疫苗的制作方法

专利名称：疫苗的制作方法
技术领域：
本发明涉及用于检测含有一种III型分泌系统的细菌致病菌株并鉴定所述菌株染色体中毒力基因所在区的一种基本方法。更具体地说，本发明涉及适用于病原体百日咳博德特氏菌(Bordetella pertussis)的方法。此外，本发明涉及这些区内新鉴定的多核苷酸、由它们编码的毒力多肽，并涉及这类多核苷酸和多肽的应用，还涉及它们的产生。
背景技术：
III型分泌系统病原菌侵入广泛宿主范围中的许多不同的生态位，并引起各种各样的综合征。这是由于先前认为每种疾病是由独特的分子机制诱发的。然而，这类机制的范围没有像最初想象地那样广；而是，细菌利用许多共同分子工具达到各种各样目的。在这些工具中有III型分泌系统，所述系统给细菌提供将毒力因子直接靶向宿主细胞的工具。然后这些因子干预宿主细胞的功能以对病原体有益。
III型输出系统负责分泌沙门氏菌属(Salmonella)和志贺氏菌属(Shigella)侵入和毒力因子、致肠病大肠杆菌(EPEC)信号转导分子、数种植物病原体(例如野油菜黄单胞菌辣椒斑点病致病变种(Xanthomonas campestris pv.vesicatoria)[Fenselau等，1992])中的毒力因子以及耶尔森氏菌属(Yersinia)中的Yop蛋白。Yop输出机制已经是研究最透彻的III型分泌器(参见例如Allaoui等，1994；Bergman等，1994)。在该系统中，推测超过20种不同的Ysc/Lcr蛋白(所有均由毒力质粒pYV编码)组成一种跨越耶尔森氏菌属胞外被膜的分泌通道。除包括在所述分泌器中的这些元件外，pYV质粒还编码Yop蛋白，所述Yop蛋白是所分泌的底物并且看来是毒力的实际效应物。
源自不同物种的III型分泌系统的比较研究表明，分泌器的组分是保守的(Gygi等，1995；Bogdanove等，1996)。另外，已经在参与鞭毛装配的决定子中发现了同系物，表明该分泌途径可能涉及表面细胞器的生物合成(Ramakrishnan等，1991)。
然而，相比之下，所分泌的底物没有共享相似性，只是在少数情况下例外。因此，被放弃的对应于每种疾病的独特分子机制的概念可能在效应蛋白水平上再现。致病性岛(pathogenicity island)致病性岛已经成为细菌毒力领域中的一个新课题。虽然它们可以包含III型分泌系统，但是它们不仅仅如此。
毒力基因研究的早期，观察到这些基因中的许多在质粒上存在。然而，在染色体上也发现了大量的毒力基因。意想不的是，染色体毒力基因在功能相关组群中还常常聚集成簇。这样的毒力基因群产生致病性岛(Pai)的概念，所述致病性岛可以被定义为携带毒力基因的密集的、独特的遗传单位。通常侧翼为同向重复序列的这些单位占据大的染色体区(通常＞30kb)并存在于致病菌株中，而不存在或偶尔分布于细菌菌种的较低致病性(或无致病性)的菌株中。这些DNA区段时常与位于其边界的tRNA基因和/或插入序列(IS)元件相关。另外，它们的G+C含量通常与宿主细菌DNA的G+C含量不同，说明一种异种起源。
已经在越来越多的细菌病原体中发现了致病性岛，所述病原体包括不同类别的大肠杆菌、鼠伤寒沙门氏菌(Salmonella typhimurium)、耶尔森氏菌(Yersinia spp)、幽门螺杆菌(Helicobacter pylori)、霍乱弧菌(Vibrio cholera)等。
最初深入细致研究的致病性岛是Pai I和Pai II，其编码致尿路病大肠杆菌的溶血素决定子。这两种Pai邻接同向重复序列，并可以以频率10-4从染色体中缺失，产生无毒性的突变菌株。另一35kb致病性岛最近已经在致肠病大肠杆菌(EPEC)的染色体上鉴定出，并发现其编码所有已知涉及所谓“粘附与消除(attaching and effacing)”(AE)损害形成的决定子。因此，将该区称为“肠细胞消除基因座”(LEE)。尽管事实上致尿路病大肠杆菌和致肠病大肠杆菌引起完全不同的传染病，但是致尿路病菌株的Pai I和EPEC的LEE基因座在完全相同的位置插入至大肠杆菌染色体中。
当某些研究者支持必需包括其染色体位置的致病性岛定义时，其它研究者已经将所述概念延伸至多区组(block)毒力基因，而与其在染色体、质粒或噬菌体中的位置无关。一方面，噬菌体和质粒可以容易地插入染色体中和从染色体中切除，并且另一方面，隐蔽性质粒复制起点或噬菌体相关序列在Pai中被检测到，这一事实支持后一较少限制的定义。
编码III型分泌系统的致病性岛(PAI)包括的基因分为两类I类和II类。I类包括编码分泌器组分及其表达的调节物的基因，II类包括编码分泌的效应蛋白的基因。耶尔森氏菌属的lcrD和yscU均属于I类。I类决定子的精确功能还不大了解。虽然有时不能明确地清楚区分I类组分和II类组分，但是I类基因可以鉴定为存在于许多不同种中，并且它们相应的基因序列的比较表明，等效基因共享显著水平(yscI、yscO)或甚至高水平(lcrD、yscU、yscN)的序列相似性(Hueck，1998)。
第二类基因(II类)编码构成由易位子分泌的底物的蛋白。看来这些蛋白为真正的毒力效应物并被称为靶蛋白、毒力效应蛋白或简称为效应物。与I类基因产物中普遍的情况相反，所述效应物在种之间没有共享相似性或共享的相似性非常低地。效应蛋白是呈现最大生物、疫苗和诊断潜能的那些蛋白。
本发明人已经发现，单个致病性岛内的I类和II类基因的聚簇，通过靶向可以采用其大量的直向同源基因其中一种的已知序列鉴定的I类基因，提供方便地发现和鉴定未知II类基因的机会百日咳博德特氏菌百日咳是一种由百日咳博德特氏菌感染引起的疾病，并且是一种严重衰弱的人类疾病，尤其是在婴幼儿中。虽然可获得有效抵抗该疾病的全细胞和无细胞疫苗，但是仍需要鉴定进一步高度纯化的、可被用于更有效的百日咳疫苗中的百日咳蛋白。
虽然知道许多百日咳毒力相关因子，例如百日咳毒素、丝状血细胞凝集素、pertactin，它们已经包括在多种无细胞疫苗中，但是没有方便的应用百日咳基因组鉴定其它毒力因子的基因方法(除了麻烦地对完整基因组测序之外)。虽然最近已经显示I类III型分泌系统毒力基因存在于支气管炎博德特氏菌(B.bronchiseptica)和百日咳博德特氏菌(Yuk等，1998)，但是没有全面地分析博德特氏菌属中的致病性岛，直到本发明才了解效应物基因在这种致病性岛中的身份和表征。
发明概述一方面，本发明涉及鉴定含有III型分泌系统的细菌菌株中的新毒力基因的方法。具体来讲，本发明允许鉴定与含有III型分泌系统基因的致病性岛相关的效应物毒力基因。本发明的另一方面是含有III型分泌系统的致病细菌菌株的鉴定方法。本发明的另一方面涉及百日咳博德特氏菌BopN、Orf1、Orf2、Orf3、Orf4、Orf5、Orf6、Orf7、Orf8、Orf9、Orf10、Orf11、Orf12、Orf13、Orf14、Orf15效应蛋白以及编码它们的相应的多核苷酸序列。
虽然已经报道了III型分泌系统和致病性岛的基本概念，但是迄今为止仍没有解决如何简便而可靠地鉴定是否任何特定的生物都具有这种细胞机器的问题。这种方法极其有用，可用于确定特定菌株是否在致病性岛内具有III型分泌系统，表征致病性岛内的未知毒力基因，以及用于测定含有III型分泌系统的培养的细菌菌株是否是致病性细菌的快速诊断方法。
在本发明中，描述了一种新的、基本方法可达到上述目的。更具体地讲，本发明利用使用根据作为靶序列的毒性小肠结肠炎耶尔森氏菌(Yersinia enterocolitica)lcrD基因的序列而具体设计的理想合适引物的方法。发现III型分泌系统存在于百日咳博德特氏菌中的致病性岛内，并且鉴定了致病性岛内的每一基因。
附图简述

图1克隆的152 bp扩增子的核苷酸序列和推断的氨基酸序列。涉及最初扩增、随后嵌套式PCR和筛选基因文库的引物都衍生自该序列，并且具体列于表1中。
图2来自与耶尔森氏菌属LcrD同源的推断的氨基酸序列的PileUp图。缩写BbuF1hA＝布氏疏螺旋体(Borrelia burgdorferi)F1hA；TpaF1hA＝苍白密螺旋体(Treponema pallidum)FlhA；BsuF1hA＝枯草芽孢杆菌(Bacillus subtilis)F1hA；CjeFlbA＝空肠弯曲杆菌(Campylobacter jejuni)F1bA；HpyF1hA＝幽门螺杆菌(Helicobacterpylori)F1hA；EcoF1hA-大肠杆菌F1hA；StyF1hA＝鼠伤寒沙门氏菌(Salmonella typhimurium)F1hA；YenF1hA＝小肠结肠炎耶尔森氏菌F1hA；PmiF1hA＝奇异变形菌(Proteus mirabilis)F1hA；CcrF1bF＝新月柄杆菌(Caulobacter crescentus)F1bF；EcoFhiA＝大肠杆菌FhiA；EamHrpI＝解淀粉欧文氏菌(Erwinia amylovora)HrpI；PsyHrpI＝丁香假单胞菌(Pseudomonas syringae)HrpI；ECEPSepA＝致肠病大肠杆菌SepA；StySsaV＝鼠伤寒沙门氏菌SsaV；RsoHrpO＝茄科罗尔斯通氏菌(Ralstonia solanacearum)HrpO；XcaHrpC2＝野油菜黄单胞菌(Xanthomonas campestris)HrpC2；Sf1MxiA＝弗氏志贺氏菌(ShigellaFlexneri)MxiA；StyInvA＝鼠伤寒沙门氏菌InvA；PaePcrD＝铜绿假单胞菌(Pseudomonas aeruginosa)PcrD；YenLcrD＝小肠结肠炎耶尔森氏菌LcrD；BpeBcrD＝百日咳博德特氏菌BcrD；CpsTtsB＝鹦鹉热衣原体(Chlamydia psittaci)TtsB。
图3百日咳博德特氏菌致病性岛(Pai)的组构。Pai周围有4个管家基因(阴影线框)和IS481转座酶基因(黑框)。Pai由编码涉及分泌器和其调节的决定子的基因(I类基因，灰色框中)以及推断编码效应蛋白的ORF(II类基因，白色框中)组成。字母表示相应的I类bsc基因，而数字对应于列于表3中的II类ORF。
图4来自与耶尔森氏菌属YscU同源的推断的氨基酸序列的PileUp图。缩写BbuF1hB＝布氏疏螺旋体F1hB；TpaF1hB＝苍白密螺旋体F1hB；EcoF1hB＝大肠杆菌F1hB；StyF1hB＝鼠伤寒沙门氏菌F1hB；PmiF1hBpart＝部分奇异变形菌F1hB；YenF1hB＝小肠结肠炎耶尔森氏菌F1hB；BsuF1hB＝枯草芽孢杆菌F1hB；HpyF1hB＝幽门螺杆菌F1hB；AtuF1hB＝根癌土壤杆菌(Agrobacterium tumefaciens)F1hB；CcrPodW＝新月柄杆菌PodW；Sf1Spa40＝弗氏志贺氏菌Spa40；StySpaS＝鼠伤寒沙门氏菌SpaS；EcoEscU-大肠杆菌EscU；StySsaU＝鼠伤寒沙门氏菌SsaU；BpeBscU＝百日咳博德特氏菌BscU；YenYscU＝小肠结肠炎耶尔森氏菌YscU；RsoHrpN＝茄科罗尔斯通氏菌HrpN；XcaOrf0part＝部分野油菜黄单胞菌Orf0；EamHrcU＝解淀粉欧文氏菌HrcU；EheHrcUpart＝部分草生欧文氏菌(Erwiniaherbicola)HrcU；PsyHrpY＝丁香假单胞菌HrpY；CpsOrf1＝鹦鹉热衣原体Orf1。
图5包含III型分泌系统致病性岛的百日咳博德特氏菌基因组的DNA序列。关于可读框的信息，应参考表2、表3和表4以及图3。
图6经亲和层析纯化MBP-Orf2、-4、-6和-10。经SDS-PAGE分析并经考马斯蓝染色显示每种裂解液的超速离心上清液(板的左部分)和从亲和柱洗脱的产物(板的右部分)。
本发明的描述迄今为止鉴定的III型分泌系统或者由染色体致病性岛基因或者由质粒致病性岛基因编码。然而，现有技术没有认识到编码III型分泌系统I类组分的基因的保守性和这些基因与效应蛋白编码序列成簇聚集提供了检测涉及宿主定居中未鉴定的靶蛋白的机会。这类蛋白可能在疫苗领域和诊断领域均具有潜在价值。
虽然，可以用编码任何保守的(I类)III型分泌器蛋白的基因的已知序列实施本发明，但是优选lcrD基因。所选定的基因将用作检测相关细菌菌种中未鉴定的致病性岛的靶。优选来自耶尔森氏菌属的lcrD基因，因为它编码最近鉴定的蛋白LcrD/FlbF家族的原祖型。该家族的成员涉及宿主细胞侵入、数种致植物病细菌中的毒力或鞭毛装配。优选lcrD，因为LcrD蛋白以及因此编码它的基因是所述分泌器的最保守的决定子之一。另外，多重氨基酸比较已经显示，可以将LcrD家族成员的分类分为两个主要的亚家族，有趣的是，它们与确定给每个亚家族的这些蛋白的功能相关。一个亚家族包括所有涉及游动性的蛋白，而另一亚家族包括所有毒力相关决定子。这种观察在图2中说明(并在Gyri等(1995)和Bogdanove等(1996)中提及)。因此，如果鉴定出一个未知lcrD同源基因，则它可以在常规测序之后，被分类为毒力基因或鞭毛基因。一旦鉴定出致病性岛，则这种简单试验就因此可以决定是否应该开始搜索所述致病性岛上的其它毒力基因。
鉴定包含III型分泌系统的未知致病性岛的优选方法是通过i) 鉴定靶蛋白序列(最好是LcrD)的两个高度保守区。最好是，两个区应均含有由最少数目密码子可能性例如甲硫氨酸(ATG是唯一可能性)或色氨酸(TGG是唯一可能性)编码的保守氨基酸。这将在所述方法的下一步骤设计的两种简并引物组中的排列数目减至最小，因此确保每种引物组会特异性地与未知LcrD等效基因退火的较高概率(由此将背景非特异性相互作用减至最低)。最优选是，选定的区应与存在于所有有鞭毛细菌菌株的平行进化同源物flhA鞭毛基因明显地区分开。ii) 设计一组用于所选定的两个区的简并引物，以致a)所述引物为至少15个碱基长，优选为20-30个碱基长，而再更优选为21-23个碱基长；b)它们在可以为超过一种类型的核苷酸的碱基上是简并的，但仍编码相同氨基酸(由于氨基酸密码子选择的简并性)，但并不比所需要的有更多的简并，以覆盖所选定的氨基酸区的所有排列，和c)编码所选定蛋白更多N端区的引物组应对应于其相应的双链DNA序列的编码链，而编码更多C端区的引物组应对应于相应的双链DNA序列的互补链。iii)采用本领域众所周知的常规DNA合成法，合成步骤ii)的简并引物组。iv) 纯化步骤iii)的引物组vi) 以合适的量并在合适的缓冲液中，将两个引物组和含有来自细菌菌株核酸的样品(最好是所述细菌菌种自身的细胞样品)混合在一起，以便进行聚合酶链式反应(PCR)vi) 进行PCR反应，以便扩增两种引物之间的基因区(采用本领域众所周知的技术，可以将进行该PCR反应的条件最优化)vii)观察凝胶(最好是琼脂糖凝胶)上预期大小的扩增产物的所述反应产物；如果没有这样的产物存在，则所述细菌菌株不太可能使用III型分泌系统；如果有这样的产物存在，则所述细菌菌株可能具有III型分泌系统，并可能是病原菌。
确证所述扩增产物实际上对应于毒力基因的优选方法是通过实施上述步骤i)-vii)(其中所述靶蛋白是LcrD)并且然后viii)任选地通过从所述凝胶中取出正确条带，而将正确大小的产物与不正确大小的任何背景产物分离开，通过常规方法纯化所述产物，并在另一PCR反应(最好是在更严格的PCR条件下)中用所述两种简并引物组将所述产物再扩增一次[假如步骤vii)的产物对于直接克隆而言还不够纯，则需要该步骤]ix) 通过常规方法将所述DNA片段插入能够被测序的载体中，并对所述片段测序。x) 将ix)的推断的氨基酸序列与已知LcrD/FlbF蛋白家族成员的氨基酸序列进行比较，以将所述扩增产物联想(associate)为或者毒力基因或者是鞭毛基因的一部分。
并且任选地xi) 采用所述片段的内部序列设计引物，所述引物是所述未知LcrD等效基因的实际序列并且对所述未知LcrD等效基因的序列是特异性的。xii) 首先采用xi)的引物筛选阳性克隆生物的基因组文库xiii)分离xii)的克隆，并将一个或更多个所述克隆测序xiv) 扫描一个克隆的序列(和其它克隆的重叠序列)以搜索可读框，所述可读框大约与LcrD(约2100 bp)大小相同并编码与LcrD同源的一种蛋白xv) 确定所述LcrD等效蛋白是与flbF(鞭毛蛋白分泌)基因家族更同源，还是与LcrD(III型分泌系统致病性岛)基因家族更同源。
鉴定完整的致病性岛并确定未鉴定毒力效应的基因的优选方法是通过实施上述步骤i)-xv)(其中所述靶蛋白是LcrD)并且然后xvi) 如果所述序列与LcrD基因家族更同源，则设计已确定的所述基因序列每端的引物，对基因组文库扫描和测序(采用标准染色体步查策略—其中将最初克隆的插入片段边界用作筛选和克隆相邻区的探针)，以最终对所述致病性岛(其两个边界均将根据同向或反向重复序列或插入序列的存在、或者管家基因的存在而确定)的全部进行测序xvii) 确定所述测序的致病性岛内未鉴定的毒力效应物基因xviii)克隆、表达和特征鉴定编码所述生物的毒力效应蛋白的xvii)的毒力基因定义“博德特氏菌属致病性蛋白”总的来讲是指具有由表2和表3中限定的基因编码的氨基酸序列的多肽或其等位基因变异体。这些蛋白是BcrD、BcrH、BscC、BscD、BscE、BscF、BscI、BscJ、BscK、BscL、BscN、BscO、BscP、BscQ、BscR、BscS、BscT、BscU、BscV、BrpL、BopN、Orf1、Orf2、Orf3、Orf4、Orf5、Orf6、Orf7、Orf8、Orf9、Orf10、Orf11、Orf12、Orf13、Orf14、Orf15。
“博德特氏菌属致病性基因”是指具有表2和表3中限定的核苷酸序列的多核苷酸或其等位基因变异体和/或其互补物。这些基因是bcrD、bcrH、bscC、bscD、bscE、bscF、bscI、bscJ、bscK、bscL、bscN、bscO、bscP、bscQ、bscR、bscS、bscT、bscU、bscV、brpL、bopN、orf1、orf2、orf3、orf4、orf5、orf6、orf7、orf8、orf9、orf10、orf11、orf12、orf13、orf14、orf15。
“多肽”是指包含由肽键或修饰肽键(即肽同配物)相互连接的两个或更多个氨基酸的任何肽或蛋白。“多肽”指两条短链时，通常称为肽、寡肽或寡聚体；而指较长链时，一般称为蛋白。多肽可以含有20个基因编码的氨基酸以外的氨基酸。“多肽”包括或者通过天然加工例如翻译后加工修饰、或者通过本领域众所周知的化学修饰技术修饰的氨基酸序列。这样的修饰全面地描述于基本教科书中和更详细描述于专著中以及在大量的研究文献中。修饰可以发生在多肽中的任何位置，包肽肽主链、氨基酸侧链以及氨基端或羧基端。人们会认识到，相同类型的修饰可以以相同或不同程度在特定的多肽中的数个位点上存在。此外，特定多肽可以包含许多类型的修饰。多肽可以通过遍在蛋白化而分支，它们可以是环状的，具有或不具有分支。环状多肽、分支多肽和分支环状多肽可以产生自翻译后天然加工或可以经合成方法制备。修饰包括乙酰化、酰化、ADP-核糖基化、酰胺化、共价连接黄素、共价连接血红素部分、共价连接核苷酸或核苷酸衍生物、共价连接脂质或脂质衍生物、共价连接磷脂酰肌醇、交朕、环化、二硫键形成、脱甲基、形成共价交朕、形成胱氨酸、形成焦谷氨酸、甲酰化、γ-羧化、糖基化、GPI锚钩形成、羟化、碘化、甲基化、豆蔻酰化、氧化、蛋白酶解加工、磷酸化、异戊二烯化、外消旋化、硒酰化(selenoylation)、硫酸化、转移RNA介导的使氨基酸添加至蛋白例如精氨酰化和遍在蛋白化。参见，例如，PROTEINS-STRUCTURE ANDMOLECULAR PROPERTIES，第二版，T.E.Creighton，W.H.Freemanand Company，New York，1993和Wold，F.，翻译后蛋白修饰回顾和展望，第1-12页，载于POSTTRANSLATIONAL COVALENTMODIFICATION OF PROTEINS，B.C.Johnson编著，Academic Press，New York，1983；Seifter等，“蛋白修饰和非蛋白辅因子的分析”，MethEnzymol(1990)182626-646和Rattan等，“蛋白质合成翻译后修饰和老化”，Ann NY Acad Sci(1992)66348-62。
“多核苷酸”一般是指任何多核糖核苷酸或多脱氧核糖核苷酸，它可以是未修饰的RNA或DNA或者修饰的RNA或DNA。“多核苷酸”包括但不限于单链DNA和双链DNA、单链区和双链区混合的DNA、单链RNA和双链RNA、单链区和双链区混合的RNA、包含DNA和RNA的杂种分子，所述杂种分子可以是单链、或更通常是双链、或者是单链区和双链区的混合物。另外，“多核苷酸”是指包含RNA或DNA或者RNA和DNA两种的三链区。术语多核苷酸也包括含有一个或更多个修饰碱基的DNA或RNA和用于稳定或为了其它原因具有主链修饰的DNA或RNA。“修饰”碱基包括例如三苯甲基化碱基和稀有碱基诸如肌苷。已经对DNA和RNA进行了各种修饰；因此，“多核苷酸”包括化学、酶学或代谢修饰形式的多核苷酸如通常在自然界发现的，以及具有病毒和细胞特征性的化学形式的DNA和RNA。“多核苷酸”也包括相对短的多核苷酸，通常是指寡核苷酸。
本文所用的术语“变异体”是指分别与参比多核苷酸或多肽不同、但保留基本特性的多核苷酸或多肽。一种典型的多核苷酸的变异体与另一参比多核苷酸在核苷酸序列方面不同。变异体核苷酸序列中的变化可以改变或不改变由参比多核苷酸编码的多肽的氨基酸序列。核苷酸变化可以导致如下文所讨论的由参比序列编码的多肽中氨基酸取代、添加、缺失、融合和截短。一种典型的多肽的变异体与另一参比多肽在氨基酸序列方面不同。一般而言，差异是有限的，以便参比多肽的序列和变异体的序列总的来讲是非常相似的，并且在许多区中是相同的。由于以任何组合的一个或更多个取代(最好是保守取代)、添加、缺失，变异体和参比多肽可以在氨基酸序列方面不同。取代或插入的氨基酸残基可以是或不是由遗传密码编码的残基。多核苷酸或多肽的变异体可以是天然存在例如等位基因变异体，或它可以是已知天然不存在的变异体。多核苷酸和多肽非天然存在的变异体可以通过诱变技术或通过直接合成制备。变异体应保留参比多肽的一种或更多种生物活性。例如，它们应具有与参比多肽相似(最好是相同)的抗原性或免疫原性活性。可以采用标准免疫印迹实验，最好是采用抗参比多肽的多克隆血清，测试抗原性。可以通过测量在标准ELISA试验中的抗纯化参比多肽的抗体应答(采用针对变异体多肽产生的多克隆血清)，测试免疫原性。最好是，变异体将保留所有的上述生物活性。
