp Phe Leu Glu Gly Glu Gln Val
1265 1270 1275 1280
Glu Tyr Ser Arg Val Lys Ile Ala Asn Arg Glu Leu Glu Ala Asn Gly
1285 1290 1295
Lys Val Gly Ala Thr Tyr Ser Arg Asp Leu Leu Gly Ile Thr Lys Ala
1300 1305 1310
Ser Leu Ala Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe Gln Glu Thr
1315 1320 1325
Thr Arg Val Leu Thr Glu Ala Ala Val Ala Gly Lys Arg Asp Glu Leu
1330 1335 1340
Arg Gly Leu Lys Glu Asn Val Ile Val Gly Arg Leu Ile Pro Ala Gly
1345 1350 1355 1360
Thr Gly Tyr Ala Tyr His Gln Asp Arg Met Arg Arg Arg Ala Ala Gly
1365 1370 1375
Glu Ala Pro Ala Ala Pro Gln Val Thr Ala Glu Asp Ala Ser Ala Ser
1380 1385 1390
Leu Ala Glu Leu Leu Asn Ala Gly Leu Gly Gly Ser Asp Asn Glu
1395 1400 1405
<210> 27
<211> 1407
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体修饰的RpoC
<400> 27
Met Lys Asp Leu Leu Lys Phe Leu Lys Ala Gln Thr Lys Thr Glu Glu
1 5 10 15
Phe Asp Ala Ile Lys Ile Ala Leu Ala Ser Pro Asp Met Ile Arg Ser
20 25 30
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Cys Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Arg Ile Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Val Thr Gln Thr Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ser Pro Thr Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Leu
115 120 125
Asp Met Pro Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Ser Tyr
130 135 140
Val Val Ile Glu Gly Gly Met Thr Asn Leu Glu Arg Gln Gln Ile Leu
145 150 155 160
Thr Glu Glu Gln Tyr Leu Asp Ala Leu Glu Glu Phe Gly Asp Glu Phe
165 170 175
Asp Ala Lys Met Gly Ala Glu Ala Ile Gln Ala Leu Leu Lys Ser Met
180 185 190
Asp Leu Glu Gln Glu Cys Glu Gln Leu Arg Glu Glu Leu Asn Glu Thr
195 200 205
Asn Ser Glu Thr Lys Arg Lys Lys Leu Thr Lys Arg Ile Lys Leu Leu
210 215 220
Glu Ala Phe Val Gln Ser Gly Asn Lys Pro Glu Trp Met Ile Leu Thr
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Asp Leu Ala Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ser Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Thr Val Gly Pro Tyr Leu Arg Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Tyr Gly Lys
370 375 380
Leu Glu Leu Arg Gly Leu Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Glu Ala Val Val Trp Asp Ile Leu Asp Glu Val Ile
405 410 415
Arg Glu His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Ala Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Asn Gly
485 490 495
Glu Pro Ile Ile Val Pro Ser Gln Asp Val Val Leu Gly Leu Tyr Tyr
500 505 510
Met Thr Arg Asp Cys Val Asn Ala Lys Gly Glu Gly Met Val Leu Thr
515 520 525
Gly Pro Lys Glu Ala Glu Arg Leu Tyr Arg Ser Gly Leu Ala Ser Leu
530 535 540
His Ala Arg Val Lys Val Arg Ile Thr Glu Tyr Glu Lys Asp Ala Asn
545 550 555 560
Gly Glu Leu Val Ala Lys Thr Ser Leu Lys Asp Thr Thr Val Gly Arg
565 570 575
Ala Ile Leu Trp Met Ile Val Pro Lys Gly Leu Pro Tyr Ser Ile Val
580 585 590
Asn Gln Ala Leu Gly Lys Lys Ala Ile Ser Lys Met Leu Asn Thr Cys
595 600 605
Tyr Arg Ile Leu Gly Leu Lys Pro Thr Val Ile Phe Ala Asp Gln Ile
610 615 620
Met Tyr Thr Gly Phe Ala Tyr Ala Ala Arg Ser Gly Ala Ser Val Gly
625 630 635 640
Ile Asp Asp Met Val Ile Pro Glu Lys Lys His Glu Ile Ile Ser Glu
645 650 655
Ala Glu Ala Glu Val Ala Glu Ile Gln Glu Gln Phe Gln Ser Gly Leu
660 665 670
Val Thr Ala Gly Glu Arg Tyr Asn Lys Val Ile Asp Ile Trp Ala Ala
675 680 685
Ala Asn Asp Arg Val Ser Lys Ala Met Met Asp Asn Leu Gln Thr Glu
690 695 700
Thr Val Ile Asn Arg Asp Gly Gln Glu Glu Lys Gln Val Ser Phe Asn
705 710 715 720
Ser Ile Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln
725 730 735
Ile Arg Gln Leu Ala Gly Met Arg Gly Leu Met Ala Lys Pro Asp Gly
740 745 750
Ser Ile Ile Glu Thr Pro Ile Thr Ala Asn Phe Arg Glu Gly Leu Asn
755 760 765
Val Leu Gln Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Leu Val Val Thr Glu Asp Asp Cys Gly Thr
805 810 815
His Glu Gly Ile Met Met Thr Pro Val Ile Glu Gly Gly Asp Val Lys
820 825 830
Glu Pro Leu Arg Asp Arg Val Leu Gly Arg Val Thr Ala Glu Asp Val
835 840 845
Leu Lys Pro Gly Thr Ala Asp Ile Leu Val Pro Arg Asn Thr Leu Leu
850 855 860
His Glu Gln Trp Cys Asp Leu Leu Glu Glu Asn Ser Val Asp Ala Val
865 870 875 880
Lys Val Arg Ser Val Val Ser Cys Asp Thr Asp Phe Gly Val Cys Ala
885 890 895
His Cys Tyr Gly Arg Asp Leu Ala Arg Gly His Ile Ile Asn Lys Gly
900 905 910
Glu Ala Ile Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
915 920 925
Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser Arg Ala
930 935 940
Ala Ala Glu Ser Ser Ile Gln Val Lys Asn Lys Gly Ser Ile Lys Leu
945 950 955 960
Ser Asn Val Lys Ser Val Val Asn Ser Ser Gly Lys Leu Val Ile Thr
965 970 975
Ser Arg Asn Thr Glu Leu Lys Leu Ile Asp Glu Phe Gly Arg Thr Lys
980 985 990
Glu Ser Tyr Lys Val Pro Tyr Gly Ala Val Leu Ala Lys Gly Asp Gly
995 1000 1005
Glu Gln Val Ala Gly Gly Glu Thr Val Ala Asn Trp Asp Pro His Thr
1010 1015 1020
Met Pro Val Ile Thr Glu Val Ser Gly Phe Val Arg Phe Thr Asp Met
1025 1030 1035 1040
Ile Asp Gly Gln Thr Ile Thr Arg Gln Thr Asp Glu Leu Thr Gly Leu
1045 1050 1055
Ser Ser Leu Val Val Leu Asp Ser Ala Glu Arg Thr Ala Gly Gly Lys
1060 1065 1070
Asp Leu Arg Pro Ala Leu Lys Ile Val Asp Ala Gln Gly Asn Asp Val
1075 1080 1085
Leu Ile Pro Gly Thr Asp Met Pro Ala Gln Tyr Phe Leu Pro Gly Lys
1090 1095 1100
Ala Ile Val Gln Leu Glu Asp Gly Val Gln Ile Ser Ser Gly Asp Thr
1105 1110 1115 1120
Leu Ala Arg Ile Pro Gln Glu Ser Gly Gly Thr Lys Asp Ile Thr Gly
1125 1130 1135
Gly Leu Pro Arg Val Ala Asp Leu Phe Glu Ala Arg Arg Pro Lys Glu
1140 1145 1150
Pro Ala Ile Leu Ala Glu Ile Ser Gly Ile Val Ser Phe Gly Lys Glu
1155 1160 1165
Thr Lys Gly Lys Arg Arg Leu Val Ile Thr Pro Val Asp Gly Ser Asp
1170 1175 1180
Pro Tyr Glu Glu Met Ile Pro Lys Trp Arg Gln Leu Asn Val Phe Glu
1185 1190 1195 1200
Gly Glu Arg Val Glu Arg Gly Asp Val Ile Ser Asp Gly Pro Glu Ala
1205 1210 1215
Pro His Asp Ile Leu Arg Leu Arg Gly Val His Ala Val Thr Arg Tyr
1220 1225 1230
Ile Val Asn Glu Val Gln Asp Val Tyr Arg Leu Gln Gly Val Lys Ile
1235 1240 1245
Asn Asp Lys His Ile Glu Val Ile Val Arg Gln Met Leu Arg Lys Ala
1250 1255 1260
Thr Ile Val Asn Ala Gly Ser Ser Asp Phe Leu Glu Gly Glu Gln Val
1265 1270 1275 1280
Glu Tyr Ser Arg Val Lys Ile Ala Asn Arg Glu Leu Glu Ala Asn Gly
1285 1290 1295
Lys Val Gly Ala Thr Tyr Ser Arg Asp Leu Leu Gly Ile Thr Lys Ala
1300 1305 1310
Ser Leu Ala Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe Gln Glu Thr
1315 1320 1325
Thr Arg Val Leu Thr Glu Ala Ala Val Ala Gly Lys Arg Asp Glu Leu
1330 1335 1340
Arg Gly Leu Lys Glu Asn Val Ile Val Gly Arg Leu Ile Pro Ala Gly
1345 1350 1355 1360
Thr Gly Tyr Ala Tyr His Gln Asp Arg Met Arg Arg Arg Ala Ala Gly
1365 1370 1375
Glu Ala Pro Ala Ala Pro Gln Val Thr Ala Glu Asp Ala Ser Ala Ser
1380 1385 1390
Leu Ala Glu Leu Leu Asn Ala Gly Leu Gly Gly Ser Asp Asn Glu
1395 1400 1405
<210> 28
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体修饰RpoC N端结构域
<400> 28
Lys Pro Glu Thr Ile Asn Tyr Cys Thr
1 5
<210> 29
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体修饰RpoC中央结构域
<400> 29
Asn Ala Asp Phe Asp Gly Asp
1 5
<210> 30
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体修饰RpoC C端结构域
<400> 30
Ser Ala Ala Ser Phe Gln
1 5
<210> 31
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> 延伸的合成构建体修饰RpoC N端结构域
<400> 31
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Cys Thr
1 5 10 15
Phe Lys Pro Glu Arg Asp Gly Leu Phe
20 25
<210> 32
<211> 23
<212> PRT
<213> 人工序列
<220>
<223> 延伸的合成构建体修饰RpoC中央结构域
<400> 32
His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp Gln
1 5 10 15
Met Ala Val His Val Pro Leu
20
<210> 33
<211> 55
<212> PRT
<213> 人工序列
<220>
<223> 延伸的合成构建体修饰RpoC C端结构域
<400> 33
Gly Ile Thr Lys Ala Ser Leu Ala Thr Glu Ser Phe Ile Ser Ala Ala
1 5 10 15
Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Glu Ala Ala Val Ala Gly
20 25 30
Lys Arg Asp Glu Leu Arg Gly Leu Lys Glu Asn Val Ile Val Gly Arg
35 40 45
Leu Ile Pro Ala Gly Thr Gly
50 55
<210> 34
<211> 1524
<212> PRT
<213> 嗜热栖热菌（Thermus thermophilus）
<400> 34
Met Lys Lys Glu Val Arg Lys Val Arg Ile Ala Leu Ala Ser Pro Glu
1 5 10 15
Lys Ile Arg Ser Trp Ser Tyr Gly Glu Val Glu Lys Pro Glu Thr Ile
20 25 30
Asn Tyr Arg Thr Leu Lys Pro Glu Arg Asp Gly Leu Phe Asp Glu Arg
35 40 45
Ile Phe Gly Pro Ile Lys Asp Tyr Glu Cys Ala Cys Gly Lys Tyr Lys
50 55 60
Arg Gln Arg Phe Glu Gly Lys Val Cys Glu Arg Cys Gly Val Glu Val
65 70 75 80
Thr Lys Ser Ile Val Arg Arg Tyr Arg Met Gly His Ile Glu Leu Ala
85 90 95
Thr Pro Ala Ala His Ile Trp Phe Val Lys Asp Val Pro Ser Lys Ile
100 105 110
Gly Thr Leu Leu Asp Leu Ser Ala Thr Glu Leu Glu Gln Val Leu Tyr
115 120 125
Phe Ser Lys Tyr Ile Val Leu Asp Pro Lys Gly Ala Ile Leu Asn Gly
130 135 140
Val Pro Val Glu Lys Arg Gln Leu Leu Thr Asp Glu Glu Tyr Arg Glu
145 150 155 160
Leu Arg Tyr Gly Lys Gln Glu Thr Tyr Pro Leu Pro Pro Gly Val Asp
165 170 175
Ala Leu Val Lys Asp Gly Glu Glu Val Val Lys Gly Gln Glu Leu Ala
180 185 190
Pro Gly Val Val Ser Arg Leu Asp Gly Val Ala Leu Tyr Arg Phe Pro
195 200 205
Arg Arg Val Arg Val Glu Tyr Val Lys Lys Glu Arg Ala Gly Leu Arg
210 215 220
Leu Pro Leu Ala Ala Trp Val Glu Lys Glu Ala Tyr Lys Pro Gly Glu
225 230 235 240
Ile Leu Ala Glu Leu Pro Glu Pro Tyr Leu Phe Arg Ala Glu Glu Glu
245 250 255
Gly Val Val Glu Leu Lys Glu Leu Glu Glu Gly Ala Phe Leu Val Leu
260 265 270
Arg Arg Glu Asp Glu Pro Val Ala Thr Tyr Phe Leu Pro Val Gly Met
275 280 285
Thr Pro Leu Val Val His Gly Glu Ile Val Glu Lys Gly Gln Pro Leu
290 295 300
Ala Glu Ala Lys Gly Leu Leu Arg Met Pro Arg Gln Val Arg Ala Ala
305 310 315 320
Gln Val Glu Ala Glu Glu Glu Gly Glu Thr Val Tyr Leu Thr Leu Phe
325 330 335
Leu Glu Trp Thr Glu Pro Lys Asp Tyr Arg Val Gln Pro His Met Asn
340 345 350
Val Val Val Pro Glu Gly Ala Arg Val Glu Ala Gly Asp Lys Ile Val
355 360 365
Ala Ala Ile Asp Pro Glu Glu Glu Val Ile Ala Glu Ala Glu Gly Val
370 375 380
Val His Leu His Glu Pro Ala Ser Ile Leu Val Val Lys Ala Arg Val
385 390 395 400
Tyr Pro Phe Glu Asp Asp Val Glu Val Ser Thr Gly Asp Arg Val Ala
405 410 415
Pro Gly Asp Val Leu Ala Asp Gly Gly Lys Val Lys Ser Asp Val Tyr
420 425 430
Gly Arg Val Glu Val Asp Leu Val Arg Asn Val Val Arg Val Val Glu
435 440 445
Ser Tyr Asp Ile Asp Ala Arg Met Gly Ala Glu Ala Ile Gln Gln Leu
450 455 460
Leu Lys Glu Leu Asp Leu Glu Ala Leu Glu Lys Glu Leu Leu Glu Glu
465 470 475 480
Met Lys His Pro Ser Arg Ala Arg Arg Ala Lys Ala Arg Lys Arg Leu
485 490 495
Glu Val Val Arg Ala Phe Leu Asp Ser Gly Asn Arg Pro Glu Trp Met
500 505 510
Ile Leu Glu Ala Val Pro Val Leu Pro Pro Asp Leu Arg Pro Met Val
515 520 525
Gln Val Asp Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr
530 535 540
Arg Arg Leu Ile Asn Arg Asn Asn Arg Leu Lys Lys Leu Leu Ala Gln
545 550 555 560
Gly Ala Pro Glu Ile Ile Ile Arg Asn Glu Lys Arg Met Leu Gln Glu
565 570 575
Ala Val Asp Ala Leu Leu Asp Asn Gly Arg Arg Gly Ala Pro Val Thr
580 585 590
Asn Pro Gly Ser Asp Arg Pro Leu Arg Ser Leu Thr Asp Ile Leu Ser
595 600 605
Gly Lys Gln Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp
610 615 620
Tyr Ser Gly Arg Ser Val Ile Val Val Gly Pro Gln Leu Lys Leu His
625 630 635 640
Gln Cys Gly Leu Pro Lys Arg Met Ala Leu Glu Leu Phe Lys Pro Phe
645 650 655
Leu Leu Lys Lys Met Glu Glu Lys Gly Ile Ala Pro Asn Val Lys Ala
660 665 670
Ala Arg Arg Met Leu Glu Arg Gln Arg Asp Ile Lys Asp Glu Val Trp
675 680 685
Asp Ala Leu Glu Glu Val Ile His Gly Lys Val Val Leu Leu Asn Arg
690 695 700
Ala Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe Gln Pro Val Leu
705 710 715 720
Val Glu Gly Gln Ser Ile Gln Leu His Pro Leu Val Cys Glu Ala Phe
725 730 735
Asn Ala Asp Phe Asp Gly Asp Gln Met Ala Val His Val Pro Leu Ser
740 745 750
Ser Phe Ala Gln Ala Glu Ala Arg Ile Gln Met Leu Ser Ala His Asn
755 760 765
Leu Leu Ser Pro Ala Ser Gly Glu Pro Leu Ala Lys Pro Ser Arg Asp
770 775 780
Ile Ile Leu Gly Leu Tyr Tyr Ile Thr Gln Val Arg Lys Glu Lys Lys
785 790 795 800
Gly Ala Gly Leu Glu Phe Ala Thr Pro Glu Glu Ala Leu Ala Ala His
805 810 815
Glu Arg Gly Glu Val Ala Leu Asn Ala Pro Ile Lys Val Ala Gly Arg
820 825 830
Glu Thr Ser Val Gly Arg Leu Lys Tyr Val Phe Ala Asn Pro Asp Glu
835 840 845
Ala Leu Leu Ala Val Ala His Gly Ile Val Asp Leu Gln Asp Val Val
850 855 860
Thr Val Arg Tyr Met Gly Lys Arg Leu Glu Thr Ser Pro Gly Arg Ile
865 870 875 880
Leu Phe Ala Arg Ile Val Ala Glu Ala Val Glu Asp Glu Lys Val Ala
885 890 895
Trp Glu Leu Ile Gln Leu Asp Val Pro Gln Glu Lys Asn Ser Leu Lys
900 905 910
Asp Leu Val Tyr Gln Ala Phe Leu Arg Leu Gly Met Glu Lys Thr Ala
915 920 925
Arg Leu Leu Asp Ala Leu Lys Tyr Tyr Gly Phe Thr Phe Ser Thr Thr
930 935 940
Ser Gly Ile Thr Ile Gly Ile Asp Asp Ala Val Ile Pro Glu Glu Lys
945 950 955 960
Lys Gln Tyr Leu Glu Glu Ala Asp Arg Lys Leu Leu Gln Ile Glu Gln
965 970 975
Ala Tyr Glu Met Gly Phe Leu Thr Asp Arg Glu Arg Tyr Asp Gln Ile
980 985 990
Leu Gln Leu Trp Thr Glu Thr Thr Glu Lys Val Thr Gln Ala Val Phe
995 1000 1005
Lys Asn Phe Glu Glu Asn Tyr Pro Phe Asn Pro Leu Tyr Val Met Ala
1010 1015 1020
Gln Ser Gly Ala Arg Gly Asn Pro Gln Gln Ile Arg Gln Leu Cys Gly
1025 1030 1035 1040
Leu Arg Gly Leu Met Gln Lys Pro Ser Gly Glu Thr Phe Glu Val Pro
1045 1050 1055
Val Arg Ser Ser Phe Arg Glu Gly Leu Thr Val Leu Glu Tyr Phe Ile
1060 1065 1070
Ser Ser His Gly Ala Arg Lys Gly Gly Ala Asp Thr Ala Leu Arg Thr
1075 1080 1085
Ala Asp Ser Gly Tyr Leu Thr Arg Lys Leu Val Asp Val Thr His Glu
1090 1095 1100
Ile Val Val Arg Glu Ala Asp Cys Gly Thr Thr Asn Tyr Ile Ser Val
1105 1110 1115 1120
Pro Leu Phe Gln Pro Asp Glu Val Thr Arg Ser Leu Arg Leu Arg Lys
1125 1130 1135
Arg Ala Asp Ile Glu Ala Gly Leu Tyr Gly Arg Val Leu Ala Arg Glu
1140 1145 1150
Val Glu Val Leu Gly Val Arg Leu Glu Glu Gly Arg Tyr Leu Ser Met
1155 1160 1165
Asp Asp Val His Leu Leu Ile Lys Ala Ala Glu Ala Gly Glu Ile Gln
1170 1175 1180
Glu Val Pro Val Arg Ser Pro Leu Thr Cys Gln Thr Arg Tyr Gly Val
1185 1190 1195 1200
Cys Gln Lys Cys Tyr Gly Tyr Asp Leu Ser Met Ala Arg Pro Val Ser
1205 1210 1215
Ile Gly Glu Ala Val Gly Ile Val Ala Ala Gln Ser Ile Gly Glu Pro
1220 1225 1230
Gly Thr Gln Leu Thr Met Arg Thr Phe His Thr Gly Gly Val Ala Gly
1235 1240 1245
Ala Ala Asp Ile Thr Gln Gly Leu Pro Arg Val Ile Glu Leu Phe Glu
1250 1255 1260
Ala Arg Arg Pro Lys Ala Lys Ala Val Ile Ser Glu Ile Asp Gly Val
1265 1270 1275 1280
Val Arg Ile Glu Glu Thr Glu Glu Lys Leu Ser Val Phe Val Glu Ser
1285 1290 1295
Glu Gly Phe Ser Lys Glu Tyr Lys Leu Pro Lys Glu Ala Arg Leu Leu
1300 1305 1310
Val Lys Asp Gly Asp Tyr Val Glu Ala Gly Gln Pro Leu Thr Arg Gly
1315 1320 1325
Ala Ile Asp Pro His Gln Leu Leu Glu Ala Lys Gly Pro Glu Ala Val
1330 1335 1340
Glu Arg Tyr Leu Val Glu Glu Ile Gln Lys Val Tyr Arg Ala Gln Gly
1345 1350 1355 1360
Val Lys Leu His Asp Lys His Ile Glu Ile Val Val Arg Gln Met Met
1365 1370 1375
Lys Tyr Val Glu Val Thr Asp Pro Gly Asp Ser Arg Leu Leu Glu Gly
1380 1385 1390
Gln Val Leu Glu Lys Trp Asp Val Glu Ala Leu Asn Glu Arg Leu Ile
1395 1400 1405
Ala Glu Gly Lys Thr Pro Val Ala Trp Lys Pro Leu Leu Met Gly Val
1410 1415 1420
Thr Lys Ser Ala Leu Ser Thr Lys Ser Trp Leu Ser Ala Ala Ser Phe
1425 1430 1435 1440
Gln Asn Thr Thr His Val Leu Thr Glu Ala Ala Ile Ala Gly Lys Lys
1445 1450 1455
Asp Glu Leu Ile Gly Leu Lys Glu Asn Val Ile Leu Gly Arg Leu Ile
1460 1465 1470
Pro Ala Gly Thr Gly Ser Asp Phe Val Arg Phe Thr Gln Val Val Asp
1475 1480 1485
Gln Lys Thr Leu Lys Ala Ile Glu Glu Ala Arg Lys Glu Ala Val Glu
1490 1495 1500
Ala Lys Glu Arg Pro Ala Ala Arg Arg Gly Val Lys Arg Glu Gln Pro
1505 1510 1515 1520
Gly Lys Gln Ala
<210> 35
<211> 1391
<212> PRT
<213> 巴氏醋杆菌（Acetobacter pasteurianus）
<400> 35
Met Asn Glu Leu Met Lys Ile Leu Gly Gln Thr Gly Gln Ala Met Thr
1 5 10 15
Phe Asp Gln Ile Lys Ile Gln Leu Ala Ser Pro Glu Gln Ile Arg Ser
20 25 30
Trp Ser Tyr Gly Glu Ile Lys Lys Pro Glu Thr Ile Asn Tyr Arg Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Arg Ile Phe Gly Pro
50 55 60
Ile Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Met Lys Phe
65 70 75 80
Arg Gly Ile Ile Cys Glu Lys Cys Gly Val Glu Val Thr Leu Ala Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Gln Leu Ala Ser Pro Val Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Met Val
115 120 125
Asp Met Thr Leu Lys Asp Leu Glu Lys Val Leu Tyr Phe Glu Ser Tyr
130 135 140
Leu Val Leu Glu Pro Gly Thr Ser Pro Leu Lys Gln Tyr Ser Leu Leu
145 150 155 160
Thr Glu Glu Gln Tyr Leu Asp Ala Met Asp Glu Tyr Gly Asp Glu Gly
165 170 175
Val Glu Val Gly Ile Gly Ala Glu Ala Ile Lys Lys Val Leu Glu Arg
180 185 190
Ile Asp Cys Asp Ala Glu Lys Val Glu Leu Arg Gln Glu Leu Lys Glu
195 200 205
Thr Thr Ser Glu Ala Lys Arg Lys Lys Leu Val Lys Arg Leu Lys Leu
210 215 220
Ile Glu Ala Phe Ala Glu Ser Gly Ser Arg Pro Glu Trp Met Ile Leu
225 230 235 240
Asp Leu Val Pro Val Ile Pro Pro Asp Leu Arg Pro Leu Val Pro Leu
245 250 255
Asp Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg
260 265 270
Val Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Met Glu Leu Arg Ala
275 280 285
Pro Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala Val
290 295 300
Asp Ala Leu Phe Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ala
305 310 315 320
Asn Lys Arg Pro Leu Lys Ser Leu Ser Asp Met Leu Lys Gly Lys Gln
325 330 335
Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly
340 345 350
Arg Ser Val Ile Val Val Gly Pro Glu Leu Lys Leu His Gln Cys Gly
355 360 365
Leu Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Tyr Ala
370 375 380
Lys Leu Glu Lys Tyr Gly His Ala Thr Thr Ile Lys Ala Ala Lys Arg
385 390 395 400
Met Val Glu Lys Glu Arg Pro Glu Val Trp Asp Ile Leu Glu Glu Val
405 410 415
Ile Arg Glu His Pro Val Met Leu Asn Arg Ala Pro Thr Leu His Arg
420 425 430
Leu Gly Ile Gln Ala Phe Glu Pro Val Leu Val Glu Gly Lys Ala Ile
435 440 445
Gln Leu His Pro Leu Val Cys Thr Ala Phe Asn Ala Asp Phe Asp Gly
450 455 460
Asp Gln Met Ala Val His Val Pro Leu Ser Leu Glu Ala Gln Leu Glu
465 470 475 480
Ala Arg Val Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Asn
485 490 495
Gly Lys Pro Ile Ile Val Pro Ser Gln Asp Ile Val Leu Gly Leu Tyr
500 505 510
Tyr Leu Ser Leu Glu Thr Pro Glu Phe Lys Val Thr Pro Asp Arg Cys
515 520 525
Glu Tyr Asp Glu Thr Thr Gly Ala Leu Thr Lys Glu Gly Ala Pro Ser
530 535 540
Phe Ser Ser Ile Gly Glu Val Glu Tyr Ala Leu Ser Ala Gly Ala Leu
545 550 555 560
Lys Leu His Asp Lys Ile Arg Ala Arg Phe Gln Lys Ile Gly Ala Asp
565 570 575
Gly Lys Val Thr Tyr Glu Thr Ala Val Thr Thr Pro Gly Arg Val Leu
580 585 590
Ile Ala Gln Ile Leu Pro Gln His Glu Ala Val Pro Phe Ser Leu Ile
595 600 605
Asn Arg Gln Leu Thr Lys Lys Ala Val Ser Asp Val Ile Asp Thr Val
610 615 620
Tyr Arg His Cys Gly Gln Lys Glu Ala Val Ile Phe Cys Asp Arg Leu
625 630 635 640
Met Ala Leu Gly Phe Arg His Ala Ala Lys Ala Gly Ile Ser Phe Gly
645 650 655
Lys Asp Asp Met Ile Ile Pro Pro Glu Lys Lys Glu Leu Val Asp Arg
660 665 670
Thr Ala Ala Glu Val Lys Glu Phe Glu Gln Gln Tyr Gln Asp Gly Leu
675 680 685
Ile Thr Ala Gly Glu Arg Tyr Asn Lys Val Val Asp Ala Trp Ser Arg
690 695 700
Cys Thr Asp Glu Val Gln Ala Ala Met Thr Lys Glu Ile Ser Arg Gln
705 710 715 720
Glu Val Gly Lys Gln Ile Asn Ser Val Trp Met Met Ser His Ser Gly
725 730 735
Ala Arg Gly Ser Pro Ala Gln Met Lys Gln Leu Ala Gly Met Arg Gly
740 745 750
Leu Met Ala Lys Pro Ser Gly Glu Ile Ile Glu Gln Pro Ile Ile Ala
755 760 765
Asn Phe Lys Glu Gly Leu Ser Val Leu Asp Tyr Phe Thr Ser Thr His
770 775 780
Gly Ala Arg Lys Gly Leu Ala Asp Thr Ala Leu Lys Thr Ala Asn Ser
785 790 795 800
Gly Tyr Leu Thr Arg Arg Leu Val Asp Val Ala Gln Asp Ser Ile Ile
805 810 815
Ile Glu Glu Asp Cys Gly Ser Glu Arg Gly Leu Thr Val Arg Ala Val
820 825 830
Met Asp Gly Gly Glu Val Val Ala Ser Leu Ser Glu Arg Ile Leu Gly
835 840 845
Arg Thr Val Ala Ser Asp Val Val Val Pro Gly Thr Gly Glu Val Ile
850 855 860
Val Pro Arg Asn His Leu Ile Asp Glu Ala Asp Ala Glu Arg Ile Glu
865 870 875 880
Lys Ser Gly Val Glu Thr Val His Ile Arg Ser Val Leu Thr Cys Asp
885 890 895
Ser Arg Val Gly Val Cys Gly Arg Cys Tyr Gly Arg Asp Leu Ala Arg
900 905 910
Gly Thr Pro Val Asn Ile Gly Glu Ala Val Gly Val Ile Ala Ala Gln
915 920 925
Ser Ile Gly Glu Pro Gly Thr Gln Leu Thr Met Arg Thr Phe His Ile
930 935 940
Gly Gly Ala Ala Gln Arg Gly Ala Glu Gln Ser Met Ile Glu Ala Ser
945 950 955 960
Arg Asp Gly His Val Val Ile Arg Asn Arg Asn Val Val His Asn Ser
965 970 975
Gln Asn Val Pro Ile Val Met Ala Arg Asn Cys Glu Ile Leu Leu Ser
980 985 990
Asp Asp Asn Gly Val Glu Lys Ala Arg Tyr Arg Val Pro Tyr Gly Ala
995 1000 1005
Arg Leu Leu Thr Glu Glu Gly Ala Lys Val Ala Arg Gly Gln Lys Leu
1010 1015 1020
Ala Glu Trp Asp Pro Tyr Thr Leu Pro Ile Ile Thr Glu Lys Ala Gly
1025 1030 1035 1040
Lys Val Glu Tyr Leu Asp Leu Ile Asp Ser Ile Thr Leu Val Glu Arg
1045 1050 1055
Met Asp Glu Val Thr Gly Leu Thr Ser Lys Val Val Val Asp Tyr Lys
1060 1065 1070
Gln Ala Gly Lys Gly Val Asp Leu Arg Pro Arg Leu Gln Leu Lys Asp
1075 1080 1085
Ala Asn Gly Asp Val Val Lys Leu Asp Asn Gly Ala Asp Ala Arg Tyr
1090 1095 1100
Phe Leu Ser Pro Glu Thr Leu Leu Ser Val Glu Asn Gly Thr Glu Val
1105 1110 1115 1120
Asn Ala Gly Asp Val Leu Ala Arg Leu Pro Arg Glu Gly Ser Lys Thr
1125 1130 1135
Arg Asp Ile Thr Gly Gly Leu Pro Arg Val Ala Glu Leu Phe Glu Ala
1140 1145 1150
Arg Arg Pro Lys Asp His Ala Ile Ile Ala Glu Met Glu Gly Arg Val
1155 1160 1165
Glu Phe Gly Lys Asp Tyr Lys Ser Lys Arg Arg Val Ile Val Lys Asn
1170 1175 1180
Asp Glu Thr Gly Glu Glu Gln Glu Tyr Leu Ile Pro Lys Gly Lys His
1185 1190 1195 1200
Ile Ser Val Gln Glu Gly Asp Phe Val Glu Lys Gly Asp Pro Leu Val
1205 1210 1215
Asp Gly Pro Arg Val Pro His Asp Ile Leu Lys Val Met Gly Val Glu
1220 1225 1230
Ala Leu Ser Asp Tyr Leu Ile Asn Glu Ile Gln Asp Val Tyr Arg Leu
1235 1240 1245
Gln Gly Val Lys Ile Asn Asp Lys His Ile Glu Val Ile Val Arg Gln
1250 1255 1260
Met Leu Gln Lys Val Glu Ile Leu Glu Pro Gly Asp Thr Thr Tyr Leu
1265 1270 1275 1280
Ile Gly Glu Thr Val Asp Arg Ile Glu Phe Glu Ala Glu Asn Ala Lys
1285 1290 1295
Cys Leu Lys Ala Gly Glu Arg Pro Ala Gln Gly Met Pro Val Leu Gln
1300 1305 1310
Gly Ile Thr Lys Ala Ser Leu Gln Thr Gln Ser Phe Ile Ser Ala Ala
1315 1320 1325
Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Glu Ala Ala Thr Ala Gly
1330 1335 1340
Lys Val Asp Lys Leu Met Gly Leu Lys Glu Asn Val Ile Val Gly Arg
1345 1350 1355 1360
Leu Ile Pro Ala Gly Thr Gly Ser Val Met Lys Arg Leu Arg Ala Ile
1365 1370 1375
Ala Ala Glu Gln Asp Arg Gln Arg Val Gly Arg Ser Ala Ala Glu
1380 1385 1390
<210> 36
<211> 1391
<212> PRT
<213> 淋病奈瑟氏菌（Neisseria gonorrhoeae）
<400> 36
Met Asn Leu Leu Asn Leu Phe Asn Pro Leu Gln Thr Ala Gly Met Glu
1 5 10 15
Glu Glu Phe Asp Ala Ile Lys Ile Gly Ile Ala Ser Pro Glu Thr Ile
20 25 30
Arg Ser Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr
35 40 45
Arg Thr Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Lys Ile Phe
50 55 60
Gly Pro Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu
65 70 75 80
Lys Phe Lys Gly Val Thr Cys Glu Lys Cys Gly Val Glu Val Thr Leu
85 90 95
Ser Lys Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ala Pro
100 105 110
Val Ala His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Leu Gly Met
115 120 125
Val Leu Asn Met Thr Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu
130 135 140
Ala Phe Val Val Thr Asp Pro Gly Met Thr Pro Leu Gln Arg Arg Gln
145 150 155 160
Leu Leu Thr Glu Asp Asp Tyr Tyr Asn Lys Leu Asp Glu Tyr Gly Asp
165 170 175
Asp Phe Asp Ala Lys Met Gly Ala Glu Gly Ile Arg Glu Leu Leu Arg
180 185 190
Thr Leu Asp Val Ala Gly Glu Ile Glu Ile Leu Arg Gln Glu Leu Glu
195 200 205
Ser Thr Gly Ser Asp Thr Lys Ile Lys Lys Ile Ala Lys Arg Leu Lys
210 215 220
Val Leu Glu Ala Phe His Arg Ser Gly Met Lys Leu Glu Trp Met Ile
225 230 235 240
Met Asp Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro
245 250 255
Leu Asp Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg
260 265 270
Arg Val Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Glu Leu His
275 280 285
Ala Pro Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala
290 295 300
Val Asp Ser Leu Leu Asp Asn Gly Arg Arg Gly Lys Ala Met Thr Gly
305 310 315 320
Ala Asn Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys
325 330 335
Gly Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser
340 345 350
Gly Arg Ser Val Ile Thr Val Gly Pro Tyr Leu Arg Leu His Gln Cys
355 360 365
Gly Leu Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Phe
370 375 380
His Lys Leu Glu Lys Gln Gly Leu Ala Ser Thr Val Lys Ala Ala Lys
385 390 395 400
Lys Leu Val Glu Gln Glu Val Pro Glu Val Trp Asp Ile Leu Glu Glu
405 410 415
Val Ile Arg Glu His Pro Ile Met Leu Asn Arg Ala Pro Thr Leu His
420 425 430
Arg Leu Gly Ile Gln Ala Phe Glu Pro Ile Leu Ile Glu Gly Lys Ala
435 440 445
Ile Gln Leu His Pro Leu Val Cys Ala Ala Phe Asn Ala Asp Phe Asp
450 455 460
Gly Asp Gln Met Ala Val His Val Pro Leu Ser Leu Glu Ala Gln Met
465 470 475 480
Glu Ala Arg Thr Leu Met Leu Ala Ser Asn Asn Val Leu Ser Pro Ala
485 490 495
Asn Gly Glu Pro Ile Ile Val Pro Ser Gln Asp Ile Val Leu Gly Leu
500 505 510
Tyr Tyr Met Thr Arg Asp Arg Ile Asn Ala Lys Gly Glu Gly Ser Leu
515 520 525
Phe Ala Asp Val Lys Glu Val His Arg Ala Tyr His Thr Lys Gln Val
530 535 540
Glu Leu Gly Thr Lys Ile Thr Val Arg Leu Arg Glu Trp Val Lys Asn
545 550 555 560
Glu Ala Gly Glu Phe Glu Pro Val Val Asn Arg Tyr Glu Thr Thr Val
565 570 575
Gly Arg Ala Leu Leu Ser Glu Ile Leu Pro Lys Gly Leu Pro Phe Glu
580 585 590
Tyr Val Asn Lys Ala Leu Lys Lys Lys Glu Ile Ser Lys Leu Ile Asn
595 600 605
Ala Ser Phe Arg Leu Cys Gly Leu Arg Asp Thr Val Ile Phe Ala Asp
610 615 620
His Leu Met Tyr Thr Gly Phe Gly Phe Ala Ala Lys Gly Gly Ile Ser
625 630 635 640
Ile Ala Val Asp Asp Met Glu Ile Pro Lys Glu Lys Ala Ala Leu Leu
645 650 655
Ala Glu Ala Asn Ala Glu Val Lys Glu Ile Glu Asp Gln Tyr Arg Gln
660 665 670
Gly Leu Val Thr Asn Gly Glu Arg Tyr Asn Lys Val Val Asp Ile Trp
675 680 685
Gly Arg Ala Gly Asp Lys Ile Ala Lys Ala Met Met Asp Asn Leu Ser
690 695 700
Lys Gln Lys Val Ile Asp Arg Asp Gly Asn Glu Val Asp Gln Glu Ser
705 710 715 720
Phe Asn Ser Ile Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala
725 730 735
Ala Gln Ile Lys Gln Leu Ser Gly Met Arg Gly Leu Met Ala Lys Pro
740 745 750
Asp Gly Ser Ile Ile Glu Thr Pro Ile Thr Ser Asn Phe Arg Glu Gly
755 760 765
Leu Thr Val Leu Gln Tyr Phe Ile Ala Thr His Gly Ala Arg Lys Gly
770 775 780
Leu Ala Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg
785 790 795 800
Arg Leu Val Asp Val Thr Gln Asp Leu Val Val Val Glu Asp Asp Cys
805 810 815
Gly Thr Ser Asp Gly Phe Val Met Lys Ala Val Val Gln Gly Gly Asp
820 825 830
Val Ile Glu Ala Leu Arg Asp Arg Ile Leu Gly Arg Val Thr Ala Ser
835 840 845
Asp Val Val Asp Pro Ser Ser Gly Glu Thr Leu Val Glu Ala Gly Thr
850 855 860
Leu Leu Thr Glu Lys Leu Val Asp Met Ile Asp Gln Ser Gly Val Asp
865 870 875 880
Glu Val Lys Val Arg Thr Pro Ile Thr Cys Lys Thr Arg His Gly Leu
885 890 895
Cys Ala His Cys Tyr Gly Arg Asp Leu Ala Arg Gly Lys Leu Val Asn
900 905 910
Ala Gly Glu Ala Val Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro
915 920 925
Gly Thr Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser
930 935 940
Arg Ala Ala Ala Ala Ser Gln Val Glu Ala Lys Ser Asn Gly Thr Ala
945 950 955 960
Arg Phe Ser Ser Gln Met Arg Tyr Val Ala Asn Asn Lys Gly Glu Leu
965 970 975
Val Val Ile Gly Arg Ser Cys Glu Val Val Ile His Asp Asp Ile Gly
980 985 990
Arg Glu Arg Glu Arg His Lys Val Pro Tyr Gly Ala Ile Leu Leu Val
995 1000 1005
Gln Asp Gly Met Ala Ile Lys Ala Gly Gln Thr Leu Ala Thr Trp Asp
1010 1015 1020
Pro His Thr Arg Pro Met Ile Thr Glu His Ala Gly Met Val Lys Phe
1025 1030 1035 1040
Glu Asn Met Glu Glu Gly Val Thr Val Ala Lys Gln Thr Asp Asp Val
1045 1050 1055
Thr Gly Leu Ser Thr Leu Val Val Ile Asp Gly Lys Arg Arg Ser Ser
1060 1065 1070
Ser Ala Ser Lys Leu Leu Arg Pro Thr Val Lys Leu Leu Asp Glu Asn
1075 1080 1085
Gly Val Glu Ile Cys Ile Pro Gly Thr Ser Thr Pro Val Ser Met Ala
1090 1095 1100
Phe Pro Val Gly Ala Val Ile Thr Val Arg Glu Gly Gln Glu Ile Gly
1105 1110 1115 1120
Lys Gly Asp Val Leu Ala Arg Ile Pro Gln Ala Ser Ser Lys Thr Arg
1125 1130 1135
Asp Ile Thr Gly Gly Leu Pro Arg Val Ala Glu Leu Phe Glu Ala Arg
1140 1145 1150
Val Pro Lys Asp Ala Gly Met Leu Ala Glu Ile Thr Gly Thr Val Ser
1155 1160 1165
Phe Gly Lys Glu Thr Lys Gly Lys Gln Arg Leu Ile Ile Thr Asp Val
1170 1175 1180
Asp Gly Val Ala Tyr Glu Thr Leu Ile Ser Lys Glu Lys Gln Ile Leu
1185 1190 1195 1200
Val His Asp Gly Gln Val Val Asn Arg Gly Glu Thr Ile Val Asp Gly
1205 1210 1215
Ala Val Asp Pro His Asp Ile Leu Arg Leu Gln Gly Ile Glu Ala Leu
1220 1225 1230
Ala Arg Tyr Ile Val Gln Glu Val Gln Glu Val Tyr Arg Leu Gln Gly
1235 1240 1245
Val Lys Ile Ser Asp Lys His Ile Glu Val Ile Ile Arg Gln Met Leu
1250 1255 1260
Arg Arg Val Asn Ile Ala Asp Ala Gly Glu Thr Gly Phe Ile Thr Gly
1265 1270 1275 1280
Glu Gln Val Glu Arg Gly Asp Val Met Ala Ala Asn Glu Lys Ala Leu
1285 1290 1295
Glu Glu Gly Lys Glu Pro Ala Arg Tyr Glu Asn Ile Leu Leu Gly Ile
1300 1305 1310
Thr Lys Ala Ser Leu Ser Thr Asp Ser Phe Ile Ser Ala Ala Ser Phe
1315 1320 1325
Gln Glu Thr Thr Arg Val Leu Thr Glu Ala Ala Ile Met Gly Lys Gln
1330 1335 1340
Asp Glu Leu Arg Gly Leu Lys Glu Asn Val Ile Val Gly Arg Leu Ile
1345 1350 1355 1360
Pro Ala Gly Thr Gly Leu Thr Tyr His Arg Ser Arg His Gln Gln Trp
1365 1370 1375
Gln Gly Val Glu Gln Glu Thr Ala Glu Thr Gln Val Thr Asp Glu
1380 1385 1390
<210> 37
<211> 1401
<212> PRT
<213> 嗜肺性军团杆菌（Legionella pneumophila）
<400> 37
Met Ser Asp Leu Leu Gly Ile Leu Lys Gln Gln Gly Gln Ser Glu Glu
1 5 10 15
Phe Asp Ala Ile Lys Ile Ala Leu Ala Ser Pro Glu Leu Ile Arg Ser
20 25 30
Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Arg Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Lys Thr Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Leu Ala Leu Ala Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ser Pro Val Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Leu
115 120 125
Asp Met Thr Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Ala Phe
130 135 140
Val Val Val Asp Pro Gly Met Thr Glu Leu Glu Arg Gly Gln Leu Leu
145 150 155 160
Asn Asp Glu Ala Tyr Leu Asp Ala Met Glu Gln Tyr Gly Asp Glu Phe
165 170 175
Asp Ala Arg Met Gly Ala Glu Ala Ile Arg Asp Leu Leu Arg Gln Ile
180 185 190
Asp Leu Glu Asp Glu Ile Arg Asn Leu Arg Glu Glu Leu Pro Thr Thr
195 200 205
Asn Ser Glu Thr Lys Ile Lys Lys Ile Thr Lys Arg Leu Lys Leu Leu
210 215 220
Glu Ala Phe Tyr Glu Ser Gly Asn Lys Pro Glu Trp Met Ile Met Asp
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Asp Leu Asn Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ser Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Thr Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Val Val Gly Pro Thr Leu Lys Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Phe Ser Lys
370 375 380
Leu Glu Phe Arg Gly Leu Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Glu Ser Val Val Trp Asp Ile Leu Asp Asp Val Ile
405 410 415
Arg Glu His Pro Ile Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Thr Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Ser Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Ser Gly
485 490 495
Glu Pro Ile Ile Val Pro Ser Gln Asp Val Val Leu Gly Leu Tyr Tyr
500 505 510
Leu Thr Arg Glu Lys Val Asn Ala Leu Gly Glu Gly Lys Ile Tyr Ser
515 520 525
Ser Ala Gln Glu Ala Gln Asn Phe Tyr Glu Ala Gly His Leu Asp Ile