“同一性”是核苷酸序列或氨基酸序列的一致性的度量。一般而言，将所述序列进行序列对比，以便获得最高有序匹配。“同一性”自身具有本领域已知的含义并可以采用已公布的技术进行计算。参见，例如(COMPUTATIONAL MOLECULAR BIOLOGY，Lesk，A.M.编著，Oxford University Press，New York，1988；BIOCOMPUTINGINFORMATICS AND GENOME PROJECTS，Smith，D.W.编著，Academic Press，New York，1993；COMPUTER ANALYSIS OFSEQUENCE DATA，PART I，Griffin，A.M.和Griffin，H.G.编著，HumanaPress，New Jersey，1994；SEQUENCE ANALYSIS IN MOLECULARBIOLOGY，von Heijne，G.，Academic Press，1987；和SEQUENCEANALYSIS PRIMER，Gribskov，M.和Devereux，J.编著，M StocktonPress，New York，1991)。虽然存在许多测定两种多核苷酸或多肽序列之间同一性的方法，但术语“同一性”是本领域技术人员熟知的(Carillo，H和Lipton，D.，SIAMJ Applied Math(1988)481073)。测定两种序列之间同一性或相似性的常用方法包括但不限于公开于Guide to HugeComputers，Martin J.Bishop编著，Academic Press，San Diego，1994以及Carillo，H.和Lipton，D.，SIAMJ Applied Math(1988)481073中的那些方法。测定同一性和相似性的方法编纂在计算机程序中。测定两种序列之间同一性和相似性的优选计算机程序方法包括但不限于GCG程序包(Devereux，J.等，Nucleic Acids Research(1984)12(1)387)、BLASTP、BLASTN、FASTA(Atschul，S.F.等，J Molec Biol(1990)215403)。最优选的是，用于测定同一性水平的程序是GCG 9软件包，如下文实施例中所用的。
作为说明，所谓与参比核苷酸序列具有至少例如95％“同一性”的核苷酸序列的多核苷酸，是指所述多核苷酸的核苷酸序列与所述参比序列相同，只是所述多核苷酸序列可以包括平均至多5个点突变/参比核苷酸序列的每100个核苷酸。换句话说，为了获得核苷酸序列与参比核苷酸序列至少95％相同的多核苷酸，在参比序列中可以缺失或用另一核苷酸取代高达5％的核苷酸，或可以将参比序列中总核苷酸的至多5％的多个核苷酸插入参比序列中。参比序列的这些突变可以发生在参比核苷酸序列的5’或3’端位置或者在那些末端位置之间的任何地方，或者单个散布在参比序列中的核苷酸中或者在参比序列中的一个或更多个连续组内。本发明的多肽一方面，本发明涉及博德特氏菌属致病性蛋白(或多肽)。所述博德特氏菌属致病性多肽包括由表2和表3中限定的基因编码的多肽；以及包含由表2和表3中限定的基因编码的氨基酸序列的多肽；和包含在其全长范围内内与由表2和表3中限定的基因编码的氨基酸序列至少有75％同一性的氨基酸序列的多肽，优选至少80％同一性，更优选至少90％同一性。高度优选具有95-99％同一性的那些多肽。
博德特氏菌属致病性多肽(或其片段)可以是“成熟”蛋白的形式或可以是较大蛋白例如融合蛋白的一部分。最好是包括含有分泌序列或前导序列的另一氨基酸序列、前序列、有助于纯化的序列例如多个组氨基残基或麦芽糖结合蛋白(MBP)、或用于重组产生期间稳定性的另一序列。此外，也考虑添加外源多肽或脂尾或多核苷酸序列以增加最终分子的免疫原性潜能。
本发明中还包括博德特氏菌属致病性多肽的片段。一种片段是氨基酸序列与前述博德特氏菌属致病性多肽的氨基酸序列部分相同而不是全部相同的多肽。与博德特氏菌属致病性多肽一样，片段可以是“独立的”或包含在较大多肽内，它们形成所述较大多肽的一个部分或区域，最优选作为单一连续区。本发明多肽片段的代表性实例包括例如，来自博德特氏菌属致病性多肽末端的大约氨基酸号1-20、21-40、41-60、61-80、81-100和101的片段。在本文中“大约”包括在任一端或两端比所述具体列举范围增加或减少数个、5、4、3、2或1个氨基酸。所述片段应根据特定序列包含来自所述序列的至少7个连续氨基酸，例如8、10、12、14、18、20个或更多个氨基酸。最好是所述片段包含一个来自所述序列的表位。
优选片段包括例如具有博德特氏菌属致病性多肽的氨基酸序列的截短多肽，只是缺失一系列连续的包括氨基端的残基，或缺失一系列连续的包括羧基端和/或跨膜区的残基，或缺失两个连续系列的残基，一个包括氨基端而另一个包括羧基端。也优选的是特征为结构或功能标志的片段，例如包含α螺旋和α螺旋形成区、β折叠和β折叠形成区、转角和转角形成区、螺旋和螺旋形成区、亲水区、疏水区、α两亲性区、β两亲性区、柔性区、表面形成区、底物结合区和高抗原性指数区的片段。其它优选片段是生物活性片段。生物活性片段是介导博德特氏菌属致病性蛋白活性的那些片段，包括具有相似活性或活性提高或不需要的活性降低的那些片段。还包括在动物尤其是在人类中具有抗原性或免疫原性的那些片段。
最好是，所有这些多肽片段保留包括抗原活性在内的博德特氏菌属致病性蛋白的生物活性(例如抗原性或免疫原性)。所确定的序列和片段的变异体也构成本发明的一部分。优选变异体是通过保守氨基酸取代，即一个残基用另一特征相似的残基取代的那些取代，与讨论的变异体不同的那些变异体。通常这样的取代是在Ala、Val、Leu和Ile中；在Ser和Thr中；在酸性残基Asp和Glu中；在Asn和Gln中；以及在碱性残基Lys和Arg中；或芳族残基Phe和Tyr中。特别优选的是其中数个、5-10、1-5或1-2个氨基酸以任何组合被取代、缺失或添加的变异体。最优选的变异体是存在于百日咳博德特氏菌菌株中的博德特氏菌属致病性多肽的天然存在的等位基因变异体。
可以将蛋白化学缀合，或作为重组融合蛋白表达，使其与非融合蛋白相比，在表达系统中产生的水平增加。融合配偶体可以有助于提供T辅助表位(免疫融合配偶体)，最好是被人类识别的T辅助表位，或有助于与天然重组蛋白相比以更高得率表达所述蛋白(表达增强子)。最好是所述融合配偶体将既是免疫融合配偶体又是表达增强配偶体。
可以以任何合适方式制备本发明博德特氏菌属致病性多肽。这类多肽包括分离的天然存在的多肽、重组产生的多肽、合成产生的多肽或通过这些方法的组合产生的多肽。这类多肽的制备方法是本领域众所周知的。
最优选本发明的多肽衍生自百日咳博德特氏菌，然而，它也可以最好是得自同一分类属的其它生物。本发明的一种多肽也可以得自例如同一分类科或目的生物，例如副百日咳博德特氏菌(Bordetellaparapertussis)或支气管炎博德特氏菌(Bordetella bronchiseptica)。
本发明的再一方面是基本纯化的本发明博德特氏菌属致病性多肽。当用于蛋白或肽的范围时“基本纯化的”是指所述分子已是大部分但不必完全从其它细胞和非细胞组分分离和纯化。如果蛋白至少约60％(重量)无其它天然存在的有机分子，则其通常为基本纯的。优选纯度是至少约75％，更优选至少约90％，最优选至少约99％(重量)纯的。本发明的多核苷酸本发明的另一方面涉及博德特氏菌属致病性多核苷酸。博德特氏菌属致病性多核苷酸包括分别编码博德特氏菌属致病性多肽和片段的分离的多核苷酸、以及与其密切相关的多核苷酸或其变异体。更具体地说，本发明博德特氏菌属致病性多核苷酸包括包含表2或表3中限定的基因的核苷酸序列、编码一种博德特氏菌属致病性多肽的多核苷酸。博德特氏菌属致病性多核苷酸还包括其包含的核苷酸序列与编码由表2和表3中限定的基因编码博德特氏菌属致病性多肽的核苷酸序列在其全长范围内具有至少75％同一性的多核苷酸，和包含与表2和表3中限定的基因的核苷酸序列至少75％相同的核苷酸序列的多核苷酸。在这方面，特别优选多核苷酸至少80％相同，尤其优选具有至少90％相同的那些多核苷酸。此外，高度优选具有至少95％的那些多核苷酸，最高度优选具有至少98％-99％的那些多核苷酸，最优选是具有至少99％的那些多核苷酸。博德特氏菌属致病性多核苷酸还包括与表2和表3中限定的基因的核苷酸序列具有足够同一性的核苷酸序列，所述核苷酸序列在可用来扩增、或用作探针或标记的条件下杂交。本发明还提供与这类博德特氏菌属致病性多核苷酸互补的多核苷酸。
采用本文提供的信息，例如具体的博德特氏菌属致病性基因和多肽序列，采用标准的克隆和筛选方法，例如，采用百日咳博德特氏菌细胞作为原始材料，从细菌中克隆和测序染色体DNA片段，然后获得全长克隆的那些方法，可以获得编码博德特氏菌属致病性多肽的本发明的多核苷酸。例如，为了获得本发明的多核苷酸序列，通常用衍生自部分序列的放射标记的寡核苷酸(最好是17-mer或更长)探测大肠杆菌或某些其它合适宿主中的百日咳博德特氏菌的染色体DNA克隆文库。然后采用严格杂交条件，可以鉴别携带与所述探针的DNA相同的DNA的克隆。通过与根据原始多肽或多核苷酸序列设计的测序引物杂交，对如此鉴定的单个克隆测序，则可能在两个方向延伸所述多核苷酸序列，以确定全长基因序列。方便的是，例如采用由质粒克隆制备的变性双链DNA进行这样的测序。合适的技术由Maniatis，T.，Fritsch，E.F.和Sambrook等，MOLECULAR CLONING，A LABORATORYMANUAL，第二版；Cold Spring Harbor Laboratory Press，Cold SpringHarbor，New York(1989)描述。(具体来说，参见，通过杂交筛选(Screening By Hybridization)1.90和变性双链DNA模板的测序(Sequencing Denatured Double-Stranded DNA Templates)13.70)。为了获得全长基因序列，也可以进行直接基因组DNA测序。
通过包括以下步骤的方法，可以获得编码本发明多肽的多核苷酸，包括来自非百日咳博德特氏菌菌种的同系物和直向同源物，所述步骤为在严格杂交条件(例如，采用的温度范围为45-65℃和SDS浓度为0.1-1％)下，用包含表2或表3中限定的序列或其片段或由所述序列或片段组成的标记探针或可检测探针，筛选合适的文库；并且分离含有所述多核苷酸序列的全长基因和/或基因组克隆。
本发明还提供包含通过下述方法获得的多核苷酸序列或由所述多苷酸序列组成的多核苷酸在严格杂交条件下，用具有表2或表3中限定的所述多核苷酸序列或其片段的序列的探针，筛选含有表2和表3中限定的多核苷酸序列的完整基因的合适文库；然后分离所述多核苷酸序列。可用于获得这种多核苷酸的片段包括例如本文其它部分描述的探针和引物。
编码由表2和表3中限定的基因编码的博德特氏菌属致病性多肽的核苷酸序列可以与在表2或表3中限定的基因含有的多肽编码序列相同，或它可是由于遗传密码的丰余性(简并性)也编码分别由表2和表3中限定的基因编码的多肽的序列。
如果将本发明的多核苷酸用于重组产生博德特氏菌属致病性多肽，那么所述多核苷酸可以包括成熟多肽或其片段的编码序列本身；符合读框地具有其它编码序列诸如编码前导序列或分泌序列、前蛋白序列、或原蛋白序列或前原蛋白序列、或其它融合肽部分的那些序列的所述成熟多肽或片段的编码序列。例如，可以编码有助于纯化融合多肽的标记序列。在本发明该方面的某些优选实施方案中，所述标记序列是一种六组氨酸肽(如在pQE载体(Qiagen，Inc.)中提供的并描述于Gentz等，Proc Natl Acad Sci USA(1989)86821-824中)，或是HA标记，或是谷胱甘肽-s-转移酶，或是MBP。所述多核苷酸还可以含有非编码5’序列和3’序列，诸如转录非翻译序列、剪接信号和聚腺苷酸化信号、核糖体结合位点和稳定mRNA的序列。
也提供包含本发明序列的片段的核酸。这些应包含来自所述序列的至少10个连续核苷酸(取决于特定的序列，例如12、14、15、18、20、25、30、35、40或更多个)。这类片段最好可以与上述序列在严格条件下杂交。
再优选的实施方案是所编码的博德特氏菌属致病性蛋白变异体包含分别由表2和表3限定的基因编码的博德特氏菌属致病性多肽的氨基酸序列的多核苷酸，其中以任何组合形式，取代、缺失或添加数个、10-25、5-10、1-5、1-3、1-2或1个氨基酸残基。最优选的变异多核苷酸是天然存在的编码博德特氏菌属菌株(最好是百日咳博德特氏菌)中的博德特氏菌属致病性蛋白的等位基因变异体的那些百日咳博德特氏菌序列。
本发明还涉及与本文上述序列杂交的多核苷酸。在这方面，本发明尤其涉及在严格条件下与本文上述多核苷酸杂交的多核苷酸。本文所用的术语“严格条件”是指只有在所述序列之间有至少80％、优选至少90％、更优选至少95％、再甚至更优选97-99％同一性时才会发生的杂交。
与表2和表3中限定的任何基因的核苷酸序列或其片段相同或足够相同的本发明多核苷酸，可以用作cDNA和基因组DNA的杂交探针，以分别分离编码博德特氏菌属致病性多肽的全长cDNA和基因组克隆，并分离与所述博德特氏菌属致病性基因具有高度序列相似性的其它基因(包括编码来自非百日咳博德特氏菌的菌种的同系物和直向同源物的基因在内)的cDNA和基因组克隆。这样的杂交技术是本领域技术人员已知的。通常，这些核苷酸序列与本文讨论的序列80％相同，优选90％相同，更优选95％相同。所述探针一般会包含至少15个核苷酸。最好是，这类探针会具有至少30个核苷酸，可以具有至少50个核苷酸。特别优选的探针的范围将在30-50个核苷酸之间。在一个实施方案中，为了获得包括编码来自非百日咳博德特氏菌的菌种的同系物和直向同源物在内的编码博德特氏菌属致病性多肽的多核苷酸，包括以下步骤在严格杂交条件下，用具有在表2和表3中限定的基因序列之一含有的核苷酸序列或其片段的标记探针，筛选合适的文库；然后分离含有所述多核苷酸序列的全长cDNA和基因组克隆。因此，另一方面，本发明的博德特氏菌属致病性多核苷酸还包括包含在严格条件下与具有表2和表3中限定的基因之一中含有的核苷酸序列或其片段的核苷酸序列杂交的核苷酸序列的核苷酸序列。博德特氏菌属致病性多肽也包括包含由通过上述杂交条件获得的核苷酸序列编码的氨基酸序列的多肽。这样的杂交技术是本领域技术人员众所周知的。严格杂交条件是上文所限定的条件，或者是这样的的条件在包含50％甲酰胺、5xSSC(150mM NaCl、15mM柠檬酸三钠)、50mM磷酸钠(pH7.6)、5x Denhardt溶液、10％硫酸葡聚糖和20μg/ml变性剪切的鲑精DNA的溶液中于42℃温育过夜，然后于大约65℃在0.1xSSC中洗涤滤膜。
采用表2或表3中限定的DNA序列合成寡核苷酸探针，可以通过筛选分离博德特氏菌属致病性基因的编码区。然后用具有与本发明基因的序列互补的序列的标记寡核苷酸，筛选cDNA、基因组DNA或mRNA的文库，以测定所述探针与哪个文库成员杂交。
有数种可利用并且是本领域技术人员众所周知的方法获得全长DNA或延伸短DNA，例如根据快速扩增cDNA末端(RACE)的方法(参见，例如，Frohman等，PNAS USA 858998-9002，1988)的那些方法。所述技术的最新改进，例如由MarathonTM例举的技术(ClontechLaboratories Inc.)，例如已经显著简化了更长cDNA的搜索。在MarathonTM技术中，已经由从选定组织提取的mRNA制备cDNA，并将“连接物”序列连接至每个末端。然后采用基因特异性寡核苷酸引物和连接物特异性寡核苷酸引物的组合，进行核酸扩增(PCR)，以扩增所述DNA的“缺少的”5’末端。然后采用“嵌套”引物，即设计在扩增产物内退火的引物(通常在连接序列中更远的3’退火的连接特异性引物和在选定的基因序列中更远的5’退火的基因特异性引物)，重复进行所述PCR反应。然后可以通过DNA测序分析该反应的产物，并或者通过将所述产物直接连接至现有DNA以得到完整序列，或者采用所述5’引物设计的新序列信息进行独立的全长PCR，从而构建全长DNA。
在本文所述方法，但最好是PCR中，可以用衍生自表2或表3中限定序列的本发明多核苷酸，测定此中鉴定的多核苷酸是否在感染组织的细菌中全部或部分被转录。认为这类序列也将在所述病原体已经达到的感染阶段和感染类型的诊断方面有效用。
可以采用本发明多核苷酸和多肽作为发现治疗和诊断动物和人类疾病的研究试剂和材料。诊断分析本发明还涉及应用博德特氏菌属致病性多肽或博德特氏菌属致病性多核苷酸作为诊断试剂。博德特氏菌属致病性多肽的检测尤其将提供可以增加或确定百日咳博德特氏菌疾病诊断的一种诊断工具。
诊断材料可以得自受治疗者的细胞，诸如来自血液、尿、唾液、组织活的细胞。
因此，另一方面，本发明涉及疾病或疾病易感性特别是百日咳博德特氏菌疾病的诊断试剂盒，所述试剂盒包括(a)一种博德特氏菌属致病性多核苷酸或其片段，所述多核苷酸最好是由表2和表3限定的基因序列之一的核苷酸序列；(b)一种与(a)的核苷酸序列互补的核苷酸序列；(c)一种博德特氏菌属致病性多肽或其片段，所述多肽最好是由表2和表3中限定的基因序列之一编码的多肽；(d)一种抗博德特氏菌属致病性多肽的抗体，最好是抗由表2和表3中限定的基因序列之一编码的多肽的抗体；或(e)一种呈现抗博德特氏菌属致病性多肽的抗体的噬菌体，所述多肽最好是由表2和表3中限定的基因序列之一编码的多肽。
人们会认识到在任何这样的试剂盒中，(a)、(b)、(c)、(d)或(e)可以包含一种基本组分。
用于预测、诊断或其它分析的多肽和多核苷酸可以得自推断受感染和/或受感染的个体材料。来自任何这些来源的多核苷酸，尤其是DNA或RNA可以直接用于检测，或可以通过采用PCR或任何其它扩增技术进行酶促扩增，然后进行分析。也可以以相同的方式使用RNA特别是mRNA、cDNA和基因组DNA。采用扩增法，对存在于个体的感染性生物或定居生物的菌种和菌株的鉴定，可以通过分析所述生物选定多核苷酸的基因型来进行。可通过将扩增产物的大小与选自相关生物的参比序列的基因型相比的变化，检测缺失和插入，所述生物最好是相同属的不同种或相同种的不同株系。通过将扩增的DNA与标记的博德特氏菌属致病性多核苷酸序列杂交，可以鉴定点突变。通过对于DNA或RNA分别用DNA酶或RNA酶消化，，或通过检测在解链温度或复性动力学方面的差异，可以将完全匹配或大部分匹配的序列与缺陷或更明显错配的双链体区分开。也可通过多核苷酸片段在凝胶中的电泳移动率与参比序列比较的变化，检测多核苷酸序列的差异。这可以用或不用变性剂进行。也可以通过直接DNA或RNA测序检测多核苷酸差异。参见，例如，Myers等，Science，2301242(1985)。在特定位置上的序列变异可以通过核酸酶保护测定，例如RNA酶、V1和S1保护测定或化学切割法来揭示。参见，例如，Cotton等，Proc.Natl.Acad.Sci.，USA，854397-4401(1985)。
本发明还涉及应用本发明多核苷酸作为诊断试剂。与疾病或致病性相关的突变形式的本发明多核苷酸的检测，将提供一种诊断工具，该诊断工具可以增加或确定疾病的诊断、病程的预测、疾病阶段的确定、或对疾病的易感性，这些起因于所述多核苷酸的表达不足、超量表达或表达变化。在多核苷酸水平上通过多种技术例如本文其它地方描述的那些技术，可以检测携带这种多核苷酸中突变的生物、尤其是感染性生物。
本发明还提供用于诊断疾病，优选细菌(特别是博德特氏菌属)感染，更优选由百日咳博德特氏菌引起的感染的方法，所述方法包括测定得自个体例如机体材料的样品中，具有表2或表3中限定的序列的多核苷酸表达水平增加。采用任一种本领域熟知的、定量多核苷酸的方法，例如，扩增法、PCR、RT-PCR、RNA酶保护法、RNA印迹法、光谱测定和其它杂交方法，可以测定多核苷酸表达的增加或减少。载体、宿主细胞、表达系统本发明还涉及包含本发明的一种多核苷酸或多种多核苷酸的载体、用本发明的载体基因工程改造的宿主细胞和通过重组技术产生本发明的多肽。采用得自本发明DNA构建体的RNA，也可以用无细胞翻译系统产生这类蛋白。
可以通过本领域技术人员熟知的方法，从包含表达系统的基因工程宿主细胞，制备本发明的重组多肽。因此，再一方面，本发明涉及包含本发明的一种多核苷酸或多种多核苷酸的表达系统，涉及用这样的表达系统基因工程改造的宿主细胞，还涉及通过重组技术产生本发明的多肽。
对于重组产生本发明多肽，可以基因工程改造宿主细胞以掺入表达系统或其部分或本发明的多核苷酸。可以通过许多标准实验手册中描述的方法，实现将多核苷酸导入宿主细胞，例如，Davis等，BASICMETHODS IN MOLECULAR BIOLOGy，(1986)和Sambrook等，MOLECULAR CLONINGA LABORATORY MANUAL，第二版，ColdSpring Harbor Laboratory Press，Cold Spring Harbor，N.Y.(1989)，诸如，磷酸钙转染、DEAE-葡聚糖介导的转染、转位、微注射、阳离子脂质介导的转染、电穿孔、转导、擦伤加样(scrape loading)、基因枪引入(ballistic introduction)和感染。
合适宿主的代表性实例包括细菌细胞，例如以下细菌的细胞链球菌、葡萄球菌、肠球菌、大肠杆菌、链霉菌、蓝细菌、枯草芽孢杆菌、粘膜炎莫拉氏菌属(Moraxella catarrhalis)、流感嗜血菌(Haemophilus influenzae)和脑膜炎奈瑟氏球菌(Neisseria meningitidis)；真菌细胞，例如以下真菌的细胞酵母、克鲁维酵母属(Kluveromyces)、酵母属(Saccharomyces)、担子菌、白假丝酵母(Candida albicans)和曲霉属(Aspergillus)；昆虫细胞例如果蝇属(Drosophila)S2的细胞和贪夜蛾属(Spodoptera)Sf9的细胞；动物细胞例如CHO、COS、HeLa、C127、3T3、BHK、293、CV-1和Bowes黑素瘤细胞；和植物细胞例如裸子植物的细胞或被子植物的细胞。
可以用各种各样的表达系统产生本发明的多肽。这样的载体其中包括染色体、附加体和病毒衍生的载体，例如，衍生自以下的载体细菌质粒、噬菌体、转座子、酵母附加体、插入元件、酵母染色体元件；衍生自以下病毒的载体杆状病毒、乳多空病毒、诸如SV40、痘苗病毒、腺病毒、禽痘病毒、假狂犬病病毒、微小RNA病毒、反转录病毒和甲病毒属以及衍生自其组合的载体，例如衍生自质粒和噬菌体遗传元件的那些载体，诸如粘粒和噬菌粒。所述表达系统构建体可以含有调节以及产生表达的控制区。一般而言，在宿主中适合于保持、增殖或表达多核苷酸和/或适合于表达多肽的任何系统或载体都可以用来在这方面进行表达。通过多种众所周知的常规技术的任一种(例如陈述于Sambrook等，MOLECULAR CLONING，A LABORATORYMANUAL，(参见上文)的技术)，可以将合适的DNA序列插入所述表达系统中。
在真核生物的重组表达系统中，为了将翻译的蛋白分泌至内质网腔、分泌至壁膜间隙或分泌至胞外环境中，可以将合适的分泌信号掺入表达的多肽中。这些信号对所述多肽而言可以是内源的，或者它们可以是异源信号。
通过众所周知的方法，可以从重组细胞培养物中回收并纯化本发明多肽，所述方法包括硫酸铵或乙醇沉淀、酸提取、阴离子或阳离子交换层析、磷酸纤维素层析、疏水作用层析、亲和层析、羟基磷灰石层析和凝集素层析。最优选为用离子金属亲和层析(IMAC)进行纯化。如果所述多肽在胞内合成、分离和/或纯化期间变性，则可以用熟知的重折叠蛋白的技术再产生活性构象。