530 535 540
His Ala Lys Ile Lys Ile Arg Met Pro Lys Glu Asp Gly Glu Thr Gly
545 550 555 560
Tyr His Leu Val Glu Thr Thr Val Gly Arg Ala Ile Leu Ala Glu Ile
565 570 575
Leu Pro Lys Gly Met Pro Phe Asp Tyr Ile Asn Arg Thr Met Thr Lys
580 585 590
Lys Val Ile Ser Lys Val Ile Asp Ser Cys Tyr Arg Lys Phe Gly Leu
595 600 605
Lys Glu Thr Val Ile Phe Ala Asp Gln Leu Met Tyr Thr Gly Phe Lys
610 615 620
Tyr Ala Thr Arg Ser Gly Ala Ser Ile Gly Ile Glu Asp Met Glu Ile
625 630 635 640
Pro Asp Asp Lys Ser Ser Ile Ile Glu His Ala Asp Asn Glu Val Arg
645 650 655
Glu Ile Glu Ser Gln Phe Arg Ser Gly Leu Val Thr Asn Gly Glu Arg
660 665 670
Tyr Asn Lys Val Ile Asp Ile Trp Ser Arg Thr Asn Glu Leu Val Ala
675 680 685
Lys Ser Met Met Ser Lys Ile Ala Thr Glu Glu Val Thr Asp Ala Lys
690 695 700
Gly Asn Lys Val Arg Gln Glu Ser Phe Asn Pro Ile Phe Met Met Ala
705 710 715 720
Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln Ile Arg Gln Leu Ala Gly
725 730 735
Met Arg Gly Leu Met Ala Ala Pro Asp Gly Ser Ile Ile Glu Thr Pro
740 745 750
Ile Thr Ala Asn Phe Arg Glu Gly Leu Asn Val Phe Gln Tyr Phe Ile
755 760 765
Ser Thr His Gly Ala Arg Lys Gly Leu Ala Asp Thr Ala Leu Lys Thr
770 775 780
Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val Ala Gln Asp
785 790 795 800
Val Val Ile Thr Glu Asp Asp Cys Gly Thr Asp Thr Gly Ile Leu Met
805 810 815
Gln Pro Leu Ile Glu Gly Gly Asp Ile Val Glu Pro Leu His Glu Arg
820 825 830
Val Leu Gly Arg Val Val Ala Ser Asp Val Tyr Ile Pro Thr Gln Thr
835 840 845
Glu Pro Val Val Lys Ala Gly Thr Leu Leu Asp Glu Glu Trp Val Glu
850 855 860
Lys Leu Glu Lys His Gly Val Asp Gln Val Met Val Arg Ser Pro Ile
865 870 875 880
Thr Cys Gln Thr Arg Phe Gly Leu Cys Ala Lys Cys Tyr Gly Arg Asp
885 890 895
Leu Ala Arg Gly His Leu Val Asn Thr Gly Glu Ala Val Gly Ile Ile
900 905 910
Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr Gln Leu Thr Met Arg Thr
915 920 925
Phe His Ile Gly Gly Ala Ala Ser Arg Ala Thr Ala Ala Asn Asn Ile
930 935 940
Gln Ile Lys Thr Lys Gly Val Ile Arg Leu His Asn Ile Lys Thr Val
945 950 955 960
Thr His Glu Asn Lys Asn Leu Val Ala Val Ser Arg Ser Gly Glu Val
965 970 975
Thr Ile Val Asp Glu Phe Gly Arg Glu Arg Glu Arg Tyr Lys Val Pro
980 985 990
Tyr Gly Ala Val Ile Ser Ala Gln Asp Asn Ser Pro Val Glu Ala Gly
995 1000 1005
Gln Val Ile Ala Thr Trp Asp Pro His Thr His Pro Val Ile Ser Glu
1010 1015 1020
Val Ser Gly Arg Leu Lys Phe Val Asp Leu Ile Asp Gly Ile Thr Met
1025 1030 1035 1040
Asn Arg Gln Thr Asp Glu Leu Thr Gly Leu Ser Asn Ile Val Ile Ile
1045 1050 1055
Asp Ala Lys Gln Arg Ser Ala Ala Gly Arg Asp Leu Arg Pro Met Val
1060 1065 1070
Lys Leu Val Thr Asp Glu Gly Asp Asp Ile Tyr Leu Ala Gly Thr Asn
1075 1080 1085
Val Pro Ala Gln Tyr Tyr Leu Pro Val Asp Ala Ile Val Asn Phe Glu
1090 1095 1100
Asp Gly Ser Leu Val Gly Ile Gly Asp Val Ile Ala Arg Ile Pro Gln
1105 1110 1115 1120
Glu Arg Ser Lys Thr Arg Asp Ile Thr Gly Gly Leu Pro Arg Val Ala
1125 1130 1135
Asp Leu Phe Glu Ala Arg Lys Pro Lys Asp Ser Ala Val Met Ala Glu
1140 1145 1150
Val Ser Gly Leu Val Asn Phe Gly Lys Glu Thr Lys Gly Lys Arg Arg
1155 1160 1165
Leu Ile Ile Asn Val Ser Glu Asp Gln Cys His Glu Glu Leu Ile Pro
1170 1175 1180
Lys Trp Arg His Ile Ser Val Phe Glu Gly Glu His Val Glu Arg Gly
1185 1190 1195 1200
Glu Ile Ile Ala Glu Gly Ala Leu Asn Pro His Asp Ile Leu Arg Leu
1205 1210 1215
Leu Gly Val Gly Ala Leu Ala Asn Tyr Ile Val Asn Glu Val Gln Asp
1220 1225 1230
Val Tyr Arg Leu Gln Gly Val Lys Ile Asn Asp Lys His Ile Glu Val
1235 1240 1245
Ile Val Arg Gln Met Leu Arg Lys Arg Val Ile Thr Phe Ala Gly Asp
1250 1255 1260
Ser Lys Phe Leu Val Gly Glu Gln Val Glu Glu Ser Ala Met Leu Gln
1265 1270 1275 1280
Glu Asn Asp Lys Leu Leu Ala Glu Gly Lys Gln Ile Ala Arg Gly Thr
1285 1290 1295
Pro Ile Leu Leu Gly Ile Thr Lys Ala Ser Leu Ala Thr Glu Ser Phe
1300 1305 1310
Ile Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Glu Ala
1315 1320 1325
Ala Val Ser Gly Lys Val Asp Glu Leu Arg Gly Leu Lys Glu Asn Val
1330 1335 1340
Met Val Gly Arg Leu Ile Pro Ala Gly Thr Gly Tyr Thr Tyr His Gln
1345 1350 1355 1360
Ser Arg Lys Ala Lys Arg Ala Arg Ala Ala Ala Gly Gly Asp Ser Ser
1365 1370 1375
Ala Thr His Thr Val Thr Ala Ser Asp Val Glu His Ala Leu Ser Glu
1380 1385 1390
Ala Leu Asn Ala Asp Asn His Glu His
1395 1400
<210> 38
<211> 1399
<212> PRT
<213> 铜绿假单胞菌（Pseudomonas aeruginosa）
<400> 38
Met Lys Asp Leu Leu Asn Leu Leu Lys Asn Gln Gly Gln Ile Glu Glu
1 5 10 15
Phe Asp Ala Ile Arg Ile Gly Leu Ala Ser Pro Glu Met Ile Arg Ser
20 25 30
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Arg Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Lys Ile Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Val Ala Leu Ala Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ser Pro Val Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Leu
115 120 125
Asp Met Thr Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Ser Tyr
130 135 140
Val Val Ile Asp Pro Gly Met Thr Thr Leu Glu Lys Gly Gln Leu Leu
145 150 155 160
Asn Asp Glu Gln Tyr Phe Glu Ala Leu Glu Glu Phe Gly Asp Asp Phe
165 170 175
Asp Ala Arg Met Gly Ala Glu Ala Val His Glu Leu Leu Asn Ala Ile
180 185 190
Asp Leu Glu His Glu Ile Gly Arg Leu Arg Glu Glu Ile Pro Gln Thr
195 200 205
Asn Ser Glu Thr Lys Ile Lys Lys Leu Ser Lys Arg Leu Lys Leu Met
210 215 220
Glu Ala Phe Gln Gly Ser Gly Asn Lys Pro Glu Trp Met Val Leu Thr
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Asp Leu Ala Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ser Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Thr Val Gly Pro Thr Leu Arg Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Phe Gly Lys
370 375 380
Leu Glu Gly Arg Gly Met Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Leu Pro Glu Val Trp Asp Val Leu Ala Glu Val Ile
405 410 415
Arg Glu His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Ala Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Asn Gly
485 490 495
Glu Pro Ile Ile Val Pro Ser Gln Asp Val Val Met Gly Leu Tyr Tyr
500 505 510
Met Thr Arg Glu Ala Ile Asn Ala Lys Gly Glu Gly Met Ala Phe Ala
515 520 525
Asp Leu Gln Glu Val Asp Arg Ala Tyr Arg Ser Gly Gln Ala Ser Leu
530 535 540
His Ala Arg Val Lys Val Arg Ile Asn Glu Lys Ile Lys Gly Glu Asp
545 550 555 560
Gly Gln Leu Thr Ala Asn Thr Arg Ile Val Asp Thr Thr Val Gly Arg
565 570 575
Ala Leu Leu Phe Gln Val Val Pro Ala Gly Leu Pro Phe Asp Val Val
580 585 590
Asn Gln Ser Met Lys Lys Lys Ala Ile Ser Lys Leu Ile Asn His Cys
595 600 605
Tyr Arg Val Val Gly Leu Lys Asp Thr Val Ile Phe Ala Asp Gln Leu
610 615 620
Met Tyr Thr Gly Phe Ala Tyr Ser Thr Ile Ser Gly Val Ser Ile Gly
625 630 635 640
Val Asn Asp Phe Val Ile Pro Asp Glu Lys Ala Arg Ile Ile Asn Ala
645 650 655
Ala Thr Asp Glu Val Lys Glu Ile Glu Ser Gln Tyr Ala Ser Gly Leu
660 665 670
Val Thr Gln Gly Glu Lys Tyr Asn Lys Val Ile Asp Leu Trp Ser Lys
675 680 685
Ala Asn Asp Glu Val Ser Lys Ala Met Met Ala Asn Leu Ser Lys Glu
690 695 700
Lys Val Val Asp Arg Glu Gly Lys Glu Val Asp Gln Glu Ser Phe Asn
705 710 715 720
Ser Met Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln
725 730 735
Ile Arg Gln Leu Ala Gly Met Arg Gly Leu Met Ala Lys Pro Asp Gly
740 745 750
Ser Ile Ile Glu Thr Pro Ile Thr Ala Asn Phe Arg Glu Gly Leu Asn
755 760 765
Val Leu Gln Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Leu Val Val Thr Glu Ile Asp Cys Gly Thr
805 810 815
Glu His Gly Leu Leu Met Ser Pro His Ile Glu Gly Gly Asp Val Val
820 825 830
Glu Pro Leu Gly Glu Arg Val Leu Gly Arg Val Ile Ala Arg Asp Val
835 840 845
Phe Lys Pro Gly Ser Asp Glu Val Ile Val Pro Ala Gly Thr Leu Ile
850 855 860
Asp Glu Lys Trp Val Asp Phe Leu Glu Val Met Ser Val Asp Glu Val
865 870 875 880
Val Val Arg Ser Pro Ile Thr Cys Glu Thr Arg His Gly Ile Cys Ala
885 890 895
Met Cys Tyr Gly Arg Asp Leu Ala Arg Gly His Arg Val Asn Ile Gly
900 905 910
Glu Ala Val Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
915 920 925
Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser Arg Thr
930 935 940
Ser Ala Ala Asp Asn Val Gln Val Lys Asn Gly Gly Thr Ile Arg Leu
945 950 955 960
His Asn Leu Lys His Val Val Arg Ala Asp Gly Ala Leu Val Ala Val
965 970 975
Ser Arg Ser Gly Glu Leu Ala Val Ala Asp Asp Phe Gly Arg Glu Arg
980 985 990
Glu Arg Tyr Lys Leu Pro Tyr Gly Ala Val Ile Ser Val Lys Glu Gly
995 1000 1005
Asp Lys Val Asp Pro Gly Ala Ile Val Ala Lys Trp Asp Pro His Thr
1010 1015 1020
His Pro Ile Val Thr Glu Val Asp Gly Thr Val Ala Phe Val Gly Met
1025 1030 1035 1040
Glu Glu Gly Ile Thr Val Lys Arg Gln Thr Asp Glu Leu Thr Gly Leu
1045 1050 1055
Thr Asn Ile Glu Val Met Asp Pro Lys Asp Arg Pro Ala Ala Gly Lys
1060 1065 1070
Asp Ile Arg Pro Ala Val Lys Leu Ile Asp Ala Ala Gly Lys Asp Leu
1075 1080 1085
Leu Leu Pro Gly Thr Asp Val Pro Ala Gln Tyr Phe Leu Pro Ala Asn
1090 1095 1100
Ala Leu Val Asn Leu Thr Asp Gly Ala Lys Val Ser Ile Gly Asp Val
1105 1110 1115 1120
Val Ala Arg Ile Pro Gln Glu Thr Ser Lys Thr Arg Asp Ile Thr Gly
1125 1130 1135
Gly Leu Pro Arg Val Ala Asp Leu Phe Glu Ala Arg Arg Pro Lys Glu
1140 1145 1150
Pro Ser Ile Leu Ala Glu Ile Ser Gly Thr Ile Ser Phe Gly Lys Glu
1155 1160 1165
Thr Lys Gly Lys Arg Arg Leu Val Ile Thr Pro Asn Asp Gly Ser Asp
1170 1175 1180
Pro Tyr Glu Glu Leu Ile Pro Lys Trp Arg His Leu Asn Val Phe Glu
1185 1190 1195 1200
Gly Glu Gln Val Asn Arg Gly Glu Val Ile Ser Asp Gly Pro Ser Asn
1205 1210 1215
Pro His Asp Ile Leu Arg Leu Leu Gly Val Ser Ser Leu Ala Lys Tyr
1220 1225 1230
Ile Val Asn Glu Ile Gln Asp Val Tyr Arg Leu Gln Gly Val Lys Ile
1235 1240 1245
Asn Asp Lys His Ile Glu Thr Ile Leu Arg Gln Met Leu Arg Lys Val
1250 1255 1260
Glu Val Ser Glu Ser Gly Asp Ser Ser Phe Ile Lys Gly Asp Gln Val
1265 1270 1275 1280
Glu Leu Thr Gln Val Leu Glu Glu Asn Glu Gln Leu Gly Thr Glu Asp
1285 1290 1295
Lys Phe Pro Ala Lys Tyr Glu Arg Val Leu Leu Gly Ile Thr Lys Ala
1300 1305 1310
Ser Leu Ser Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe Gln Glu Thr
1315 1320 1325
Thr Arg Val Leu Thr Glu Ala Ala Val Thr Gly Lys Arg Asp Phe Leu
1330 1335 1340
Arg Gly Leu Lys Glu Asn Val Val Val Gly Arg Leu Ile Pro Ala Gly
1345 1350 1355 1360
Thr Gly Leu Ala Tyr His Ser Glu Arg Lys Arg Gln Arg Asp Leu Gly
1365 1370 1375
Lys Pro Gln Arg Val Ser Ala Ser Glu Ala Glu Ala Ala Leu Thr Glu
1380 1385 1390
Ala Leu Asn Ser Ser Gly Asn
1395
<210> 39
<211> 1401
<212> PRT
<213> 霍乱弧菌（Vibrio cholerae）
<400> 39
Met Lys Asp Leu Leu Asn Phe Leu Lys Ala Gln His Lys Thr Glu Glu
1 5 10 15
Phe Asp Ala Ile Lys Ile Gly Leu Ala Ser Pro Asp Met Ile Arg Ser
20 25 30
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Arg Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Arg Ile Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Val Thr Gln Thr Lys
85 90 95
Val Arg Arg Asp Arg Met Gly His Ile Glu Leu Ala Ser Pro Val Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Met
115 120 125
Asp Met Pro Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Met Tyr
130 135 140
Val Val Thr Glu Pro Gly Met Thr Asp Leu Glu Arg Gly Gln Met Leu
145 150 155 160
Thr Glu Glu Glu Tyr Leu Asp Arg Leu Glu Glu Trp Gly Asp Glu Phe
165 170 175
Thr Ala Lys Met Gly Ala Glu Ala Ile Lys Asp Leu Leu Ala Ser Met
180 185 190
Asp Leu Pro Ala Glu Ala Glu Gln Met Arg Glu Glu Leu Asp Thr Thr
195 200 205
Asn Ser Glu Thr Lys Arg Lys Lys Leu Thr Lys Arg Leu Lys Leu Val
210 215 220
Glu Ala Phe Val Ala Ser Gly Asn Lys Pro Glu Trp Met Ile Leu Thr
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Glu Leu Ala Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ser Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ser Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Thr Val Gly Pro Tyr Leu Arg Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Tyr Ser Lys
370 375 380
Leu Glu Thr Arg Gly Leu Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Glu Ala Val Val Trp Asp Ile Leu Asp Glu Val Ile
405 410 415
Arg Glu His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Thr Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Ser Gly
485 490 495
Asp Pro Ile Ile Val Pro Ser Gln Asp Val Val Leu Gly Leu Tyr Tyr
500 505 510
Met Thr Arg Glu Lys Ile Asn Ala Lys Gly Glu Gly Met Tyr Leu Thr
515 520 525
Gly Pro Ala Glu Ala Glu Lys Ala Tyr Arg Thr Lys Thr Ala Glu Leu
530 535 540
His Ala Arg Val Lys Val Arg Ile Thr Glu Thr Ile Lys His Glu Asn
545 550 555 560
Gly Lys Leu Thr Thr Glu Thr Lys Met Ile Asp Thr Thr Val Gly Arg
565 570 575
Ala Met Leu Trp Gln Ile Val Pro Lys Gly Leu Pro Tyr Ser Leu Val
580 585 590
Asn Gln Lys Leu Gly Lys Lys Gln Ile Ser Asn Leu Leu Asn Glu Ala
595 600 605
Tyr Arg Lys Leu Gly Leu Lys Asp Thr Val Ile Phe Ala Asp Gln Ile
610 615 620
Met Tyr Thr Gly Phe Ala Tyr Ala Ala Leu Ser Gly Val Ser Val Gly
625 630 635 640
Ile Asp Asp Met Val Val Pro Ala Ala Lys Tyr Thr Glu Ile Ala Glu
645 650 655
Ala Glu Glu Glu Val Arg Glu Ile Gln Glu Gln Phe Gln Ser Gly Leu
660 665 670
Val Thr Ala Gly Glu Arg Tyr Asn Lys Val Ile Asp Ile Trp Ala Ser
675 680 685
Thr Asn Asp Arg Val Ala Lys Ala Met Met Glu Asn Leu Ser Ser Glu
690 695 700
Gln Val Ile Asn Arg Gln Gly Glu Gln Glu Lys Gln Glu Ser Phe Asn
705 710 715 720
Ser Ile Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln
725 730 735
Ile Arg Gln Leu Ala Gly Met Arg Gly Leu Met Ala Arg Pro Asp Gly
740 745 750
Ser Ile Ile Glu Thr Pro Ile Thr Ala Asn Phe Lys Glu Gly Leu Asn
755 760 765
Val Leu Gln Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Val Val Val Thr Glu His Asp Cys Gly Thr
805 810 815
Leu Glu Gly Val Val Met Thr Pro His Ile Glu Gly Gly Asp Val Lys
820 825 830
Val Ala Leu Thr Glu Leu Ala Leu Gly Arg Val Val Ser Glu Asp Ile
835 840 845
Leu Lys Pro Gly Thr Asp Glu Val Leu Ile Pro Arg Asn Thr Leu Leu
850 855 860
Asp Glu Lys Trp Cys Lys Val Ile Asn Asp Asn Ser Val Asp Gln Ile
865 870 875 880
Lys Val Arg Ser Val Val Thr Cys Asp Ser Asp Phe Gly Cys Cys Ala
885 890 895
Gln Cys Tyr Gly Arg Asp Leu Ala Arg Gly His Leu Val Asn Gln Gly
900 905 910
Glu Ala Val Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
915 920 925
Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser Thr Ala
930 935 940
Ala Ala Glu Asn Ser Ile Gln Ala Lys Asn Asn Gly Ser Val Lys Leu
945 950 955 960
His Asn Ala Lys Phe Val Thr Asn Lys Asp Gly Lys Leu Val Ile Thr
965 970 975
Ser Arg Ala Ser Glu Leu Thr Ile Ile Asp Glu Phe Gly Arg Thr Lys
980 985 990
Glu Lys His Lys Leu Pro Tyr Gly Ser Met Leu Ser Lys Ala Asp Gly
995 1000 1005
Asp Ala Val Ala Ala Gly Glu Thr Val Ala Asn Trp Glu Ala His Thr
1010 1015 1020
Met Pro Ile Ile Thr Glu Val Ala Gly Arg Val Gln Phe Val Asp Met
1025 1030 1035 1040
Ile Asp Gly Val Thr Val Ser Arg Gln Thr Asp Asp Leu Thr Gly Leu
1045 1050 1055
Ser Ser Ser Glu Val Thr Glu Ala Ala Ala Arg Pro Ala Ala Gly Lys
1060 1065 1070
Asp Met Arg Pro Ala Ile Lys Leu Val Asp Ala Asn Gly Lys Asp Val
1075 1080 1085
Leu Ile Pro Gly Thr Asp Met Pro Ala Gln Tyr Phe Leu Pro Gly Lys
1090 1095 1100
Ala Ile Val Asn Leu Asp Asp Gly Ala Glu Val Asn Val Gly Asp Thr
1105 1110 1115 1120
Leu Ala Arg Ile Pro Gln Lys Ser Gly Gly Asn Lys Asp Ile Thr Gly
1125 1130 1135
Gly Leu Pro Arg Val Ala Asp Leu Phe Glu Ala Arg Lys Pro Lys Glu
1140 1145 1150
Pro Ala Ile Leu Ala Glu His Ser Gly Thr Val Ser Phe Gly Lys Glu
1155 1160 1165
Thr Lys Gly Lys Arg Arg Leu Ile Ile Thr Arg Asp Ser Gly Asp Thr
1170 1175 1180
Tyr Glu Glu Met Ile Pro Lys His Arg Gln Leu Asn Val Phe Glu Gly
1185 1190 1195 1200
Glu Arg Ile Glu Arg Gly Asp Val Ile Ala Asp Gly Pro Glu Ser Pro
1205 1210 1215
His Asp Ile Leu Arg Leu Arg Gly Ile His Ala Val Thr Thr Tyr Ile
1220 1225 1230
Ala Asn Glu Val Gln Glu Val Tyr Arg Leu Gln Gly Val Lys Ile Asn
1235 1240 1245
Asp Lys His Ile Glu Thr Ile Val Arg Gln Met Leu Arg Lys Cys Thr