表达系统也可以是重组活的微生物，例如病毒或细菌。可以将目的基因插入活的重组病毒或细菌的基因组中。用该活的载体接种和体内感染会导致体内表达抗原并诱发免疫应答。用于该目的的病毒和细菌例如是痘病毒(例如，痘苗病毒、禽痘病毒、金丝雀痘病毒)、甲病毒(新培斯病毒、西门利克森林甲病毒、委内瑞拉马脑炎病毒)、腺病毒、腺伴随病毒、微小RNA病毒(脊髓灰质炎病毒、鼻病毒)、疱疹病毒(水痘-带状疱疹病毒等)、李斯特氏菌属(Listeria)、沙门氏菌属、志贺氏菌属、奈瑟氏球菌属、BCG。这些病毒和细菌可以是毒性的或是以各种方式减毒的，以便获得活疫苗。这样的活疫苗也构成本发明的一部分。抗体根据再一方面，本发明提供与本发明多肽特异性结合的抗体。这些抗体可以是多克隆抗体或是单克隆抗体，并且可以通过本领域技术人员熟知的任何合适的方法产生。
通常，将小鼠或大鼠用蛋白免疫(最好用弗氏完全佐剂辅助)和注射(足够的剂量通常为50-200μg/注射)。通过将所述动物放血以提取血清，可以分离多克隆抗体。或者，通过取出脾脏(或大淋巴结)并将其离解成单细胞，可以产生单克隆抗体(Kohler和Milstein，(1975)Nature，256495-497)。然后诱导这些细胞与骨髓瘤细胞一起融合形成杂交瘤，并将其在选择性培养基(例如次黄嘌呤、氨基蝶呤、胸苷培养基，“HAT”)中培养。将产生的杂交瘤通过有限稀释进行接种，并分析与所述免疫抗原特异性结合(并且不与不相关抗原结合)的抗体的产生。然后将选定的分泌单克隆的杂交瘤或者在体外(例如在组织培养瓶或中空纤维反应器)或者在体内(如在小鼠的腹水中)培养。
单链抗体的产生技术(美国专利第4,946,778号)可以适合于产生抗本发明多肽或多核苷酸的单链抗体。此外，可以用转基因的小鼠、或其它生物或动物例如其它哺乳动物表达抗本发明多肽或多核苷酸的免疫专一性人源化抗体。
另一方面，可以应用噬菌体展示技术，或者从来自经筛选具有抗博德特氏菌属致病性多肽的人类淋巴细胞的PCR扩增的v-基因的所有组成成分，或者从首次用于实验的文库(naive libraries)，选择具有与本发明多肽结合活性的抗体基因(McCafferty等，(1990)，Nature 348，552-554；Marks等，(1992)Biotechnology 10，779-783)。也可以通过例如链改组改善这些抗体的亲和性(Clackson等，(1991)Nature 352628)。
可以利用上述抗体分离或鉴定表达本发明多肽或多核苷酸的克隆，以通过例如亲和层析纯化所述多肽或多核苷酸。
可以利用抗博德特氏菌属致病性多肽或多核苷酸的抗体治疗感染、特别是细菌感染。
多肽变异体包括抗原性、表位或免疫学方面的等效变异体，构成本发明的一个特别的方面。
最好是，将所述抗体或其变异体修饰，使其在个体中较少免疫原性。例如，如果所述个体是人，则抗体可以最优选经“人源化”，其中已经将得自杂交瘤的抗体的一个或多个互补决定区或移植至人单克隆抗体中，例如在Jones等(1986)，Nature 321，522-525或Tempest等，(1991)Biotechnology 9，266-273中描述的。疫苗本发明的另一方面涉及在哺乳动物中诱发免疫应答的方法，所述方法包括用足以产生抗体和/或T细胞免疫应答的博德特氏菌属致病性多肽或表位携带片断、类似物、外膜小泡或细胞(减毒的或非减毒的)接种哺乳动物，以保护所述动物尤其免患博德特氏菌属(特别是百日咳博德特氏菌)疾病。这类药物可以单独使用或与另一改善其免疫能的分子结合。具体来说，本发明涉及由表3中限定的基因编码的博德特氏菌属致病性多肽—效应蛋白的应用。本发明的再一方面涉及在哺乳动物中诱发免疫应答的方法，所述方法包括通过在体内指导表达博德特氏菌属致病性多核苷酸的载体传递博德特氏菌属致病性多肽，以便诱发这样的免疫应答，以产生保护所述动物免患疾病的抗体。
本发明的再一方面涉及免疫组合物或疫苗制剂，当将所述免疫组合物或疫苗制剂导入哺乳动物宿主后，在该哺乳动物中诱发对博德特氏菌属致病性多肽(特别是由表3中限定的基因编码的多肽)的免疫应答，其中所述组合物包含博德特氏菌属致病性基因、或博德特氏菌属致病性多肽或表位携带片段、类似物、外膜小泡或细胞(减毒的或非减毒的)。所述疫苗制剂还可以包含一种合适的载体。最好是口服或胃肠外(包括皮下、肌内、静脉内、皮内等注射)给予博德特氏菌属致病性多肽疫苗组合物。适合于胃肠外给药的制剂包括无菌注射水溶液或非水溶液，所述溶液可以含有抗氧化剂、缓冲剂、抑菌剂和赋予所述制剂与受体血液等渗的溶质；以及可以包括悬浮剂或增稠剂的无菌含水和非水悬浮液。所述制剂可以存在于单位剂量容器或多剂量容器中，例如，密封的安瓿和管形瓶中，并可以在冷冻干燥条件下贮藏，只需要恰好在使用之前加入无菌液体载体。所述疫苗制剂还可以包括增强所述制剂免疫原性的佐剂系统，例如本领域已知的水包油系统和其它系统。所述剂量取决于所述疫苗的比活，并可以通过常规实验法容易地确定。
本发明的疫苗制剂还可以包含其它已知是合适疫苗试剂的博德特氏菌属抗原，例如，百日咳类毒素、pertactin、凝集素1和2、FHA(丝状血细胞凝集素)、和腺苷酸环化酶/溶血素(AC/HLY)、或其免疫原性片段(Locht等，NAR(1986)143251-3261；Relman等，PNAS USA(1989)862637-2641；Roberts等，Mol.Microbiol.(1991)51393-1404；Mooi等，Microb.Pathog.(1992)12127-135；Hewlett和Gordon，载于Pathogenesis and Immunity in Pertussis(1988)，New York，Wiley &Sons，第193-209页)。
本发明的再另一方面涉及包含本发明多核苷酸的免疫制剂/疫苗制剂。这类技术是本领域已知的，参见例如Wolff等，Science，(1990)2471465-8。
疫苗组合物可以包含本发明的多肽、抗体或多核苷酸。所述药用组合物包含治疗有效量的本发明要求保护的多肽、抗体或者多核苷酸。
本文所用的术语“治疗有效量”是指治疗、改善或预防所需疾病或病症(在这种情况下是博德特氏菌属疾病，尤其是百日咳博德特氏菌疾病)，或表现出可检测治疗或预防效应的治疗药物的量。所述效应可以通过例如抗原水平进行检测。治疗效应也包括在物理体征方面的减轻，例如体温降低。用作疫苗的免疫原性组合物包含免疫有效量的抗原性多肽或免疫原性多肽。所谓“免疫有效量”，它是指或者以单剂量或作为系列的一部分给予个体以有效治疗或预防的量。
实施例采用标准技术实施以下实施例，所述标准技术是本领域技术人员熟知并且是常规技术，只是在其它地方详细描述。所述实施例说明但不限制本发明。实施例1在百日咳博德特氏菌的致病性岛中存在III型分泌系统通过聚合酶链式反应(PCR)，研究百日咳博德特氏菌基因组中的lcrD同源基因的存在。所用的引物(示于表1中的寡核苷酸95080和95081)是对应于LcrD/FlbF蛋白家族的氨基酸序列高度保守区的简并寡核苷酸。也设计这些引物有利于扩增毒力基因，以取代存在于有鞭毛细菌菌株中的其平行进化同源flhA或flbF鞭毛基因。寡核苷酸95081中3’三联体CAT的存在是一个决定子—实际上当采用已知同源序列进行多序列分析时(用或者GCG9软件包的FASTA和TFASTA程序或者用BLASTN、BLASTP和BLASTX程序进行数据库检索，以及用得自GCG9软件包的PILEUP程序进行序列对比)，可以观察到CAT三联体编码只存在于毒力序列而不存在于鞭毛序列中的甲硫氨酸。
当在琼脂糖凝胶上分析时，看来所述PCR产物作为多种片段的异质混合物，其中一种呈现预期大小(约150 bp)。采用大约150 bp DNA为模板的第二轮扩增产生一个在pCRII(得自Invitrogen)中克隆的单扩增子，将其用于进一步鉴定。它看来为152 bp片段，其核苷酸序列(图1)虽然类似于所有lcrD/flbF同源基因，但与毒力(lcrD样)基因共享较高同一性水平。表1
1采用IUB(命名委员会，1985，Eur.J.Biochem.，1501-5)提出的核苷酸双关性的字母代码。2Plano等(1991)公布了用于该研究的来自小肠结肠炎耶尔森氏菌的lcrD基因的DNA序列。
为确保所述克隆片段实际上是百日咳博德特氏菌序列，在严格条件下，用百日咳博德特氏菌DNA的10倍系列稀释液，进行PCR。最优化严格PCR条件需要模板和引物之间的完全匹配。然而，可能是由于原始引物的简并性，最初获得的152 bp序列与实际的百日咳博德特氏菌lcrD样(下文称为bcrD)序列比较在其边界具有几个碱基对差异。因此优选采用内部引物(表1的寡核苷酸95363和95364)的嵌套式PCR方法，使用已知为正确百日咳博德特氏菌序列的引物。在10倍稀释的百日咳博德特氏菌模板DNA和嵌套式PCR产物之间观察剂量-反应关系，表明152 bp扩增子确实源自博德特氏菌属基因组。
152 bp序列与lcrD/flbF基因的比较，让我们确定特定DNA序列段(表1中的寡核苷酸96110)，所述序列段用作筛选质粒载体pBR327中构建的百日咳博德特氏菌的基因组文库(Delisse-Gathoye等，1990，Infect-Immun.582895-905)的探针。分离了几个阳性克隆，其驻留(resident)质粒的限制酶切分析显示它们含有重叠插入片段。测定一个插入片段的全部核苷酸序列，揭示一个大可读框(ORF)。该2100 bp ORF编码一种75 kDa、与耶尔森氏菌蛋白LcrD和FlhA分别有59％和47％相同的多肽。包括百日咳博德特氏菌BcrD推断的氨基酸序列在内的所有已知LcrD/FlbF蛋白家族成员的多重氨基酸比较，显示该序列清楚地分类在毒力相关决定子内(图2)。这些数据强烈地表明，百日咳博德特氏菌具有III型输出系统，涉及毒力效应物的分泌。
百日咳博德特氏菌lcrD样核苷酸序列(bcrD)已提交至EMBL并指定登录号Y13383。
该基本技术已经可用于测定其它细菌菌株中是否存在III型分泌系统。采用该技术，深入细致地对人类病原体布氏疏螺旋体和幽门螺杆菌筛选这种系统。没有发现III型分泌系统的证据。随后公布的这些微生物基因组序列已经证实在这些菌种内缺乏类似系统。相比之下，所述方法使得可以扩增来自植物病原体皱纹假单胞菌(Pseudomonascorrugata)的DNA片段，所述片段清楚地分类在毒力序列之中。可以将该技术应用于医学或农学上重要的任何革兰氏阴性病原体例如奈瑟氏球菌、粘膜炎莫拉氏菌、霍乱弧菌；任何肠杆菌科，假单胞菌、流感嗜血菌、布鲁氏菌(Brucella spp)、土拉热弗朗西丝氏菌(Francisellatularensis)、巴斯德氏菌(Pasteurella spp)、嗜肺军团菌(Legionellapneumophila)。甚至在已经全部测序的菌株中，可以将该技术用作检查相同种的交替型(alternate type)或菌株的简单方法。例如，某些类型的致病性大肠杆菌含有III型分泌系统，而另一些类型的大肠杆菌却没有。实施例2分析百日咳博德特氏菌bcrD侧翼序列以鉴定其中编码的致病性岛和毒力相关蛋白致病性岛内的III型编码基因的系统成簇的倾向性促使分析百日咳博德特氏菌bcrD侧翼序列。由于注意到这样一个事实在至少两个独立克隆中必须显示每个致病性岛区，因此通过染色体步查对含致病性岛的全区测序，以避免可能由于嵌合DNA插入造成的假象。这揭示可能分为3类的成簇ORFI类ORF型(表2)；II类ORF型(表3)—具有最佳疫苗和诊断特性的效应蛋白；以及插入序列和与其它种的管家基因同源的ORF(表4)。虽然没有确定致病性岛边界的一般规律，但它们可以在一个或另一边界用同向或反向重复序列进行定界，然而边界的绝对定界可能只有真正在序列的末端通过检测管家基因来进行。在本文情况下，插入序列(图3中的IS)存在于所述岛屿的5’端(将毒力ORF与管家基因中分开)，而在3’端不存在。另外，根据序列数据，包括许多毒力序列的基因座周围的管家基因(greA和ICFG样)的存在是所述岛屿边界的一种好的指示。所述致病性岛的全部基因组构图示于图3中。PAI边界的准确确定需要进一步实验数据，例如对缺乏III型分泌系统的博德特氏菌属菌株的相应染色体区的特征鉴定。表2
表3
表4
紧接bcrD基因之后，有一个可读框(ORF)和其它已知的YscU同系物，所述可读框的推断氨基酸序列与耶尔森氏菌(Yersinia spp)的YscU蛋白共享显著相似性(39％同一性和51％相似性)(图4)。与LcrD一样，YscU也是涉及所述细菌的毒力机制的耶尔森氏菌属III型分泌器的组分。百日咳博德特氏菌因此具有一个最有可能涉及致病性的典型的III型分泌系统。这后一点可以通过突变体的表型分析来研究(参见下文)。
所述Pai的全长约为30-40kb。完整区的DNA序列示于图5中，并在表2、表3和表4中指出。在脉冲场凝胶电泳上的限制酶切分析使需作图的III型基因座定位至Tohama I菌株染色体上的坐标位置1,590kb。
在百日咳博德特氏菌II类PaiDNA序列和在GenEMBL数据库中报道的序列之间没能发现同系物(只是表3中陈述的那些除外)。这些未知基因在Pai内的表达产物引起毒力，将可用于开发抵抗病原体百日咳博德特氏菌的疫苗制剂。
为了研究所述Pai的准确功能，通过等位基因交换工程改造bcrD突变体。在所获得的突变体中，bcrD基因被提供卡那霉素抗性的aphA-3盒破坏。将该盒以不中断翻译的这样一种方式插入，避免对推断的下游顺反子表达的任何极性效应。已经将突变体分离出，目前正在对其相关表型进行分析。实施例3致病性岛基因的原位表达的分析基因构建为了产生III型分泌缺陷的突变体，从bcrD编码序列中缺失255-bp片段(密码子363-445)，然后用含有提供卡那霉素抗性的aphA-3基因的盒取代(Menard等，J.Bacteriol.(1993)1755899-5906)。aphA-3盒通过EcoRI-PstI消化从pUC18K中切下并引入bcrD EcoRI-Sse8387I位点中。这种构建成体在bcrD翻译中产生早期终止，并提供符合读框翻译突变基因的其余3’端，避免对下游顺反子表达的可能的极性效应。所述突变的bcrD基因与其侧翼序列通过BglII-NotI切割切下，随后由于DNA连接物而将其插入至自杀质粒pSS1129的XbaI-EcoRI位点(Stibitz，Methods Enzymol.(1994)235458-465)。将获得的构建体命名为pAF214。pAF218是含有另外两个唯一SpeII和PacI位点的pAF214的衍生物。包括在一对互补寡核苷酸内的这些位点，被引入至pAF214的BamHI位点。其它构建体包括pAF245和pAF246。产生覆盖bcrD 5’区和前4个密码子的831 bp片段的PCR扩增。将该扩增子再引入BamHI-HinDIII线性化的pNM480(Minton，Gene(1984)31269-273)中，其引入方式使得将bcrD起始密码子与用作报道基因的lacZ符合读框地放置。将获得的构建体命名为pAF245。同样，设计引物，以将含有包括其前3个密码子在内的上游bscN序列的849 bp片段置于lacZ下游。通过将该片段克隆在pNM480中获得pAF246。转化和等位基因交换洗涤来自10ml SS培养基中的新鲜饱和培养物的百日咳博德特氏菌细胞，将其再悬浮于100μl冷10％(v/v)甘油溶液中。将最多20μl水中的多达10μg超螺旋纯化DNA加入至100μl所述细菌悬浮液中。将细胞和DNA转移至预冷的0.2cm电穿孔小池(Bio-Rad)，然后置于基因脉冲仪(Gene Pulser apparatus)(Bio-Rad)中。用25μF、2.5kV和600Ω的设定，得到时间常数范围为11-14ms，进行脉冲处理。
根据描述的方法(Stibitz，参见上文)，在BG加上庆大霉素上的最初分离后，在链霉素上选择经过一次第二个重组步骤的pAF214和pAF248转化子。无效bcrD突变体最后通过其获得的对卡那霉素的抗性与回复体区别。通过DNA印迹分析，确定正确整合的aphA-3。相比之下，pAF245和pAF246的引入只需在BG加上氨苄青霉素上选择的一次交换。该重组步骤导致将lacZ编码序列置于分别操纵bcrD和bscN转录的信号控制之下。小鼠模型在BG琼脂平板上生长2天后，回收野生型和突变型细菌，以浓度为108PFU/ml的浓度再悬浮于PBS中。在戊巴比妥麻醉小鼠的每个鼻孔中注射25μl所述悬浮液。4小时、3天、7天、14天、26天、39天和45天后，通过将每只小鼠的两个肺在Ultraturax研磨机中处理并在BG琼脂平板上滴定(titrating)所述再悬浮的细菌，分析肺部定居(lungs colonization)。β-半乳糖苷酶分析根据先前所述(Miller，(1972)“分子遗传学实验(Experiments inmolecular genetics)”Cold Spring Harbor Laboratory，Cold Spring Harbor，N.Y.)，分析0.5ml来自生长至对数期(OD＝0.2)的液体培养物的细菌悬浮液。我们使用Sigma的显色底物对硝基苯基-β-D半乳糖苷(ONPG)。看来bcrD和bscN两种转录物的转录受bvg基因座的控制大多数博德特氏菌属毒力功能受bvg基因座的控制。Bvg+相的特征为毒力因子的表达并且是动物模型的定居所必需的。相反，所述细菌在可以被烟酸或MgSO4诱导的Bvg-相中是无毒性的。我们通过采用在这些基因中的lacZ转录融合物，研究了属于不同转录单位的两个基因即bcrD和bscN的表达水平。为此，我们分离出分别整合pAF245和pAF246的突变体NIVh86和NIVh87。在前一突变体中，一个重组步骤导致放置lacZ以代替bcrD编码序列，而在后者中，lacZ取代bscN。或者在Bvg+相中或者在Bvg-相中，评估bcrD和bscN两种转录物的表达水平。两种百日咳博德特氏菌基因均在体外弱表达。另外，然而，看来这些表达水平明显地受Bvg系统的调节。实际上，在Bvg+条件下可以测定β-半乳糖苷酶，但在Bvg-相中没有检测到酶活性(表5)。表5.当将lacZ置于指导bcrD或bscN表达的控制下时，以Miller单位(Miller，参见上文)计的β-半乳糖苷酶活性。
实施例4效应蛋白疫苗候选物的重组表达在所发现的序列中，7个ORF(orf2至-8)尤其满足某些标准，使它们成作为效应蛋白的好的候选物和疫苗候选物。首先，看来它们被典型的III型分泌(I类)基因包围，并因此不容置疑地属于III型分泌基因座。此外，它们没有表现出与其它生物的相关III型系统中存在的基因有显著相似性，因此可能是对博德特氏菌属特异性的效应蛋白。除了这些ORF外，bopN、orf9和orf10也是特别重要的疫苗候选物。尽管实际上这些序列没有达到上述第二个标准(它们与铜绿假单胞菌的popN、pcrH和pcr4具有某些相似性)，但这些产物也可能由所述特化的易位子输出。为此，挑选10个ORF，即orf2至orf10和bopN用于进一步分析。为此，设计10对引物(表6)用于扩增其相应的ORF。然后在pCR-TOPOT/A克隆系统(Invitrogen)中，将扩增的ORF克隆，并检查该序列中推断由Taq DNA聚合酶诱导的错误。通过EcoRI和BamHI(或BglII-参见表6)切割，挽回正确的插入片段，并将其转移至pMAL载体(New England Biolabs；Maina等，Gene(1988)74365-373)中，并通过EcoRI和BamHI限制酶切打开。在这些载体中，所克隆的插入片段的表达产生融合至大肠杆菌麦芽糖结合蛋白(MBP)的重组蛋白。所述融合蛋白的MBP结构域提供用于检测表达产物和将其通过亲和层析纯化的工具。
已经将4个ORF，即一方面将orf2、-4和-10，而另一方面将orf6，分别克隆至pMAL-c2E和pMAL-2E中。在300ml培养基中生长的转化细菌用IPTG(300μM)诱导，并在弗氏压碎器中裂解。通过超速离心沉淀不溶性物质并将其弃去，而将获得的上清液应用于直链淀粉树脂。通过应用麦芽糖10mM，进一步洗脱通过其MBP结构域与直链淀粉特异性结合的融合蛋白。该方法使我们回收10-50mg每种融合蛋白(图6)。通过利用博德特氏菌属多肽和MBP之间的肠激酶切割位点，可以从MBP中分离表达的博德特氏菌属产物。采用同样方法，其它ORF应是可表达的。
采用标准技术分析分泌的蛋白，以确定其功能特性和免疫学特性。首先，通过研究针对受感染患者血清中的这些蛋白的抗体的存在，将评价所述分泌蛋白的免疫原性，另外，它们作为保护性抗原的推断鉴定将基于在小鼠模型中完成的攻击实验。其次，通过分析效应蛋白的催化活性，将评价其生物学特性。例如，预期其中一种分泌的蛋白会表现出酪氨酸磷酸酶活性。最后，通过将所述蛋白微注射至真核细胞的胞质中，研究所述效应蛋白的功能。这将使我们表明推断的肌动蛋白聚合作用的抑制活性、细胞毒性或诱导编程性细胞死亡，即已经归因于由在其它种中发现的III型分泌系统分泌的效应蛋白的那些类型的活性。表6.用于扩增编码疫苗候选物的ORF的PCR引物
文献目录Allaoui，A.，Woestyn，S.，Sluiters，C.和Comelis，G.R.(1994)YscU-一种涉及Yop分泌的小肠结肠炎耶尔森氏菌内膜蛋白(YscU，AYersinia enterocolitica inner membrane protein involved in Yopsecretion.)J Bacteriol 1764534-4542。Bergman，T.，Erickson，K.，Galyov，E.，Persson，C.和Wolf-Watz，H.(1994)假结核耶尔森氏菌lcrB(yscN/U)基因簇涉及Yop分泌并显示与弗氏志贺氏菌和鼠伤寒沙门氏菌的spa基因簇高度同源(The lcrB(yscN/U)gene cluster of Yersinia pseudotuberculosis is involved inYop secretion and shows high homology to the spa gene clusters ofShigella flexneri and Salmonella typhimurium)。J Bacteriol1762619-2626。Bogdanove，A.J.，Wei，Z.-M.，Zhao，L.和Beer，S.V.，解淀粉欧文氏菌通过III型途径分泌一种Harpin并含有一种耶尔森氏菌yopN的同系物(Erwinia amylovora secretes a Harpin via a type III pathway andcontains a homolog of yopN of Yersimia spp.)。J Bacteriol1781720-1730。Fenselau，S.，Balbo，I.和Bonas，U(1992)野油菜黄单胞菌辣椒斑点病致病变种中的致病性决定子与动物的细菌病原体中涉及分泌的蛋白相关(Determinants of pathogenicity in Xanthomonas campestris pv.