1250 1255 1260
Ile Thr Phe Ala Gly Asp Ser Glu Phe Leu Pro Gly Glu Thr Val Glu
1265 1270 1275 1280
Tyr Ser Gln Val Lys Ile Ala Asn Arg Lys Leu Val Glu Glu Gly Lys
1285 1290 1295
Glu Pro Ala Arg Phe Glu Arg Glu Leu Leu Gly Ile Thr Lys Ala Ser
1300 1305 1310
Leu Ala Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe Gln Glu Thr Thr
1315 1320 1325
Arg Val Leu Thr Glu Ala Ala Val Ser Gly Lys Arg Asp Asp Leu Arg
1330 1335 1340
Gly Leu Lys Glu Asn Val Ile Val Gly Arg Leu Ile Pro Ala Gly Thr
1345 1350 1355 1360
Gly Phe Ala Tyr His Gln Asp Arg Gln Ala Lys Arg Ala Gln Glu Gln
1365 1370 1375
Gln Gly Pro Ser Ala Glu Gln Ala Thr Asp Asn Leu Ala Ala Leu Leu
1380 1385 1390
Asn Ala Gly Phe Ser Ser Asp Asp Glu
1395 1400
<210> 40
<211> 1407
<212> PRT
<213> 肠道沙门氏菌（Salmonella enterica）
<400> 40
Met Lys Asp Leu Leu Lys Phe Leu Lys Ala Gln Thr Lys Thr Glu Glu
1 5 10 15
Phe Asp Ala Ile Lys Ile Ala Leu Ala Ser Pro Asp Met Ile Arg Ser
20 25 30
Trp Ser Phe Gly Glu Val Lys Lys Pro Glu Thr Ile Asn Tyr Arg Thr
35 40 45
Phe Lys Pro Glu Arg Asp Gly Leu Phe Cys Ala Arg Ile Phe Gly Pro
50 55 60
Val Lys Asp Tyr Glu Cys Leu Cys Gly Lys Tyr Lys Arg Leu Lys His
65 70 75 80
Arg Gly Val Ile Cys Glu Lys Cys Gly Val Glu Val Thr Gln Thr Lys
85 90 95
Val Arg Arg Glu Arg Met Gly His Ile Glu Leu Ala Ser Pro Thr Ala
100 105 110
His Ile Trp Phe Leu Lys Ser Leu Pro Ser Arg Ile Gly Leu Leu Leu
115 120 125
Asp Met Pro Leu Arg Asp Ile Glu Arg Val Leu Tyr Phe Glu Ser Tyr
130 135 140
Val Val Ile Glu Gly Gly Met Thr Asn Leu Glu Arg Gln Gln Ile Leu
145 150 155 160
Thr Glu Glu Gln Tyr Leu Asp Ala Leu Glu Glu Phe Gly Asp Glu Phe
165 170 175
Asp Ala Lys Met Gly Ala Glu Ala Ile Gln Ala Leu Leu Lys Ser Met
180 185 190
Asp Leu Glu Gln Glu Cys Glu Thr Leu Arg Glu Glu Leu Asn Glu Thr
195 200 205
Asn Ser Glu Thr Lys Arg Lys Lys Leu Thr Lys Arg Ile Lys Leu Leu
210 215 220
Glu Ala Phe Val Gln Ser Gly Asn Lys Pro Glu Trp Met Ile Leu Thr
225 230 235 240
Val Leu Pro Val Leu Pro Pro Asp Leu Arg Pro Leu Val Pro Leu Asp
245 250 255
Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val
260 265 270
Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu Asp Leu Ala Ala Pro
275 280 285
Asp Ile Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu Ala Val Asp
290 295 300
Ala Leu Leu Asp Asn Gly Arg Arg Gly Arg Ala Ile Thr Gly Ser Asn
305 310 315 320
Lys Arg Pro Leu Lys Ser Leu Ala Asp Met Ile Lys Gly Lys Gln Gly
325 330 335
Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg
340 345 350
Ser Val Ile Thr Val Gly Pro Tyr Leu Arg Leu His Gln Cys Gly Leu
355 360 365
Pro Lys Lys Met Ala Leu Glu Leu Phe Lys Pro Phe Ile Tyr Gly Lys
370 375 380
Leu Glu Leu Arg Gly Leu Ala Thr Thr Ile Lys Ala Ala Lys Lys Met
385 390 395 400
Val Glu Arg Glu Glu Ala Val Val Trp Asp Ile Leu Asp Glu Val Ile
405 410 415
Arg Glu His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu
420 425 430
Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys Ala Ile Gln
435 440 445
Leu His Pro Leu Val Cys Ala Ala Tyr Asn Ala Asp Phe Asp Gly Asp
450 455 460
Gln Met Ala Val His Val Pro Leu Thr Leu Glu Ala Gln Leu Glu Ala
465 470 475 480
Arg Ala Leu Met Met Ser Thr Asn Asn Ile Leu Ser Pro Ala Asn Gly
485 490 495
Glu Pro Ile Ile Val Pro Ser Gln Asp Val Val Leu Gly Leu Tyr Tyr
500 505 510
Met Thr Arg Asp Cys Val Asn Ala Lys Gly Glu Gly Met Val Leu Thr
515 520 525
Gly Pro Lys Glu Ala Glu Arg Ile Tyr Arg Ala Gly Leu Ala Ser Leu
530 535 540
His Ala Arg Val Lys Val Arg Ile Thr Glu Tyr Glu Lys Asp Glu Asn
545 550 555 560
Gly Glu Phe Val Ala His Thr Ser Leu Lys Asp Thr Thr Val Gly Arg
565 570 575
Ala Ile Leu Trp Met Ile Val Pro Lys Gly Leu Pro Phe Ser Ile Val
580 585 590
Asn Gln Ala Leu Gly Lys Lys Ala Ile Ser Lys Met Leu Asn Thr Cys
595 600 605
Tyr Arg Ile Leu Gly Leu Lys Pro Thr Val Ile Phe Ala Asp Gln Thr
610 615 620
Met Tyr Thr Gly Phe Ala Tyr Ala Ala Arg Ser Gly Ala Ser Val Gly
625 630 635 640
Ile Asp Asp Met Val Ile Pro Glu Lys Lys His Glu Ile Ile Ser Glu
645 650 655
Ala Glu Ala Glu Val Ala Glu Ile Gln Glu Gln Phe Gln Ser Gly Leu
660 665 670
Val Thr Ala Gly Glu Arg Tyr Asn Lys Val Ile Asp Ile Trp Ala Ala
675 680 685
Ala Asn Asp Arg Val Ser Lys Ala Met Met Asp Asn Leu Gln Thr Glu
690 695 700
Thr Val Ile Asn Arg Asp Gly Gln Glu Glu Gln Gln Val Ser Phe Asn
705 710 715 720
Ser Ile Tyr Met Met Ala Asp Ser Gly Ala Arg Gly Ser Ala Ala Gln
725 730 735
Ile Arg Gln Leu Ala Gly Met Arg Gly Leu Met Ala Lys Pro Asp Gly
740 745 750
Ser Ile Ile Glu Thr Pro Ile Thr Ala Asn Phe Arg Glu Gly Leu Asn
755 760 765
Val Leu Gln Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Leu Val Val Thr Glu Asp Asp Cys Gly Thr
805 810 815
His Glu Gly Ile Leu Met Thr Pro Val Ile Glu Gly Gly Asp Val Lys
820 825 830
Glu Pro Leu Arg Asp Arg Val Leu Gly Arg Val Thr Ala Glu Asp Val
835 840 845
Leu Lys Pro Gly Thr Ala Asp Ile Leu Val Pro Arg Asn Thr Leu Leu
850 855 860
His Glu Gln Trp Cys Asp Leu Leu Glu Ala Asn Ser Val Asp Ala Val
865 870 875 880
Lys Val Arg Ser Val Val Ser Cys Asp Thr Asp Phe Gly Val Cys Ala
885 890 895
His Cys Tyr Gly Arg Asp Leu Ala Arg Gly His Ile Ile Asn Lys Gly
900 905 910
Glu Ala Ile Gly Val Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
915 920 925
Gln Leu Thr Met Arg Thr Phe His Ile Gly Gly Ala Ala Ser Arg Ala
930 935 940
Ala Ala Glu Ser Ser Ile Gln Val Lys Asn Lys Gly Ser Ile Lys Leu
945 950 955 960
Ser Asn Val Lys Ser Val Val Asn Ser Ser Gly Lys Leu Val Ile Thr
965 970 975
Ser Arg Asn Thr Glu Leu Lys Leu Ile Asp Glu Phe Gly Arg Thr Lys
980 985 990
Glu Ser Tyr Lys Val Pro Tyr Gly Ala Val Met Ala Lys Gly Asp Gly
995 1000 1005
Glu Gln Val Ala Gly Gly Glu Thr Val Ala Asn Trp Asp Pro His Thr
1010 1015 1020
Met Pro Val Ile Thr Glu Val Ser Gly Phe Ile Arg Phe Thr Asp Met
1025 1030 1035 1040
Ile Asp Gly Gln Thr Ile Thr Arg Gln Thr Asp Glu Leu Thr Gly Leu
1045 1050 1055
Ser Ser Leu Val Val Leu Asp Ser Ala Glu Arg Thr Thr Gly Gly Lys
1060 1065 1070
Asp Leu Arg Pro Ala Leu Lys Ile Val Asp Ala Gln Gly Asn Asp Val
1075 1080 1085
Leu Ile Pro Gly Thr Asp Met Pro Ala Gln Tyr Phe Leu Pro Gly Lys
1090 1095 1100
Ala Ile Val Gln Leu Glu Asp Gly Val Gln Ile Ser Ser Gly Asp Thr
1105 1110 1115 1120
Leu Ala Arg Ile Pro Gln Glu Ser Gly Gly Thr Lys Asp Ile Thr Gly
1125 1130 1135
Gly Leu Pro Arg Val Ala Asp Leu Phe Glu Ala Arg Arg Pro Lys Glu
1140 1145 1150
Pro Ala Ile Leu Ala Glu Ile Ala Gly Ile Val Ser Phe Gly Lys Glu
1155 1160 1165
Thr Lys Gly Lys Arg Arg Leu Val Ile Thr Pro Val Asp Gly Ser Asp
1170 1175 1180
Pro Tyr Glu Glu Met Ile Pro Lys Trp Arg Gln Leu Asn Val Phe Glu
1185 1190 1195 1200
Gly Glu Arg Val Glu Arg Gly Asp Val Ile Ser Asp Gly Pro Glu Ala
1205 1210 1215
Pro His Asp Ile Leu Arg Leu Arg Gly Val His Ala Val Thr Arg Tyr
1220 1225 1230
Ile Val Asn Glu Val Gln Asp Val Tyr Arg Leu Gln Gly Val Lys Ile
1235 1240 1245
Asn Asp Lys His Ile Glu Val Ile Val Arg Gln Met Leu Arg Lys Ala
1250 1255 1260
Thr Ile Glu Ser Ala Gly Ser Ser Asp Phe Leu Glu Gly Glu Gln Val
1265 1270 1275 1280
Glu Tyr Ser Arg Val Lys Ile Ala Asn Arg Glu Leu Glu Ala Asn Gly
1285 1290 1295
Lys Val Gly Ala Thr Phe Ser Arg Asp Leu Leu Gly Ile Thr Lys Ala
1300 1305 1310
Ser Leu Ala Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe Gln Glu Thr
1315 1320 1325
Thr Arg Val Leu Thr Glu Ala Ala Val Ala Gly Lys Arg Asp Glu Leu
1330 1335 1340
Arg Gly Leu Lys Glu Asn Val Ile Val Gly Arg Leu Ile Pro Ala Gly
1345 1350 1355 1360
Thr Gly Tyr Ala Tyr His Gln Asp Arg Met Arg Arg Arg Ala Ala Gly
1365 1370 1375
Glu Gln Pro Ala Thr Pro Gln Val Thr Ala Glu Asp Ala Ser Ala Ser
1380 1385 1390
Leu Ala Glu Leu Leu Asn Ala Gly Leu Gly Gly Ser Asp Asn Glu
1395 1400 1405
<210> 41
<211> 1300
<212> PRT
<213> 溶齿放线菌（Actinomyces odontolyticus）
<400> 41
Met Leu Asp Ala Lys Thr Phe Asp Ser Leu Lys Ile Thr Leu Ala Thr
1 5 10 15
Gly Asp Asp Ile Ala Glu Trp Ser His Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Arg Asp Gly Leu Phe Gly
35 40 45
Glu Gln Ile Phe Gly Pro Thr Arg Asp Trp Glu Cys Ala Cys Gly Lys
50 55 60
Tyr Lys Arg Val Arg Tyr Lys Gly Ile Val Cys Glu Lys Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ser Arg Val Arg Arg Glu Arg Met Gly His Ile Asp
85 90 95
Leu Ala Ala Pro Val Thr His Ile Trp Tyr Phe Lys Gly Val Pro Ser
100 105 110
Arg Leu Gly Tyr Val Leu Asn Leu Ala Pro Lys Asp Leu Glu Lys Val
115 120 125
Ile Tyr Phe Ala Ala Tyr Met Ile Thr Glu Val Asp Glu Lys Gly Arg
130 135 140
His Glu Asp Leu Ala Glu Leu Arg Ala Glu Leu Glu Val Gln Lys Lys
145 150 155 160
Gln Met Glu Asn Asn Arg Asp Ala Thr Ile Asn Asp Phe Ala Glu Gln
165 170 175
Leu Glu Ser Asp Met Ala Ala Leu Glu Lys Asp Gly Ala Ser Gln Ala
180 185 190
Glu Arg Glu Arg Ala Arg Lys Gln Gly Glu Arg Glu Met Ala Lys Ile
195 200 205
Arg Arg Arg Phe Asp Gly Asp Ile Glu Gly Leu Glu Ala Val Trp Glu
210 215 220
Arg Phe Lys Asp Leu Lys Val Gly Asp Leu Glu Gly Asp Glu Arg Leu
225 230 235 240
Tyr Arg Ala Met Val Ala Arg Tyr Gly Thr Tyr Phe Lys Gly Asp Met
245 250 255
Gly Ala Ala Ala Ile Gln Lys Arg Leu Glu Thr Phe Asp Leu Glu Ala
260 265 270
Glu Val Ala Ala Leu Arg Gln Thr Ile Gly Ser Asp Ser Gly Pro Arg
275 280 285
Lys Ala Arg Ala Ile Lys Arg Leu Lys Val Ile Asn Ala Phe Val Ala
290 295 300
Thr Gly Asn Ser Pro Ala Ser Met Val Leu Thr Lys Ile Pro Val Ile
305 310 315 320
Pro Pro Asp Leu Arg Pro Met Val Gln Leu Asp Gly Gly Arg Phe Ala
325 330 335
Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Thr
340 345 350
Arg Leu Lys Arg Leu Leu Glu Leu Gly Ala Pro Glu Ile Ile Val Asn
355 360 365
Asn Glu Lys Arg Met Leu Gln Glu Ser Val Asp Ala Leu Phe Asp Asn
370 375 380
Gly Arg Arg Gly Arg Pro Val Ala Gly Pro Gly Asn Arg Pro Leu Lys
385 390 395 400
Ser Ile Ser Asp Met Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn
405 410 415
Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg Ser Val Ile Val Val
420 425 430
Gly Pro Gln Leu Gln Leu His Gln Cys Gly Leu Pro Lys Gln Met Ala
435 440 445
Leu Glu Leu Phe Lys Pro Phe Val Met Lys Arg Leu Val Glu Lys Asn
450 455 460
Tyr Ala Gln Asn Val Lys Ala Ala Lys Arg Lys Val Glu Arg Gln Arg
465 470 475 480
Pro Glu Val Trp Asp Val Leu Asp Asp Val Ile Arg Glu His Pro Val
485 490 495
Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe
500 505 510
Glu Pro Gln Leu Ile Glu Gly Lys Ala Ile Gln Leu His Pro Leu Ala
515 520 525
Cys Gly Ala Phe Asn Ala Asp Phe Asp Gly Asp Gln Met Ala Val His
530 535 540
Leu Pro Leu Gly Ala Glu Ala Gln Ala Glu Ala Arg Ile Leu Met Leu
545 550 555 560
Ser Thr Asn Asn Ile Leu Lys Pro Ser Asp Gly Arg Pro Val Ala Met
565 570 575
Pro Ser Gln Asp Met Ile Ile Gly Leu Phe His Leu Thr Ser Thr Pro
580 585 590
Asp Pro Ser Val Pro Val Glu Lys Asp Glu Asp Gly Asn Pro Val Ile
595 600 605
Pro Tyr Phe Ser Ser Gln Ala Glu Ala Gln Met Ala Tyr Asp Ala Gly
610 615 620
Asn Leu His Leu Asn Ala Thr Ala Arg Ile Arg Phe Ala Asp Gly Thr
625 630 635 640
Val Pro Pro Glu Gly Trp Glu Ala Pro Glu Gly Trp Glu Pro Gly Asp
645 650 655
Glu Leu Ile Leu Glu Thr Ser Leu Gly Arg Ala Ile Phe Asn Glu Gln
660 665 670
Leu Pro Thr Asp Tyr Pro Phe Ile Asn Glu Val Val Gly Lys Lys Gln
675 680 685
Leu Gly Asn Ile Val Asn Thr Leu Thr Gln Arg Tyr Pro Asn Val Leu
690 695 700
Val Ala Asp Cys Leu Asp Ala Leu Lys Ser Ala Gly Phe His Trp Ser
705 710 715 720
Thr Trp Ser Gly Ile Thr Ile Ala Phe Ser Asp Ile Gln Ala Ser Pro
725 730 735
Arg Lys Arg Glu Ile Leu Ala Arg Tyr Glu Ala Lys Ala Ala Glu Ile
740 745 750
Val Glu Gln Phe Glu Thr Gly Ile Ile Leu Glu Glu Thr Arg Tyr Glu
755 760 765
Glu Leu Val Lys Leu Trp Leu Gln Cys Thr Glu Glu Val Ala Asp Asp
770 775 780
Met Arg Ala Asn Phe Asp Glu Arg Asn Thr Val Tyr Arg Met Val Asn
785 790 795 800
Ser Gly Ala Arg Gly Asn Trp Ser Gln Val Gln Gln Ile Ala Gly Met
805 810 815
Arg Gly Leu Val Ser Asp Pro Lys Gln Lys Leu Ile Glu Gln Pro Ile
820 825 830
Lys Ala Asn Tyr Arg Glu Gly Leu Thr Val Leu Glu Tyr Phe Ile Ala
835 840 845
Thr His Gly Ala Arg Lys Gly Leu Val Asp Thr Ala Leu Arg Thr Ala
850 855 860
Glu Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val Ser Gln Asp Val
865 870 875 880
Ile Val Arg Glu Gly Asp Cys Gly Thr Arg Ala Gly Leu Lys Ile Asp
885 890 895
Ile Ala His Lys Asn Glu Phe Gly Glu Trp Glu Ala Ser Glu Thr Ile
900 905 910
Glu Thr Thr Ala Tyr Ala Arg Asn Leu Ala Arg Asp Ala Val Asn Glu
915 920 925
Ala Gly Glu Val Val Met Pro Ala Gly Thr Asp Leu Gly Asp Asp Gln
930 935 940
Leu Ala Glu Leu Val Ala Ala Gly Val Glu Gln Ile Val Cys Arg Ser
945 950 955 960
Val Leu Thr Cys Glu Ser Gln Val Gly Thr Cys Ala Ala Cys Tyr Gly
965 970 975
Arg Ser Leu Ala Thr Gly Lys Gln Val Asp Ile Gly Glu Ala Val Gly
980 985 990
Ile Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr Gln Leu Thr Met
995 1000 1005
Arg Thr Phe His Thr Gly Gly Ala Ala Ser Ala Ala Asp Ile Thr Gln
1010 1015 1020
Gly Leu Pro Arg Val Gln Glu Leu Phe Glu Ala Arg Ser Pro Lys Val
1025 1030 1035 1040
Glu Ala Lys Met Asn Glu Ala Ala Gly Arg Val His Ile Asp Asp Glu
1045 1050 1055
Asp Pro Ser Ala Arg Lys Val Val Ile Thr Arg Asp Asp Gly Lys Glu
1060 1065 1070
Asp Leu Val Ile Glu Val Ser Arg Arg Gln Lys Leu Leu Val Ser Glu
1075 1080 1085
Gly Gln His Ile Glu Ala Gly Thr Pro Leu Thr Glu Gly Gln Leu Asp
1090 1095 1100
Pro Lys Glu Ile Leu Arg Ile Met Gly Arg Asn Val Ser Gln Lys Met
1105 1110 1115 1120
Leu Val Asp Glu Val Gln Lys Val Tyr Arg Asp Gln Gly Val Gly Ile
1125 1130 1135
His Ala Lys His Ile Glu Val Ile Val Arg Gln Met Leu Arg Arg Val
1140 1145 1150
Thr Ile Leu Glu Pro Gly Asp Thr Thr Phe Met Pro Gly Glu Leu Val
1155 1160 1165
Asp Arg Met Ala Tyr Leu Thr Gln Asn Arg Arg Val Ala Ala Glu Gly
1170 1175 1180
Gly Gln Pro Ala Ser Gly Arg Gln Met Leu Met Gly Ile Thr Lys Ala
1185 1190 1195 1200
Ser Leu Ala Thr Asp Ser Trp Leu Ser Ala Ala Ser Phe Gln Glu Thr
1205 1210 1215
Thr Lys Val Leu Thr Glu Ala Ala Met Asn Gly Lys Ser Asp Ser Leu
1220 1225 1230
Val Gly Leu Lys Glu Asn Val Ile Leu Gly Lys Leu Ile Pro Ala Gly
1235 1240 1245
Thr Gly Leu Ser Arg Tyr Asn Asp Val Ile Val Glu Pro Thr Ala Glu
1250 1255 1260
Ala Met Ala Asn Ser Asn Tyr Ser Glu Ala Asp Phe Gly Asp Gly Ser
1265 1270 1275 1280
Val Ser Glu Asp Phe Leu Asp Ala Leu Gly Ala Ile Asp Phe Gly Met
1285 1290 1295
Asn Phe Arg Glu
1300
<210> 42
<211> 1299
<212> PRT
<213> 天蓝色链霉菌（Streptomyces coelicolor）
<400> 42
Met Leu Asp Val Asn Phe Phe Asp Glu Leu Arg Ile Gly Leu Ala Thr
1 5 10 15
Ala Asp Asp Ile Arg Gln Trp Ser His Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Lys Asp Gly Leu Phe Cys
35 40 45
Glu Lys Ile Phe Gly Pro Thr Arg Asp Trp Glu Cys Tyr Cys Gly Lys
50 55 60
Tyr Lys Arg Val Arg Phe Lys Gly Ile Ile Cys Glu Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ala Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Thr His Ile Trp Tyr Phe Lys Gly Val Pro Ser
100 105 110
Arg Leu Gly Tyr Leu Leu Asp Leu Ala Pro Lys Asp Leu Glu Lys Val
115 120 125
Ile Tyr Phe Ala Ala Tyr Met Ile Thr Phe Val Asp Glu Glu Arg Arg
130 135 140
Thr Arg Asp Leu Pro Ser Leu Glu Ala His Val Ser Val Glu Arg Gln
145 150 155 160
Gln Ile Glu Gln Arg Arg Asp Ser Asp Leu Glu Ala Arg Ala Lys Lys
165 170 175
Leu Glu Thr Asp Leu Ala Glu Leu Glu Ala Glu Gly Ala Lys Ala Asp
180 185 190
Val Arg Arg Lys Val Arg Glu Gly Ala Glu Arg Glu Met Lys Gln Leu
195 200 205
Arg Asp Arg Ala Gln Arg Glu Ile Asp Arg Leu Asp Glu Val Trp Asn
210 215 220
Arg Phe Lys Asn Leu Lys Val Gln Asp Leu Glu Gly Asp Glu Leu Leu
225 230 235 240
Tyr Arg Glu Leu Arg Asp Arg Phe Gly Thr Tyr Phe Asp Gly Ser Met
245 250 255
Gly Ala Ala Ala Leu Gln Lys Arg Leu Glu Ser Phe Asp Leu Asp Glu
260 265 270
Glu Ala Glu Arg Leu Arg Glu Ile Ile Arg Thr Gly Lys Gly Gln Lys
275 280 285
Lys Thr Arg Ala Leu Lys Arg Leu Lys Val Val Ser Ala Phe Leu Gln
290 295 300
Thr Ser Asn Ser Pro Lys Gly Met Val Leu Asp Cys Val Pro Val Ile
305 310 315 320
Pro Pro Asp Leu Arg Pro Met Val Gln Leu Asp Gly Gly Arg Phe Ala
325 330 335
Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn
340 345 350
Arg Leu Lys Arg Leu Leu Asp Leu Gly Ala Pro Glu Ile Ile Val Asn
355 360 365
Asn Glu Lys Arg Met Leu Gln Glu Ala Val Asp Ala Leu Phe Asp Asn
370 375 380
Gly Arg Arg Gly Arg Pro Val Thr Gly Pro Gly Asn Arg Pro Leu Lys
385 390 395 400
Ser Leu Ser Asp Met Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn
405 410 415
Leu Leu Gly Lys Arg Val Asp Tyr Ser Ala Arg Ser Val Ile Val Val
420 425 430
Gly Pro Gln Leu Lys Leu His Gln Cys Gly Leu Pro Lys Ala Met Ala
435 440 445
Leu Glu Leu Phe Lys Pro Phe Val Met Lys Arg Leu Val Asp Leu Asn
450 455 460
His Ala Gln Asn Ile Lys Ser Ala Lys Arg Met Val Glu Arg Gly Arg
465 470 475 480
Thr Val Val Tyr Asp Val Leu Glu Glu Val Ile Ala Glu His Pro Val
485 490 495
Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe
500 505 510
Glu Pro Gln Leu Val Glu Gly Lys Ala Ile Gln Ile His Pro Leu Val
515 520 525
Cys Thr Ala Phe Asn Ala Asp Phe Asp Gly Asp Gln Met Ala Val His
530 535 540
Leu Pro Leu Ser Ala Glu Ala Gln Ala Glu Ala Arg Ile Leu Met Leu
545 550 555 560
Ser Ser Asn Asn Ile Leu Lys Pro Ala Asp Gly Arg Pro Val Thr Met
565 570 575
Pro Thr Gln Asp Met Val Leu Gly Leu Phe Phe Leu Thr Thr Asp Ser
580 585 590
Glu Gly Arg Ser Pro Lys Gly Glu Gly Arg Ala Phe Gly Ser Ser Ala
595 600 605
Glu Ala Ile Met Ala Phe Asp Ala Gly Asp Leu Thr Leu Gln Ala Lys
610 615 620
Ile Asp Ile Arg Phe Pro Val Gly Thr Ile Pro Pro Arg Gly Phe Glu
625 630 635 640
Pro Pro Ala Arg Glu Glu Gly Glu Pro Glu Trp Gln Gln Gly Asp Thr
645 650 655
Phe Thr Leu Lys Thr Thr Leu Gly Arg Ala Leu Phe Asn Glu Leu Leu
660 665 670
Pro Glu Asp Tyr Pro Phe Val Asp Tyr Glu Val Gly Lys Lys Gln Leu
675 680 685
Ser Glu Ile Val Asn Asp Leu Ala Glu Arg Tyr Pro Lys Val Ile Val
690 695 700
Ala Ala Thr Leu Asp Asn Leu Lys Ala Ala Gly Phe Phe Trp Ala Thr
705 710 715 720
Arg Ser Gly Val Thr Val Ala Ile Ser Asp Ile Val Val Pro Asp Ala
725 730 735
Lys Lys Glu Ile Val Lys Gly Tyr Glu Gly Gln Asp Glu Lys Val Gln
740 745 750
Lys Gln Tyr Glu Arg Gly Leu Ile Thr Lys Glu Glu Arg Thr Gln Glu
755 760 765
Leu Ile Ala Ile Trp Thr Lys Ala Thr Asn Glu Val Ala Glu Ala Met
770 775 780
Asn Asp Asn Phe Pro Lys Thr Asn Pro Val Ser Met Met Val Asn Ser
785 790 795 800
Gly Ala Arg Gly Asn Met Met Gln Met Arg Gln Ile Ala Gly Met Arg
805 810 815
Gly Leu Val Ser Asn Ala Lys Asn Glu Thr Ile Pro Arg Pro Ile Lys
820 825 830
Ala Ser Phe Arg Glu Gly Leu Ser Val Leu Glu Tyr Phe Ile Ser Thr
835 840 845
His Gly Ala Arg Lys Gly Leu Ala Asp Thr Ala Leu Arg Thr Ala Asp
850 855 860
Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val Ser Gln Asp Val Ile
865 870 875 880
Ile Arg Glu Glu Asp Cys Gly Thr Glu Arg Gly Leu Lys Leu Pro Ile
885 890 895
Ala Thr Arg Asp Ala Asp Gly Thr Leu Arg Lys Ala Glu Asp Val Glu
900 905 910
Thr Ser Val Tyr Ala Arg Met Leu Ala Glu Asp Val Val Ile Asp Gly
915 920 925
Lys Val Ile Ala Pro Ala Asn Val Asp Leu Gly Asp Val Leu Ile Asp
930 935 940
Ala Leu Val Ala His Gly Val Glu Glu Val Lys Thr Arg Ser Ile Leu
945 950 955 960
Thr Cys Glu Ser Gln Val Gly Thr Cys Ala Met Cys Tyr Gly Arg Ser
965 970 975
Leu Ala Thr Gly Lys Leu Val Asp Ile Gly Glu Ala Val Gly Ile Ile
980 985 990
Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr Gln Leu Thr Met Arg Thr
995 1000 1005
Phe His Thr Gly Gly Val Ala Gly Asp Asp Ile Thr Gln Gly Leu Pro
1010 1015 1020
Arg Val Val Glu Leu Phe Glu Ala Arg Thr Pro Lys Gly Val Ala Pro
1025 1030 1035 1040
Ile Ser Glu Ala Ser Gly Arg Val Arg Ile Glu Glu Thr Glu Lys Thr
1045 1050 1055
Lys Lys Ile Val Val Thr Pro Asp Asp Gly Ser Asp Glu Thr Ala Phe
1060 1065 1070
Pro Ile Ser Lys Arg Ala Arg Leu Leu Val Gly Glu Gly Asp His Val
1075 1080 1085
Glu Val Gly Gln Lys Leu Thr Val Gly Ala Thr Asn Pro His Asp Val
1090 1095 1100
Leu Arg Ile Leu Gly Gln Arg Ala Val Gln Val His Leu Val Gly Glu
1105 1110 1115 1120
Val Gln Lys Val Tyr Asn Ser Gln Gly Val Ser Ile His Asp Lys His
1125 1130 1135
Ile Glu Ile Ile Ile Arg Gln Met Leu Arg Arg Val Thr Ile Ile Glu
1140 1145 1150
Ser Gly Asp Ala Glu Leu Leu Pro Gly Glu Leu Val Glu Arg Thr Lys
1155 1160 1165
Phe Glu Thr Glu Asn Arg Arg Val Val Gln Glu Gly Gly His Pro Ala
1170 1175 1180
Ser Gly Arg Pro Gln Leu Met Gly Ile Thr Lys Ala Ser Leu Ala Thr
1185 1190 1195 1200
Glu Ser Trp Leu Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg Val Leu
1205 1210 1215
Thr Asp Ala Ala Ile Asn Ala Lys Ser Asp Ser Leu Ile Gly Leu Lys
1220 1225 1230
Glu Asn Val Ile Ile Gly Lys Leu Ile Pro Ala Gly Thr Gly Leu Ser
1235 1240 1245
Arg Tyr Arg Asn Ile Arg Val Glu Pro Thr Glu Glu Ala Lys Ala Ala
1250 1255 1260
Met Tyr Ser Ala Val Gly Tyr Asp Asp Ile Asp Tyr Ser Pro Phe Gly
1265 1270 1275 1280
Thr Gly Ser Gly Gln Ala Val Pro Leu Glu Asp Tyr Asp Tyr Gly Pro
1285 1290 1295
Tyr Asn Gln
<210> 43
<211> 1336
<212> PRT
<213> 白喉棒状杆菌（Corynebacterium diphtheriae）
<400> 43
Met Ile Asp Val Asn Phe Phe Asp Glu Leu Arg Ile Gly Leu Ala Thr
1 5 10 15
Ala Asp Asp Ile Arg Arg Trp Ser Lys Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Lys Asp Gly Leu Phe Cys
35 40 45
Glu Arg Ile Phe Gly Pro Thr Arg Asp Trp Glu Cys Ala Cys Gly Lys
50 55 60
Tyr Lys Arg Val Arg Tyr Lys Gly Ile Ile Cys Glu Arg Cys Gly Val
65 70 75 80
Glu Val Thr Lys Ser Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Thr His Ile Trp Tyr Phe Lys Gly Val Pro Ser
100 105 110
Arg Leu Gly Tyr Leu Leu Asp Leu Ala Pro Lys Asp Leu Glu Arg Ile
115 120 125
Ile Tyr Phe Ala Ala Asn Ile Ile Thr Ser Val Asp Asp Glu Ala Arg
130 135 140
His Ala Asp Gln Thr Thr Leu Glu Ala Glu Met Leu Leu Glu Lys Lys
145 150 155 160
Asp Val Glu Ala Asp Met Glu Ser Glu Ile Ala Glu Arg Ala Ala Lys
165 170 175
Leu Glu Glu Asp Leu Ala Glu Leu Glu Ala Ala Gly Ala Lys Ala Asp
180 185 190
Ala Arg Asn Lys Val Lys Lys Ala Ala Glu Lys Glu Met Gln His Ile
195 200 205
Arg Glu Arg Ala Glu Arg Glu Ile Asp Arg Leu Glu Glu Ile Trp Gln
210 215 220
Thr Phe Ile Lys Leu Ala Pro Lys Gln Met Ile Ile Asp Glu Thr Ile
225 230 235 240
Tyr Glu Glu Leu Val Asp Arg Tyr Glu Asp Tyr Phe Thr Gly Gly Met
245 250 255
Gly Ala Glu Ala Ile Gln Thr Leu Ile Arg Asn Phe Asp Leu Asp Ser
260 265 270
Glu Ala Glu Glu Leu Arg Glu Ile Ile Asn Asn Gly Lys Gly Gln Lys
275 280 285
Lys Met Arg Ala Leu Lys Arg Leu Lys Val Val Ala Ala Phe Gln Arg
290 295 300
Ser Gly Asn Asp Pro Ala Gly Met Val Leu Asp Cys Ile Pro Val Ile
305 310 315 320
Pro Pro Glu Leu Arg Pro Met Val Gln Leu Asp Gly Gly Arg Phe Ala
325 330 335
Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn
340 345 350
Arg Leu Lys Arg Met Ile Glu Leu Gly Ala Pro Glu Ile Ile Val Asn
355 360 365
Asn Glu Lys Arg Met Leu Gln Glu Ser Val Asp Ala Leu Phe Asp Asn
370 375 380
Gly Arg Arg Gly Arg Pro Val Thr Gly Pro Gly Asn Arg Pro Leu Lys
385 390 395 400
Ser Leu Ser Asp Leu Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn
405 410 415
Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg Ser Val Ile Ile Val
420 425 430
Gly Pro Gln Leu Lys Leu His Glu Cys Gly Leu Pro Lys Leu Met Ala
435 440 445
Leu Glu Leu Phe Lys Pro Phe Val Met Lys Arg Leu Val Glu Asn Asp
450 455 460
Tyr Ala Gln Asn Ile Lys Ser Ala Lys Arg Met Val Glu Arg Gln Arg
465 470 475 480
Pro Glu Val Trp Asp Val Leu Glu Glu Ala Ile Ser Glu His Pro Val
485 490 495
Met Leu Asn Arg Ala Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe
500 505 510
Glu Pro Lys Leu Val Glu Gly Lys Ala Ile Gln Leu His Pro Leu Ala
515 520 525
Cys Glu Ala Phe Asn Ala Asp Phe Asp Gly Asp Gln Met Ala Val His
530 535 540
Leu Pro Leu Ser Ala Glu Ala Gln Ala Glu Ala Arg Ile Leu Met Leu
545 550 555 560
Ala Ser Asn Asn Ile Leu Ser Pro Ala Ser Gly Lys Pro Leu Ala Met
565 570 575
Pro Arg Leu Asp Met Val Thr Gly Leu Tyr Tyr Leu Thr Met Asp Lys
580 585 590
Asn Glu Asn Glu Ile Gly Gly Gln Gly Ala Tyr Ala Ser Ala Thr Glu
595 600 605
Glu Gly Pro Ala Gln Gly Val Tyr Ser Ser Tyr Ala Glu Ala Ile Met
610 615 620
Ala Arg Asp Arg Gly Val Leu Gly Leu Gln Ala Lys Ile Lys Val Arg
625 630 635 640
Ile Ser His Leu Arg Pro Pro Val Asp Ile Glu Ala Glu Gln Phe Pro
645 650 655
Glu Gly Trp Asn Lys Gly Asp Val Trp Leu Ala Asp Thr Thr Leu Gly
660 665 670
Arg Ile Met Phe Asn Glu Leu Leu Pro Trp Asn Tyr Pro Tyr Leu Glu
675 680 685
Gly Val Met Val Arg Lys Gly Gly Gly Thr Gly Lys Ile Met Leu Gly
690 695 700
Asp Val Ile Asn Asp Leu Ala Ala Thr Tyr Pro Met Ile Thr Val Ala
705 710 715 720
Gln Thr Met Asp Lys Met Lys Asp Ala Gly Phe Tyr Trp Ala Thr Arg
725 730 735
Ser Gly Val Thr Ile Thr Met Ser Asp Val Leu Val Leu Pro Asn Lys
740 745 750
Glu Glu Ile Leu Asp Arg Tyr Glu Ala Glu Ala Arg Lys Ile Glu Arg
755 760 765
Lys Tyr Trp Glu Gln Gly Ala Leu Thr Glu Arg Glu Arg Tyr Asp Arg
770 775 780
Leu Val Glu Leu Trp Lys Asp Ala Thr Asp Glu Val Gly Asn Ala Val
785 790 795 800
Glu Lys Leu Tyr Pro Asp Asp Asn Pro Ile Pro Met Ile Val Lys Ser
805 810 815
Gly Ala Ala Gly Asn Met Arg Gln Ile Trp Thr Leu Ala Gly Met Lys
820 825 830
Gly Met Val Val Asn Ser Lys Gly Asp Tyr Ile Thr Arg Pro Ile Lys
835 840 845
Thr Ser Phe Arg Glu Gly Leu Ser Val Leu Glu Tyr Phe Asn Asn Ser
850 855 860
His Gly Ser Arg Lys Gly Leu Ala Asp Thr Ala Leu Arg Thr Ala Asp
865 870 875 880
Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val Ala Gln Asp Val Ile
885 890 895
Val Arg Glu Asp Asp Cys Gly Thr Lys Gln Gly Ile Arg Val Pro Val
900 905 910
Ala Val Glu Val Lys Asp Ala Glu Gly Asn Val Thr Gly Tyr Thr Gly
915 920 925
His Ser Leu Ile Glu Thr Ser Val Ala Gly Arg Val Ala Ala Thr Ala
930 935 940
Val Lys Asp Ala Glu Gly Asn Val Met Val Glu Pro Gly Glu Asn Leu
945 950 955 960
Thr Asp Gln Leu Ile Asp Glu Leu Ile Ala Ala Gly Val Lys Glu Val
965 970 975
Lys Val Arg Ser Val Leu Thr Cys Gln Thr Pro Thr Gly Val Cys Ala
980 985 990
Lys Cys Tyr Gly Lys Ser Met Ala Thr Gly Lys Leu Val Asp Ile Gly
995 1000 1005
Glu Ala Val Gly Ile Val Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
1010 1015 1020
Gln Leu Thr Met Arg Thr Phe His Gln Gly Gly Val Gly Gly Asp Ile
1025 1030 1035 1040
Thr Gly Gly Leu Pro Arg Val Gln Glu Leu Phe Glu Ala Arg Val Pro
1045 1050 1055
Lys Asn Arg Ala Pro Ile Ala Ser Val Ala Gly Thr Val His Leu Asp
1060 1065 1070
Asp Glu Gly Asn Phe Tyr Thr Leu Thr Ile Asn Pro Asp Asp Gly Ser
1075 1080 1085
Asp Val Val Val Tyr Glu Lys Leu Ser Lys Arg Gln Gly Leu Ala Thr
1090 1095 1100
Val Arg Val Pro Met Glu Ser Asn Pro Gly Ala Met Ile Glu Arg Thr
1105 1110 1115 1120
Leu Ala Glu Gly Asp His Val Glu Val Gly Asp Arg Leu Leu Arg Gly
1125 1130 1135
Pro Ala Asp Pro His Asp Val Leu Glu Val Leu Gly Arg Arg Gly Val
1140 1145 1150
Glu Gln His Leu Val Asp Glu Val Gln Asp Val Tyr Arg Ala Gln Gly
1155 1160 1165
Val Ala Ile His Asp Lys His Ile Glu Ile Ile Ile Arg Gln Met Leu
1170 1175 1180
Arg Arg Gly Thr Val Ile Glu Ser Gly Ser Thr Glu Phe Leu Pro Gly
1185 1190 1195 1200
Thr Leu Val Asp Leu Ser Glu Ala Lys Ala Ala Asn Ala Glu Ala Leu
1205 1210 1215
Ala Asn Gly Gly Gln Pro Ala Glu Leu Arg Ser Glu Ile Met Gly Ile
1220 1225 1230
Thr Lys Ala Ser Leu Ala Thr Glu Ser Trp Leu Ser Ala Ala Ser Phe
1235 1240 1245
Gln Glu Thr Thr Arg Val Leu Thr Asp Ala Ala Ile Asn Lys Arg Ser
1250 1255 1260
Asp Lys Leu Ile Gly Leu Lys Glu Asn Val Ile Ile Gly Lys Leu Ile
1265 1270 1275 1280
Pro Ala Gly Thr Gly Ile Ser Arg Tyr Arg Asn Ile Ser Val Lys Pro
1285 1290 1295
Thr Glu Ala Ala Arg Asn Ala Ala Tyr Ser Ile Pro Thr Tyr Gly Asp
1300 1305 1310
Ser Ile Tyr Gly Asp Asp Gly Tyr Gly Glu Phe Thr Gly Ala Ser Val
1315 1320 1325
Pro Leu Asp Glu Ala Tyr Asp Leu
1330 1335
<210> 44
<211> 1316
<212> PRT
<213> 结核分支杆菌（Mycobacterium tuberculosis）
<400> 44
Met Leu Asp Val Asn Phe Phe Asp Glu Leu Arg Ile Gly Leu Ala Thr
1 5 10 15
Ala Glu Asp Ile Arg Gln Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Lys Asp Gly Leu Phe Cys
35 40 45
Glu Lys Ile Phe Gly Pro Thr Arg Asp Trp Glu Cys Tyr Cys Gly Lys
50 55 60
Tyr Lys Arg Val Arg Phe Lys Gly Ile Ile Cys Glu Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ala Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Thr His Ile Trp Tyr Phe Lys Gly Val Pro Ser
100 105 110
Arg Leu Gly Tyr Leu Leu Asp Leu Ala Pro Lys Asp Leu Glu Lys Ile
115 120 125
Ile Tyr Phe Ala Ala Tyr Val Ile Thr Ser Val Asp Glu Glu Met Arg
130 135 140
His Asn Glu Leu Ser Thr Leu Glu Ala Glu Met Ala Val Glu Arg Lys
145 150 155 160
Ala Val Glu Asp Gln Arg Asp Gly Glu Leu Glu Ala Arg Ala Gln Lys
165 170 175
Leu Glu Ala Asp Leu Ala Glu Leu Glu Ala Glu Gly Ala Lys Ala Asp
180 185 190
Ala Arg Arg Lys Val Arg Asp Gly Gly Glu Arg Glu Met Arg Gln Ile
195 200 205
Arg Asp Arg Ala Gln Arg Glu Leu Asp Arg Leu Glu Asp Ile Trp Ser
210 215 220
Thr Phe Thr Lys Leu Ala Pro Lys Gln Leu Ile Val Asp Glu Asn Leu
225 230 235 240
Tyr Arg Glu Leu Val Asp Arg Tyr Gly Glu Tyr Phe Thr Gly Ala Met
245 250 255
Gly Ala Glu Ser Ile Gln Lys Leu Ile Glu Asn Phe Asp Ile Asp Ala
260 265 270
Glu Ala Glu Ser Leu Arg Asp Val Ile Arg Asn Gly Lys Gly Gln Lys
275 280 285
Lys Leu Arg Ala Leu Lys Arg Leu Lys Val Val Ala Ala Phe Gln Gln
290 295 300
Ser Gly Asn Ser Pro Met Gly Met Val Leu Asp Ala Val Pro Val Ile
305 310 315 320
Pro Pro Glu Leu Arg Pro Met Val Gln Leu Asp Gly Gly Arg Phe Ala
325 330 335
Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn
340 345 350
Arg Leu Lys Arg Leu Ile Asp Leu Gly Ala Pro Glu Ile Ile Val Asn
355 360 365
Asn Glu Lys Arg Met Leu Gln Glu Ser Val Asp Ala Leu Phe Asp Asn
370 375 380
Gly Arg Arg Gly Arg Pro Val Thr Gly Pro Gly Asn Arg Pro Leu Lys
385 390 395 400
Ser Leu Ser Asp Leu Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn
405 410 415
Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg Ser Val Ile Val Val
420 425 430
Gly Pro Gln Leu Lys Leu His Gln Cys Gly Leu Pro Lys Leu Met Ala
435 440 445
Leu Glu Leu Phe Lys Pro Phe Val Met Lys Arg Leu Val Asp Leu Asn
450 455 460
His Ala Gln Asn Ile Lys Ser Ala Lys Arg Met Val Glu Arg Gln Arg
465 470 475 480
Pro Gln Val Trp Asp Val Leu Glu Glu Val Ile Ala Glu His Pro Val
485 490 495
Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe
500 505 510
Glu Pro Met Leu Val Glu Gly Lys Ala Ile Gln Leu His Pro Leu Val
515 520 525
Cys Glu Ala Phe Asn Ala Asp Phe Asp Gly Asp Gln Met Ala Val His
530 535 540
Leu Pro Leu Ser Ala Glu Ala Gln Ala Glu Ala Arg Ile Leu Met Leu
545 550 555 560
Ser Ser Asn Asn Ile Leu Ser Pro Ala Ser Gly Arg Pro Leu Ala Met
565 570 575
Pro Arg Leu Asp Met Val Thr Gly Leu Tyr Tyr Leu Thr Thr Glu Val
580 585 590
Pro Gly Asp Thr Gly Glu Tyr Gln Pro Ala Ser Gly Asp His Pro Glu
595 600 605
Thr Gly Val Tyr Ser Ser Pro Ala Glu Ala Ile Met Ala Ala Asp Arg
610 615 620
Gly Val Leu Ser Val Arg Ala Lys Ile Lys Val Arg Leu Thr Gln Leu
625 630 635 640
Arg Pro Pro Val Glu Ile Glu Ala Glu Leu Phe Gly His Ser Gly Trp
645 650 655
Gln Pro Gly Asp Ala Trp Met Ala Glu Thr Thr Leu Gly Arg Val Met
660 665 670
Phe Asn Glu Leu Leu Pro Leu Gly Tyr Pro Phe Val Asn Lys Gln Met
675 680 685
His Lys Lys Val Gln Ala Ala Ile Ile Asn Asp Leu Ala Glu Arg Tyr
690 695 700
Pro Met Ile Val Val Ala Gln Thr Val Asp Lys Leu Lys Asp Ala Gly
705 710 715 720
Phe Tyr Trp Ala Thr Arg Ser Gly Val Thr Val Ser Met Ala Asp Val
725 730 735
Leu Val Pro Pro Arg Lys Lys Glu Ile Leu Asp His Tyr Glu Glu Arg
740 745 750
Ala Asp Lys Val Glu Lys Gln Phe Gln Arg Gly Ala Leu Asn His Asp
755 760 765
Glu Arg Asn Glu Ala Leu Val Glu Ile Trp Lys Glu Ala Thr Asp Glu
770 775 780
Val Gly Gln Ala Leu Arg Glu His Tyr Pro Asp Asp Asn Pro Ile Ile
785 790 795 800
Thr Ile Val Asp Ser Gly Ala Thr Gly Asn Phe Thr Gln Thr Arg Thr
805 810 815
Leu Ala Gly Met Lys Gly Leu Val Thr Asn Pro Lys Gly Glu Phe Ile
820 825 830
Pro Arg Pro Val Lys Ser Ser Phe Arg Glu Gly Leu Thr Val Leu Glu
835 840 845