vesicatoria are related to proteins involved in secretion in bacterialpathogens of animals)。Mol Plant Interact 5390-396。Gyri(1995).Mol.Microbiol.15761-769。Hueck，C.J.(1998)动物及植物的细菌病原体中III型蛋白分泌系统(Type III protein secretion systems in Bacterial Pathogens of Animalsand Plants)。Microb.Mol.Biol.Rev.62379-433Plano(1991).J.Bact.1737293-7303。Ramakrishnan，G.，Zhao，J.L.和Newton，A.(1991)新月柄杆菌细胞周期调节的鞭毛基因f1bF与鼠疫耶尔森氏菌的一个毒力基因座(lcrD)同源(The cell cycle-regulated flagellar gene flbF of Caulobactercrescentus is homologous to a virulence locus(lcrD)of Yersiniapestis)。J Bacteriol 1737283-7292。
Yuk，M.H.，Harvill，E.T.和Miller，J.F.(1998)BvgAS毒力控制系统调节支气管炎博德特氏菌中的III型分泌(The BvgAS virulencecontrol system regulates type III secretion in Bordetellabronchiseptica)。Mol.Microbiol.28945-959。CPCH0161819P 说明书的核苷酸和氨基酸序列表序列表<110>Alex BollenAlain FauconnierEdmond Godfroid<120>疫苗<130>B45168<160>82<170>FastSEQ for Windows Version 3.0<210>1<211>2100<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(2100)<400>1atg acg agc aag aaa tcc att cgc cgc ctg caa cgc gcg gtg gcg ctg48Met Thr Ser Lys Lys Ser Ile Arg Arg Leu Gln Arg Ala Val Ala Leu1 5 10 15gcc acc agc cgc aac gac atc gta ctg gcc gtg ctc atc gtg gcg atc96Ala Thr Ser Arg Asn Asp Ile Val Leu Ala Val Leu Ile Val Ala Ile20 25 30gtc ttc atg atg atc ctg ccg ttg ccc aca acg ctg gtc gac gtg ctg144Val Phe Met Met Ile Leu Pro Leu Pro Thr Thr Leu Val Asp Val Leu35 40 45atc ggt gcg aac atg acg ctg tcg gca gtc ctg ctg atg gtc gcg atg192Ile Gly Ala Asn Met Thr Leu Ser Ala Val Leu Leu Met Val Ala Met50 55 60tac ctg cct tcg ccc ctg gcg ttt tcc tcg ttc cct tcg gtc ctg ctg240Tyr Leu Pro Ser Pro Leu Ala Phe Ser Ser Phe Pro Ser Val Leu Leu65 70 75 80gtc acc acg ctg ttc cgg ctg ggc atc tcc atc gcg acc acg cgg ctg288Val Thr Thr Leu Phe Arg Leu Gly Ile Ser Ile Ala Thr Thr Arg Leu85 90 95atc ctg ctg caa ggc gat gcc ggc cac atc atc gag acc ttc ggc aac 336Ile Leu Leu Gln Gly Asp Ala Gly His Ile Ile Glu Thr Phe Gly Asn100 105 110ttc gtg gtg ggc ggc aac ctg atc gtc ggc ctg gtg gtt ttc ctc atc 384Phe Val Val Gly Gly Asn Leu Ile Val Gly Leu Val Val Phe Leu Ile115 120 125ctc acg atc gtg cag ttc gtg gtc atc acc aaa ggc gcg gag cgg gtg 432Leu Thr Ile Val Gln Phe Val Val Ile Thr Lys Gly Ala Glu Arg Val130 135 140gcc gaa gtc gcc gcg cgc ttc tcg ctg gac gcc atg ccc ggc aag cag 480Ala Glu Val Ala Ala Arg Phe Ser Leu Asp Ala Met Pro Gly Lys Gln145 150155 160atg tcc atc gac gcg gac ttg cgc gcg ggc acc ata gac atg gac gaa 528Met Ser Ile Asp Ala Asp Leu Arg Ala Gly Thr Ile Asp Met Asp Glu165 170 175gcc cga cgc cga cgc cgt acg gtc gag aag gaa agc caa ctg tat ggc 576Ala Arg Arg Arg Arg Arg Thr Val Glu Lys Glu Ser Gln Leu Tyr Gly180 185 190gcc atg gac ggc gcg atg aag ttc gtc aag ggc gat gcc atc gcc ggc 624Ala Met Asp Gly Ala Met Lys Phe Val Lys Gly Asp Ala Ile Ala Gly195 200 205ctg atc atc gtt gcc gtc aac ctg ctt ggc ggc atg ctg gtc ggc gtg 672Leu Ile Ile Val Ala Val Asn Leu Leu Gly Gly Met Leu Val Gly Val210 215 220ctg cag cgc ggc ctg agc gcc ggc gag gcc gtg cag aca tat gcc atc 720Leu Gln Arg Gly Leu Ser Ala Gly Glu Ala Val Gln Thr Tyr Ala Ile225 230 235 240ctg acc ata ggc gac ggg ctc atc gcg cag atc ccg gcg ctg ttc atc 768Leu Thr Ile Gly Asp Gly Leu Ile Ala Gln Ile Pro Ala Leu Phe Ile245 250 255gcc atc tgc gcc gga atc atc gtg acg cgg gtg cag acc ggg gat ggc 816Ala Ile Cys Ala Gly Ile Ile Val Thr Arg Val Gln Thr Gly Asp Gly260 265 270ccc tcc aac gta ggc acc gac atc ggc gca caa gtg ctg gcg cag cct 864Pro Ser Asn Val Gly Thr Asp Ile Gly Ala Gln Val Leu Ala Gln Pro275 280 285cgc gcc ctg gtc att gcc ggc gcg atc tcg gca ggc ctg ggc ctc att 912Arg Ala Leu Val Ile Ala Gly Ala Ile Ser Ala Gly Leu Gly Leu Ile290295 300ccc ggc atg ccc acg ctg gtc ttc ttc gcc ctg gcc gcc gcg gtg ggc 960Pro Gly Met Pro Thr Leu Val Phe Phe Ala Leu Ala Ala Ala Val Gly305 310 315 320acc atc ggt ttc gta ctg ctg cgc gca tcc cag cgt ccg ccc gaa ggc 1008Thr Ile Gly Phe Val Leu Leu Arg Ala Ser Gln Arg Pro Pro Glu Gly325 330 335gcc gag ccc gcg ctc gcc ggc atg gct gcc gac ggc cag ccc cgc acc 1056Ala Glu Pro Ala Leu Ala Gly Met ala Ala Asp Gly Gln Pro Arg Thr340 345 350cgc gcg ccg gcg gat ggg cag gcg gaa ttc gcc ccc acc gtc ccg ctg 1104Arg Ala Pro Ala Asp Gly Gln Ala Glu Phe Ala Pro Thr Val Pro Leu355 360 365atc atc gac gta gcc gcg cgg ctg cag ccc cgg ttc gag ccg gcc acc 1152Ile Ile Asp Val Ala Ala Arg Leu Gln Pro Arg Phe Glu Pro Ala Thr370 375 380ctc acc gac gat ctg ctg cag atc cgg cgg gcg ctc tat ttc gac ctg 1200Leu Thr Asp Asp Leu Leu Gln Ile Arg Arg Ala Leu Tyr Phe Asp Leu385 390 395 400ggc gtg ccg ttt ccc ggc atc cag ttg cgc ttc acc gaa gcg ctg gcc 1248Gly Val Pro Phe Pro Gly Ile Gln Leu Arg Phe Thr Glu Ala Leu Ala405 410 415gcc aat acc tac acc atc gtg ctg tcg gag atc ccg gtg gcg caa gga 1296Ala Asn Thr Tyr Thr Ile Val Leu Ser Glu Ile Pro Val Ala Gln Gly420 425 430atg ttg cgc gac gat gcc gtg ctg gtg cgg gac acc gag cag aac ctg 1344Met Leu Arg Asp Asp Ala Val Leu Val Arg Asp Thr Glu Gln Asn Leu435 440 445cag gcc ctg cgg atc gca tac gaa acg ggc gcg gcc ttt ctg ccc gat 1392Gln Ala Leu Arg Ile Ala Tyr Glu Thr Gly Ala Ala Phe Leu Pro Asp450455 460acg ccc acg atc tgg gtt gcg gcc agt ctg acc ggc gcc ttg cgc gat 1440Thr Pro Thr Ile Trp Val Ala Ala Ser Leu Thr Gly Ala Leu Arg Asp465 470 475 480gca ggt att cct tac ctg ggt atc agc cag atc ctg act tgg cac ttg 1488Ala Gly Ile Pro Tyr Leu Gly Ile Ser Gln Ile Leu Thr Trp His Leu485 490 495gca tat gta ttg aaa aaa tat tca gcc gat ttc atc ggc atc cag gaa 1536Ala Tyr Val Leu Lys Lys Tyr Ser Ala Asp Phe Ile Gly Ile Gln Glu
500 505 510acc cgg ttt ctg ctt tcg gcc atg gaa gaa cga ttt ccc gat ctg gtc 1584Thr Arg Phe Leu Leu Ser Ala Met Glu Glu Arg Phe Pro Asp Leu Val515 520 525aag gag tgc ctg cgc gtc atg ccg gtg cag aag att gcc gaa atc ctg 1632Lys Glu Cys Leu Arg Val Met Pro Val Gln Lys Ile Ala Glu Ile Leu530 535 540cag cgc ctt gtt tcc gaa gaa gtg tcg ata cgc aac ctg cgc gcc gtc 1680Gln Arg Leu Val Ser Glu Glu Val Ser Ile Arg Asn Leu Arg Ala Val545 550 555 560ctg gaa gcg ctg gtc gaa tgg ggc cag aag gaa aag gat acc gtc ctg 1728Leu Glu Ala Leu Val Glu Trp Gly Gln Lys Glu Lys Asp Thr Val Leu565 570 575ctt acg gag tat gtc cga atc gca ctc aag cgc tat atc agc cac aag 1776Leu Thr Glu Tyr Val Arg Ile Ala Leu Lys Arg Tyr Ile Ser His Lys580 585 590tac acc agc ggc cac aat atc ctg ccc gcc tac ctg ctg gcc ccc aag 1824Tyr Thr Ser Gly His Asn Ile Leu Pro Ala Tyr Leu Leu Ala Pro Lys595 600 605gtc gag gaa acc gtg cgc gcc gcc atc cgg cag acc gcc gcc ggc agt 1872Val Glu Glu Thr Val Arg Ala Ala Ile Arg Gln Thr Ala Ala Gly Ser610 615 620tat ctc gcc ctc gat ccg gac acg aca cgc cga ctg gtc gag cac atc 1920Tyr Leu Ala Leu Asp Pro Asp Thr Thr Arg Arg Leu Val Glu His Ile625 630 635 640cgt caa tgt gtc ggc gat ctg gcc gcc ggc gcg agc cgt ccc gtc ttg 1968Arg Gln Cys Val Gly Asp Leu Ala Ala Gly Ala Ser Arg Pro Val Leu645 650 655ctg acg tcg atg gac atc cgg cgc tac acg cgc aag atg ata gaa gcc 2016Leu Thr Ser Met Asp Ile Arg Arg Tyr Thr Arg Lys Met Ile Glu Ala660 665 670gat ctc tac gcc ctg ccg gtg ctg tcc tac cag gaa ctg acg ccg gag 2064Asp Leu Tyr Ala Leu Pro Val Leu Ser Tyr Gln Glu Leu Thr Pro Glu675 680 685atc aat gta cag ccc ctg ggc agg gtg gat cta tga 2100Ile Asn Val Gln Pro Leu Gly Arg Val Asp Leu *690 695
<210>2<211>699<212>PRT<213>百日咳博德特氏菌<400>2Met Thr Ser Lys Lys Ser Ile Arg Arg Leu Gln Arg Ala Val Ala Leu1 5 10 15Ala Thr Ser Arg Asn Asp Ile Val Leu Ala Val Leu Ile Val Ala Ile20 25 30Val Phe Met Met Ile Leu Pro Leu Pro Thr Thr Leu Val Asp Val Leu35 40 45Ile Gly Ala Asn Met Thr Leu Ser Ala Val Leu Leu Met Val Ala Met50 55 60Tyr Leu Pro Ser Pro Leu Ala Phe Ser Ser Phe Pro Ser Val Leu Leu65 70 75 80Val Thr Thr Leu Phe Arg Leu Gly Ile Ser Ile Ala Thr Thr Arg Leu85 90 95Ile Leu Leu Gln Gly Asp Ala Gly His Ile Ile Glu Thr Phe Gly Asn100 105 110Phe Val Val Gly Gly Asn Leu Ile Val Gly Leu Val Val Phe Leu Ile115 120 125Leu Thr Ile Val Gln Phe Val Val Ile Thr Lys Gly Ala Glu Arg Val130 135 140Ala Glu Val Ala Ala Arg Phe Ser Leu Asp Ala Met Pro Gly Lys Gln145 150 155 160Met Ser Ile Asp Ala Asp Leu Arg Ala Gly Thr Ile Asp Met Asp Glu165 170 175Ala Arg Arg Arg Arg Arg Thr Val Glu Lys Glu Ser Gln Leu Tyr Gly180 185 190Ala Met Asp Gly Ala Met Lys Phe Val Lys Gly Asp Ala Ile Ala Gly195 200 205Leu Ile Ile Val Ala Val Asn Leu Leu Gly Gly Met Leu Val Gly Val210 215 220Leu Gln Arg Gly Leu Ser Ala Gly Glu Ala Val Gln Thr Tyr Ala Ile225 230 235 240Leu Thr Ile Gly Asp Gly Leu Ile Ala Gln Ile Pro Ala Leu Phe Ile245 250 255Ala Ile Cys Ala Gly Ile Ile Val Thr Arg Val Gln Thr Gly Asp Gly260 265 270Pro Ser Asn Val Gly Thr Asp Ile Gly Ala Gln Val Leu Ala Gln Pro275 280 285Arg Ala Leu Val Ile Ala Gly Ala Ile Ser Ala Gly Leu Gly Leu Ile290 295 300Pro Gly Met Pro Thr Leu Val Phe Phe Ala Leu Ala Ala Ala Val Gly305 310 315 320ThrIle Gly Phe Val Leu Leu Arg Ala Ser Gln Arg Pro Pro Glu Gly325 330 335Ala Glu Pro Ala Leu Ala Gly Met ala Ala Asp Gly Gln Pro Arg Thr340 345 350Arg Ala Pro Ala Asp Gly Gln Ala Glu Phe Ala Pro Thr Val Pro Leu
355 360 365Ile Ile Asp Val Ala Ala Arg Leu Gln Pro Arg Phe Glu Pro Ala Thr370 375 380Leu Thr Asp Asp Leu Leu Gln Ile Arg Arg Ala Leu Tyr Phe Asp Leu385 390 395 400Gly Val Pro Phe Pro Gly Ile Gln Leu Arg Phe Thr Glu Ala Leu Ala405 410 415Ala Asn Thr Tyr Thr Ile Val Leu Ser Glu Ile Pro Val Ala Gln Gly420 425 430Met Leu Arg Asp Asp Ala Val Leu Val Arg Asp Thr Glu Gln Asn Leu435 440 445Gln Ala Leu Arg Ile Ala Tyr Glu Thr Gly Ala Ala Phe Leu Pro Asp450 455 460Thr Pro Thr Ile Trp Val Ala Ala Ser Leu Thr Gly Ala Leu Arg Asp465 470 475 480Ala Gly Ile Pro Tyr Leu Gly Ile Ser Gln Ile Leu Thr Trp His Leu485 490 495Ala Tyr Val Leu Lys Lys Tyr Ser Ala Asp Phe Ile Gly Ile Gln Glu500 505 510Thr Arg Phe Leu Leu Ser Ala Met Glu Glu Arg Phe Pro Asp Leu Val515 520 525Lys Glu Cys Leu Arg Val Met Pro Val Gln Lys Ile Ala Glu Ile Leu530 535 540Gln Arg Leu Val Ser Glu Glu Val Ser Ile Arg Asn Leu Arg Ala Val545 550 555 560Leu Glu Ala Leu Val Glu Trp Gly Gln Lys Glu Lys Asp Thr Val Leu565 570 575Leu Thr Glu Tyr Val Arg Ile Ala Leu Lys Arg Tyr Ile Ser His Lys580 585 590Tyr Thr Ser Gly His Asn Ile Leu Pro Ala Tyr Leu Leu Ala Pro Lys595 600 605Val Glu Glu Thr Val Arg Ala Ala Ile Arg Gln Thr Ala Ala Gly Ser610 615 620Tyr Leu Ala Leu Asp Pro Asp Thr Thr Arg Arg Leu Val Glu His Ile625 630 635 640Arg Gln Cys Val Gly Asp Leu Ala Ala Gly Ala Ser Arg Pro Val Leu645 650 655Leu Thr Ser Met Asp Ile Arg Arg Tyr Thr Arg Lys Met Ile Glu Ala660 665 670Asp Leu Tyr Ala Leu Pro Val Leu Ser Tyr Gln Glu Leu Thr Pro Glu675 680 685Ile Asn Val Gln Pro Leu Gly Arg Val Asp Leu690 695<210>3<211>486<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS
<222>(1)...(486)<400>3atg cca aag tca gcc gac cag ggc ggc tcc ccg gcg tca gct tcg cat 48Met Pro Lys Ser Ala Asp Gln Gly Gly Ser Pro Ala Ser Ala Ser His1 5 10 15gag gcg ttg cgc cat att ctc gac gca ggc gct tcg atg ggg ggc ttg 96Glu Ala Leu Arg His Ile Leu Asp Ala Gly Ala Ser Met Gly Gly Leu20 25 30cag ggg ttg gac gag gcg cag cag cag gcg ttg tac gcg atc ggt cat 144Gln Gly Leu Asp Glu Ala Gln Gln Gln Ala Leu Tyr Ala Ile Gly His35 40 45ggc gcc tac gaa cag ggg cgc tat gcc gac gcg ttg aaa atg ttc tgc 192Gly Ala Tyr Glu Gln Gly Arg Tyr Ala Asp Ala Leu Lys Met Phe Cys50 55 60ctg ctg gtc gcg tgc gat ccg ctg gaa gcc cgt tat ctg ctg gcc ctg 240Leu Leu Val Ala Cys Asp Pro Leu Glu Ala Arg Tyr Leu Leu Ala Leu65 70 75 80ggc gcc gcg gcc cag gag ctg ggg ctg tac gag cat gcc ttg cag caa 288Gly Ala Ala Ala Gln Glu Leu Gly Leu Tyr Glu His Ala Leu Gln Gln85 90 95tac gcg gcc gcg gcg gct ttg cag ttg gac tcc ccc agg ccc ctg ttg 336Tyr Ala Ala Ala Ala Ala Leu Gln Leu Asp Ser Pro Arg Pro Leu Leu100 105 110cat ggc gcc gag tgc ctg tat gcg ttg ggt cgt cgc cgc gac gcc ctg 384His Gly Ala Glu Cys Leu Tyr Ala Leu Gly Arg Arg Arg Asp Ala Leu115 120 125gat acg ctc gac atg gtg ctt gag ttg tgc ggc tcg ccg gag cgt gcg 432Asp Thr Leu Asp Met Val Leu Glu Leu Cys Gly Ser Pro Glu Arg Ala130 135 140gcc ctg cgc gaa cgg gcc gag ttg ctg cgc agg agc tat gca cgt gcc 480Ala Leu Arg Glu Arg Ala Glu Leu Leu Arg Arg Ser Tyr Ala Arg Ala145 150 155 160gac tga 486Asp *<210>4<211>161<212>PRT
<213>百日咳博德特氏菌<400>4Met Pro Lys Ser Ala Asp Gln Gly Gly Ser Pro Ala Ser Ala Ser His1 5 10 15Glu Ala Leu Arg His Ile Leu Asp Ala Gly Ala Ser Met Gly Gly Leu20 25 30Gln Gly Leu Asp Glu Ala Gln Gln Gln Ala Leu Tyr Ala Ile Gly His35 40 45Gly Ala Tyr Glu Gln Gly Arg Tyr Ala Asp Ala Leu Lys Met Phe Cys50 55 60Leu Leu Val Ala Cys Asp Pro Leu Glu Ala Arg Tyr Leu Leu Ala Leu65 70 75 80Gly Ala Ala Ala Gln Glu Leu Gly Leu Tyr Glu His Ala Leu Gln Gln85 90 95Tyr Ala Ala Ala Ala Ala Leu Gln Leu Asp Ser Pro Arg Pro Leu Leu100 105 110His Gly Ala Glu Cys Leu Tyr Ala Leu Gly Arg Arg Arg Asp Ala Leu115 120 125Asp Thr Leu Asp Met Val Leu Glu Leu Cys Gly Ser Pro Glu Arg Ala130 135 140Ala Leu Arg Glu Arg Ala Glu Leu Leu Arg Arg Ser Tyr Ala Arg Ala145 150 155 160Asp<210>5<211>1803<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1803)<400>5atg gca ata ggt cgg ctt ggg tat ctt gtc cgc ggc gca tgg gcc ggg48Met ala Ile Gly Arg Leu Gly Tyr Leu Val Arg Gly Ala Trp Ala Gly1 5 10 15ggt gtc atg ctg ttg gcg gcc ggt agc gcc tgg gcg gcg ccg aac tgg96Gly Val Met Leu Leu Ala Ala Gly Ser Ala Trp Ala Ala Pro Asn Trp20 25 30cct ttg gcg ccg tat agc tac tac gcg cag cag cag agc ctg tcc gat144Pro Leu Ala Pro Tyr Ser Tyr Tyr Ala Gln Gln Gln Ser Leu Ser Asp35 40 45gtg ctg cgc gag ttc gcc gca ggc ttc agc ctg gcg ttg caa cag ggc192Val Leu Arg Glu Phe Ala Ala Gly Phe Ser Leu Ala Leu Gln Gln Gly50 55 60aaa ggg gtg caa ggc gtg gtc aat ggg cgt ttc aat gcg cgc aca ccc 240Lys Gly Val Gln Gly Val Val Asn Gly Arg Phe Asn Ala Arg Thr Pro65 70 75 80acg gag ttc atc gag cgt ctc agc ggc atc tat ggg ttc aac tgg ttc 288Thr Glu Phe Ile Glu Arg Leu Ser Gly Ile Tyr Gly Phe Asn Trp Phe85 90 95gtg cat gcc ggc acg ctg tat gtc agc cgc acc agc gac gtg gtt acc 336Val His Ala Gly Thr Leu Tyr Val Ser Arg Thr Ser Asp Val Val Thr100 105 110cgc gcg gtg gat gca gcc ggc gct tcg ccg tcg gcg ttg cgc cag gcc 384Arg Ala Val Asp Ala Ala Gly Ala Ser Pro Ser Ala Leu Arg Gln Ala115 120 125ttg ctg caa ctg ggc atc ctg gac gaa cgc ttc gga tgg gga gag ctg 432Leu Leu Gln Leu Gly Ile Leu Asp Glu Arg Phe Gly Trp Gly Glu Leu130 135 140ccg gcg caa ggc gtg gcc atg gtg tca ggg ccg ccg gcc tat gtc gcg 480Pro Ala Gln Gly Val Ala Met Val Ser Gly Pro Pro Ala Tyr Val Ala145 150 155 160ctg gtc gag cag gcg gta gcg gcg ttg ccc aag ggg gcc ggc aat cag 528Leu Val Glu Gln Ala Val Ala Ala Leu Pro Lys Gly Ala Gly Asn Gln165 170 175cag gtg gcg gtg ttt cgc ctc aag cat gct tcc gtg agc gac cgg gtg 576Gln Val Ala Val Phe Arg Leu Lys His Ala Ser Val Ser Asp Arg Val180 185 190atc cgt tat cga gac cag cag gta gtt acg ccg ggg atg gcc acc atg 624Ile Arg Tyr Arg Asp Gln Gln Val Val Thr Pro Gly Met ala Thr Met195 200 205ctg cgc caa ttg atc ctg ggg gcg ggg ccg ggc aac gac gcg gcg ctg 672Leu Arg Gln Leu Ile Leu Gly Ala Gly Pro Gly Asn Asp Ala Ala Leu210 215 220gcc gcg gtg gcg gcg ccg ctg cgg gaa aat ccg ccg gtg ttc ggc gat 720Ala Ala Val Ala Ala Pro Leu Arg Glu Asn Pro Pro Val Phe Gly Asp225 230 235 240gcg gca gct gac ggg aac gcg ccg ctc gct ggc gca gcc cag gca gcc 768Ala Ala Ala Asp Gly Asn Ala Pro Leu Ala Gly Ala Ala Gln Ala Ala245 250 255ggc cgg cgc ctg agc gag ccc agc gtg cag gcc gac acg cgc ctc aat 816Gly Arg Arg Leu Ser Glu Pro Ser Val Gln Ala Asp Thr Arg Leu Asn