Tyr Phe Ile Asn Thr His Gly Ala Arg Lys Gly Leu Ala Asp Thr Ala
850 855 860
Leu Arg Thr Ala Asp Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val
865 870 875 880
Ser Gln Asp Val Ile Val Arg Glu His Asp Cys Gln Thr Glu Arg Gly
885 890 895
Ile Val Val Glu Leu Ala Glu Arg Ala Pro Asp Gly Thr Leu Ile Arg
900 905 910
Asp Pro Tyr Ile Glu Thr Ser Ala Tyr Ala Arg Thr Leu Gly Thr Asp
915 920 925
Ala Val Asp Glu Ala Gly Asn Val Ile Val Glu Arg Gly Gln Asp Leu
930 935 940
Gly Asp Pro Glu Ile Asp Ala Leu Leu Ala Ala Gly Ile Thr Gln Val
945 950 955 960
Lys Val Arg Ser Val Leu Thr Cys Ala Thr Ser Thr Gly Val Cys Ala
965 970 975
Thr Cys Tyr Gly Arg Ser Met Ala Thr Gly Lys Leu Val Asp Ile Gly
980 985 990
Glu Ala Val Gly Ile Val Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr
995 1000 1005
Gln Leu Thr Met Arg Thr Phe His Gln Gly Gly Val Gly Glu Asp Ile
1010 1015 1020
Thr Gly Gly Leu Pro Arg Val Gln Glu Leu Phe Glu Ala Arg Val Pro
1025 1030 1035 1040
Arg Gly Lys Ala Pro Ile Ala Asp Val Thr Gly Arg Val Arg Leu Glu
1045 1050 1055
Asp Gly Glu Arg Phe Tyr Lys Ile Thr Ile Val Pro Asp Asp Gly Gly
1060 1065 1070
Glu Glu Val Val Tyr Asp Lys Ile Ser Lys Arg Gln Arg Leu Arg Val
1075 1080 1085
Phe Lys His Glu Asp Gly Ser Glu Arg Val Leu Ser Asp Gly Asp His
1090 1095 1100
Val Glu Val Gly Gln Gln Leu Met Glu Gly Ser Ala Asp Pro His Glu
1105 1110 1115 1120
Val Leu Arg Val Gln Gly Pro Arg Glu Val Gln Ile His Leu Val Arg
1125 1130 1135
Glu Val Gln Glu Val Tyr Arg Ala Gln Gly Val Ser Ile His Asp Lys
1140 1145 1150
His Ile Glu Val Ile Val Arg Gln Met Leu Arg Arg Val Thr Ile Ile
1155 1160 1165
Asp Ser Gly Ser Thr Glu Phe Leu Pro Gly Ser Leu Ile Asp Arg Ala
1170 1175 1180
Glu Phe Glu Ala Glu Asn Arg Arg Val Val Ala Glu Gly Gly Glu Pro
1185 1190 1195 1200
Ala Ala Gly Arg Pro Val Leu Met Gly Ile Thr Lys Ala Ser Leu Ala
1205 1210 1215
Thr Asp Ser Trp Leu Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg Val
1220 1225 1230
Leu Thr Asp Ala Ala Ile Asn Cys Arg Ser Asp Lys Leu Asn Gly Leu
1235 1240 1245
Lys Glu Asn Val Ile Ile Gly Lys Leu Ile Pro Ala Gly Thr Gly Ile
1250 1255 1260
Asn Arg Tyr Arg Asn Ile Ala Val Gln Pro Thr Glu Glu Ala Arg Ala
1265 1270 1275 1280
Ala Ala Tyr Thr Ile Pro Ser Tyr Glu Asp Gln Tyr Tyr Ser Pro Asp
1285 1290 1295
Phe Gly Ala Ala Thr Gly Ala Ala Val Pro Leu Asp Asp Tyr Gly Tyr
1300 1305 1310
Ser Asp Tyr Arg
1315
<210> 45
<211> 1319
<212> PRT
<213> 马红球菌（Rhodococcus equi）
<400> 45
Met Leu Asp Val Asn Phe Phe Asp Glu Leu Arg Ile Gly Leu Ala Thr
1 5 10 15
Ala Glu Asp Ile Arg Asn Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Lys Asp Gly Leu Phe Cys
35 40 45
Glu Lys Ile Phe Gly Pro Thr Arg Asp Trp Glu Cys Tyr Cys Gly Lys
50 55 60
Tyr Lys Arg Val Arg Phe Lys Gly Ile Ile Cys Glu Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ala Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Thr His Ile Trp Tyr Phe Lys Gly Val Pro Ser
100 105 110
Arg Leu Gly Tyr Leu Leu Asp Leu Ala Pro Lys Asp Leu Glu Lys Ile
115 120 125
Ile Tyr Phe Ala Ala Tyr Val Ile Val Gly Val Asp Glu Glu Leu Arg
130 135 140
His Asn Glu Leu Ser Thr Leu Glu Ala Glu Met Glu Val Glu Lys Lys
145 150 155 160
Thr Val Ala Asp Gln Arg Asp Ala Asp Leu Glu Ala Arg Ala Gln Lys
165 170 175
Leu Glu Ala Asp Ile Ala Glu Leu Glu Ala Glu Gly Ala Lys Ser Asp
180 185 190
Val Arg Arg Lys Val Lys Asp Gly Gly Glu Arg Glu Met Arg Gln Leu
195 200 205
Arg Asp Arg Ala Gln Arg Glu Leu Asp Arg Leu Asp Glu Ile Trp Thr
210 215 220
Thr Phe Thr Lys Leu Ser Val Lys Gln Leu Ile Val Asp Glu Ser Leu
225 230 235 240
Tyr Arg Glu Leu Val Asp Arg Tyr Gly Glu Tyr Phe Thr Gly Ala Met
245 250 255
Gly Ala Glu Ser Ile Gln Lys Leu Met Glu Asn Phe Asp Ile Glu Ala
260 265 270
Glu Ala Glu Ser Leu Arg Glu Thr Ile Arg Ser Gly Lys Gly Gln Lys
275 280 285
Lys Leu Arg Ala Leu Lys Arg Leu Lys Val Val Ala Ala Phe Gln Gln
290 295 300
Ser Gly Asn Ser Pro Met Gly Met Val Leu Asp Ala Val Pro Val Ile
305 310 315 320
Pro Pro Glu Leu Arg Pro Met Val Gln Leu Asp Gly Gly Arg Phe Ala
325 330 335
Thr Ser Asp Leu Asn Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn
340 345 350
Arg Leu Lys Arg Leu Ile Asp Leu Gly Ala Pro Glu Ile Ile Val Asn
355 360 365
Asn Glu Lys Arg Met Leu Gln Glu Ser Val Asp Ala Leu Phe Asp Asn
370 375 380
Gly Arg Arg Gly Arg Pro Val Thr Gly Pro Gly Asn Arg Pro Leu Lys
385 390 395 400
Ser Leu Ser Asp Leu Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn
405 410 415
Leu Leu Gly Lys Arg Val Asp Tyr Ser Gly Arg Ser Val Ile Val Val
420 425 430
Gly Pro Gln Leu Lys Leu His Gln Cys Gly Leu Pro Lys Leu Met Ala
435 440 445
Leu Glu Leu Phe Lys Pro Phe Val Met Lys Arg Leu Val Asp Leu Asn
450 455 460
His Ala Gln Asn Ile Lys Ser Ala Lys Arg Met Val Glu Arg Gln Arg
465 470 475 480
Pro Gln Val Trp Asp Val Leu Glu Glu Val Ile Asn Glu His Pro Val
485 490 495
Leu Leu Asn Arg Ala Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe
500 505 510
Glu Pro Gln Leu Val Glu Gly Lys Ala Ile Gln Leu His Pro Leu Val
515 520 525
Cys Glu Ala Phe Asn Ala Asp Phe Asp Gly Asp Gln Met Ala Val His
530 535 540
Leu Pro Leu Ser Ala Glu Ala Gln Ala Glu Ala Arg Ile Leu Met Leu
545 550 555 560
Ser Ser Asn Asn Ile Leu Ser Pro Ala Ser Gly Arg Pro Leu Ala Met
565 570 575
Pro Arg Leu Asp Met Val Thr Gly Leu Phe His Leu Thr Arg Glu Val
580 585 590
Glu Gly Ala Ile Gly Ala Tyr Gln Pro Ala Ala Asp Gly Gln Pro Glu
595 600 605
Gln Gly Val Tyr Ser Ser Pro Ala Glu Ala Gln Met Ala Val Asp Arg
610 615 620
Gly Val Leu Ser Val Gln Ala Lys Ile Lys Val Arg Leu Thr His Gln
625 630 635 640
Arg Pro Pro Arg Glu Ile Glu Ala Glu Leu Phe Pro Glu Gly Trp Asn
645 650 655
Phe Gly Asp Gly Trp Met Val Glu Thr Thr Leu Gly Arg Val Met Phe
660 665 670
Asn Asp Leu Leu Pro Ala Asp Tyr Pro Phe Ile Asn Glu Gln Met Pro
675 680 685
Lys Lys Arg Gln Ala Thr Ile Ile Asn Asp Leu Ala Glu Arg Tyr Pro
690 695 700
Met Ile Val Val Ala Gln Thr Val Asp Lys Met Lys Asp Thr Gly Phe
705 710 715 720
Tyr Trp Ala Thr Arg Ser Gly Val Thr Val Ser Ile Ser Asp Val Leu
725 730 735
Val Pro Pro Glu Lys Ala Gln Ile Met Glu Gln Phe Glu Ala Gln Ala
740 745 750
Asp Gln Ile Glu Lys Lys Tyr Gln Arg Gly Ala Leu Asn His Thr Glu
755 760 765
Arg Asn Ser Ala Leu Val Lys Ile Trp Ser Glu Ala Thr Asp Glu Val
770 775 780
Gly Lys Ala Met Glu Ala His Phe Pro Asp Asp Asn Pro Ile Pro Met
785 790 795 800
Ile Val Lys Ser Gly Ala Ala Gly Asn Met Thr Gln Val Arg Ser Leu
805 810 815
Ala Gly Met Lys Gly Leu Val Thr Asn Pro Lys Gly Glu Phe Ile Pro
820 825 830
Arg Pro Ile Lys Ser Ser Phe Lys Glu Gly Leu Thr Val Leu Glu Tyr
835 840 845
Phe Ile Asn Thr His Gly Ala Arg Lys Gly Leu Ala Asp Thr Ala Leu
850 855 860
Arg Thr Ala Asp Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val Ser
865 870 875 880
Gln Asp Val Ile Val Arg Glu Val Asp Cys Gly Thr Glu Arg Gly Ile
885 890 895
Leu Thr Thr Ile Ala Glu Lys Ala Ala Asp Gly Thr Met Ile Arg Asp
900 905 910
Ala His Val Glu Thr Ser Thr Tyr Ala Arg Thr Leu Ala Ala Asp Ala
915 920 925
Ile Asp Glu Asn Gly Asn Val Val Val Glu Arg Gly His Asp Leu Gly
930 935 940
Asp Pro Ala Ile Asp Ala Leu Leu Ala Ala Gly Ile Thr Gln Val Lys
945 950 955 960
Val Arg Ser Val Leu Thr Cys Thr Thr Ala Thr Gly Val Cys Ala Thr
965 970 975
Cys Tyr Gly Arg Ser Met Ala Thr Gly Lys Leu Val Asp Ile Gly Glu
980 985 990
Ala Val Gly Ile Val Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr Gln
995 1000 1005
Leu Thr Met Arg Thr Phe His Gln Gly Gly Ala Ala Gly Ala Ala Asp
1010 1015 1020
Ile Thr Gly Gly Leu Pro Arg Val Gln Glu Leu Phe Glu Ala Arg Val
1025 1030 1035 1040
Pro Lys Gly Lys Ala Pro Ile Thr Glu Val Ser Gly Arg Val Gln Leu
1045 1050 1055
Asp Asp Asp Asp Arg Phe Tyr Lys Ile Thr Val Val Pro Asp Asp Gly
1060 1065 1070
Gly Glu Glu Val Val Tyr Asp Lys Leu Ser Lys Arg Gln Arg Leu Arg
1075 1080 1085
Val Phe Lys His Asp Asp Gly Ser Glu Arg Leu Leu Ser Asp Gly Asp
1090 1095 1100
His Val Asp Val Gly Gln Gln Leu Leu Glu Gly Ala Ala Asp Pro His
1105 1110 1115 1120
Asp Val Leu Arg Val Met Gly Pro Arg Gln Val Gln Ile His Leu Val
1125 1130 1135
Asn Glu Val Gln Glu Val Tyr Arg Ser Gln Gly Val Ser Ile His Asp
1140 1145 1150
Lys His Ile Glu Val Ile Val Arg Gln Met Leu Arg Arg Val Thr Ile
1155 1160 1165
Ile Asp Ser Gly Ser Thr Glu Phe Leu Pro Gly Ser Leu Val Glu Arg
1170 1175 1180
Ala Glu Phe Glu Ala Ser Asn Arg Arg Val Val Ala Glu Gly Gly Glu
1185 1190 1195 1200
Pro Ala Ala Gly Arg Pro Val Leu Met Gly Ile Thr Lys Ala Ser Leu
1205 1210 1215
Ala Thr Asp Ser Trp Leu Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg
1220 1225 1230
Val Leu Thr Asp Ala Ala Ile Asn Cys Arg Ser Asp Lys Leu Ile Gly
1235 1240 1245
Leu Lys Glu Asn Val Ile Ile Gly Lys Leu Ile Pro Ala Gly Thr Gly
1250 1255 1260
Ile Asn Arg Tyr Arg Asn Ile Gln Val Gln Pro Thr Glu Glu Ala Arg
1265 1270 1275 1280
Ala Ala Ala Tyr Ala Val Pro Ser Tyr Asp Asp Gln Tyr Tyr Ser Pro
1285 1290 1295
Glu Gly Phe Gly Thr Gly Thr Gly Ala Ala Val Pro Leu Asp Asp Tyr
1300 1305 1310
Gly Phe Gly Ser Asp Tyr Arg
1315
<210> 46
<211> 1396
<212> PRT
<213> 沙眼衣原体（Chlamydia trachomatis）
<400> 46
Met Phe Arg Glu Gly Ser Arg Asp Asp Ala Ala Leu Val Lys Glu Gly
1 5 10 15
Leu Phe Asp Lys Leu Glu Ile Gly Ile Ala Ser Asp Val Thr Ile Arg
20 25 30
Asp Lys Trp Ser Cys Gly Glu Ile Lys Lys Pro Glu Thr Ile Asn Tyr
35 40 45
Arg Thr Phe Lys Pro Glu Lys Gly Gly Leu Phe Cys Glu Lys Ile Phe
50 55 60
Gly Pro Thr Lys Asp Trp Glu Cys Tyr Cys Gly Lys Tyr Lys Lys Ile
65 70 75 80
Lys His Lys Gly Ile Val Cys Asp Arg Cys Gly Val Glu Val Thr Leu
85 90 95
Ser Lys Val Arg Arg Glu Arg Met Ala His Ile Glu Leu Ala Val Pro
100 105 110
Ile Val His Ile Trp Phe Phe Lys Thr Thr Pro Ser Arg Ile Gly Asn
115 120 125
Val Leu Gly Met Thr Ala Ser Asp Leu Glu Arg Val Ile Tyr Tyr Glu
130 135 140
Glu Tyr Val Val Ile Asp Pro Gly Asn Thr Asp Leu Val Lys Lys Gln
145 150 155 160
Leu Leu Asn Asp Ala Lys Tyr Arg Glu Val Val Glu Lys Trp Gly Lys
165 170 175
Asp Ala Phe Val Ala Lys Met Gly Gly Glu Ala Val Tyr Asp Leu Leu
180 185 190
Lys Ser Glu Asp Leu Glu Ser Leu Leu Gly Glu Leu Lys Glu Arg Leu
195 200 205
Arg Lys Thr Lys Ser Gln Gln Ala Arg Met Lys Leu Ala Lys Arg Leu
210 215 220
Lys Ile Val Glu Gly Phe Val Ser Ser Ser Asn Arg Pro Glu Trp Met
225 230 235 240
Val Leu Lys Asn Ile Pro Val Val Pro Pro Asp Leu Arg Pro Leu Val
245 250 255
Pro Leu Asp Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp Leu Tyr
260 265 270
Arg Arg Val Ile Asn Arg Asn Asn Arg Leu Lys Ala Ile Leu Arg Leu
275 280 285
Lys Thr Pro Glu Val Ile Val Arg Asn Glu Lys Arg Met Leu Gln Glu
290 295 300
Ala Val Asp Ala Leu Phe Asp Asn Gly Arg His Gly His Pro Val Met
305 310 315 320
Gly Ala Gly Asn Arg Pro Leu Lys Ser Leu Ser Glu Met Leu Lys Gly
325 330 335
Lys Asn Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val Asp Tyr
340 345 350
Ser Gly Arg Ser Val Ile Ile Val Gly Pro Glu Leu Lys Phe Asn Gln
355 360 365
Cys Gly Leu Pro Lys Glu Met Ala Leu Glu Leu Phe Glu Pro Phe Ile
370 375 380
Ile Lys Arg Leu Lys Asp Gln Gly Ser Val Tyr Thr Ile Arg Ser Ala
385 390 395 400
Lys Lys Met Ile Gln Arg Gly Ala Pro Glu Val Trp Asp Val Leu Glu
405 410 415
Glu Ile Ile Lys Gly His Pro Val Leu Leu Asn Arg Ala Pro Thr Leu
420 425 430
His Arg Leu Gly Ile Gln Ala Phe Glu Pro Val Leu Ile Glu Gly Lys
435 440 445
Ala Ile Arg Val His Pro Leu Val Cys Ala Ala Phe Asn Ala Asp Phe
450 455 460
Asp Gly Asp Gln Met Ala Val His Val Pro Leu Ser Ile Glu Ala Gln
465 470 475 480
Leu Glu Ala Lys Val Leu Met Met Ala Pro Asp Asn Ile Phe Leu Pro
485 490 495
Ser Ser Gly Lys Pro Val Ala Thr Pro Ser Lys Asp Met Thr Leu Gly
500 505 510
Ile Tyr Tyr Leu Met Ala Asp Pro Thr Tyr Phe Pro Glu Glu His Gly
515 520 525
Gly Lys Thr Lys Ala Phe Lys Asp Glu Val Glu Val Leu Arg Ala Leu
530 535 540
Asn Ala Gly Gly Phe Ile Leu Lys Asp Glu Ile Cys Gly Ser Arg Arg
545 550 555 560
Asp Glu Thr Gly Arg Gly Ile His Ile His Glu Lys Ile Lys Val Arg
565 570 575
Ile Asp Gly Gln Ile Ile Glu Thr Thr Pro Gly Arg Val Phe Phe Asn
580 585 590
Thr Ile Val Pro Lys Glu Leu Gly Phe Gln Asn Tyr Ser Met Pro Ser
595 600 605
Lys Arg Ile Ser Glu Leu Ile Leu Gln Cys Tyr Lys Lys Val Gly Leu
610 615 620
Glu Ala Thr Val Arg Phe Leu Asp Asp Leu Lys Glu Leu Gly Phe Val
625 630 635 640
Gln Ser Thr Lys Ala Ala Ile Ser Met Gly Leu Lys Asp Val Lys Ile
645 650 655
Pro Glu Ile Lys Lys Glu Ile Leu Lys Asp Ala Tyr Asp Lys Val Ala
660 665 670
Val Val Lys Lys Gln Tyr Glu Asp Gly Ile Ile Thr Asp Gly Glu Arg
675 680 685
His Ser Lys Thr Ile Ser Ile Trp Thr Glu Val Ser Asp Leu Leu Ser
690 695 700
Asn Ala Leu Tyr Ser Glu Ile Lys Lys Gln Thr Asn Ser Lys His Asn
705 710 715 720
Pro Leu Phe Leu Met Ile Asp Ser Gly Ala Arg Gly Asn Lys Ser Gln
725 730 735
Leu Lys Gln Leu Gly Ala Leu Arg Gly Leu Met Ala Lys Pro Asn Gly
740 745 750
Ala Ile Ile Glu Ser Pro Ile Thr Ser Asn Phe Arg Glu Gly Leu Thr
755 760 765
Val Leu Glu Tyr Ser Ile Ser Ser His Gly Ala Arg Lys Gly Leu Ala
770 775 780
Asp Thr Ala Leu Lys Thr Ala Asp Ser Gly Tyr Leu Thr Arg Arg Leu
785 790 795 800
Val Asp Val Ala Gln Asp Val Ile Ile Thr Glu Arg Asp Cys Gly Thr
805 810 815
Leu Asn His Ile Glu Val Ser Thr Ile Arg Gln Gly Ser Glu Glu Leu
820 825 830
Leu Pro Leu Lys Asp Arg Val Tyr Gly Arg Thr Val Ser Glu Asn Ile
835 840 845
Tyr Gln Pro Gly Asp Lys Ser Asn Val Leu Ala Tyr Ala Gly Asp Val
850 855 860
Leu Thr Ser Ala Gln Ala Glu Ala Ile Asp Asp Ala Gly Ile Glu Ser
865 870 875 880
Val Lys Ile Arg Ser Thr Leu Thr Cys Glu Ser Arg Arg Gly Val Cys
885 890 895
Ala Lys Cys Tyr Gly Leu Asn Leu Ala Asn Gly Arg Leu Ile Gly Leu
900 905 910
Gly Glu Ala Val Gly Ile Ile Ala Ala Gln Ser Ile Gly Glu Pro Gly
915 920 925
Thr Gln Leu Thr Met Arg Thr Phe His Leu Gly Gly Ile Ala Ala Thr
930 935 940
Ser Ser Thr Pro Glu Ile Val Ala Glu Cys Asp Gly Ile Leu Val Tyr
945 950 955 960
Leu Asp Leu Arg Val Val Val Asp Gln Glu Gly Asn Asn Leu Val Leu
965 970 975
Asn Lys Met Gly Ala Leu His Leu Val Gln Asp Glu Gly Arg Ser Leu
980 985 990
Ser Glu Tyr Lys Lys Leu Leu Ser Thr Lys Ser Ile Glu Ser Leu Ala
995 1000 1005
Thr Phe Pro Val Glu Leu Gly Ala Lys Ile Leu Val Asn Asp Gly Ala
1010 1015 1020
Ala Val Ala Ala Gly Gln Arg Ile Ala Glu Val Glu Leu His Asn Ile
1025 1030 1035 1040
Pro Ile Ile Cys Asp Lys Pro Gly Phe Val His Tyr Glu Asp Leu Val
1045 1050 1055
Glu Gly Val Ser Thr Glu Lys Val Thr Asn Lys Asn Thr Gly Leu Val
1060 1065 1070
Glu Leu Ile Val Lys Gln His Arg Gly Glu Leu His Pro Gln Ile Ala
1075 1080 1085
Ile Tyr Ala Asp Ala Asn Met Lys Glu Leu Val Gly Thr Tyr Ala Ile
1090 1095 1100
Pro Ser Gly Ala Ile Ile Ser Val Glu Glu Gly Gln Arg Ile Ala Pro
1105 1110 1115 1120
Gly Met Leu Leu Ala Arg Leu Pro Arg Gly Ala Ile Lys Thr Lys Asp
1125 1130 1135
Ile Thr Gly Gly Leu Pro Arg Val Ala Glu Leu Val Glu Ala Arg Lys
1140 1145 1150
Pro Glu Asp Ala Ala Asp Ile Ala Lys Ile Asp Gly Val Val Asp Phe
1155 1160 1165
Lys Gly Ile Gln Lys Asn Lys Arg Ile Leu Val Val Arg Asp Glu Ile
1170 1175 1180
Thr Gly Met Glu Glu Glu His Leu Ile Ser Leu Thr Lys His Leu Ile
1185 1190 1195 1200
Val Gln Arg Gly Asp Ser Val Ile Lys Gly Gln Gln Leu Thr Asp Gly
1205 1210 1215
Leu Val Val Pro His Glu Ile Leu Glu Ile Cys Gly Val Arg Glu Leu
1220 1225 1230
Gln Lys Tyr Leu Val Asn Glu Val Gln Glu Val Tyr Arg Leu Gln Gly
1235 1240 1245
Val Asp Ile Asn Asp Lys His Val Glu Ile Ile Val Arg Gln Met Leu
1250 1255 1260
Gln Lys Val Arg Ile Thr Asp Pro Gly Asp Thr Thr Leu Leu Phe Gly
1265 1270 1275 1280
Glu Asp Val Asp Lys Lys Glu Phe Tyr Glu Glu Asn Arg Arg Thr Glu
1285 1290 1295
Glu Asp Gly Gly Lys Pro Ala Gln Ala Val Pro Val Leu Leu Gly Ile
1300 1305 1310
Thr Lys Ala Ser Leu Gly Thr Glu Ser Phe Ile Ser Ala Ala Ser Phe
1315 1320 1325
Gln Asp Thr Thr Arg Val Leu Thr Asp Ala Ala Cys Ser Ser Lys Thr
1330 1335 1340
Asp Tyr Leu Leu Gly Phe Lys Glu Asn Val Ile Met Gly His Met Ile