260 265 270gcc ttg atc gtg cag gat att ccc gaa cgg atg cca atc tac cgt gcc 864Ala Leu Ile Val Gln Asp Ile Pro Glu Arg Met Pro Ile Tyr Arg Ala275 280 285ctg atc gag cag ttg gat gtg ccc agc acc ctg atc gaa ata gag gcc 912Leu Ile Glu Gln Leu Asp Val Pro Ser Thr Leu Ile Glu Ile Glu Ala290 295 300atg atc gtg gac gtc aat acc gat ctg gtc aac gag ctg ggt gtc acc 960Met Ile Val Asp Val Asn Thr Asp Leu Val Asn Glu Leu Gly Val Thr305 310 315 320tgg ggg gcg cag atc gga acc acc agc ctg ggc tat ggc gat ctg ggg 1008Trp Gly Ala Gln Ile Gly Thr Thr Ser Leu Gly Tyr Gly Asp Leu Gly325 330 335ctg cgt ccc ggc aac ggc ctg ccc gtg gac ggc gcg gcg gcc gac ctg 1056Leu Arg Pro Gly Asn Gly Leu Pro Val Asp Gly Ala Ala Ala Asp Leu340 345 350gcg ccc gga acc ttg ggg atc agt gtc agt acc cgg ctg gcg gcg cgc 1104Ala Pro Gly Thr Leu Gly Ile Ser Val Ser Thr Arg Leu Ala Ala Arg355 360 365ttg cgt gcg ttg gag tcg gac ggg cag gcc aat atc ctg tct cag ccg 1152Leu Arg Ala Leu Glu Ser Asp Gly Gln Ala Asn Ile Leu Ser Gln Pro370 375 380tcc atc ctg acc gcc gac aac ctc ggc gcc atg ata gac ctg tcg gat 1200Ser Ile Leu Thr Ala Asp Asn Leu Gly Ala Met Ile Asp Leu Ser Asp385 390 395 400acc ttc tac att cgc acc ctg ggc gag cgc gta gcg aca gtc acg cct 1248Thr Phe Tyr Ile Arg Thr Leu Gly Glu Arg Val Ala Thr Val Thr Pro405 410 415gtc acg gtg ggt acg tcg ttg cgt gtg acg ccg cgc tat atc gcc gcc 1296Val Thr Val Gly Thr Ser Leu Arg Val Thr Pro Arg Tyr Ile Ala Ala420 425 430aag gga gga cgc cag gtg gaa ttg gcg atc gat atc gag gac gga cgg 1344Lys Gly Gly Arg Gln Val Glu Leu Ala Ile Asp Ile Glu Asp Gly Arg435 440 445gtc ttg cag gag tat ccc atc gat ggt ctg ccc cgg gtt cgg aaa agc 1392Val Leu Gln Glu Tyr Pro Ile Asp Gly Leu Pro Arg Val Arg Lys Ser450 455 460agc atc agc acg ctg gcg gtg gtg ggg gac gag cag acg ctg ctg atc 1440Ser Ile Ser Thr Leu Ala Val Val Gly Asp Glu Gln Thr Leu Leu Ile465 470 475 480ggc ggc tac aac aat cgc cgt gac gaa gag cag gtc gag aaa gtg ccg1488Gly Gly Tyr Asn Asn Arg Arg Asp Glu Glu Gln Val Glu Lys Val Pro485 490 495ctg ctg gga gat atc ccc ggc ctg ggg ttc ttg ttc tcg agc aag tcc1536Leu Leu Gly Asp Ile Pro Gly Leu Gly Phe Leu Phe Ser Ser Lys Ser500 505 510cgg gcg gta cag cgc cgc gag cgg ctg ttc ctg atc cgg ccg cgt gtc1584Arg Ala Val Gln Arg Arg Glu Arg Leu Phe Leu Ile Arg Pro Arg Val515 520 525gtg gct atc gag ggc aag ccg gtc ttc agc ccc gtt gcg ggc acg tcg1632Val Ala Ile Glu Gly Lys Pro Val Phe Ser Pro Val Ala Gly Thr Ser530 535 540cag gtg ttc atg agc acg ggt tgg ggc ggg cat ggc agc agc ctg agc1680Gln Val Phe Met Ser Thr Gly Trp Gly Gly His Gly Ser Ser Leu Ser545 550 555 560att gca ccc ggc gag ggc ggg cat aca caa gtg cgt cat gat gcc cgg1728Ile Ala Pro Gly Glu Gly Gly His Thr Gln Val Arg His Asp Ala Arg565 570 575gcg ggc agg ccg gtc cgg ctg gtg ccg gat tca ttg cat gtg gag tat1776Ala Gly Arg Pro Val Arg Leu Val Pro Asp Ser Leu His Val Glu Tyr580 585 590ggc gag gcg ggg gag gcg tcg ccc tga1803Gly Glu Ala Gly Glu Ala Ser Pro *595 600<210>6<211>600<212>PRT<213>百日咳博德特氏菌<400>6Met ala Ile Gly Arg Leu Gly Tyr Leu Val Arg Gly Ala Trp Ala Gly1 5 10 15Gly Val Met Leu Leu Ala Ala Gly Ser Ala Trp Ala Ala Pro Asn Trp20 25 30Pro Leu Ala Pro Tyr Ser Tyr Tyr Ala Gln Gln Gln Ser Leu Ser Asp35 40 45Val Leu Arg Glu Phe Ala Ala Gly Phe Ser Leu Ala Leu Gln Gln Gly50 55 60Lys Gly Val Gln Gly Val Val Asn Gly Arg Phe Asn Ala Arg Thr Pro65 70 75 80Thr Glu Phe Ile Glu Arg Leu Ser Gly Ile Tyr Gly Phe Asn Trp Phe85 90 95Val His Ala Gly Thr Leu Tyr Val Ser Arg Thr Ser Asp Val Val Thr100 105 110Arg Ala Val Asp Ala Ala Gly Ala Ser Pro Ser Ala Leu Arg Gln Ala115 120 125Leu Leu Gln Leu Gly Ile Leu Asp Glu Arg Phe Gly Trp Gly Glu Leu130 135 140Pro Ala Gln Gly Val Ala Met Val Ser Gly Pro Pro Ala Tyr Val Ala145 150 155 160Leu Val Glu Gln Ala Val Ala Ala Leu Pro Lys Gly Ala Gly Asn Gln165 170 175Gln Val Ala Val Phe Arg Leu Lys His Ala Ser Val Ser Asp Arg Val180 185 190Ile Arg Tyr Arg Asp Gln Gln Val Val Thr Pro Gly Met ala Thr Met195 200 205Leu Arg Gln Leu Ile Leu Gly Ala Gly Pro Gly Asn Asp Ala Ala Leu210 215 220Ala Ala Val Ala Ala Pro Leu Arg Glu Asn Pro Pro Val Phe Gly Asp225 230 235 240Ala Ala Ala Asp Gly Asn Ala Pro Leu Ala Gly Ala Ala Gln Ala Ala245 250 255Gly Arg Arg Leu Ser Glu Pro Ser Val Gln Ala Asp Thr Arg Leu Asn260 265 270Ala Leu Ile Val Gln Asp Ile Pro Glu Arg Met Pro Ile Tyr Arg Ala275 280 285Leu Ile Glu Gln Leu Asp Val Pro Ser Thr Leu Ile Glu Ile Glu Ala290 295 300Met Ile Val Asp Val Asn Thr Asp Leu Val Asn Glu Leu Gly Val Thr305 310 315 320Trp Gly Ala Gln Ile Gly Thr Thr Ser Leu Gly Tyr Gly Asp Leu Gly325 330 335Leu Arg Pro Gly Asn Gly Leu Pro Val Asp Gly Ala Ala Ala Asp Leu340 345 350Ala Pro Gly Thr Leu Gly Ile Ser Val Ser Thr Arg Leu Ala Ala Arg355 360 365Leu Arg Ala Leu Glu Ser Asp Gly Gln Ala Asn Ile Leu Ser Gln Pro370 375 380Ser Ile Leu Thr Ala Asp Asn Leu Gly Ala Met Ile Asp Leu Ser Asp385 390 395 400Thr Phe Tyr Ile Arg Thr Leu Gly Glu Arg Val Ala Thr Val Thr Pro405 410 415Val Thr Val Gly Thr Ser Leu Arg Val Thr Pro Arg Tyr Ile Ala Ala420 425 430Lys Gly Gly Arg Gln Val Glu Leu Ala Ile Asp Ile Glu Asp Gly Arg435 440 445Val Leu Gln Glu Tyr Pro Ile Asp Gly Leu Pro Arg Val Arg Lys Ser450 455 460Ser Ile Ser Thr Leu Ala Val Val Gly Asp Glu Gln Thr Leu Leu Ile465 470 475 480Gly Gly Tyr Asn Asn Arg Arg Asp Glu Glu Gln Val Glu Lys Val Pro485 490 495Leu Leu Gly Asp Ile Pro Gly Leu Gly Phe Leu Phe Ser Ser Lys Ser500 505 510Arg Ala Val Gln Arg Arg Glu Arg Leu Phe Leu Ile Arg Pro Arg Val515 520 525Val Ala Ile Glu Gly Lys Pro Val Phe Ser Pro Val Ala Gly Thr Ser530 535 540Gln Val Phe Met Ser Thr Gly Trp Gly Gly His Gly Ser Ser Leu Ser545 550 555 560Ile Ala Pro Gly Glu Gly Gly His Thr Gln Val Arg His Asp Ala Arg565 570 575Ala Gly Arg Pro Val Arg Leu Val Pro Asp Ser Leu His Val Glu Tyr580 585 590Gly Glu Ala Gly Glu Ala Ser Pro595 600<210>7<211>1281<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1281)<400>7atg acg acg gcg ctg gaa ttc cgc gtg ctt tca ggc gca cag tgc atg48Met Thr Thr Ala Leu Glu Phe Arg Val Leu Ser Gly Ala Gln Cys Met1 5 10 15gcg cgc tgc ccg gcc gtg cat ggc gcg cgc gtg ggc gcc aat ccg cat96Ala Arg Cys Pro Ala Val His Gly Ala Arg Val Gly Ala Asn Pro His20 25 30tgc gat atc gtc ctg acc ggc gag gac atg ccc gaa gtg gcg gga tgg144Cys Asp Ile Val Leu Thr Gly Glu Asp Met Pro Glu Val Ala Gly Trp35 40 45ctg gag atc gac cag tcc ggc tgg cgg ttg gcc ggc gcc gtg acg ccc192Leu Glu Ile Asp Gln Ser Gly Trp Arg Leu Ala Gly Ala Val Thr Pro50 55 60ggc ctg gac gcc cag gcg ccg tgt ccg ccc gcg gcc ttc aac gaa ccc240Gly Leu Asp Ala Gln Ala Pro Cys Pro Pro Ala Ala Phe Asn Glu Pro65 70 75 80gta gag ctg gga gcc gcc tgg atc acc gtg gcc gcc cct tcc gcg ccg288Val Glu Leu Gly Ala Ala Trp Ile Thr Val Ala Ala Pro Ser Ala Pro85 90 95tgg ccc gcg ccg ccg gag ccg tgc ggc ccg gac ggc agc gac aca gcc 336Trp Pro Ala Pro Pro Glu Pro Cys Gly Pro Asp Gly Ser Asp Thr Ala100 105 110ttg cac gac gtc cct ggc tcg aca agc ccg ccg tcc gtc gct gcc ctc 384Leu His Asp Val Pro Gly Ser Thr Ser Pro Pro Ser Val Ala Ala Leu115 120 125atg ccg cgc cga cgt gca gga cgg ccc tgg ctg gcg ctg ggc gcg gcc 432Met Pro Arg Arg Arg Ala Gly Arg Pro Trp Leu Ala Leu Gly Ala Ala130 135 140gcg gcc gtc ctg ctg gtc ggc ctg gcc acg gcg ctg gtt tcc gtg acc 480Ala Ala Val Leu Leu Val Gly Leu Ala Thr Ala Leu Val Ser Val Thr145 150 155 160aca ccc gcc acg ccg ccg gcc gcg ccg ccc cca acg ccc acc gcg ccg 528Thr Pro Ala Thr Pro Pro Ala Ala Pro Pro Pro Thr Pro Thr Ala Pro165 170 175ctg gtc cgc gcc gcg gcg ctc atc gac agc ctg ggc ctt acc gag caa 576Leu Val Arg Ala Ala Ala Leu Ile Asp Ser Leu Gly Leu Thr Glu Gln180 185 190tta caa gcg gcc tac ggc cgt ggc ggc gtg ctc acc gtg acc gga tgg 624Leu Gln Ala Ala Tyr Gly Arg Gly Gly Val Leu Thr Val Thr Gly Trp195 200 205gtg cac gac gag acg gaa ttc gct cgg gtc gcc agg gcg ttg gcg caa 672Val His Asp Glu Thr Glu Phe Ala Arg Val Ala Arg Ala Leu Ala Gln210 215 220ctt gcg cca cgg cct gcc atg cag gta agc agg cag gac gag gcc agg 720Leu Ala Pro Arg Pro Ala Met Gln Val Ser Arg Gln Asp Glu Ala Arg225 230 235 240gcc ctg gcc tgc gat gtc ctg gcg aca ttc ggg gtg cgc tac atg gcg 768Ala Leu Ala Cys Asp Val Leu Ala Thr Phe Gly Val Arg Tyr Met ala245 250 255cgc ccg tac ggc aat ggc cgc ctg gcg atc tcg ggc atc gcc agc gat 816Arg Pro Tyr Gly Asn Gly Arg Leu Ala Ile Ser Gly Ile Ala Ser Asp260 265 270gcg cac gaa cgc gcc gcg gcg ctg cat gcg gtg cgc atg cgc ctg ccg 864Ala His Glu Arg Ala Ala Ala Leu His Ala Val Arg Met Arg Leu Pro275 280 285ggc atg acg atc ctc ggt cgc gat gta cgc ctg gcc gac gag gtc tcg 912Gly Met Thr Ile Leu Gly Arg Asp Val Arg Leu Ala Asp Glu Val Ser290 295 300gcc cag ttc gcg gcc cag ctg gcc gac gaa cgc ctc gac ggc gtc aag960Ala Gln Phe Ala Ala Gln Leu Ala Asp Glu Arg Leu Asp Gly Val Lys305 310 315 320ctc agc tgg cac gcc gac cgc ctg gac gca gat ccc ggc gga ttg gcg1008Leu Ser Trp His Ala Asp Arg Leu Asp Ala Asp Pro Gly Gly Leu Ala325 330 335gca ggc cgc atg gcg cgc ctg cgc gag ctg gtg gcc gcg ttc aac cag1056Ala Gly Arg Met ala Arg Leu Arg Glu Leu Val Ala Ala Phe Asn Gln340 345 350cgc aac tac gac gtc gtc cgg ctg ccg gcc acc gcc gcg cgc gcg acg1104Arg Asn Tyr Asp Val Val Arg Leu Pro Ala Thr Ala Ala Arg Ala Thr355 360 365cgg gat cac gtg ccg ttc gag ata cgc agt gtc gtg agc ggc ccg caa1152Arg Asp His Val Pro Phe Glu Ile Arg Ser Val Val Ser Gly Pro Gln370 375 380ccg tac ctg atg ctg gcc gat ggc agc cgc ctc ctg gtg ggc gga ctg1200Pro Tyr Leu Met Leu Ala Asp Gly Ser Arg Leu Leu Val Gly Gly Leu385 390 395 400cgg gac cag tat cgc ctt acc gcc atc gaa tcc ggc cgc ctg gtc ttc1248Arg Asp Gln Tyr Arg Leu Thr Ala Ile Glu Ser Gly Arg Leu Val Phe405 410 415gat ggt ccc gaa ccg gtc atc gtg acg cga tga1281Asp Gly Pro Glu Pro Val Ile Val Thr Arg *420 425<210>8<211>426<212>PRT<213>百日咳博德特氏菌<400>8Met Thr Thr Ala Leu Glu Phe Arg Val Leu Ser Gly Ala Gln Cys Met1 5 10 15Ala Arg Cys Pro Ala Val His Gly Ala Arg Val Gly Ala Asn Pro His20 25 30Cys Asp Ile Val Leu Thr Gly Glu Asp Met Pro Glu Val Ala Gly Trp35 40 45Leu Glu Ile Asp Gln Ser Gly Trp Arg Leu Ala Gly Ala Val Thr Pro50 55 60Gly Leu Asp Ala Gln Ala Pro Cys Pro Pro Ala Ala Phe Asn Glu Pro65 70 75 80Val Glu Leu Gly Ala Ala Trp Ile Thr Val Ala Ala Pro Ser Ala Pro
85 90 95Trp Pro Ala Pro Pro Glu Pro Cys Gly Pro Asp Gly Ser Asp Thr Ala100 105 110Leu His Asp Val Pro Gly Ser Thr Ser Pro Pro Ser Val Ala Ala Leu115 120 125Met Pro Arg Arg Arg Ala Gly Arg Pro Trp Leu Ala Leu Gly Ala Ala130 135 140Ala Ala Val Leu Leu Val Gly Leu Ala Thr Ala Leu Val Ser Val Thr145 150 155 160Thr Pro Ala Thr Pro Pro Ala Ala Pro Pro Pro Thr Pro Thr Ala Pro165 170 175Leu Val Arg Ala Ala Ala Leu Ile Asp Ser Leu Gly Leu Thr Glu Gln180 185 190Leu Gln Ala Ala Tyr Gly Arg Gly Gly Val Leu Thr Val Thr Gly Trp195 200 205Val His Asp Glu Thr Glu Phe Ala Arg Val Ala Arg Ala Leu Ala Gln210 215 220Leu Ala Pro Arg Pro Ala Met Gln Val Ser Arg Gln Asp Glu Ala Arg225 230 235 240Ala Leu Ala Cys Asp Val Leu Ala Thr Phe Gly Val Arg Tyr Met ala245 250 255Arg Pro Tyr Gly Asn Gly Arg Leu Ala Ile Ser Gly Ile Ala Ser Asp260 265 270Ala His Glu Arg Ala Ala Ala Leu His Ala Val Arg Met Arg Leu Pro275 280 285Gly Met Thr Ile Leu Gly Arg Asp Val Arg Leu Ala Asp Glu Val Ser290 295 300Ala Gln Phe Ala Ala Gln Leu Ala Asp Glu Arg Leu Asp Gly Val Lys305 310 315 320Leu Ser Trp His Ala Asp Arg Leu Asp Ala Asp Pro Gly Gly Leu Ala325 330 335Ala Gly Arg Met ala Arg Leu Arg Glu Leu Val Ala Ala Phe Asn Gln340 345 350Arg Asn Tyr Asp Val Val Arg Leu Pro Ala Thr Ala Ala Arg Ala Thr355 360 365Arg Asp His Val Pro Phe Glu Ile Arg Ser Val Val Ser Gly Pro Gln370 375 380Pro Tyr Leu Met Leu Ala Asp Gly Ser Arg Leu Leu Val Gly Gly Leu385 390 395 400Arg Asp Gln Tyr Arg Leu Thr Ala Ile Glu Ser Gly Arg Leu Val Phe405 410 415Asp Gly Pro Glu Pro Val Ile Val Thr Arg420 425<210>9<211>300<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS
<222>(1)...(300)<400>9atg agt aca tct gtt cta gcc ctg acc gaa ctg gaa gtg cgc ctg gca48Met Ser Thr Ser Val Leu Ala Leu Thr Glu Leu Glu Val Arg Leu Ala1 5 10 15tcg ccg ggc ggt tcc gcc ttg cgc gac acc ttg ctg tcg cag ctt ggc96Ser Pro Gly Gly Ser Ala Leu Arg Asp Thr Leu Leu Ser Gln Leu Gly20 25 30gaa ctg gag aca cgt ttg cgc gtc cgc ctg cac gat ggc gtg ggg cgc144Glu Leu Glu Thr Arg Leu Arg Val Arg Leu His Asp Gly Val Gly Arg35 40 45gac acc tat ccc gta tgg cgc gac gcg ctg gcg gcc gcc acc gcg gcc192Asp Thr Tyr Pro Val Trp Arg Asp Ala Leu Ala Ala Ala Thr Ala Ala50 55 60cgg cag gta ttg ctg cag cgc ccg acc ggg ccg gac aac cct ccg gcg240Arg Gln Val Leu Leu Gln Arg Pro Thr Gly Pro Asp Asn Pro Pro Ala65 70 75 80tca gtc ttg acg cgc ctg agc aat gaa caa tgc gcc gaa gga gac aag288Ser Val Leu Thr Arg Leu Ser Asn Glu Gln Cys Ala Glu Gly Asp Lys85 90 95cat ggc cat taa300His Gly His *<210>10<211>99<212>PRT<213>百日咳博德特氏菌<400>10Met Ser Thr Ser Val Leu Ala Leu Thr Glu Leu Glu Val Arg Leu Ala1 5 10 15Ser Pro Gly Gly Ser Ala Leu Arg Asp Thr Leu Leu Ser Gln Leu Gly20 25 30Glu Leu Glu Thr Arg Leu Arg Val Arg Leu His Asp Gly Val Gly Arg35 40 45Asp Thr Tyr Pro Val Trp Arg Asp Ala Leu Ala Ala Ala Thr Ala Ala50 55 60Arg Gln Val Leu Leu Gln Arg Pro Thr Gly Pro Asp Asn Pro Pro Ala65 70 75 80Ser Val Leu Thr Arg Leu Ser Asn Glu Gln Cys Ala Glu Gly Asp Lys85 90 95His Gly His
<210>11<211>267<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(267)<400>11atg gcc att aac ctg gga ggc gac gca ggc cga gtg acc atg cag agc48Met ala Ile Asn Leu Gly Gly Asp Ala Gly Arg Val Thr Met Gln Ser1 5 10 15gtc aac cag gcg gtc aat acg cgg ctg aac gct cac gaa cgc gac ctg96Val Asn Gln Ala Val Asn Thr Arg Leu Asn Ala His Glu Arg Asp Leu20 25 30cgc agc cgc ctg gag gcg ctc agc gcg cgc gga gac ggc gcg gtc agc144Arg Ser Arg Leu Glu Ala Leu Ser Ala Arg Gly Asp Gly Ala Val Ser35 40 45acg tcc gac ctg ctg atc gtg caa cag gaa atg caa tcg tgg gtc gtg192Thr Ser Asp Leu Leu Ile Val Gln Gln Glu Met Gln Ser Trp Val Val50 55 60atg atc gat cta cag agc acg gtg gtc aag cag gtc gcg gat tcg ctc240Met Ile Asp Leu Gln Ser Thr Val Val Lys Gln Val Ala Asp Ser Leu65 70 75 80aag ggc gtc ata cag aag gcg agt tga267Lys Gly Val Ile Gln Lys Ala Ser *85<210>12<211>88<212>PRT<213>百日咳博德特氏菌<400>12Met ala Ile Asn Leu Gly Gly Asp Ala Gly Arg Val Thr Met Gln Ser1 5 10 15Val Asn Gln Ala Val Asn Thr Arg Leu Asn Ala His Glu Arg Asp Leu20 25 30Arg Ser Arg Leu Glu Ala Leu Ser Ala Arg Gly Asp Gly Ala Val Ser35 40 45Thr Ser Asp Leu Leu Ile Val Gln Gln Glu Met Gln Ser Trp Val Val50 55 60Met Ile Asp Leu Gln Ser Thr Val Val Lys Gln Val Ala Asp Ser Leu65 70 75 80Lys Gly Val Ile Gln Lys Ala Ser85<210>13<211>327<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(327)<400>13atg gcg gat cag gcg cgc ttt gaa ttg gcg ctg ggc gag atg ccc ggc48Met ala Asp Gln Ala Arg Phe Glu Leu Ala Leu Gly Glu Met Pro Gly1 5 10 15gca tcg gcc ccg aac ggg gcg atc gcc ctg gcg ccg gtc gcg ctc gac96Ala Ser Ala Pro Asn Gly Ala Ile Ala Leu Ala Pro Val Ala Leu Asp20 25 30gag ccg ctg ggc cgt cgc att ctt gga cag ttg cgc ggc ggc ctg gcc144Glu Pro Leu Gly Arg Arg Ile Leu Gly Gln Leu Arg Gly Gly Leu Ala35 40 45gat gtg gca gga aaa tgg cgg gcg gtg cag acg ggc ttg gcc gag gtg192Asp Val Ala Gly Lys Trp Arg Ala Val Gln Thr Gly Leu Ala Glu Val50 55 60agc cag gcg cct acc gtg gtg ggt atg ctc gat ctg cag gcc agg ttg240Ser Gln Ala Pro Thr Val Val Gly Met Leu Asp Leu Gln Ala Arg Leu65 70 75 80cta cag gca tcc gtg gag tac gag ttg gtg ggc aag gca ata ggg cgc288Leu Gln Ala Ser Val Glu Tyr Glu Leu Val Gly Lys Ala Ile Gly Arg85 90 95gcc acc caa aac gtc gat acg ctg gcg aga atg tca tga327Ala Thr Gln Asn Val Asp Thr Leu Ala Arg Met Ser *100 105<210>14<211>108<212>PRT<213>百日咳博德特氏菌<400>14Met ala Asp Gln Ala Arg Phe Glu Leu Ala Leu Gly Glu Met Pro Gly1 5 10 15Ala Ser Ala Pro Asn Gly Ala Ile Ala Leu Ala Pro Val Ala Leu Asp20 25 30Glu Pro Leu Gly Arg Arg Ile Leu Gly Gln Leu Arg Gly Gly Leu Ala35 40 45Asp Val Ala Gly Lys Trp Arg Ala Val Gln Thr Gly Leu Ala Glu Val50 55 60Ser Gln Ala Pro Thr Val Val Gly Met Leu Asp Leu Gln Ala Arg Leu65 70 75 80Leu Gln Ala Ser Val Glu Tyr Glu Leu Val Gly Lys Ala Ile Gly Arg85 90 95Ala Thr Gln Asn Val Asp Thr Leu Ala Arg Met Ser100 105<210>15<211>825<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(825)<400>15atg aac gcc atc ggg gcg atc caa cgg tat cgg cgc ggc gcg gga tgg48Met Asn Ala Ile Gly Ala Ile Gln Arg Tyr Arg Arg Gly Ala Gly Trp1 5 10 15gcg gcc ctg gtg ctc gcc ctg gcg ctg ctg gcc ggc tgc ggt gcc cgc96Ala Ala Leu Val Leu Ala Leu Ala Leu Leu Ala Gly Cys Gly Ala Arg20 25 30gtc gag ctg ttg ggc gcg gcg ccc gag aac gaa gcc aac gaa gta ttg144Val Glu Leu Leu Gly Ala Ala Pro Glu Asn Glu Ala Asn Glu Val Leu35 40 45gcg gcg ctg ctc gag gca ggc atc gct gcg cag aag cag tcc ggc aag192Ala Ala Leu Leu Glu Ala Gly Ile Ala Ala Gln Lys Gln Ser Gly Lys50 55 60gcc ggc tac gcg gtt tcg gtg ccg gcc gag gcg gtg gcc cgg tcg ctg240Ala Gly Tyr Ala Val Ser Val Pro Ala Glu Ala Val Ala Arg Ser Leu65 70 75 80gag atc ctg cgc gca agc ggc ctg ccc cgc gag cag ttc gac gga atg288Glu Ile Leu Arg Ala Ser Gly Leu Pro Arg Glu Gln Phe Asp Gly Met85 90 95gga cgc ata ttc cgc aag gaa ggc ctg gtt tca tct ccg ctc gaa gag336Gly Arg Ile Phe Arg Lys Glu Gly Leu Val Ser Ser Pro Leu Glu Glu100 105 110cgc gcc cgc tac att tat gcg ctg tct cag gaa ttg gcc gsc acc ctg384Arg Ala Arg Tyr Ile Tyr Ala Leu Ser Gln Glu Leu Ala Asp Thr Leu115 120 125tcg cag atc gac ggc gtg ctc agc gcc cgc gtg cac gtg gtg ctt ccc432Ser Gln Ile Asp Gly Val Leu Ser Ala Arg Val His Val Val Leu Pro130 135 140gaa cgc ggc gcg gtc ggc gag ccg gcc acc cct tcg acg gca ggg gtg480Glu Arg Gly Ala Val Gly Glu Pro Ala Thr Pro Ser Thr Ala Gly Val145 150 155 160ttt ctc aag tac cgc gac gga cag agc ctc gac gcg ctc gtg ccc gag528Phe Leu Lys Tyr Arg Asp Gly Gln Ser Leu Asp Ala Leu Val Pro Glu165 170 175atc cgc aag ctg gtc acg cat gcc atc ccg ggc ctg gcc gag gac cgt576Ile Arg Lys Leu Val Thr His Ala Ile Pro Gly Leu Ala Glu Asp Arg180 185 190gta tcg gtt gcc ctg gtg gtg gcc cag ccc gtt cag gcc gca ccc gcg624Val Ser Val Ala Leu Val Val Ala Gln Pro Val Gln Ala Ala Pro Ala195 200 205ccg gtc gcg tgg cgc cgc gtg ctt ggc gta cag gtc gcg gac gga tcg672Pro Val Ala Trp Arg Arg Val Leu Gly Val Gln Val Ala Asp Gly Ser210 215 220gtc ctg aga ttt tcg ctg ttg ctg ctg ttg ttg ccg gtg ctg tgc ctg720Val Leu Arg Phe Ser Leu Leu Leu Leu Leu Leu Pro Val Leu Cys Leu225 230 235 240ata gtg gcg ggg gcc gcg ctc tac gtc tgg cgc acg cgc tgg tcc cgc768Ile Val Ala Gly Ala Ala Leu Tyr Val Trp Arg Thr Arg Trp Ser Arg245 250 255ggc gaa ggg cgc ggc ggc gct ggc gcc ggc gcc acg gaa gga gcc ggg816Gly Glu Gly Arg Gly Gly Ala Gly Ala Gly Ala Thr Glu Gly Ala Gly260 265 270cat gac tga825His Asp *<210>16<21l>274<212>PRT<213>百日咳博德特氏菌
<400>16Met Asn Ala Ile Gly Ala Ile Gln Arg Tyr Arg Arg Gly Ala Gly Trp1 5 10 15Ala Ala Leu Val Leu Ala Leu Ala Leu Leu Ala Gly Cys Gly Ala Arg20 25 30Val Glu Leu Leu Gly Ala Ala Pro Glu Asn Glu Ala Asn Glu Val Leu35 40 45Ala Ala Leu Leu Glu Ala Gly Ile Ala Ala Gln Lys Gln Ser Gly Lys50 55 60Ala Gly Tyr Ala Val Ser Val Pro Ala Glu Ala Val Ala Arg Ser Leu65 70 75 80Glu Ile Leu Arg Ala Ser Gly Leu Pro Arg Glu Gln Phe Asp Gly Met85 90 95Gly Arg Ile Phe Arg Lys Glu Gly Leu Val Ser Ser Pro Leu Glu Glu100 105 110Arg Ala Arg Tyr Ile Tyr Ala Leu Ser Gln Glu Leu Ala Asp Thr Leu115 120 125Ser Gln Ile Asp Gly Val Leu Ser Ala Arg Val His Val Val Leu Pro130 135 140Glu Arg Gly Ala Val Gly Glu Pro Ala Thr Pro Ser Thr Ala Gly Val145 150 155 160Phe Leu Lys Tyr Arg Asp Gly Gln Ser Leu Asp Ala Leu Val Pro Glu165 170 175Ile Arg Lys Leu Val Thr His Ala Ile Pro Gly Leu Ala Glu Asp Arg180 185 190Val Ser Val Ala Leu Val Val Ala Gln Pro Val Gln Ala Ala Pro Ala195 200 205Pro Val Ala Trp Arg Arg Val Leu Gly Val Gln Val Ala Asp Gly Ser210 215 220Val Leu Arg Phe Ser Leu Leu Leu Leu Leu Leu Pro Val Leu Cys Leu225 230 235 240Ile Val Ala Gly Ala Ala Leu Tyr Val Trp Arg Thr Arg Trp Ser Arg245 250 255Gly Glu Gly Arg Gly Gly Ala Gly Ala Gly Ala Thr Glu Gly Ala Gly260 265 270His Asp<210>17<211>663<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(663)<400>17atg act gag gcg agc gtg ctg ctt tcc gag cgg ctc atg ata ttc aat48Met Thr Glu Ala Ser Val Leu Leu Ser Glu Arg Leu Met Ile Phe Asn1 5 10 15ctc ctg ccc agc ctg acc ctg cat gcc agt cgc cac gac gag atg ttt 96Leu Leu Pro Ser Leu Thr Leu His Ala Ser Arg His Asp Glu Met Phe20 25 30cca gcc gat tgg gtg cgc gcg ttg tgc aat gcc gac gcg gcg ttg gcc 144Pro Ala Asp Trp Val Arg Ala Leu Cys Asn Ala Asp Ala Ala Leu Ala35 40 45aac gcg tgg cat cgc cat tgg tcg cgc tgg atc ttg tgc gaa ttg ggc 192Asn Ala Trp His Arg His Trp Ser Arg Trp Ile Leu Cys Glu Leu Gly50 55 60ctg ctg aac cag ccg gtc ctg agc ctc gat ccg ccg cag ttg aag gtc 240Leu Leu Asn Gln Pro Val Leu Ser Leu Asp Pro Pro Gln Leu Lys Val65 70 75 80gcg cta ttg tcc acg gac gcc ttg cgg acc tgc gcc gcc cat gcg gga 288Ala Leu Leu Ser Thr Asp Ala Leu Arg Thr Cys Ala Ala His Ala Gly85 90 95gcg ctg ctg tgc gcg ccg cgc ctg cga cgc gcg ata gac ggc gcc gag 336Ala Leu Leu Cys Ala Pro Arg Leu Arg Arg Ala Ile Asp Gly Ala Glu100 105 110gtc cgt acc ttg cat gcc gcg ctc ggg cgc gat gtg atg aat ttc gcc 384Val Arg Thr Leu His Ala Ala Leu Gly Arg Asp Val Met Asn Phe Ala115 120 125gtg tct tcc gcg gcg cgg gcc ctg cat gac ggg ctc gcc gcc agt tcg 432Val Ser Ser Ala Ala Arg Ala Leu His Asp Gly Leu Ala Ala Ser Ser130 135 140gac tgg acc ctg gcc gcc acg gtc cag gcg gcg cag aaa ctg ggc tgg 480Asp Trp Thr Leu Ala Ala Thr Val Gln Ala Ala Gln Lys Leu Gly Trp145 150 155 160gcc ctg ctg cgc gac gcc gtg cag ggc gcc gcc gac gag ata gcg ctg 528Ala Leu Leu Arg Asp Ala Val Gln Gly Ala Ala Asp Glu Ile Ala Leu165 170 175cgt tgc gcg ctg aag ttg ccg cgc gac ctt gat ccc gcg ccc gtc ctg 576Arg Cys Ala Leu Lys Leu Pro Arg Asp Leu Asp Pro Ala Pro Val Leu180 185 190ccg ccc gag gcg gcg ctt gcg ctg gtg ctg tcc atg ctc gaa atc ctg 624Pro Pro Glu Ala Ala Leu Ala Leu Val Leu Ser Met Leu Glu Ile Leu195 200 205gat gca gaa tgg ctt tcc tcg ttc ccc gcc caa gcc tga 663Asp Ala Glu Trp Leu Ser Ser Phe Pro Ala Gln Ala *
210 215 220<210>18<211>220<212>PRT<213>百日咳博德特氏菌<400>18Met Thr Glu Ala Ser Val Leu Leu Ser Glu Arg Leu Met Ile Phe Asn1 5 10 15Leu Leu Pro Ser Leu Thr Leu His Ala Ser Arg His Asp Glu Met Phe20 25 30Pro Ala Asp Trp Val Arg Ala Leu Cys Asn Ala Asp Ala Ala Leu Ala35 40 45Asn Ala Trp His Arg His Trp Ser Arg Trp Ile Leu Cys Glu Leu Gly50 55 60Leu Leu Asn Gln Pro Val Leu Ser Leu Asp Pro Pro Gln Leu Lys Val65 70 75 80Ala Leu Leu Ser Thr Asp Ala Leu Arg Thr Cys Ala Ala His Ala Gly85 90 95Ala Leu Leu Cys Ala Pro Arg Leu Arg Arg Ala Ile Asp Gly Ala Glu100 105 110Val Arg Thr Leu His Ala Ala Leu Gly Arg Asp Val Met Asn Phe Ala115 120 125Val Ser Ser Ala Ala Arg Ala Leu His Asp Gly Leu Ala Ala Ser Ser130 135 140Asp Trp Thr Leu Ala Ala Thr Val Gln Ala Ala Gln Lys Leu Gly Trp145 150 155 160Ala Leu Leu Arg Asp Ala Val Gln Gly Ala Ala Asp Glu Ile Ala Leu165 170 175Arg Cys Ala Leu Lys Leu Pro Arg Asp Leu Asp Pro Ala Pro Val Leu180 185 190Pro Pro Glu Ala Ala Leu Ala Leu Val Leu Ser Met Leu Glu Ile Leu195 200 205Asp Ala Glu Trp Leu Ser Ser Phe Pro Ala Gln Ala210 215 220<210>19<211>639<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(639)<400>19atg gct ttc ctc gtt ccc cgc cca agc ctg atc cag gcg gta cgg ccc48Met ala Phe Leu Val Pro Arg Pro Ser Leu Ile Gln Ala Val Arg Pro1 5 10 15ggc cgt gcg gat ccc gcg acc gac gtc ttg cgc gct gaa gac tac gcc 96Gly Arg Ala Asp Pro Ala Thr Asp Val Leu Arg Ala Glu Asp Tyr Ala20 25 30gag ctg ctc agc gcc gcg cag atc gtt gcc cag gca cac cgg cgg gcc 144Glu Leu Leu Ser Ala Ala Gln Ile Val Ala Gln Ala His Arg Arg Ala35 40 45ggc gaa atc gtg gcc gag gcg cga gag gag ttc gag cgc gag cgc agg 192Gly Glu Ile Val Ala Glu Ala Arg Glu Glu Phe Glu Arg Glu Arg Arg50 55 60cga ggc tat gag gag ggg cgc cgc gaa gcg ctt acg gat cag gcg gag 240Arg Gly Tyr Glu Glu Gly Arg Arg Glu Ala Leu Thr Asp Gln Ala Glu65 70 75 80aag atg ata gaa acc gta agc cgc acg atc gac tac ttc gcg ggt atc 288Lys Met Ile Glu Thr Val Ser Arg Thr Ile Asp Tyr Phe Ala Gly Ile85 90 95gag aac gag atg atc gaa ctg gtc atg agt gcg gtc cgc aag atc gtc 336Glu Asn Glu Met Ile Glu Leu Val Met Ser Ala Val Arg Lys Ile Val100 105 110gac ggt tac gac gac cgc gag cgc acc gtg atc gcc gtg cgc aac gca 384Asp Gly Tyr Asp Asp Arg Glu Arg Thr Val Ile Ala Val Arg Asn Ala115 120 125ttg gcg gtc gtg cgc aat cag cgc cag atg acc ttg cgc ctg cac cca 432Leu Ala Val Val Arg Asn Gln Arg Gln Met Thr Leu Arg Leu His Pro130 135 140gac gag gtg gat gtg ctc cgg gaa ggc atg aac cag ctt ctg gcg gcc 480Asp Glu Val Asp Val Leu Arg Glu Gly Met Asn Gln Leu Leu Ala Ala145 150 155 160tat ccg ggc gtg ggc tac ctg gac ctg ctg ccc gac gcc agg ctg gcg 528Tyr Pro Gly Val Gly Tyr Leu Asp Leu Leu Pro Asp Ala Arg Leu Ala165 170 175ccg gga gcc tgc ata ctg gag agc gag ata ggc atg gtc gag gcc agc 576Pro Gly Ala Cys Ile Leu Glu Ser Glu Ile Gly Met Val Glu Ala Ser180 185 190ctc gag gac cag ctg tgc gcc ttg cgg gcg gcc ttc gaa cgt aca ttc 624Leu Glu Asp Gln Leu Cys Ala Leu Arg Ala Ala Phe Glu Arg Thr Phe195 200 205ggc cgg cgc gga tag 639Gly Arg Arg Gly *
210<210>20<211>212<212>PRT<213>百日咳博德特氏菌<400>20Met ala Phe Leu Val Pro Arg Pro Ser Leu Ile Gln Ala Val Arg Pro1 5 10 15Gly Arg Ala Asp Pro Ala Thr Asp Val Leu Arg Ala Glu Asp Tyr Ala20 25 30Glu Leu Leu Ser Ala Ala Gln Ile Val Ala Gln Ala His Arg Arg Ala35 40 45Gly Glu Ile Val Ala Glu Ala Arg Glu Glu Phe Glu Arg Glu Arg Arg50 55 60Arg Gly Tyr Glu Glu Gly Arg Arg Glu Ala Leu Thr Asp Gln Ala Glu65 70 75 80Lys Met Ile Glu Thr Val Ser Arg Thr Ile Asp Tyr Phe Ala Gly Ile85 90 95Glu Asn Glu Met Ile Glu Leu Val Met Ser Ala Val Arg Lys Ile Val100 105 110Asp Gly Tyr Asp Asp Arg Glu Arg Thr Val Ile Ala Val Arg Asn Ala115 120 125Leu Ala Val Val Arg Asn Gln Arg Gln Met Thr Leu Arg Leu His Pro130 135 140Asp Glu Val Asp Val Leu Arg Glu Gly Met Asn Gln Leu Leu Ala Ala145 150 155 160Tyr Pro Gly Val Gly Tyr Leu Asp Leu Leu Pro Asp Ala Arg Leu Ala165 170 175Pro Gly Ala Cys Ile Leu Glu Ser Glu Ile Gly Met Val Glu Ala Ser180 185 190Leu Glu Asp Gln Leu Cys Ala Leu Arg Ala Ala Phe Glu Arg Thr Phe195 200 205Gly Arg Arg Gly210<210>21<211>1335<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1335)<400>21atg cgt cag tac cac tac atc acg gag atg atg cgg gtg gcc ctg cag48Met Arg Gln Tyr His Tyr Ile Thr Glu Met Met Arg Val Ala Leu Gln1 5 10 15gat ctg tcc acg ctg cgg ata aag ggc cgg gtg gtg caa gtg gtg gga 96Asp Leu Ser Thr Leu Arg Ile Lys Gly Arg Val Val Gln Val Val Gly20 25 30acg atc atc aag gcc gtc gtt ccg atg gtc aag atc ggc gaa gtg tgc 144Thr Ile Ile Lys Ala Val Val Pro Met Val Lys Ile Gly Glu Val Cys35 40 45ctg ctg cgc aat ccc ggc gag gac ttc gag atg cac ggc gaa gtg gtg 192Leu Leu Arg Asn Pro Gly Glu Asp Phe Glu Met His Gly Glu Val Val50 55 60ggc ttt gtc cgc gac gcc gcc ttg ctc acg cct atc ggc gac atg tac 240Gly Phe Val Arg Asp Ala Ala Leu Leu Thr Pro Ile Gly Asp Met Tyr65 70 75 80ggg att tcc tcg gcg acc gag gtg ata ccg acc gga cgc acg cat atg 288Gly Ile Ser Ser Ala Thr Glu Val Ile Pro Thr Gly Arg Thr His Met85 90 95gtc ccc gtc ggt ccg ggc ttg ctg gga cgc gtg ctg gac ggg ctg gga 336Val Pro Val Gly Pro Gly Leu Leu Gly Arg Val Leu Asp Gly Leu Gly100 105 110cgt ccg ctg gac gcc gcc gag tca ggg ccg ctg cat gcc cac aag ttc 384Arg Pro Leu Asp Ala Ala Glu Ser Gly Pro Leu His Ala His Lys Phe115 120 125tat ccg gtc ttc gcc gat gcg cca gac ccg ctg acg cgt cgc atc atc 432Tyr Pro Val Phe Ala Asp Ala Pro Asp Pro Leu Thr Arg Arg Ile Ile130 135 140cat gct ccg ctg gag ctg ggg gtg cgc gta ctg gac ggt ttg ctt aca 480His Ala Pro Leu Glu Leu Gly Val Arg Val Leu Asp Gly Leu Leu Thr145 150 155 160tgc ggg gaa ggc cag cgt ctg gga att ttc gca gcc gcc ggc ggc ggc 528Cys Gly Glu Gly Gln Arg Leu Gly Ile Phe Ala Ala Ala Gly Gly Gly165 170 175aag tcg acc ctg ctg ggc atg ctg gtc aag ggc gcc gcg gtc gac gtg 576Lys Ser Thr Leu Leu Gly Met Leu Val Lys Gly Ala Ala Val Asp Val180 185 190acg gtg gtg gcg ctg atc ggc gag cgt ggg cgg gaa gtt cgc gag ttc 624Thr Val Val Ala Leu Ile Gly Glu Arg Gly Arg Glu Val Arg Glu Phe195 200 205ctt gag cac gaa ctc ggt ccg gag ggc aga cgc aag agc gtg atc gtc 672Leu Glu His Glu Leu Gly Pro Glu Gly Arg Arg Lys Ser Val Ile Val
210 215 220tgc gcg acc agc gac aag tcc tcg atg gag cgt gcc aag gcg gcg tac 720Cys Ala Thr Ser Asp Lys Ser Ser Met Glu Arg Ala Lys Ala Ala Tyr225 230 235 240gtc gca acc gcc atc gcc gaa tac ttc cgc gat caa ggg cag cgt gta 768Val Ala Thr Ala Ile Ala Glu Tyr Phe Arg Asp Gln Gly Gln Arg Val245 250 255ctt ttt ctg atg gac tcg gtc acc cgc ttt gcg cga gcc cag cgt gaa 816Leu Phe Leu Met Asp Ser Val Thr Arg Phe Ala Arg Ala Gln Arg Glu260 265 270atc ggc ttg gcg gca ggc gag ccg ccg acg cgg cgc ggc tat cca ccg 864Ile Gly Leu Ala Ala Gly Glu Pro Pro Thr Arg Arg Gly Tyr Pro Pro275 280 285tcg gtg ttc gcc acc ttg ccc aaa ctg atg gag cgc gcc ggc atg aac 912Ser Val Phe Ala Thr Leu Pro Lys Leu Met Glu Arg Ala Gly Met Asn290 295 300cag acg ggt tcg atc acg gcg ctg tat acg gtg ctg gtc gag ggg gac 960Gln Thr Gly Ser Ile Thr Ala Leu Tyr Thr Val Leu Val Glu Gly Asp305 310 315 320gac atg aac gaa ccg gtg gcc gac gag acg cgt tcg ata ctg gac ggc 1008Asp Met Asn Glu Pro Val Ala Asp Glu Thr Arg Ser Ile Leu Asp Gly325 330 335cac atc gtg ctc tcg cgc aag ctg gga gcg gcg aat cac tat cct gcc 1056His Ile Val Leu Ser Arg Lys Leu Gly Ala Ala Asn His Tyr Pro Ala340 345 350gtc gac gtg ctg gcc tcg gcc agc cgg gtc atg aat gcc gtg gtg tcg 1104Val Asp Val Leu Ala Ser Ala Ser Arg Val Met Asn Ala Val Val Ser355 360 365ccg cgt cac aag tac ctg gcc gga cgt atg cgc gaa ctg atg gcc aag 1152Pro Arg His Lys Tyr Leu Ala Gly Arg Met Arg Glu Leu Met ala Lys370 375 380tac cag gat gtc gag ctg ttg gtg aaa atc ggc gag tac aag cag ggc 1200Tyr Gln Asp Val Glu Leu Leu Val Lys Ile Gly Glu Tyr Lys Gln Gly385 390 395 400gcc gat gcg tcg acc gat gag gcg ata cag aag atc gga cag atc aat 1248Ala Asp Ala Ser Thr Asp Glu Ala Ile Gln Lys Ile Gly Gln Ile Asn405 410 415gcg ttt ctc aga caa cta acc gac gaa cgc gaa gca ttc gag gat acc 1296Ala Phe Leu Arg Gln Leu Thr Asp Glu Arg Glu Ala Phe Glu Asp Thr420 425 430gta ctg cgc atg gct gaa atc atc gga ccc gaa tcc taa1335Val Leu Arg Met ala Glu Ile Ile Gly Pro Glu Ser *435 440<210>22<211>444<212>PRT<213>百日咳博德特氏菌<400>22Met Arg Gln Tyr His Tyr Ile Thr Glu Met Met Arg Val Ala Leu Gln1 5 10 15Asp Leu Ser Thr Leu Arg Ile Lys Gly Arg Val Val Gln Val Val Gly20 25 30Thr Ile Ile Lys Ala Val Val Pro Met Val Lys Ile Gly Glu Val Cys35 40 45Leu Leu Arg Asn Pro Gly Glu Asp Phe Glu Met His Gly Glu Val Val50 55 60Gly Phe Val Arg Asp Ala Ala Leu Leu Thr Pro Ile Gly Asp Met Tyr65 70 75 80Gly Ile Ser Ser Ala Thr Glu Val Ile Pro Thr Gly Arg Thr His Met85 90 95Val Pro Val Gly Pro Gly Leu Leu Gly Arg Val Leu Asp Gly Leu Gly100 105 110Arg Pro Leu Asp Ala Ala Glu Ser Gly Pro Leu His Ala His Lys Phe115 120 125Tyr Pro Val Phe Ala Asp Ala Pro Asp Pro Leu Thr Arg Arg Ile Ile130 135 140His Ala Pro Leu Glu Leu Gly Val Arg Val Leu Asp Gly Leu Leu Thr145 150 155 160Cys Gly Glu Gly Gln Arg Leu Gly Ile Phe Ala Ala Ala Gly Gly Gly165 170 175Lys Ser Thr Leu Leu Gly Met Leu Val Lys Gly Ala Ala Val Asp Val180 185 190Thr Val Val Ala Leu Ile Gly Glu Arg Gly Arg Glu Val Arg Glu Phe195 200 205Leu Glu His Glu Leu Gly Pro Glu Gly Arg Arg Lys Ser Val Ile Val210 215 220Cys Ala Thr Ser Asp Lys Ser Ser Met Glu Arg Ala Lys Ala Ala Tyr225 230 235 240Val Ala Thr Ala Ile Ala Glu Tyr Phe Arg Asp Gln Gly Gln Arg Val245 250 255Leu Phe Leu Met Asp Ser Val Thr Arg Phe Ala Arg Ala Gln Arg Glu260 265 270Ile Gly Leu Ala Ala Gly Glu Pro Pro Thr Arg Arg Gly Tyr Pro Pro275 280 285Ser Val Phe Ala Thr Leu Pro Lys Leu Met Glu Arg Ala Gly Met Asn
290 295 300Gln Thr Gly Ser Ile Thr Ala Leu Tyr Thr Val Leu Val Glu Gly Asp305 310 315 320Asp Met Asn Glu Pro Val Ala Asp Glu Thr Arg Ser Ile Leu Asp Gly325330 335His Ile Val Leu Ser Arg Lys Leu Gly Ala Ala Asn His Tyr Pro Ala340 345 350Val Asp Val Leu Ala Ser Ala Ser Arg Val Met Asn Ala Val Val Ser355 360 365Pro Arg His Lys Tyr Leu Ala Gly Arg Met Arg Glu Leu Met ala Lys370 375 380Tyr Gln Asp Val Glu Leu Leu Val Lys Ile Gly Glu Tyr Lys Gln Gly385 390 395400Ala Asp Ala Ser Thr Asp Glu Ala Ile Gln Lys Ile Gly Gln Ile Asn405 410 415Ala Phe Leu Arg Gln Leu Thr Asp Glu Arg Glu Ala Phe Glu Asp Thr420 425 430Val Leu Arg Met ala Glu Ile Ile Gly Pro Glu Ser435 440<210>23<211>510<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(510)<400>23atg gac ctg gaa agc ctg ctt gcc atc aag cat ttt cgc gcc gac caa48Met Asp Leu Glu Ser Leu Leu Ala Ile Lys His Phe Arg Ala Asp Gln1 5 10 15gcc cag ctt gcg ctg aaa cgc caa cag cag gcc tgc gcg gtt gct gcc96Ala Gln Leu Ala Leu Lys Arg Gln Gln Gln Ala Cys Ala Val Ala Ala20 25 30gcg gcg cag cgt cag gcg caa ggc cgc ctc gac gat tgt cgc ctg tgg144Ala Ala Gln Arg Gln Ala Gln Gly Arg Leu Asp Asp Cys Arg Leu Trp35 40 45gcc gga cag ctc gaa aac cgt cta tat gcc gag ctg tgc cgg cgc atc192Ala Gly Gln Leu Glu Asn Arg Leu Tyr Ala Glu Leu Cys Arg Arg Ile50 55 60gtc aag aca cgc gac atc gac gag gtg ctg caa cga gtg ggc cac gcc240Val Lys Thr Arg Asp Ile Asp Glu Val Leu Gln Arg Val Gly His Ala65 70 75 80cgc gac cgc cag gcc agc ctg gcg ctg cag ctc gac gac gcc gtg cgc288Arg Asp Arg Gln Ala Ser Leu Ala Leu Gln Leu Asp Asp Ala Val Arg85 90 95cgt cac gaa cat gaa atc cag ctg ctc gcg cag cag cgc gag cag cac336Arg His Glu His Glu Ile Gln Leu Leu Ala Gln Gln Arg Glu Gln His100 105 110cgg gag tgc ttc cag gcg cag caa cgg atc gcc gag ttg gtg cgc ctg384Arg Glu Cys Phe Gln Ala Gln Gln Arg Ile Ala Glu Leu Val Arg Leu115 120 125cag cag gtc gag gcg gcg gcc ttg cgc gag agc cag gaa gat cgc gaa432Gln Gln Val Glu Ala Ala Ala Leu Arg Glu Ser Gln Glu Asp Arg Glu130 135 140att cag gaa gcc atc gaa ttg tcg gcg cgt ggg cgc gac gat gca tcg480Ile Gln Glu Ala Ile Glu Leu Ser Ala Arg Gly Arg Asp Asp Ala Ser145 150 155 160cga gcc ggc gac ggc ctg gcg cgg cta tga510Arg Ala Gly Asp Gly Leu Ala Arg Leu *165<210>24<211>169<212>PRT<213>百日咳博德特氏菌<400>24Met Asp Leu Glu Ser Leu Leu Ala Ile Lys His Phe Arg Ala Asp Gln1 5 10 15Ala Gln Leu Ala Leu Lys Arg Gln Gln Gln Ala Cys Ala Val Ala Ala20 25 30Ala Ala Gln Arg Gln Ala Gln Gly Arg Leu Asp Asp Cys Arg Leu Trp35 40 45Ala Gly Gln Leu Glu Asn Arg Leu Tyr Ala Glu Leu Cys Arg Arg Ile50 55 60Val Lys Thr Arg Asp Ile Asp Glu Val Leu Gln Arg Val Gly His Ala65 70 75 80Arg Asp Arg Gln Ala Ser Leu Ala Leu Gln Leu Asp Asp Ala Val Arg85 90 95Arg His Glu His Glu Ile Gln Leu Leu Ala Gln Gln Arg Glu Gln His100 105 110Arg Glu Cys Phe Gln Ala Gln Gln Arg Ile Ala Glu Leu Val Arg Leu115 120 125Gln Gln Val Glu Ala Ala Ala Leu Arg Glu Ser Gln Glu Asp Arg Glu130 135 140Ile Gln Glu Ala Ile Glu Leu Ser Ala Arg Gly Arg Asp Asp Ala Ser145 150 155 160Arg Ala Gly Asp Gly Leu Ala Arg Leu