1345 1350 1355 1360
Pro Gly Gly Thr Gly Phe Asp Thr His Lys Arg Ile Lys Gln His Leu
1365 1370 1375
Glu Lys Glu Gln Glu Asp Leu Val Phe Asp Phe Asp Ser Glu Phe Glu
1380 1385 1390
Ser Val Ala Gly
1395
<210> 47
<211> 1178
<212> PRT
<213> 肉毒梭状芽孢杆菌（Clostridium botulinum）
<400> 47
Met Phe Glu Leu Asn Asn Phe Asp Ala Leu Gln Ile Gly Leu Ala Ser
1 5 10 15
Pro Glu Lys Ile Arg Glu Trp Ser Arg Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Arg Asp Gly Leu Phe Cys
35 40 45
Glu Arg Ile Phe Gly Pro Met Lys Asp Trp Glu Cys His Cys Gly Lys
50 55 60
Tyr Lys Arg Ile Arg Tyr Lys Gly Ile Val Cys Asp Arg Cys Gly Val
65 70 75 80
Glu Val Thr Lys Ala Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Ser His Ile Trp Tyr Phe Lys Gly Ile Pro Ser
100 105 110
Arg Met Gly Leu Ile Leu Asp Met Ser Pro Arg Ala Leu Glu Lys Val
115 120 125
Leu Tyr Phe Ala Ser Tyr Val Val Leu Asp Pro Lys Glu Thr Pro Leu
130 135 140
Leu Lys Lys Gln Leu Leu Asn Glu Lys Glu Tyr Arg Glu Ser Ile Asp
145 150 155 160
Lys Tyr Gly Asp Asp Ser Phe Val Ala Ala Met Gly Ala Glu Ala Val
165 170 175
Lys Thr Leu Leu Asp Glu Ile Asp Leu Glu Gln Ser Ser Ile Glu Leu
180 185 190
Lys Glu Glu Leu Lys Thr Ser Thr Gly Gln Lys Lys Ile Arg Ile Ile
195 200 205
Arg Arg Leu Glu Val Val Glu Ser Phe Arg Lys Ser Gly Asn Arg Pro
210 215 220
Asp Trp Met Val Ile Asp Val Ile Pro Val Ile Pro Pro Asp Leu Arg
225 230 235 240
Pro Met Val Gln Leu Asp Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn
245 250 255
Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn Arg Leu Lys Lys Leu
260 265 270
Leu Asp Leu Gly Ala Pro Asp Ile Ile Val Arg Asn Glu Lys Arg Met
275 280 285
Leu Gln Glu Ala Val Asp Ala Leu Ile Asp Asn Gly Arg Arg Gly Arg
290 295 300
Pro Val Thr Gly Pro Gly Asn Arg Pro Leu Lys Ser Leu Ser Asp Met
305 310 315 320
Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg
325 330 335
Val Asp Tyr Ser Gly Arg Ser Val Ile Val Val Gly Pro Glu Leu Lys
340 345 350
Met Tyr Gln Cys Gly Leu Pro Lys Glu Met Ala Leu Glu Leu Phe Lys
355 360 365
Pro Phe Val Met Lys Lys Leu Val Gln Asn Gly Leu Ala His Asn Ile
370 375 380
Lys Ser Ala Lys Arg Met Val Glu Arg Val Gln Pro Gln Val Trp Asp
385 390 395 400
Val Leu Glu Glu Val Ile Ser Asp His Pro Val Leu Leu Asn Arg Ala
405 410 415
Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe Gln Pro Val Leu Val
420 425 430
Glu Gly Arg Ala Ile Lys Leu His Pro Leu Val Cys Thr Ala Tyr Asn
435 440 445
Ala Asp Phe Asp Gly Asp Gln Met Ala Val His Val Pro Leu Ser Val
450 455 460
Glu Ala Gln Ala Glu Ala Arg Phe Leu Met Leu Ala Ala His Asn Ile
465 470 475 480
Leu Lys Pro Ser Asp Gly Lys Pro Val Ser Val Pro Thr Gln Asp Met
485 490 495
Val Leu Gly Ser Tyr Tyr Leu Thr Met Asp Lys Asp Gly Val Lys Gly
500 505 510
Glu Gly Lys Val Phe Ser Cys Pro Glu Glu Val Leu Met Ala Tyr Gln
515 520 525
Cys Lys Ala Val Asp Ile His Ala Lys Ile Lys Val Arg Leu Lys Lys
530 535 540
Val Ile Asp Gly Glu Thr Ile Glu Gly Ile Ile Glu Thr Thr Pro Gly
545 550 555 560
Lys Ile Ile Phe Asn Glu Ser Ile Pro Gln Asp Leu Gly Tyr Ile Asp
565 570 575
Arg Thr Val Pro Glu Asn Lys Leu Lys Leu Glu Val Asp Phe Leu Val
580 585 590
Ser Lys Lys Thr Leu Gly Gly Ile Ile Asn Arg Cys Tyr Met Lys His
595 600 605
Gly Ala Thr Lys Thr Ser Ile Met Leu Asp Lys Ile Lys Ala Lys Gly
610 615 620
Tyr His Tyr Ser Thr Ile Gly Ala Ile Thr Ile Ser Thr Ser Asp Met
625 630 635 640
Val Val Pro Glu Ala Lys Arg Glu Leu Leu Gln Asn Thr Glu Lys Gln
645 650 655
Val Glu Lys Ile Gln Lys Met Tyr Arg Arg Gly Phe Ile Ser Glu Glu
660 665 670
Glu Arg Tyr Glu Lys Val Ile Asp Leu Trp Thr Lys Thr Thr Glu Asp
675 680 685
Val Ala Asn Ala Leu Met Ala Ser Leu Asp Ser Phe Asn Pro Ile Tyr
690 695 700
Met Met Ala Asp Ser Gly Ala Arg Gly Ser Lys Ser Gln Ile Lys Gln
705 710 715 720
Leu Ala Gly Met Arg Gly Leu Met Ala Asn Pro Ser Gly Lys Ile Ile
725 730 735
Glu Leu Pro Ile Lys Ala Ser Phe Arg Glu Gly Leu Asp Val Leu Glu
740 745 750
Tyr Phe Ile Ser Thr His Gly Ala Arg Lys Gly Asn Ala Asp Thr Ala
755 760 765
Leu Lys Thr Ala Asp Ser Gly Tyr Leu Thr Arg Arg Leu Val Asp Val
770 775 780
Ser Gln Asp Val Ile Val Arg Gln Glu Asp Cys Gly Thr Glu Glu Gly
785 790 795 800
Tyr Glu Val Ser Glu Ile Lys Glu Gly Asn Glu Val Ile Glu Pro Leu
805 810 815
Val Glu Arg Leu Ser Gly Arg Tyr Pro Ser Glu Asp Ile Ile Asn Pro
820 825 830
Thr Thr Gly Glu Val Ile Val Lys Arg Asn Thr Tyr Met Asn Glu Asp
835 840 845
Ile Ala Lys Lys Val Ser Asp Ala Gly Ile Lys Lys Val Lys Ile Arg
850 855 860
Ser Val Phe Thr Cys Lys Ser Lys His Gly Val Cys Ala Arg Cys Tyr
865 870 875 880
Gly Met Asn Met Ala Thr Ser Gln Lys Ile His Ile Gly Glu Ala Val
885 890 895
Gly Ile Val Ala Ala Gln Ser Ile Gly Glu Pro Gly Thr Gln Leu Thr
900 905 910
Met Arg Thr Phe His Thr Gly Gly Val Ala Gly Ala Asp Ile Thr Gln
915 920 925
Gly Leu Pro Arg Val Glu Glu Leu Phe Glu Ala Arg Lys Pro Lys Gly
930 935 940
Leu Ala Ile Val Ser Glu Val Ser Gly Thr Val Lys Met Glu Glu Thr
945 950 955 960
Lys Lys Lys Arg Thr Ile Ile Val Val Thr Asp Asp Gly Glu Glu Val
965 970 975
Ser Tyr Asp Ile Pro Phe Gly Ser Arg Ile Lys Val Lys Asn Gly Asp
980 985 990
Ile Ile Ser Ala Gly Asp Glu Ile Thr Glu Gly Ser Ile Asn Pro His
995 1000 1005
Asp Ile Leu Arg Ile Lys Gly Val Asp Gly Val Lys Asn Tyr Leu Leu
1010 1015 1020
Ser Glu Val Gln Lys Val Tyr Arg Leu Gln Gly Val Asp Ile Asn Asp
1025 1030 1035 1040
Lys His Leu Glu Val Val Ile Arg Gln Met Thr Arg Lys Ile Lys Ile
1045 1050 1055
Glu Asp Ser Gly Asp Thr Glu Leu Leu Pro Gly Thr Met Ile Asp Val
1060 1065 1070
Phe Asp Phe Glu Glu Ala Asn Arg Glu Ile Leu Glu Lys Gly Gly Glu
1075 1080 1085
Pro Ala Val Gly Arg Ile Ala Leu Leu Gly Ile Thr Lys Ala Ala Leu
1090 1095 1100
Ala Thr Asp Ser Phe Leu Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg
1105 1110 1115 1120
Val Leu Thr Asp Ala Ala Ile Lys Gly Lys Ile Asp Pro Leu Leu Gly
1125 1130 1135
Leu Lys Glu Asn Val Ile Ile Gly Lys Leu Ile Pro Ala Gly Thr Gly
1140 1145 1150
Met Thr Arg Tyr Arg Ser Ile Gln Ile Asn Thr Asp Asp Glu Asn Ile
1155 1160 1165
Glu Glu Asp Ser Met Asp Ser Ile Glu Val
1170 1175
<210> 48
<211> 1199
<212> PRT
<213> 枯草芽孢杆菌（Bacillus subtilis）
<400> 48
Met Leu Asp Val Asn Asn Phe Glu Tyr Met Asn Ile Gly Leu Ala Ser
1 5 10 15
Pro Asp Lys Ile Arg Ser Trp Ser Phe Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Lys Asp Gly Leu Phe Cys
35 40 45
Glu Arg Ile Phe Gly Pro Thr Lys Asp Trp Glu Cys His Cys Gly Lys
50 55 60
Tyr Lys Arg Val Arg Tyr Lys Gly Val Val Cys Asp Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ala Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Ser His Ile Trp Tyr Phe Lys Gly Ile Pro Ser
100 105 110
Arg Met Gly Leu Val Leu Asp Met Ser Pro Arg Ala Leu Glu Glu Val
115 120 125
Ile Tyr Phe Ala Ser Tyr Val Val Thr Asp Pro Ala Asn Thr Pro Leu
130 135 140
Glu Lys Lys Gln Leu Leu Ser Glu Lys Glu Tyr Arg Ala Tyr Leu Asp
145 150 155 160
Lys Tyr Gly Asn Lys Phe Gln Ala Ser Met Gly Ala Glu Ala Ile His
165 170 175
Lys Leu Leu Gln Asp Ile Asp Leu Val Lys Glu Val Asp Met Leu Lys
180 185 190
Glu Glu Leu Lys Thr Ser Gln Gly Gln Arg Arg Thr Arg Ala Ile Lys
195 200 205
Arg Leu Glu Val Leu Glu Ala Phe Arg Asn Ser Gly Asn Lys Pro Ser
210 215 220
Trp Met Ile Leu Asp Val Leu Pro Val Ile Pro Pro Glu Leu Arg Pro
225 230 235 240
Met Val Gln Leu Asp Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp
245 250 255
Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu
260 265 270
Asp Leu Gly Ala Pro Ser Ile Ile Val Gln Asn Glu Lys Arg Met Leu
275 280 285
Gln Glu Ala Val Asp Ala Leu Ile Asp Asn Gly Arg Arg Gly Arg Pro
290 295 300
Val Thr Gly Pro Gly Asn Arg Pro Leu Lys Ser Leu Ser His Met Leu
305 310 315 320
Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val
325 330 335
Asp Tyr Ser Gly Arg Ser Val Ile Val Val Gly Pro His Leu Lys Met
340 345 350
Tyr Gln Cys Gly Leu Pro Lys Glu Met Ala Leu Glu Leu Phe Lys Pro
355 360 365
Phe Val Met Lys Glu Leu Val Glu Lys Gly Leu Ala His Asn Ile Lys
370 375 380
Ser Ala Lys Arg Lys Ile Glu Arg Val Gln Pro Glu Val Trp Asp Val
385 390 395 400
Leu Glu Ser Val Ile Lys Glu His Pro Val Leu Leu Asn Arg Ala Pro
405 410 415
Thr Leu His Arg Leu Gly Ile Gln Ala Phe Glu Pro Thr Leu Val Glu
420 425 430
Gly Arg Ala Ile Arg Leu His Pro Leu Val Cys Thr Ala Tyr Asn Ala
435 440 445
Asp Phe Asp Gly Asp Gln Met Ala Val His Val Pro Leu Ser Ala Glu
450 455 460
Ala Gln Ala Glu Ala Arg Ile Leu Met Leu Ala Ala Gln Asn Ile Leu
465 470 475 480
Asn Pro Lys Asp Gly Lys Pro Val Val Thr Pro Ser Gln Asp Met Val
485 490 495
Leu Gly Asn Tyr Tyr Leu Thr Leu Glu Arg Ala Gly Ala Val Gly Glu
500 505 510
Gly Met Val Phe Lys Asn Thr Asp Glu Ala Leu Leu Ala Tyr Gln Asn
515 520 525
Gly Tyr Val His Leu His Thr Arg Val Ala Val Ala Ala Asn Ser Leu
530 535 540
Lys Asn Val Thr Phe Thr Glu Glu Gln Arg Ser Lys Leu Leu Ile Thr
545 550 555 560
Thr Val Gly Lys Leu Val Phe Asn Glu Ile Leu Pro Glu Ser Phe Pro
565 570 575
Tyr Met Asn Glu Pro Thr Lys Ser Asn Ile Glu Glu Lys Thr Pro Asp
580 585 590
Arg Phe Phe Leu Glu Lys Gly Ala Asp Val Lys Ala Val Ile Ala Gln
595 600 605
Gln Pro Ile Asn Ala Pro Phe Lys Lys Gly Ile Leu Gly Lys Ile Ile
610 615 620
Ala Glu Ile Phe Lys Arg Phe His Ile Thr Glu Thr Ser Lys Met Leu
625 630 635 640
Asp Arg Met Lys Asn Leu Gly Phe Lys Tyr Ser Thr Lys Ala Gly Ile
645 650 655
Thr Val Gly Val Ser Asp Ile Val Val Leu Asp Asp Lys Gln Glu Ile
660 665 670
Leu Glu Glu Ala Gln Ser Lys Val Asp Asn Val Met Lys Gln Phe Arg
675 680 685
Arg Gly Leu Ile Thr Glu Glu Glu Arg Tyr Glu Arg Val Ile Ser Ile
690 695 700
Trp Ser Ala Ala Lys Asp Val Ile Gln Gly Lys Leu Met Lys Ser Leu
705 710 715 720
Asp Glu Leu Asn Pro Ile Tyr Met Met Ser Asp Ser Gly Ala Arg Gly
725 730 735
Asn Ala Ser Asn Phe Thr Gln Leu Ala Gly Met Arg Gly Leu Met Ala
740 745 750
Asn Pro Ala Gly Arg Ile Ile Glu Leu Pro Ile Lys Ser Ser Phe Arg
755 760 765
Glu Gly Leu Thr Val Leu Glu Tyr Phe Ile Ser Thr His Gly Ala Arg
770 775 780
Lys Gly Leu Ala Asp Thr Ala Leu Lys Thr Ala Asp Ser Gly Tyr Leu
785 790 795 800
Thr Arg Arg Leu Val Asp Val Ala Gln Asp Val Ile Ile Arg Glu Thr
805 810 815
Asp Cys Gly Thr Asp Arg Gly Ile Leu Ala Lys Pro Leu Lys Glu Gly
820 825 830
Thr Glu Thr Ile Glu Arg Leu Glu Glu Arg Leu Ile Gly Arg Phe Ala
835 840 845
Arg Lys Gln Val Lys His Pro Glu Thr Gly Glu Val Leu Val Asn Glu
850 855 860
Asn Glu Leu Ile Asp Glu Asp Lys Ala Leu Glu Ile Val Glu Ala Gly
865 870 875 880
Ile Glu Glu Val Trp Ile Arg Ser Ala Phe Thr Cys Asn Thr Pro His
885 890 895
Gly Val Cys Lys Arg Cys Tyr Gly Arg Asn Leu Ala Thr Gly Ser Asp
900 905 910
Val Glu Val Gly Glu Ala Val Gly Ile Ile Ala Ala Gln Ser Ile Gly
915 920 925
Glu Pro Gly Thr Gln Leu Thr Met Arg Thr Phe His Thr Gly Gly Val
930 935 940
Ala Gly Asp Asp Ile Thr Gln Gly Leu Pro Arg Ile Gln Glu Leu Phe
945 950 955 960
Glu Ala Arg Asn Pro Lys Gly Gln Ala Thr Ile Thr Glu Ile Asp Gly
965 970 975
Thr Val Val Glu Ile Asn Glu Val Arg Asp Lys Gln Gln Glu Ile Val
980 985 990
Val Gln Gly Ala Val Glu Thr Arg Ser Tyr Thr Ala Pro Tyr Asn Ser
995 1000 1005
Arg Leu Lys Val Ala Glu Gly Asp Lys Ile Thr Arg Gly Gln Val Leu
1010 1015 1020
Thr Glu Gly Ser Ile Asp Pro Lys Glu Leu Leu Lys Val Thr Asp Leu
1025 1030 1035 1040
Thr Thr Val Gln Glu Tyr Leu Leu His Glu Val Gln Lys Val Tyr Arg
1045 1050 1055
Met Gln Gly Val Glu Ile Gly Asp Lys His Val Glu Val Met Val Arg
1060 1065 1070
Gln Met Leu Arg Lys Val Arg Val Ile Asp Ala Gly Asp Thr Asp Val
1075 1080 1085
Leu Pro Gly Thr Leu Leu Asp Ile His Gln Phe Thr Glu Ala Asn Lys
1090 1095 1100
Lys Val Leu Leu Glu Gly Asn Arg Pro Ala Thr Gly Arg Pro Val Leu
1105 1110 1115 1120
Leu Gly Ile Thr Lys Ala Ser Leu Glu Thr Asp Ser Phe Leu Ser Ala
1125 1130 1135
Ala Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Asp Ala Ala Ile Lys
1140 1145 1150
Gly Lys Arg Asp Glu Leu Leu Gly Leu Lys Glu Asn Val Ile Ile Gly
1155 1160 1165
Lys Leu Val Pro Ala Gly Thr Gly Met Met Lys Tyr Arg Lys Val Lys
1170 1175 1180
Pro Val Ser Asn Val Gln Pro Thr Asp Asp Met Val Pro Val Glu
1185 1190 1195
<210> 49
<211> 1225
<212> PRT
<213> 肺炎链球菌（Streptococcus pneumoniae）
<400> 49
Met Val Asp Val Asn Arg Phe Lys Ser Met Gln Ile Thr Leu Ala Ser
1 5 10 15
Pro Ser Lys Val Arg Ser Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Arg Glu Gly Leu Phe Asp
35 40 45
Glu Val Ile Phe Gly Pro Thr Lys Asp Trp Glu Cys Ala Cys Gly Lys
50 55 60
Tyr Lys Arg Ile Arg Tyr Arg Gly Ile Val Cys Asp Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Thr Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Lys Ala Pro Val Ser His Ile Trp Tyr Phe Lys Gly Ile Pro Ser
100 105 110
Arg Met Gly Leu Thr Leu Asp Met Ser Pro Arg Ala Leu Glu Glu Val
115 120 125
Ile Tyr Phe Ala Ala Tyr Val Val Ile Asp Pro Lys Asp Thr Pro Leu
130 135 140
Glu His Lys Ser Ile Met Thr Glu Arg Glu Tyr Arg Glu Arg Leu Arg
145 150 155 160
Glu Tyr Gly Tyr Gly Ser Phe Val Ala Lys Met Gly Ala Glu Ala Ile
165 170 175
Gln Asp Leu Leu Lys Gln Val Asp Leu Glu Lys Glu Ile Ala Glu Leu
180 185 190
Lys Glu Glu Leu Lys Thr Ala Thr Gly Gln Lys Arg Val Lys Ala Ile
195 200 205
Arg Arg Leu Asp Val Leu Asp Ala Phe Tyr Lys Ser Gly Asn Lys Pro
210 215 220
Glu Trp Met Ile Leu Asn Ile Leu Pro Val Ile Pro Pro Asp Leu Arg
225 230 235 240
Pro Met Leu Gln Leu Asp Gly Gly Arg Phe Ala Ser Ser Asp Leu Asn
245 250 255
Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn Arg Leu Ala Arg Leu
260 265 270
Leu Glu Leu Asn Ala Pro Gly Ile Ile Val Gln Asn Glu Lys Arg Met
275 280 285
Leu Gln Glu Ala Val Asp Ala Leu Ile Asp Asn Gly Arg Arg Gly Arg
290 295 300
Pro Ile Thr Gly Pro Gly Ser Arg Pro Leu Lys Ser Leu Ser His Met
305 310 315 320
Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg
325 330 335
Val Asp Phe Ser Gly Arg Ser Val Ile Ala Val Gly Pro Thr Leu Lys
340 345 350
Met Tyr Gln Cys Gly Val Pro Arg Glu Met Ala Ile Glu Leu Phe Lys
355 360 365
Pro Phe Val Met Arg Glu Ile Val Ala Arg Asp Ile Val Gln Asn Val
370 375 380
Lys Ala Ala Lys Arg Leu Val Glu Arg Gly Asp Glu Arg Ile Trp Asp
385 390 395 400
Ile Leu Glu Glu Val Ile Lys Glu His Pro Val Leu Leu Asn Arg Ala
405 410 415
Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe Glu Pro Val Leu Ile
420 425 430
Asp Gly Lys Ala Leu Arg Leu His Pro Leu Val Cys Glu Ala Tyr Asn
435 440 445
Ala Asp Phe Asp Gly Asp Gln Met Ala Ile His Val Pro Leu Ser Glu
450 455 460
Glu Ala Gln Ala Glu Ala Arg Ile Leu Met Leu Ala Ala Glu His Ile
465 470 475 480
Leu Asn Pro Lys Asp Gly Lys Pro Val Val Thr Pro Ser Gln Asp Met
485 490 495
Val Leu Gly Asn Tyr Tyr Leu Thr Met Glu Glu Ala Gly Arg Glu Gly
500 505 510
Glu Gly Met Val Phe Lys Asp Arg Asp Glu Ala Val Met Ala Tyr Arg
515 520 525
Asn Gly Tyr Val His Leu His Ser Arg Val Gly Ile Ala Thr Asp Ser
530 535 540
Leu Asn Lys Pro Trp Thr Glu Glu Gln Arg His Lys Val Leu Leu Thr
545 550 555 560
Thr Val Gly Lys Ile Leu Phe Asn Asp Ile Met Pro Glu Gly Leu Pro
565 570 575
Tyr Leu Gln Glu Pro Asn Asn Ala Asn Leu Thr Glu Gly Val Pro Ala
580 585 590
Lys Tyr Phe Leu Pro Leu Gly Gly Asp Ile Lys Glu Ala Ile Ser Asn
595 600 605
Leu Glu Leu Asn Pro Pro Phe Lys Lys Lys Asn Leu Gly Asn Ile Ile
610 615 620
Ala Glu Ile Phe Lys Arg Phe Arg Thr Thr Glu Thr Ser Ala Leu Leu
625 630 635 640
Asp Arg Met Lys Asn Leu Gly Tyr His His Ser Thr Leu Ala Gly Leu
645 650 655
Thr Val Gly Ile Ala Asp Ile Pro Val Val Asp Asp Lys Ala Glu Ile
660 665 670
Ile Glu Glu Ser His Lys Arg Val Glu Gln Ile Thr Lys Gln Phe Arg
675 680 685
Arg Gly Met Ile Thr Asp Asp Glu Arg Tyr Asn Ala Val Thr Ala Glu
690 695 700
Trp Arg Ala Ala Arg Glu Lys Leu Glu Lys Arg Leu Ile Ala Asn Gln
705 710 715 720
Asp Pro Lys Asn Pro Ile Val Met Met Met Asp Ser Gly Ala Arg Gly
725 730 735
Asn Ile Ser Asn Phe Ser Gln Leu Ala Gly Met Arg Gly Leu Met Ala
740 745 750