165<210>25<211>549<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(549)<400>25atg aac cag cca gac ggg ctg ggt tcg ccc atg gcc ggc ggc ggg cag48Met Asn Gln Pro Asp Gly Leu Gly Ser Pro Met ala Gly Gly Gly Gln1 5 10 15cgc atg ggc gtg gcg cgc acg ccg tat gcg cgt cag ccg gat cgg gat96Arg Met Gly Val Ala Arg Thr Pro Tyr Ala Arg Gln Pro Asp Arg Asp20 25 30gcg cag cgt gcc ttc gag cgg gaa atg gaa cag gag aaa gcg aag gaa144Ala Gln Arg Ala Phe Glu Arg Glu Met Glu Gln Glu Lys Ala Lys Glu35 40 45gaa ctg ccc ggg ccg caa cgc ctg gcg ccg ggt ccg gcc tgc gtc ggc192Glu Leu Pro Gly Pro Gln Arg Leu Ala Pro Gly Pro Ala Cys Val Gly50 55 60tgg ctg gcg tcg atg gaa cct gcc gcc ggc cgt cca ccg gcc agt ctg240Trp Leu Ala Ser Met Glu Pro Ala Ala Gly Arg Pro Pro Ala Ser Leu65 70 75 80gcc cag gcg ctg gca agc gtg gct gcg ggg ctg gcg gta ggc gac gtg288Ala Gln Ala Leu Ala Ser Val Ala Ala Gly Leu Ala Val Gly Asp Val85 90 95ctg gag ggg tat cgc gaa gcc cgt atc gtt gtg gac gat acg ctg cta336Leu Glu Gly Tyr Arg Glu Ala Arg Ile Val Val Asp Asp Thr Leu Leu100 105 110ccc gac acc acc ttg tcg gta cgg gag gac ggc ggc tgg atc gtg gtg384Pro Asp Thr Thr Leu Ser Val Arg Glu Asp Gly Gly Trp Ile Val Val115 120 125gct ttc gca tgc cga caa cgg gac gct tgc gag cgc ctg cac gcg tgc432Ala Phe Ala Cys Arg Gln Arg Asp Ala Cys Glu Arg Leu His Ala Cys130 135 140gcc gac cgg ttg gcc atg gag ctc gcg ctg gag ctg gcg cgc gac gtc480Ala Asp Arg Leu Ala Met Glu Leu Ala Leu Glu Leu Ala Arg Asp Val145 150 155 160gag gtt gcg gtg gca tgc gac ggc gag ccg cac gag cgg gtg gcg cgc528Glu Val Ala Val Ala Cys Asp Gly Glu Pro His Glu Arg Val Ala Arg165 170 175gcg cag cgg ccg tgg cga tga549Ala Gln Arg Pro Trp Arg *180<210>26<211>182<212>PRT<213>百日咳博德特氏菌<400>26Met Asn Gln Pro Asp Gly Leu Gly Ser Pro Met ala Gly Gly Gly Gln1 5 10 15Arg Met Gly Val Ala Arg Thr Pro Tyr Ala Arg Gln Pro Asp Arg Asp20 25 30Ala Gln Arg Ala Phe Glu Arg Glu Met Glu Gln Glu Lys Ala Lys Glu35 40 45Glu Leu Pro Gly Pro Gln Arg Leu Ala Pro Gly Pro Ala Cys Val Gly50 55 60Trp Leu Ala Ser Met Glu Pro Ala Ala Gly Arg Pro Pro Ala Ser Leu65 70 75 80Ala Gln Ala Leu Ala Ser Val Ala Ala Gly Leu Ala Val Gly Asp Val85 90 95Leu Glu Gly Tyr Arg Glu Ala Arg Ile Val Val Asp Asp Thr Leu Leu100 105 110Pro Asp Thr Thr Leu Ser Val Arg Glu Asp Gly Gly Trp Ile Val Val115 120 125Ala Phe Ala Cys Arg Gln Arg Asp Ala Cys Glu Arg Leu His Ala Cys130 135 140Ala Asp Arg Leu Ala Met Glu Leu Ala Leu Glu Leu Ala Arg Asp Val145 150 155 160Glu Val Ala Val Ala Cys Asp Gly Glu Pro His Glu Arg Val Ala Arg165 170 175Ala Gln Arg Pro Trp Arg180<210>27<211>1080<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1080)<400>27atg aat cga gtg gcc ggc ggg gcg gcg gcg cag gcc gct ggc atg gtg 48Met Asn Arg Val Ala Gly Gly Ala Ala Ala Gln Ala Ala Gly Met Val1 5 10 15gat ctc gcg gtt ccg cgg ttg agc gcc ggc gag gcc cat gcc ctg tcg 96Asp Leu Ala Val Pro Arg Leu Ser Ala Gly Glu Ala His Ala Leu Ser20 25 30agg att gca tgc cat ggc gcg cga ttc gac gtt cgg ctt ggc gag ccg 144Arg Ile Ala Cys His Gly Ala Arg Phe Asp Val Arg Leu Gly Glu Pro35 40 45gcc gtg cgc tgg cac tgc gcc ctg acg cct tgc gtg cac ggc gac ctt 192Ala Val Arg Trp His Cys Ala Leu Thr Pro Cys Val His Gly Asp Leu50 55 60gcc gat ggc gag atg gaa agc ctg caa ctg caa tgg gcc ggg acg tac 240Ala Asp Gly Glu Met Glu Ser Leu Gln Leu Gln Trp Ala Gly Thr Tyr65 70 75 80atc ggc ctg acg gtt ccg cgc gcg gcc gcg gcg gga tgg ctg gcg gcg 288Ile Gly Leu Thr Val Pro Arg Ala Ala Ala Ala Gly Trp Leu Ala Ala85 90 95cgc ctg ccc cgg ttt tcc ggc gtg gag ttg ccg gaa ccc att gcg gcg 336Arg Leu Pro Arg Phe Ser Gly Val Glu Leu Pro Glu Pro Ile Ala Ala100 105 110gcg gcc ctg gag gca atg ctg gag gag gtc tgt cga ggc gtg gcc gga 384Ala Ala Leu Glu Ala Met Leu Glu Glu Val Cys Arg Gly Val Ala Gly115 120 125ctc gac cag caa ggc ccg gtc cgc gtg gcg cgg caa ggc ggg acg cca 432Leu Asp Gln Gln Gly Pro Val Arg Val Ala Arg Gln Gly Gly Thr Pro130 135 140ccg gtc cag ccg cat cgc tgg acc ctg acg gta cgg gcg cct gac ggt 480Pro Val Gln Pro His Arg Trp Thr Leu Thr Val Arg Ala Pro Asp Gly145 150 155 160ggc gtc tgg cgc gcg gta ctg gcg tgc gac gca tgg gcc ttg caa gcg 528Gly Val Trp Arg Ala Val Leu Ala Cys Asp Ala Trp Ala Leu Gln Ala165 170 175gtc gcg gcg gcg ctg gat tcc gtt gcg cct gcc gat ggt cgg gtc aat 576Val Ala Ala Ala Leu Asp Ser Val Ala Pro Ala Asp Gly Arg Val Asn180 185 190ccg gag cgc gtg ccg gtc agg ttg cgt gcc gat gtc ggc gcg gcg tcc 624Pro Glu Arg Val Pro Val Arg Leu Arg Ala Asp Val Gly Ala Ala Ser195 200 205gtg acc gca ggc cag ctg cgg acg ctg cga gcg ggc gac gtc gtg ttg672Val Thr Ala Gly Gln Leu Arg Thr Leu Arg Ala Gly Asp Val Val Leu210 215 220ctc gcg cag tac cgg gtg agc gat gcc gca gaa cta tgg ttg tcg gcc720Leu Ala Gln Tyr Arg Val Ser Asp Ala Ala Glu Leu Trp Leu Ser Ala225 230 235 240gga ccc agc gcg atc cgg gta cgg gcc gag cat gcg tct ttt cgt gta768Gly Pro Ser Ala Ile Arg Val Arg Ala Glu His Ala Ser Phe Arg Val245 250 255act caa ggt tgg act ccc atc atg acg gaa ccc gcg aca cct gac cct816Thr Gln Gly Trp Thr Pro Ile Met Thr Glu Pro Ala Thr Pro Asp Pro260 265 270ggc gaa acc ccg gca cag gcc gac gcg acg ctc gat acc gat cag ata864Gly Glu Thr Pro Ala Gln Ala Asp Ala Thr Leu Asp Thr Asp Gln Ile275 280 285ccc gtg cgc ctg acg ttc gac ctg ggc gag cgc gag ttc acg ctt gcg912Pro Val Arg Leu Thr Phe Asp Leu Gly Glu Arg Glu Phe Thr Leu Ala290 295 300cag ctg cgc agc ctg cat ccg ggc tgc acg ttc gac ctc gag cgg ccc960Gln Leu Arg Ser Leu His Pro Gly Cys Thr Phe Asp Leu Glu Arg Pro305 310 315 320atc gcc gac ggg ccg gtc atg gtg cgg gcc aat ggc ctg ttg ctg ggc1008Ile Ala Asp Gly Pro Val Met Val Arg Ala Asn Gly Leu Leu Leu Gly325 330 335agc ggc cgg ctg gtc gac atc gac ggc cgc atc ggc gtg gta ttg cag1056Ser Gly Arg Leu Val Asp Ile Asp Gly Arg Ile Gly Val Val Leu Gln340 345 350tcg gtc agg cct gga ctc gca tga1080Ser Val Arg Pro Gly Leu Ala *355<210>28<211>359<212>PRT<213>百日咳博德特氏菌<400>28Met Asn Arg Val Ala Gly Gly Ala Ala Ala Gln Ala Ala Gly Met Val1 5 10 15Asp Leu Ala Val Pro Arg Leu Ser Ala Gly Glu Ala His Ala Leu Ser
20 25 30Arg Ile Ala Cys His Gly Ala Arg Phe Asp Val Arg Leu Gly Glu Pro35 40 45Ala Val Arg Trp His Cys Ala Leu Thr Pro Cys Val His Gly Asp Leu50 55 60Ala Asp Gly Glu Met Glu Ser Leu Gln Leu Gln Trp Ala Gly Thr Tyr65 70 75 80Ile Gly Leu Thr Val Pro Arg Ala Ala Ala Ala Gly Trp Leu Ala Ala85 90 95Arg Leu Pro Arg Phe Ser Gly Val Glu Leu Pro Glu Pro Ile Ala Ala100 105 110Ala Ala Leu Glu Ala Met Leu Glu Glu Val Cys Arg Gly Val Ala Gly115 120 125Leu Asp Gln Gln Gly Pro Val Arg Val Ala Arg Gln Gly Gly Thr Pro130 135 140Pro Val Gln Pro His Arg Trp Thr Leu Thr Val Arg Ala Pro Asp Gly145 150 155 160Gly Val Trp Arg Ala Val Leu Ala Cys Asp Ala Trp Ala Leu Gln Ala165 170 175Val Ala Ala Ala Leu Asp Ser Val Ala Pro Ala Asp Gly Arg Val Asn180 185 190Pro Glu Arg Val Pro Val Arg Leu Arg Ala Asp Val Gly Ala Ala Ser195 200 205Val Thr Ala Gly Gln Leu Arg Thr Leu Arg Ala Gly Asp Val Val Leu210 215 220Leu Ala Gln Tyr Arg Val Ser Asp Ala Ala Glu Leu Trp Leu Ser Ala225 230 235 240GlV Pro Ser Ala Ile Arg Val Arg Ala Glu His Ala Ser Phe Arg Val245 250 255Thr Gln Gly Trp Thr Pro Ile Met Thr Glu Pro Ala Thr Pro Asp Pro260 265 270Gly Glu Thr Pro Ala Gln Ala Asp Ala Thr Leu Asp Thr Asp Gln Ile275 280 285Pro Val Arg Leu Thr Phe Asp Leu Gly Glu Arg Glu Phe Thr Leu Ala290 295 300Gln Leu Arg Ser Leu His Pro Gly Cys Thr Phe Asp Leu Glu Arg Pro305 310 315 320Ile Ala Asp Gly Pro Val Met Val Arg Ala Asn Gly Leu Leu Leu Gly325 330 335Ser Gly Arg Leu Val Asp Ile Asp Gly Arg Ile Gly Val Val Leu Gln340 345 350Ser Val Arg Pro Gly Leu Ala355<210>29<211>672<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS
<222>(1)...(672)<400> 29atg agc gat acc gac ccc ttc agc ctg gcc ctg ttt ctg gcg ctg ctg 48Met Ser Asp Thr Asp Pro Phe Ser Leu Ala Leu Phe Leu Ala Leu Leu1 5 10 15gcg ctg gta ccg ctc atc gtc gtc atg acc acg tcg ttc ctg aag atc 96Ala Leu Val Pro Leu Ile Val Val Met Thr Thr Ser Phe Leu Lys Ile20 25 30gcc gtc gtg ctt gcc ttg gtg cgc aac gcc ctg gga gtg caa cag gta 144Ala Val Val Leu Ala Leu Val Arg Asn Ala Leu Gly Val Gln Gln Val35 40 45ccg ccc aac atg gcc ctg tac ggg ctg gcg ctt att ctt tcc gcg tat 192Pro Pro Asn Met ala Leu Tyr Gly Leu Ala Leu Ile Leu Ser Ala Tyr50 55 60gtg atg gcg ccg gtc gtt cac agg ata ggc acc gag gtc cag gcc ttg 240Val Met ala Pro Val Val His Arg Ile Gly Thr Glu Val Gln Ala Leu65 70 75 80acc gcg caa gcc ggg gag tcc ggc acc gcc gcg ccg atg gcg ctg gac 288Thr Ala Gln Ala Gly Glu Ser Gly Thr Ala Ala Pro Met ala Leu Asp85 90 95gcc gtg ctt ggc gtg gcc gag cga ggc gtg ggg ccg ctg cgg gcc ttc 336Ala Val Leu Gly Val Ala Glu Arg Gly Val Gly Pro Leu Arg Ala Phe100 105 110atg ttg cgc aac agc cag ccg gcc cag cgt gat ttc ttc ctg cgc aca 384Met Leu Arg Asn Ser Gln Pro Ala Gln Arg Asp Phe Phe Leu Arg Thr115 120 125gcg cgt cat ctc tgg ggc gag gag gca tcg cgg gac ctg tcg gaa gac 432Ala Arg His Leu Trp Gly Glu Glu Ala Ser Arg Asp Leu Ser Glu Asp130 135 140aac ctg ctg gta ttg acg ccc gca ttt ctg gtt tcg gag ctg acc gcc 480Asn Leu Leu Val Leu Thr Pro Ala Phe Leu Val Ser Glu Leu Thr Ala145 150 155 160gca ttc cag ctt ggc ttt ctg ctg tac ctg ccg ttc atc atc atc gac 528Ala Phe Gln Leu Gly Phe Leu Leu Tyr Leu Pro Phe Ile Ile Ile Asp165 170 175ctc atc gta tcg aac att ctt ctt gcc atg gga atg atg atg gtt tct 576Leu Ile Val Ser Asn Ile Leu Leu Ala Met Gly Met Met Met Val Ser180 185 190ccc gtg acg atc tcc atg ccg ttg aag ctg ttc ctg ttc gtc atg gtg624Pro Val Thr Ile Ser Met Pro Leu Lys Leu Phe Leu Phe Val Met Val195 200 205gac ggc tgg acg cgc ctg atc cag ggc ctg gtg ctt tcc tat cgg tga672Asp Gly Trp Thr Arg Leu Ile Gln Gly Leu Val Leu Ser Tyr Arg *210 215 220<210>30<211>223<212>PRT<213>百日咳博德特氏菌<400>30Met Ser Asp Thr Asp Pro Phe Ser Leu Ala Leu Phe Leu Ala Leu Leu1 5 10 15Ala Leu Val Pro Leu Ile Val Val Met Thr Thr Ser Phe Leu Lys Ile20 25 30Ala Val Val Leu Ala Leu Val Arg Asn Ala Leu Gly Val Gln Gln Val35 40 45Pro Pro Asn Met ala Leu Tyr Gly Leu Ala Leu Ile Leu Ser Ala Tyr50 55 60Val Met ala Pro Val Val His Arg Ile Gly Thr Glu Val Gln Ala Leu65 70 75 80Thr Ala Gln Ala Gly Glu Ser Gly Thr Ala Ala Pro Met ala Leu Asp85 90 95Ala Val Leu Gly Val Ala Glu Arg Gly Val Gly Pro Leu Arg Ala Phe100 105 110Met Leu Arg Asn Ser Gln Pro Ala Gln Arg Asp Phe Phe Leu Arg Thr115 120 125Ala Arg His Leu Trp Gly Glu Glu Ala Ser Arg Asp Leu Ser Glu Asp130 135 140Asn Leu Leu Val Leu Thr Pro Ala Phe Leu Val Ser Glu Leu Thr Ala145 150 155 160Ala Phe Gln Leu Gly Phe Leu Leu Tyr Leu Pro Phe Ile Ile Ile Asp165 170 175Leu Ile Val Ser Asn Ile Leu Leu Ala Met Gly Met Met Met Val Ser180 185 190Pro Val Thr Ile Ser Met Pro Leu Lys Leu Phe Leu Phe Val Met Val195 200 205Asp Gly Trp Thr Arg Leu Ile Gln Gly Leu Val Leu Ser Tyr Arg210 215 220<210>31<211>267<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS
<222>(1)...(267)<400>31atg caa acc caa gac ctg gtt tcg ttc atg aca cag gcg ttg tac ctg48Met Gln Thr Gln Asp Leu Val Ser Phe Met Thr Gln Ala Leu Tyr Leu1 5 10 15gtg ctc tgg ctg tcg ctg ccg ccc atc gcc gtg gtg gcg atc gtg gga96Val Leu Trp Leu Ser Leu Pro Pro Ile Ala Val Val Ala Ile Val Gly20 25 30acg ctg ttt tcc ctg ttg cag gcc ttg acg cag gtg cag gag cag acc144Thr Leu Phe Ser Leu Leu Gln Ala Leu Thr Gln Val Gln Glu Gln Thr35 40 45ctg tcc ttc gcc gtg aag ctg ata gcc gtg ttc gcc acg ctg atg ctg192Leu Ser Phe Ala Val Lys Leu Ile Ala Val Phe Ala Thr Leu Met Leu50 55 60gcg gcc cgg tgg ata agc gcg gaa atc tat aac ttc acg att gcg gtg240Ala Ala Arg Trp Ile Ser Ala Glu Ile Tyr Asn Phe Thr Ile Ala Val65 70 75 80ttc gat gcc ttt cat cgg atc cac tga267Phe Asp Ala Phe His Arg Ile His *85<210>32<211>88<212>PRT<213>百日咳博德特氏菌<400>32Met Gln Thr Gln Asp Leu Val Ser Phe Met Thr Gln Ala Leu Tyr Leu1 5 10 15Val Leu Trp Leu Ser Leu Pro Pro Ile Ala Val Val Ala Ile Val Gly20 25 30Thr Leu Phe Ser Leu Leu Gln Ala Leu Thr Gln Val Gln Glu Gln Thr35 40 45Leu Ser Phe Ala Val Lys Leu Ile Ala Val Phe Ala Thr Leu Met Leu50 55 60Ala Ala Arg Trp Ile Ser Ala Glu Ile Tyr Asn Phe Thr Ile Ala Val65 70 75 80Phe Asp Ala Phe His Arg Ile His85<210>33<211>801<212>DNA<213>百日咳博德特氏菌
<220>
<221>CDS<222>(1)...(801)<400>33atg cac acg gag ttc aat ttc gtc gag gcg aag gtt ttc ctg gga acg 48Met His Thr Glu Phe Asn Phe Val Glu Ala Lys Val Phe Leu Gly Thr1 5 10 15ctg gcc atg acg caa ccg cgg ata ctc acg gcc atg ctc ttt ctg ccg 96Leu Ala Met Thr Gln Pro Arg Ile Leu Thr Ala Met Leu Phe Leu Pro20 25 30atg ttc aac cgt cag ttt ctg cct ggt ccg ctg cgt tac gcc gtc ggc 144Met Phe Asn Arg Gln Phe Leu Pro Gly Pro Leu Arg Tyr Ala Val Gly35 40 45gcc tgt ctc ggg ctg atc gtg gtt ccc cag ctg gcg ccg cag tat gcc 192Ala Cys Leu Gly Leu Ile Val Val Pro Gln Leu Ala Pro Gln Tyr Ala50 55 60gcg ctg gat atc gac tgg ccc cgg ctg ctg gcg ctg ctg gcc aag gag 240Ala Leu Asp Ile Asp Trp Pro Arg Leu Leu Ala Leu Leu Ala Lys Glu65 70 75 80gcg atg gtg ggc atg ttc ctg ggt tgg ctg gct gcc ttg cca ttc tgg 288Ala Met Val Gly Met Phe Leu Gly Trp Leu Ala Ala Leu Pro Phe Trp85 90 95atc ttc gag gcc atc ggc ttc gtc ata gac aac caa cgg ggc gcc agc 336Ile Phe Glu Ala Ile Gly Phe Val Ile Asp Asn Gln Arg Gly Ala Ser100 105 110ctg ggc gct atc ctc aac ccc gcc acg ggc aac gat tcg tcg ccc atg 384Leu Gly Ala Ile Leu Asn Pro Ala Thr Gly Asn Asp Ser Ser Pro Met115 120 125ggc att ctc ttc aat ctg gga ttc atg gtg ttc ttc ctg acg gcg ggc 432Gly Ile Leu Phe Asn Leu Gly Phe Met Val Phe Phe Leu Thr Ala Gly130 135 140gga ttc ggg ttg ttc gcc acg atg ctg tat gac agc ttc ggg ttg tgg 480Gly Phe Gly Leu Phe Ala Thr Met Leu Tyr Asp Ser Phe Gly Leu Trp145 150 155 160aac atc tgg gcg tgg tgg ccg tcc atg ccc gca cag ggc gcc gtg cgg 528Asn Ile Trp Ala Trp Trp Pro Ser Met Pro Ala Gln Gly Ala Val Arg165 170 175atg ctg gac cag ttc agt ggc ttt gcc gcg cgt gtc ctg ctg ctg gcc 576Met Leu Asp Gln Phe Ser Gly Phe Ala Ala Arg Val Leu Leu Leu Ala180 185 190tcg ccg gcc atc gtg gcc atg ttc ctg gcc gag ctg ggc ctg gcc ctg624Ser Pro Ala Ile Val Ala Met Phe Leu Ala Glu Leu Gly Leu Ala Leu195 200 205atc agc cgc ttc gcg cct caa ctg cag gtg ttc ttc ctg gct ctg ccg672Ile Ser Arg Phe Ala Pro Gln Leu Gln Val Phe Phe Leu Ala Leu Pro210 215 220gta aag agc gcg ctg gtg ctg ttc gtg ctg gtg ctg tac atg gca acg720Val Lys Ser Ala Leu Val Leu Phe Val Leu Val Leu Tyr Met ala Thr225 230 235 240ttg ttc cag tat gca ggc gaa atc ctg ggt tct gtg ggc cgg atc gtg768Leu Phe Gln Tyr Ala Gly Glu Ile Leu Gly Ser Val Gly Arg Ile Val245 250 255ccg ttc ctg cat tca gcg tgg ccc ggc cca tga801Pro Phe Leu His Ser Ala Trp Pro Gly Pro *260 265<210>34<211>266<212>PRT<213>百日咳博德特氏菌<400>34Met His Thr Glu Phe Asn Phe Val Glu Ala Lys Val Phe Leu Gly Thr1 5 10 15Leu Ala Met Thr Gln Pro Arg Ile Leu Thr Ala Met Leu Phe Leu Pro20 25 30Met Phe Asn Arg Gln Phe Leu Pro Gly Pro Leu Arg Tyr Ala Val Gly35 40 45Ala Cys Leu Gly Leu Ile Val Val Pro Gln Leu Ala Pro Gln Tyr Ala50 55 60Ala Leu Asp Ile Asp Trp Pro Arg Leu Leu Ala Leu Leu Ala Lys Glu65 70 75 80Ala Met Val Gly Met Phe Leu Gly Trp Leu Ala Ala Leu Pro Phe Trp85 90 95Ile Phe Glu Ala Ile Gly Phe Val Ile Asp Asn Gln Arg Gly Ala Ser100 105 110Leu Gly Ala Ile Leu Asn Pro Ala Thr Gly Asn Asp Ser Ser Pro Met115 120 125Gly Ile Leu Phe Asn Leu Gly Phe Met Val Phe Phe Leu Thr Ala Gly130 135 140Gly Phe Gly Leu Phe Ala Thr Met Leu Tyr Asp Ser Phe Gly Leu Trp145 150 155 160Asn Ile Trp Ala Trp Trp Pro Ser Met Pro Ala Gln Gly Ala Val Arg