Ala Pro Asn Gly Arg Ile Met Glu Leu Pro Ile Leu Ser Asn Phe Arg
755 760 765
Glu Gly Leu Ser Val Leu Glu Met Phe Phe Ser Thr His Gly Ala Arg
770 775 780
Lys Gly Met Thr Asp Thr Ala Leu Lys Thr Ala Asp Ser Gly Tyr Leu
785 790 795 800
Thr Arg Arg Leu Val Asp Val Ala Gln Asp Val Ile Ile Arg Glu Asp
805 810 815
Asp Cys Gly Thr Asp Arg Gly Leu Leu Ile Arg Ser Ile Ala Glu Gly
820 825 830
Lys Glu Met Ile Glu Ser Leu Glu Glu Arg Leu Asn Gly Arg Tyr Thr
835 840 845
Lys Lys Thr Val Lys His Pro Glu Thr Gly Ala Val Ile Ile Gly Pro
850 855 860
Asn Glu Leu Ile Thr Glu Asp Lys Ala Arg Glu Ile Val Asn Ala Gly
865 870 875 880
Val Glu Glu Val Thr Ile Arg Ser Val Phe Thr Cys Asn Thr Arg His
885 890 895
Gly Val Cys Arg His Cys Tyr Gly Ile Asn Leu Ala Thr Gly Asp Ala
900 905 910
Val Glu Val Gly Glu Ala Val Gly Thr Ile Ala Ala Gln Ser Ile Gly
915 920 925
Glu Pro Gly Thr Gln Leu Thr Met Arg Thr Phe His Thr Gly Gly Val
930 935 940
Ala Ser Asn Thr Asp Ile Thr Gln Gly Leu Pro Arg Val Gln Glu Ile
945 950 955 960
Phe Glu Ala Arg Asn Pro Lys Gly Glu Ala Val Ile Thr Glu Val Lys
965 970 975
Gly Gln Val Thr Ala Ile Glu Glu Asp Ala Ser Thr Arg Thr Lys Lys
980 985 990
Val Phe Val Lys Gly Glu Thr Gly Glu Gly Glu Tyr Val Val Pro Phe
995 1000 1005
Thr Ala Arg Met Arg Val Glu Val Gly Gly Gln Val Ala Arg Gly Ala
1010 1015 1020
Ala Leu Thr Glu Gly Ser Ile Gln Pro Lys Arg Leu Leu Ala Val Arg
1025 1030 1035 1040
Asp Val Leu Ser Val Glu Thr Tyr Leu Leu Gly Glu Val Gln Lys Val
1045 1050 1055
Tyr Arg Ser Gln Gly Val Glu Ile Gly Asp Lys His Ile Glu Val Met
1060 1065 1070
Val Arg Gln Met Ile Arg Lys Val Arg Val Met Asp Pro Gly Asp Thr
1075 1080 1085
Asp Leu Leu Met Gly Thr Leu Met Asp Ile Asn Asp Phe Thr Asp Ala
1090 1095 1100
Asn Lys Asp Val Leu Ile Ala Gly Gly Val Pro Ala Thr Gly Arg Pro
1105 1110 1115 1120
Val Leu Met Gly Ile Thr Lys Ala Ser Leu Glu Thr Asn Ser Phe Leu
1125 1130 1135
Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Asp Ala Ala
1140 1145 1150
Ile Arg Gly Lys Lys Asp His Leu Leu Gly Leu Lys Glu Asn Val Ile
1155 1160 1165
Ile Gly Lys Ile Ile Pro Ala Gly Thr Gly Met Ala Arg Tyr Arg Asn
1170 1175 1180
Leu Glu Pro His Ala Val Asn Glu Glu Glu Tyr Leu Asn Pro Pro Val
1185 1190 1195 1200
Glu Glu Glu Gly Asn Glu Glu Thr Thr Glu Val Val Val Asp Thr Ala
1205 1210 1215
Val Glu Thr Val Glu Glu Thr Val Glu
1220 1225
<210> 50
<211> 1217
<212> PRT
<213> 屎肠球菌（Enterococcus faecalis）
<400> 50
Met Ile Asp Val Asn Lys Phe Glu Ser Met Gln Ile Gly Leu Ala Ser
1 5 10 15
Pro Glu Lys Ile Arg Ser Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Arg Glu Gly Leu Phe Cys
35 40 45
Glu Arg Ile Phe Gly Pro Thr Lys Asp Trp Glu Cys Ala Cys Gly Lys
50 55 60
Tyr Lys Arg Ile Arg Tyr Lys Gly Ile Val Cys Asp Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ser Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Ser His Ile Trp Tyr Phe Lys Gly Ile Pro Ser
100 105 110
Arg Met Gly Leu Val Leu Asp Met Ser Pro Arg Ala Leu Glu Glu Val
115 120 125
Ile Tyr Phe Ala Ser Tyr Val Val Ile Glu Pro Gly Asp Thr Thr Leu
130 135 140
Glu Lys Lys Gln Leu Leu Thr Glu Arg Glu Tyr Arg Glu Lys Arg Glu
145 150 155 160
Gln Tyr Gly Gln Ala Phe Lys Ala Ala Met Gly Ala Glu Ala Val Lys
165 170 175
Gln Leu Leu Asp Asn Val Asp Leu Asp Gly Glu Val Ala Gln Leu Lys
180 185 190
Glu Glu Leu Lys Thr Ala Ser Gly Gln Lys Arg Thr Arg Ala Ile Arg
195 200 205
Arg Leu Asp Ile Leu Glu Ala Phe Arg Ala Ser Gly Asn Gln Pro Ser
210 215 220
Trp Met Val Met Asp Val Ile Pro Val Ile Pro Pro Asp Leu Arg Pro
225 230 235 240
Met Val Gln Leu Glu Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn Asp
245 250 255
Leu Tyr Arg Arg Val Ile Asn Arg Asn Asn Arg Leu Lys Arg Leu Leu
260 265 270
Asp Leu Asn Ala Pro Ser Ile Ile Val Gln Asn Glu Lys Arg Met Leu
275 280 285
Gln Glu Ala Val Asp Ala Leu Ile Asp Asn Gly Arg Arg Gly Arg Pro
290 295 300
Val Thr Gly Pro Gly Asn Arg Pro Leu Lys Ser Leu Ser His Met Leu
305 310 315 320
Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg Val
325 330 335
Asp Tyr Ser Gly Arg Ser Val Ile Val Val Gly Pro Phe Leu Lys Met
340 345 350
Tyr Gln Cys Gly Leu Pro Lys Glu Met Ala Ile Glu Leu Phe Lys Pro
355 360 365
Phe Val Met Arg Glu Leu Val Gln Arg Glu Ile Ala Thr Asn Ile Lys
370 375 380
Asn Ala Lys Arg Lys Ile Glu Arg Gly Glu Asp Glu Val Trp Asp Ile
385 390 395 400
Leu Glu Glu Val Ile Gln Glu His Pro Val Leu Leu Asn Arg Ala Pro
405 410 415
Thr Leu His Arg Leu Gly Ile Gln Ala Phe Glu Pro Val Leu Val Glu
420 425 430
Gly Arg Ala Ile Arg Leu His Pro Leu Val Cys Glu Ala Tyr Asn Ala
435 440 445
Asp Phe Asp Gly Asp Gln Met Ala Val His Val Pro Leu Asn Glu Glu
450 455 460
Ala Gln Ala Glu Ala Arg Met Leu Met Leu Ala Ala Gln Asn Ile Leu
465 470 475 480
Asn Pro Lys Asp Gly Lys Pro Val Val Thr Pro Ser Gln Asp Met Val
485 490 495
Leu Gly Asn Tyr Tyr Leu Thr Met Glu Glu Glu Gly Arg Glu Gly Glu
500 505 510
Gly Met Ile Phe Arg Asp Met Asn Glu Ala Val Leu Ala Trp Gln Asn
515 520 525
Gly Tyr Val His Leu His Ser Arg Ile Gly Val Gln Thr Thr Leu Leu
530 535 540
Gly Asp Lys Pro Phe Thr Asp Trp Gln Lys Glu Arg Ile Leu Ile Thr
545 550 555 560
Thr Val Gly Lys Ile Ile Phe Asn Glu Ile Met Pro Val Glu Phe Pro
565 570 575
Tyr Leu Asn Glu Pro Thr Asp Tyr Asn Leu Thr Val Gln Thr Pro Asp
580 585 590
Lys Tyr Phe Val Glu Ala Gly Thr Asp Ile Pro Ala His Ile Lys Glu
595 600 605
Gln Glu Leu Val Leu Pro Phe Lys Lys Lys Asn Leu Gly Asn Ile Ile
610 615 620
Ala Glu Val Phe Lys Arg Phe His Ile Thr Glu Thr Ser Lys Met Leu
625 630 635 640
Asp Arg Met Lys Asp Leu Gly Tyr Lys His Ser Thr Tyr Ala Gly Met
645 650 655
Thr Val Gly Ile Ala Asp Ile Met Val Leu His Glu Lys Gln Ala Ile
660 665 670
Ile Asp Ala Ala His Lys Gln Val Glu Thr Ile Thr Lys Gln Phe Arg
675 680 685
Arg Gly Leu Ile Thr Asp Asp Glu Arg Tyr Glu Arg Val Ile Gly Val
690 695 700
Trp Asn Gly Ala Lys Asp Glu Ile Gln Gln Lys Leu Ile Glu Ser Met
705 710 715 720
Glu Ala Arg Asn Pro Ile Phe Met Met Ser Asp Ser Gly Ala Arg Gly
725 730 735
Asn Ile Ser Asn Phe Thr Gln Leu Ala Gly Met Arg Gly Leu Met Ala
740 745 750
Ala Pro Asn Gly Arg Ile Met Glu Leu Pro Ile Ile Ser Asn Phe Arg
755 760 765
Glu Gly Leu Ser Val Leu Glu Met Phe Ile Ser Thr His Gly Ala Arg
770 775 780
Lys Gly Met Thr Asp Thr Ala Leu Lys Thr Ala Asp Ser Gly Tyr Leu
785 790 795 800
Thr Arg Arg Leu Val Asp Val Ala Gln Asp Val Ile Ile Arg Glu Asp
805 810 815
Asp Cys Gly Thr Asp Arg Gly Leu Glu Ile Glu Ala Ile Arg Glu Gly
820 825 830
Asn Glu Ile Ile Glu Pro Leu Asp Glu Arg Leu Leu Gly Arg Tyr Thr
835 840 845
Arg Lys Ser Val Val His Pro Glu Thr Gly Ala Ile Ile Ile Gly Ala
850 855 860
Asp Gln Leu Ile Thr Glu Asp Leu Ala Arg Glu Ile Val Asp Ala Gly
865 870 875 880
Ile Glu Lys Val Thr Ile Arg Ser Val Phe Thr Cys Asn Thr Lys His
885 890 895
Gly Val Cys Lys His Cys Tyr Gly Arg Asn Leu Ala Thr Gly Ser Asp
900 905 910
Val Glu Val Gly Glu Ala Val Gly Thr Ile Ala Ala Gln Ser Ile Gly
915 920 925
Glu Pro Gly Thr Gln Leu Thr Met Arg Thr Phe His Thr Gly Gly Val
930 935 940
Ala Gly Asp Asp Ile Thr Gln Gly Leu Pro Arg Ile Gln Glu Ile Phe
945 950 955 960
Glu Ala Arg Asn Pro Lys Gly Gln Ala Val Ile Thr Glu Val Thr Gly
965 970 975
Glu Val Ile Asp Ile Ser Glu Asp Pro Ala Thr Arg Gln Lys Glu Val
980 985 990
Thr Ile Lys Gly Lys Thr Asp Thr Arg Thr Tyr Thr Val Pro Tyr Thr
995 1000 1005
Ala Arg Met Lys Val Ala Glu Gly Asp Ile Ile His Arg Gly Ala Pro
1010 1015 1020
Leu Thr Glu Gly Ser Ile Asp Pro Lys Gln Leu Leu Gln Val Arg Asp
1025 1030 1035 1040
Val Leu Ser Val Glu Asn Tyr Leu Leu Arg Glu Val Gln Arg Val Tyr
1045 1050 1055
Arg Met Gln Gly Val Glu Ile Gly Asp Lys His Ile Glu Val Met Val
1060 1065 1070
Arg Gln Met Leu Arg Lys Ile Arg Val Met Asp Pro Gly Asp Thr Glu
1075 1080 1085
Ile Leu Pro Gly Thr Leu Met Asp Ile Ala Glu Phe Lys Asp Arg Asn
1090 1095 1100
Tyr Asp Thr Leu Val Ala Gly Gly Val Pro Ala Thr Ser Arg Pro Val
1105 1110 1115 1120
Leu Leu Gly Ile Thr Lys Ala Ser Leu Glu Thr Asn Ser Phe Leu Ser
1125 1130 1135
Ala Ala Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Asp Ala Ala Ile
1140 1145 1150
Arg Gly Lys Lys Asp Pro Leu Leu Gly Leu Lys Glu Asn Val Ile Ile
1155 1160 1165
Gly Lys Ile Ile Pro Ala Gly Thr Gly Met Ala Arg Tyr Arg Asn Met
1170 1175 1180
Glu Pro Lys Glu Val Gly Val Ala Ser Glu Asn Val Tyr Ser Ile Ser
1185 1190 1195 1200
Asp Ile Glu Ala Gln Met Ala Ala Glu Asp Ala Met Lys Asn Ile Asn
1205 1210 1215
Lys
<210> 51
<211> 1215
<212> PRT
<213> 短乳杆菌（Lactobacillus brevis）
<400> 51
Met Val Asp Val Asn Lys Phe Glu Ser Met Gln Ile Gly Leu Ala Ser
1 5 10 15
Pro Asp Lys Ile Arg Ser Trp Ser Tyr Gly Glu Val Lys Lys Pro Glu
20 25 30
Thr Ile Asn Tyr Arg Thr Leu Lys Pro Glu Lys Asp Gly Leu Phe Asp
35 40 45
Glu Arg Ile Phe Gly Pro Thr Lys Asp Trp Glu Cys Ala Cys Gly Lys
50 55 60
Tyr Lys Arg Ile Arg Tyr Lys Gly Val Val Cys Asp Arg Cys Gly Val
65 70 75 80
Glu Val Thr Arg Ser Lys Val Arg Arg Glu Arg Met Gly His Ile Glu
85 90 95
Leu Ala Ala Pro Val Thr His Ile Trp Tyr Phe Lys Gly Ile Pro Ser
100 105 110
Arg Met Gly Leu Val Leu Asp Met Ser Pro Arg Ala Leu Glu Glu Ile
115 120 125
Ile Tyr Phe Ala Ser Tyr Val Val Leu Asp Pro Gly Asn Thr Pro Leu
130 135 140
Glu Lys Lys Gln Leu Leu Ser Glu Arg Asp Tyr Arg Asp Lys Leu Leu
145 150 155 160
Glu Tyr Gly Ser Asp Ala Phe Lys Ala Glu Met Gly Ala Glu Ala Ile
165 170 175
Lys Lys Leu Leu Met Ser Val Asp Leu Asp Lys Glu Val Thr Glu Leu
180 185 190
Lys Glu Glu Leu Lys Glu Ala Thr Gly Gln Lys Arg Thr Arg Ala Val
195 200 205
Arg Arg Leu Asp Ile Leu Glu Ala Phe Val Met Ser Gly Asn Arg Pro
210 215 220
Glu Trp Met Val Met Asp Ala Ile Pro Val Ile Pro Pro Asp Leu Arg
225 230 235 240
Pro Met Val Gln Leu Glu Gly Gly Arg Phe Ala Thr Ser Asp Leu Asn
245 250 255
Asp Leu Tyr Arg Arg Val Ile Asn Arg Asn Ser Arg Leu Lys Arg Leu
260 265 270
Leu Asp Leu Asn Ala Pro Gly Ile Ile Val Gln Asn Glu Lys Arg Met
275 280 285
Leu Gln Glu Ala Val Asp Ala Leu Ile Asp Asn Gly Arg Arg Gly Arg
290 295 300
Pro Val Ala Gly Pro Gly Asn Arg Pro Leu Lys Ser Leu Ser His Met
305 310 315 320
Leu Lys Gly Lys Gln Gly Arg Phe Arg Gln Asn Leu Leu Gly Lys Arg
325 330 335
Val Asp Tyr Ser Gly Arg Ser Val Ile Asp Val Gly Pro Ser Leu Lys
340 345 350
Phe Asn Gln Met Gly Leu Pro Val Pro Met Ala Leu Glu Leu Phe Arg
355 360 365
Pro Phe Ile Met Lys Glu Leu Val Ala Arg Gly Leu Ala Ser Asn Ile
370 375 380
Lys Asn Ala Lys Arg Gln Ile Asp Arg Glu Asp Asp Asp Val Phe Asn
385 390 395 400
Val Leu Glu Asp Val Ile Lys Glu His Pro Val Leu Leu Asn Arg Ala
405 410 415
Pro Thr Leu His Arg Leu Gly Ile Gln Ala Phe Glu Pro Val Leu Val
420 425 430
Ser Gly Lys Ala Met Arg Leu His Pro Leu Ala Cys Glu Ala Tyr Asn
435 440 445
Ala Asp Phe Asp Gly Asp Gln Met Ala Ile His Val Pro Leu Ser Asp
450 455 460
Glu Ala Gln Ala Glu Ala Arg Leu Leu Met Leu Ala Ala His His Ile
465 470 475 480
Leu Ala Pro Ala Ser Gly Lys Pro Val Val Ala Pro Ser Gln Asp Met
485 490 495
Val Ile Gly Asn Tyr Tyr Leu Thr Met Glu Glu Ala Asn Arg Glu Gly
500 505 510
Glu Gly Met Ile Phe Thr Asp Leu Asp Glu Ala Thr Leu Ala Tyr Arg
515 520 525
Asn Gly Ile Val His Trp His Thr Arg Val Gly Val Gln Val Thr Ser
530 535 540
Met Pro Asp Lys Pro Phe Thr Asp Glu Gln Arg Ser Lys Ile Met Val
545 550 555 560
Thr Thr Val Gly Lys Leu Ile Phe Asn Asn Ile Leu Pro Lys Ser Phe
565 570 575
Pro Tyr Leu Asn Glu Pro Thr Ser Thr Asn Leu Asn Gly Tyr Val Pro
580 585 590
Asp Lys Tyr Phe Leu Glu Pro Gly Glu Asp Ile His Asp Tyr Leu Gln
595 600 605
Asn Ala Glu Ile Ile Pro Pro Phe Lys Lys Gly Phe Leu Ser Asp Ile
610 615 620
Ile Ala Ala Val Tyr Gln Gln Tyr Lys Val Thr Ala Thr Ser Glu Leu
625 630 635 640
Leu Asp Arg Ile Lys Asp Leu Gly Tyr Asn Glu Ser Thr Lys Ser Gly
645 650 655
Leu Thr Val Gly Met Val Asp Val Thr Asp Leu Lys Glu Lys Pro Glu
660 665 670
Ile Ile Ala Ala Ala His Lys Gln Val Ser Thr Val Thr Lys Gln Phe
675 680 685
Arg Arg Gly Leu Ile Thr Asp His Glu Arg Tyr Glu Arg Val Ile Gly
690 695 700
Ile Trp Asn Asp Ala Lys Asp Glu Ile Gln Asn Ala Leu Ile His Ser
705 710 715 720
Phe Asp Gln Gln Asn Pro Ile Phe Met Met Ser Asp Ser Gly Ala Arg
725 730 735
Gly Asn Ile Ser Asn Phe Thr Gln Leu Ala Gly Met Arg Gly Leu Met
740 745 750
Ala Ala Pro Ser Gly Asp Ile Met Glu Leu Pro Ile Thr Ser Asn Phe
755 760 765
Arg Glu Gly Leu Thr Val Met Glu Met Phe Ile Ser Thr His Gly Ala
770 775 780
Arg Lys Gly Met Thr Asp Thr Ala Leu Lys Thr Ala Asn Ser Gly Tyr
785 790 795 800
Leu Thr Arg Arg Leu Val Asp Val Ala Gln Asp Val Ile Ile Arg Glu
805 810 815
Lys Asp Cys Gly Thr Asp Arg Gly Leu Lys Ile Arg Ala Ile Thr Asp
820 825 830
Gly Asn Glu Met Ile Glu Pro Leu Tyr Asp Arg Ile Leu Gly Arg Tyr
835 840 845
Thr Gln Lys Thr Val Tyr Asp Pro Gln Thr Gly Asp Val Ile Val Pro
850 855 860
Lys Asn Gln Met Ile Val Glu Asp Thr Ala Gln Gln Ile Val Asp Ala
865 870 875 880
Gly Val Glu Glu Val Thr Ile Arg Ser Ala Phe Thr Cys Asn Thr Glu
885 890 895
His Gly Val Cys Glu His Cys Tyr Gly Arg Asn Met Ala Thr Gly Asp
900 905 910
Glu Val Glu Val Gly Glu Ala Val Gly Thr Val Ala Ala Gln Ser Ile
915 920 925
Gly Glu Pro Gly Thr Gln Leu Thr Met Arg Asn Phe His Thr Gly Gly
930 935 940
Val Ala Gly Asn Glu Asp Ile Thr Gln Gly Leu Pro Arg Val Gln Glu
945 950 955 960
Leu Phe Glu Ser Arg Asn Pro Lys Gly Lys Ala Glu Ile Thr Glu Val
965 970 975
Thr Gly Thr Val Glu Ser Ile Glu Glu Asn Pro Ala Glu Arg Thr Lys
980 985 990
Glu Ile Thr Ile Lys Gly Glu Ala Asp Thr Arg Ser Tyr Thr Leu Pro
995 1000 1005
Ile Thr Ala Arg Met Arg Val Ser Glu Gly Asp Phe Ile His Arg Gly
1010 1015 1020
Gly Ala Leu Asn Tyr Gly Ser Val Asp Pro Lys Glu Leu Leu Arg Val
1025 1030 1035 1040
Arg Asp Val Leu Ser Thr Glu Thr Tyr Ile Leu Gly Glu Val Gln Arg
1045 1050 1055
Val Tyr Arg Met Gln Gly Val Ala Ile Ser Asp Lys His Val Glu Ile
1060 1065 1070
Met Val Arg Gln Met Leu Arg Lys Val Arg Ile Met Asp Pro Gly Asp
1075 1080 1085
Thr Asp Val Leu Pro Gly Thr Leu Met Asp Ile Gln Asp Phe Arg Arg
1090 1095 1100
Ala Asn Tyr Gln Thr Leu Ile Asp Gly Gly Ile Ala Ala Thr Ala Arg
1105 1110 1115 1120
Pro Val Ile Leu Gly Ile Thr Lys Ala Ala Leu Glu Thr Asn Ser Phe
1125 1130 1135
Leu Ser Ala Ala Ser Phe Gln Glu Thr Thr Arg Val Leu Thr Asp Ala
1140 1145 1150
Ala Ile Arg Gly Lys Asn Asp Pro Leu Val Gly Leu Lys Glu Asn Val
1155 1160 1165
Ile Ile Gly Lys Ile Ile Pro Ala Gly Thr Gly Met Pro Asp Tyr Arg
1170 1175 1180
Gln Ile Lys Pro Lys Glu Val Gly Gly Thr Ser Thr Glu Gly Val Tyr
1185 1190 1195 1200
Ser Ile Ser Asp Leu Glu Lys Gln Met Gln Glu Asp Ser Gln Ala
1205 1210 1215

Claims (15)

1.核酸分子，所述核酸分子包含变体rpoC编码序列，其中：
所述变体rpoC编码序列编码变体RpoC RNA聚合酶β'亚基蛋白；并且
所述变体RpoC包含R47C取代，其中所述R47C取代的编号基于SEQ ID NO:26的大肠埃希氏菌(Escherichia coli)的野生型RpoC RNA聚合酶β'亚基蛋白来限定。
2.根据权利要求1所述的核酸分子，其中相对于包含SEQ ID NO:26的野生型RpoC的表达，所述变体RpoC的表达使质粒的拷贝数减少。
3.根据权利要求1所述的核酸分子，其中所述变体RpoC包含：
(1)包含SEQ ID NO:28的N端结构域；
(2)包含SEQ ID NO:29的中央结构域；和
(3)包含SEQ ID NO:30的C端结构域；
其中所述R47C取代存在于所述N端结构域内。
4.根据权利要求1所述的核酸分子，其中所述变体RpoC包含：
(1)包含SEQ ID NO:31的N端结构域；
(2)包含SEQ ID NO:32的中央结构域；和
(3)包含SEQ ID NO:33的C端结构域；
其中所述R47C取代存在于所述N端结构域内。
5.根据权利要求1所述的核酸分子，其中所述变体RpoC包含SEQ ID NO:27。
6.载体，所述载体包含根据权利要求1至5中任一项所述的核酸分子。
7.重组微生物，所述重组微生物包含根据权利要求1至5中任一项所述的核酸分子。
8.根据权利要求7所述的重组微生物，其中：
所述重组微生物通过转化、接合或转导中的一种或多种而将包含所述核酸序列的rpoC编码序列载体引入前体微生物中来制备；或
所述重组微生物包含染色体，并且所述变体rpoC编码序列基于由所述变体rpoC编码序列置换内源rpoC编码序列而存在于所述染色体中。
9.根据权利要求7所述的重组微生物，其中所述重组微生物由埃希氏菌属(Escherichia)细菌制备。
10.根据权利要求7所述的重组微生物，其中所述重组微生物由大肠埃希氏菌种细菌制备。
11.用于调控根据权利要求7至10中任一项所述的重组微生物中的对象载体的拷贝数的方法，所述方法包括在足以进行所述对象载体的复制的条件下在培养基中培养所述重组微生物，从而调控所述对象载体的拷贝数。
12.通过使用权利要求7至10中任一项所述的重组微生物制备目标产物的方法，其中所述重组微生物包括目标基因载体，并且所述目标基因载体包括用于制备目标产物的目标基因，所述方法包括以下步骤：
(1)在所述重组微生物表达所述目标基因的条件下在培养基中培养所述重组微生物，从而制备所述目标产物；和
(2)从所述重组微生物和/或所述培养基回收所述目标产物。
13.根据权利要求12所述的方法，其中所述目标产物包含目标生物材料。
14.根据权利要求12所述的方法，其中所述目标产物包含目标药物产品、其前体和/或用于其生产的酶。
15.根据权利要求12所述的方法，其中所述目标产物包括目标聚合物、其前体和/或用于其生产的酶。
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