165 170 175Met Leu Asp Gln Phe Ser Gly Phe Ala Ala Arg Val Leu Leu Leu Ala180 185 190Ser Pro Ala Ile Val Ala Met Phe Leu Ala Glu Leu Gly Leu Ala Leu195 200 205Ile Ser Arg Phe Ala Pro Gln Leu Gln Val Phe Phe Leu Ala Leu Pro210 215 220Val Lys Ser Ala Leu Val Leu Phe Val Leu Val Leu Tyr Met ala Thr225 230 235 240Leu Phe Gln Tyr Ala Gly Glu Ile Leu Gly Ser Val Gly Arg Ile Val245 250 255Pro Phe Leu His Ser Ala Trp Pro Gly Pro260 265<210>35<211>1050<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1050)<400>35atg agc ggc gag aaa acc gag cgg ccc acc ccg aag cgc ctg cgc gat48Met Ser Gly Glu Lys Thr Glu Arg Pro Thr Pro Lys Arg Leu Arg Asp15 10 15tcc cgc gag aaa ggc gag gtc gca cac agc cgg gac ttt acc cag acg96Ser Arg Glu Lys Gly Glu Val Ala His Ser Arg Asp Phe Thr Gln Thr20 25 30gcg ctg ata tgc gcc ttg ttc ggg cac ttt ctg atc aat gcc ccg tcc144Ala Leu Ile Cys Ala Leu Phe Gly His Phe Leu Ile Asn Ala Pro Ser35 40 45att ctc gcg tcg ctg cga gcg ctg ata ctg gcg ccg gcg gcc ttt gcc192Ile Leu Ala Ser Leu Arg Ala Leu Ile Leu Ala Pro Ala Ala Phe Ala50 55 60gac cag ggg ttc gcc gtc gca ttg ggg ccc gtg ctg acg gaa atc ctc240Asp Gln Gly Phe Ala Val Ala Leu Gly Pro Val Leu Thr Glu Ile Leu65 70 75 80gat cag gcc gtc cgc gtg ctc gct ccg ctg att ctc atc gtg ctt ggg288Asp Gln Ala Val Arg Val Leu Ala Pro Leu Ile Leu Ile Val Leu Gly85 90 95gtg ggg atg ttc gcc gaa ttc ctg cag gta ggc gtc gtg ctg gcg ttt336Val Gly Met Phe Ala Glu Phe Leu Gln Val Gly Val Val Leu Ala Phe100 105 110cga aag ctc aag cct tcg gcg gag aaa ctg aat ccc gcc ggc aat ttg384Arg Lys Leu Lys Pro Ser Ala Glu Lys Leu Asn Pro Ala Gly Asn Leu115 120 125aag aat atc ttc tcg gcg cgc aac ctg atg gag ttc atc aag tcg gta432Lys Asn Ile Phe Ser Ala Arg Asn Leu Met Glu Phe Ile Lys Ser Val130 135 140tgc aag atc ctg ttt ctg gcg gtg ttg gtc acg ttg gtg ata cgg gat480Cys Lys Ile Leu Phe Leu Ala Val Leu Val Thr Leu Val Ile Arg Asp145 150 155 160tcc ttg cag ccg ctg atg gcc gtt ccc cat agc ggg ctg gac ggg ttg528Ser Leu Gln Pro Leu Met ala Val Pro His Ser Gly Leu Asp Gly Leu165 170 175cga acg ggc gta ggc cgc att ctg cag gtc atg gtc tgg aac atc gga576Arg Thr Gly Val Gly Arg Ile Leu Gln Val Met Val Trp Asn Ile Gly180 185 190ctg gcg tac ggg gcg att tcg ctg gcg gac ctg gcc tgg cag cgt tac624Leu Ala Tyr Gly Ala Ile Ser Leu Ala Asp Leu Ala Trp Gln Arg Tyr195 200 205cag tat cgc aaa ggc ttg cgg atg agc aag gac gaa gtg aag cag gag672Gln Tyr Arg Lys Gly Leu Arg Met Ser Lys Asp Glu Val Lys Gln Glu210 215 220tac aag gag atg gaa ggc gat ccc cat atc aag cag caa cgc aag cac720Tyr Lys Glu Met Glu Gly Asp Pro His Ile Lys Gln Gln Arg Lys His225 230 235 240ctg cac cag gag ctg atc atg cat ggc gcg gcg gcc cag gtt cgc cgg768Leu His Gln Glu Leu Ile Met His Gly Ala Ala Ala Gln Val Arg Arg245 250 255gcg acg gtg ctg gtg acc aat ccg aca cac ctg gcc gtg gcc ctg tac816Ala Thr Val Leu Val Thr Asn Pro Thr His Leu Ala Val Ala Leu Tyr260 265 270tac gcg gcg ggc gag acg ccc ttg ccg cgc gtg ctg gcc atg ggg cag864Tyr Ala Ala Gly Glu Thr Pro Leu Pro Arg Val Leu Ala Met Gly Gln275 280 285gga gcc gtg gcc gct ctc atg gtc gag gcc gcg cgc gat gcc ggc gtg912Gly Ala Val Ala Ala Leu Met Val Glu Ala Ala Arg Asp Ala Gly Val290 295 300ccg gtc atg cag aac gtc gcg ctg gcc cgc gcc ttg cac gac cag gcg960Pro Val Met Gln Asn Val Ala Leu Ala Arg Ala Leu His Asp Gln Ala305 310 315 320gag gtg gac caa tac att ccc ggc gag ttg gtg gag ccg gtg gcc gcg1008Glu Val Asp Gln Tyr Ile Pro Gly Glu Leu Val Glu Pro Val Ala Ala325 330 335gtg ttg cgg gcg gtg cgc cag gca ctc aag gag cag aca tga1050Val Leu Arg Ala Val Arg Gln Ala Leu Lys Glu Gln Thr *340 345<210>36<211>349<212>PRT<213>百日咳博德特氏菌<400>36Met Ser Gly Glu Lys Thr Glu Arg Pro Thr Pro Lys Arg Leu Arg Asp1 5 10 15Ser Arg Glu Lys Gly Glu Val Ala His Ser Arg Asp Phe Thr Gln Thr20 25 30Ala Leu Ile Cys Ala Leu Phe Gly His Phe Leu Ile Asn Ala Pro Ser35 40 45Ile Leu Ala Ser Leu Arg Ala Leu Ile Leu Ala Pro Ala Ala Phe Ala50 55 60Asp Gln Gly Phe Ala Val Ala Leu Gly Pro Val Leu Thr Glu Ile Leu65 70 75 80Asp Gln Ala Val Arg Val Leu Ala Pro Leu Ile Leu Ile Val Leu Gly85 90 95Val Gly Met Phe Ala Glu Phe Leu Gln Val Gly Val Val Leu Ala Phe100 105 110Arg Lys Leu Lys Pro Ser Ala Glu Lys Leu Asn Pro Ala Gly Asn Leu115 120 125Lys Asn Ile Phe Ser Ala Arg Asn Leu Met Glu Phe Ile Lys Ser Val130 135 140Cys Lys Ile Leu Phe Leu Ala Val Leu Val Thr Leu Val Ile Arg Asp145 150 155 160Ser Leu Gln Pro Leu Met ala Val Pro His Ser Gly Leu Asp Gly Leu165 170 175Arg Thr Gly Val Gly Arg Ile Leu Gln Val Met Val Trp Asn Ile Gly180 185 190Leu Ala Tyr Gly Ala Ile Ser Leu Ala Asp Leu Ala Trp Gln Arg Tyr195 200 205Gln Tyr Arg Lys Gly Leu Arg Met Ser Lys Asp Glu Val Lys Gln Glu210 215 220Tyr Lys Glu Met Glu Gly Asp Pro His Ile Lys Gln Gln Arg Lys His225 230 235 240Leu His Gln Glu Leu Ile Met His Gly Ala Ala Ala Gln Val Arg Arg245 250 255Ala Thr Val Leu Val Thr Asn Pro Thr His Leu Ala Val Ala Leu Tyr260 265 270Tyr Ala Ala Gly Glu Thr Pro Leu Pro Arg Val Leu Ala Met Gly Gln275 280 285Gly Ala Val Ala Ala Leu Met Val Glu Ala Ala Arg Asp Ala Gly Val290 295 300Pro Val Met Gln Asn Val Ala Leu Ala Arg Ala Leu His Asp Gln Ala305 310 315 320Glu Val Asp Gln Tyr Ile Pro Gly Glu Leu Val Glu Pro Val Ala Ala325 330 335Val Leu Arg Ala Val Arg Gln Ala Leu Lys Glu Gln Thr340 345<210>37<211>399<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(399)<400>37atg aca gca acc att cat ccc gat att gcc gat tat gcg cga cgc cat48Met Thr Ala Thr Ile His Pro Asp Ile Ala Asp Tyr Ala Arg Arg His15 10 15ggc ctc gaa ccc tcg gtc gac gcc gat ggc ggg ctt gcc gtc cgg atc96Gly Leu Glu Pro Ser Val Asp Ala Asp Gly Gly Leu Ala Val Arg Ile20 25 30gac gga cgg cat cgc gtc agg ttg atc ccc gcc gaa gac ggc atg ctg144Asp Gly Arg His Arg Val Arg Leu Ile Pro Ala Glu Asp Gly Met Leu35 40 45gtg ttg cgg gcg cgg ctg gcc gag ctg ccc gat ggg tgg cag gcg cgc192Val Leu Arg Ala Arg Leu Ala Glu Leu Pro Asp Gly Trp Gln Ala Arg50 55 60gcg gcg cag ttg cgc cgg gcg ggc ctg ctg gcc agc gcc atg gcc cct240Ala Ala Gln Leu Arg Arg Ala Gly Leu Leu Ala Ser Ala Met ala Pro65 70 75 80gcg acc gat gcg tac tgc ggc ata gac cag ggc gaa acc gcg ttg tat288Ala Thr Asp Ala Tyr Cys Gly Ile Asp Gln Gly Glu Thr Ala Leu Tyr85 90 95ctg cac cag cgc gtc gca ccg gcc ggc agt gcg ctg gcg gtg gac gag336Leu His Gln Arg Val Ala Pro Ala Gly Ser Ala Leu Ala Val Asp Glu100 105 110gcg gtg ggc gag ttc gtc aat gcc ttg gcc act tgg aaa agg gcg atg384Ala Val Gly Glu Phe Val Asn Ala Leu Ala Thr Trp Lys Arg Ala Met
115 120 125gcg caa tgg caa tag399Ala Gln Trp Gln *130<210>38<211>132<212>PRT<213>百日咳博德特氏菌<400>38Met Thr Ala Thr Ile His Pro Asp Ile Ala Asp Tyr Ala Arg Arg His1 5 10 15Gly Leu Glu Pro Ser Val Asp Ala Asp Gly Gly Leu Ala Val Arg Ile20 25 30Asp Gly Arg His Arg Val Arg Leu Ile Pro Ala Glu Asp Gly Met Leu35 40 45Val Leu Arg Ala Arg Leu Ala Glu Leu Pro Asp Gly Trp Gln Ala Arg50 55 60Ala Ala Gln Leu Arg Arg Ala Gly Leu Leu Ala Ser Ala Met ala Pro65 70 75 80Ala Thr Asp Ala Tyr Cys Gly Ile Asp Gln Gly Glu Thr Ala Leu Tyr85 90 95Leu His Gln Arg Val Ala Pro Ala Gly Ser Ala Leu Ala Val Asp Glu100 105 110Ala Val Gly Glu Phe Val Asn Ala Leu Ala Thr Trp Lys Arg Ala Met115 120 125Ala Gln Trp Gln130<210>39<211>603<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(603)<400>39atg gtt tct ccc ccg tca tcc ggt ctt ccc gct tct ctc gaa aaa ccg48Met Val Ser Pro Pro Ser Ser Gly Leu Pro Ala Ser Leu Glu Lys Pro1 5 10 15gac aac gca tat ccc gat atc gcc acc gag cga tcg gac cag cag ttg96Asp Asn Ala Tyr Pro Asp Ile Ala Thr Glu Arg Ser Asp Gln Gln Leu20 25 30ctg agc agc ctg gta gcc gaa cat gcc ggc cga tta cag aga ttc atc144Leu Ser Ser Leu Val Ala Glu His Ala Gly Arg Leu Gln Arg Phe Ile35 40 45gcc aag cac atc ggc cac agc agc gac gtc gag gac ctt gcg cag cag192Ala Lys His Ile Gly His Ser Ser Asp Val Glu Asp Leu Ala Gln Gln50 55 60gct ttc gcc gag gcg gcg cgc gcg tat caa tcg ttc cgt ggc gac tcc240Ala Phe Ala Glu Ala Ala Arg Ala Tyr Gln Ser Phe Arg Gly Asp Ser65 70 75 80cag ctt tcc acc tgg ctg tac ggc atc gcg ctc aat ctg gtc cgc aat288Gln Leu Ser Thr Trp Leu Tyr Gly Ile Ala Leu Asn Leu Val Arg Asn85 90 95cac ttg tcg cgt gcg cca gag cgc cgt tat gaa ttc acc tcc gac gcc336His Leu Ser Arg Ala Pro Glu Arg Arg Tyr Glu Phe Thr Ser Asp Ala100 105 110agc ctg ggt gtc atg cca tgc agt gcg ccc aac ccc gaa gcc gtg acc384Ser Leu Gly Val Met Pro Cys Ser Ala Pro Asn Pro Glu Ala Val Thr115 120 125gag cag cgt caa cgc atg cgc ttg cta cgc gaa gcg ctg gag cag ctc432Glu Gln Arg Gln Arg Met Arg Leu Leu Arg Glu Ala Leu Glu Gln Leu130 135 140ccc gaa agc atg cgc gac gtg atc ctc atg gtc ggc gtg gaa gaa ctc480Pro Glu Ser Met Arg Asp Val Ile Leu Met Val Gly Val Glu Glu Leu145 150 155 160tcc tat gaa gag gct gcc gca ctg ctt tcg gtt cct gta gga acc att528Ser Tyr Glu Glu Ala Ala Ala Leu Leu Ser Val Pro Val Gly Thr Ile165 170 175cgc agc cga ctt tcc cgc gcc cgc tgt gcc ttg cgc gaa gcg ctg cgc576Arg Ser Arg Leu Ser Arg Ala Arg Cys Ala Leu Arg Glu Ala Leu Arg180 185 190gaa cga ggc tac gac agc gtg ccg tag603Glu Arg Gly Tyr Asp Ser Val Pro *195 200<210>40<211>200<212>PRT<213>百日咳博德特氏菌<400>40Met Val Ser Pro Pro Ser Ser Gly Leu Pro Ala Ser Leu Glu Lys Pro1 5 10 15Asp Asn Ala Tyr Pro Asp Ile Ala Thr Glu Arg Ser Asp Gln Gln Leu20 25 30Leu Ser Ser Leu Val Ala Glu His Ala Gly Arg Leu Gln Arg Phe Ile35 40 45Ala Lys His Ile Gly His Ser Ser Asp Val Glu Asp Leu Ala Gln Gln50 55 60Ala Phe Ala Glu Ala Ala Arg Ala Tyr Gln Ser Phe Arg Gly Asp Ser65 70 75 80Gln Leu Ser Thr Trp Leu Tyr Gly Ile Ala Leu Asn Leu Val Arg Asn85 90 95His Leu Ser Arg Ala Pro Glu Arg Arg Tyr Glu Phe Thr Ser Asp Ala100 105 110Ser Leu Gly Val Met Pro Cys Ser Ala Pro Asn Pro Glu Ala Val Thr115 120 125Glu Gln Arg Gln Arg Met Arg Leu Leu Arg Glu Ala Leu Glu Gln Leu130 135 140Pro Glu Ser Met Arg Asp Val Ile Leu Met Val Gly Val Glu Glu Leu145 150 155 160Ser Tyr Glu Glu Ala Ala Ala Leu Leu Ser Val Pro Val Gly Thr Ile165 170 175Arg Ser Arg Leu Ser Arg Ala Arg Cys Ala Leu Arg Glu Ala Leu Arg180 185 190Glu Arg Gly Tyr Asp Ser Val Pro195 200<210>41<211>1098<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1098)<400>41atg act cgt atc gat gcc gcc ccc aat ccc ttc cac gcc gcc atg cag48Met Thr Arg Ile Asp Ala Ala Pro Asn Pro Phe His Ala Ala Met Gln1 5 10 15ggg cgc cac gac gcc tcg gcc aac acc tcc tcc ggc tgg ctg caa ggc96Gly Arg His Asp Ala Ser Ala Asn Thr Ser Ser Gly Trp Leu Gln Gly20 25 30cag cgc atc gca ccg gcg ccc acc ggc ata tcg ctg gcg gac gcg gcc144Gln Arg Ile Ala Pro Ala Pro Thr Gly Ile Ser Leu Ala Asp Ala Ala35 40 45gag gag ctc agc ctg cac atg gcg cag gct gcc gag gaa aag cat cac192Glu Glu Leu Ser Leu His Met ala Gln Ala Ala Glu Glu Lys His His50 55 60tcc gaa cgc aag gtc acg gcc gaa cgt ccg atg ctc tgg ctg gac gcg 240Ser Glu Arg Lys Val Thr Ala Glu Arg Pro Met Leu Trp Leu Asp Ala65 70 75 80gcg cag ctt gcg gaa ctg ttt tcc cac acc cac gac ccc gac gcg cag 288Ala Gln Leu Ala Glu Leu Phe Ser His Thr His Asp Pro Asp Ala Gln85 90 95gca aaa ctg gaa gcc ctg acc gcc gag ctg ctg cgc ggc cgg ggc gcc 336Ala Lys Leu Glu Ala Leu Thr Ala Glu Leu Leu Arg Gly Arg Gly Ala100 105 110ccc atg cag ctg gcc gcg caa gcg ttt ccc ggt gtc acg cag caa tac 384Pro Met Gln Leu Ala Ala Gln Ala Phe Pro Gly Val Thr Gln Gln Tyr115 120 125ctc gcg ctg cag cac gcg ctg cag cgc ggc gag cac gag gac gcc gcg 432Leu Ala Leu Gln His Ala Leu Gln Arg Gly Glu His Glu Asp Ala Ala130 135 140ccg cac gcg ctc gaa gcc ctg cgc gat gca ttg gcc gac ctg gag ctc 480Pro His Ala Leu Glu Ala Leu Arg Asp Ala Leu Ala Asp Leu Glu Leu145 150 155 160gcc cat ggc ccc gaa atc cgc gcc ggc atc aac acc ctg ccc acg gcc 528Ala His Gly Pro Glu Ile Arg Ala Gly Ile Asn Thr Leu Pro Thr Ala165 170 175ggc gca ttc gcg cgt tcc gct gac gag ctg gcc ggc ttc cag cac gcg 576Gly Ala Phe Ala Arg Ser Ala Asp Glu Leu Ala Gly Phe Gln His Ala180 185 190tac cgc gac atc gcc ctg ggc cag ctg tcg ttg gcg cgc acg ctg gac 624Tyr Arg Asp Ile Ala Leu Gly Gln Leu Ser Leu Ala Arg Thr Leu Asp195 200 205ctg gtg ctg gaa cgc tat ggg aac gac gac atc cac ggc gcg ctg ggc 672Leu Val Leu Glu Arg Tyr Gly Asn Asp Asp Ile His Gly Ala Leu Gly210 215 220gcg ctg att cag gcg ctg gga cac gac ctg gcc gcg gcg aca ccg tcg 720Ala Leu Ile Gln Ala Leu Gly His Asp Leu Ala Ala Ala Thr Pro Ser225 230 235 240acg gac ggc gtc agg ctg caa gtg ttg gcg agc gat ctc tat caa gtc 768Thr Asp Gly Val Arg Leu Gln Val Leu Ala Ser Asp Leu Tyr Gln Val245 250 255gag gtg gcc gcc acg gta ctg gag gaa tgc aat gcc ctg aaa caa cgg 816Glu Val Ala Ala Thr Val Leu Glu Glu Cys Asn Ala Leu Lys Gln Arg
260 265 270ttg ggc aac gca ggc tcg cag gag tgt gcg gac gcc cag ggc ctg atg864Leu Gly Asn Ala Gly Ser Gln Glu Cys Ala Asp Ala Gln Gly Leu Met275 280 285cgc gat ctt gtg gga atc agc gag gac aaa tgg att gcg ccc gcg agc912Arg Asp Leu Val Gly Ile Ser Glu Asp Lys Trp Ile Ala Pro Ala Arg290 295 300ttc gag aag ctg gcc gag cgc cac ggt gcg aat gcc ctc tcc gag cgc960Phe Glu Lys Leu Ala Glu Arg His Gly Ala Asn Ala Leu Ser Glu Arg305 310 315 320atc gca ttc ctc ggt ggc gta cgc cag att ctc aaa gac ctg ccc acg1008Ile Ala Phe Leu Gly Gly Val Arg Gln Ile Leu Lys Asp Leu Pro Thr325 330 335cag atc tac gcc gac atg gac gtg cgc gcc acc gtc ctg gcg gcc gcg1056Gln Ile Tyr Ala Asp Met Asp Val Arg Ala Thr Val Leu Ala Ala Ala340 345 350cag gac gcg ctg gac aac gcg ata gca atg gag aac gca tga1098Gln Asp Ala Leu Asp Asn Ala Ile Ala Met Glu Asn Ala *355 360 365<210>42<211>365<212>PRT<213>百日咳博德特氏菌<400>42Met Thr Arg Ile Asp Ala Ala Pro Asn Pro Phe His Ala Ala Met Gln1 5 10 15Gly Arg His Asp Ala Ser Ala Asn Thr Ser Ser Gly Trp Leu Gln Gly20 25 30Gln Arg Ile Ala Pro Ala Pro Thr Gly Ile Ser Leu Ala Asp Ala Ala35 40 45Glu Glu Leu Ser Leu His Met ala Gln Ala Ala Glu Glu Lys His His50 55 60Ser Glu Arg Lys Val Thr Ala Glu Arg Pro Met Leu Trp Leu Asp Ala65 70 75 80Ala Gln Leu Ala Glu Leu Phe Ser His Thr His Asp Pro Asp Ala Gln85 90 95Ala Lys Leu Glu Ala Leu Thr Ala Glu Leu Leu Arg Gly Arg Gly Ala100 105 110Pro Met Gln Leu Ala Ala Gln Ala Phe Pro Gly Val Thr Gln Gln Tyr115 120 125Leu Ala Leu Gln His Ala Leu Gln Arg Gly Glu His Glu Asp Ala Ala130 135 140Pro nis Ala Leu Glu Ala Leu Arg Asp Ala Leu Ala Asp Leu Glu Leu145 150 155 160Ala His Gly Pro Glu Ile Arg Ala Gly Ile Asn Thr Leu Pro Thr Ala165 170 175Gly Ala Phe Ala Arg Ser Ala Asp Glu Leu Ala Gly Phe Gln His Ala180 185 190Tyr Arg Asp Ile Ala Leu Gly Gln Leu Ser Leu Ala Arg Thr Leu Asp195 200 205Leu Val Leu Glu Arg Tyr Gly Asn Asp Asp Ile His Gly Ala Leu Gly210 215 220Ala Leu Ile Gln Ala Leu Gly His Asp Leu Ala Ala Ala Thr Pro Ser225 230 235 240Thr Asp Gly Val Arg Leu Gln Val Leu Ala Ser Asp Leu Tyr Gln Val245 250 255Glu Val Ala Ala Thr Val Leu Glu Glu Cys Asn Ala Leu Lys Gln Arg260 265 270Leu Gly Asn Ala Gly Ser Gln Glu Cys Ala Asp Ala Gln Gly Leu Met275 280 285Arg Asp Leu Val Gly Ile Ser Glu Asp Lys Trp Ile Ala Pro Ala Arg290 295 300Phe Glu Lys Leu Ala Glu Arg His Gly Ala Asn Ala Leu Ser Glu Arg305 310 315 320Ile Ala Phe Leu Gly Gly Val Arg Gln Ile Leu Lys Asp Leu Pro Thr325 330 335Gln Ile Tyr Ala Asp Met Asp Val Arg Ala Thr Val Leu Ala Ala Ala340 345 350Gln Asp Ala Leu Asp Asn Ala Ile Ala Met Glu Asn Ala355 360 365<210>43<211>588<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(588)<400>43atg ccc ttg gcg ggg tcc tgc ggc cgg acg tcc gaa aac cgg gat att48Met Pro Leu Ala Gly Ser Cys Gly Arg Thr Ser Glu Asn Arg Asp Ile1 5 10 15ccg aga agc aat cgg ctg gcg cct tcc aga ctg tgc gca tac cat tgc96Pro Arg Ser Asn Arg Leu Ala Pro Ser Arg Leu Cys Ala Tyr His Cys20 25 30cct ctt ttg cca cgc att tcg agc gta tgg ttt ccc ttg cgc ccg acg144Pro Leu Leu Pro Arg Ile Ser Ser Val Trp Phe Pro Leu Arg Pro Thr35 40 45cag gct cgc ctc gcc atg acc gac acg gca tac cac caa ctc atc gcc192Gln Ala Arg Leu Ala Met Thr Asp Thr Ala Tyr His Gln Leu Ile Ala50 55 60gat ttc ggc cgc ctc atc ggc atc gac tcg ctc aac ccc ggt gcc ggc240Asp Phe Gly Arg Leu Ile Gly Ile Asp Ser Leu Asn Pro Gly Ala Gly65 70 75 80ggc ctg tgt cag ttg att ttc gaa ccg tgc gca ccg gtc ttc atc gca288Gly Leu Cys Gln Leu Ile Phe Glu Pro Cys Ala Pro Val Phe Ile Ala85 90 95ccg gtg cac gcc cgg acg gaa atc atg att tcc tgc gtg ctg ggc acg336Pro Val His Ala Arg Thr Glu Ile Met Ile Ser Cys Val Leu Gly Thr100 105 110gcg gac gcg gcc aac ccg gca agc atg gcc cga gcc aac ttc atg cag384Ala Asp Ala Ala Asn Pro Ala Ser Met ala Arg Ala Asn Phe Met Gln115 120 125gcc ggc agc ggc gtc gtg gcc tgc atc ggc ggc gat ggg ttg ttc tat432Ala Gly Ser Gly Val Val Ala Cys Ile Gly Gly Asp Gly Leu Phe Tyr130 135 140ctg cag cag gcc ata ccc ctg tcg cgc gcc acg ccc gca atc ctg ctc480Leu Gln Gln Ala Ile Pro Leu Ser Arg Ala Thr Pro Ala Ile Leu Leu145 150 155 160gat cac tgt gag cgt ctg ctg cag gaa gcc tcg cgc tgg cgc gtc ggc528Asp His Cys Glu Arg Leu Leu Gln Glu Ala Ser Arg Trp Arg Val Gly165 170 175gac cac gac ggc tgc gcc acc tcg gcc ccg aat atc gcc gcg ctg acg576Asp His Asp Gly Cys Ala Thr Ser Ala Pro Asn Ile Ala Ala Leu Thr180 185 190cgc ggc gtc tag588Arg Gly Val *195<210>44<211>195<212>PRT<213>百日咳博德特氏菌<400>44Met Pro Leu Ala Gly Ser Cys Gly Arg Thr Ser Glu Asn Arg Asp Ile1 5 10 15Pro Arg Ser Asn Arg Leu Ala Pro Ser Arg Leu Cys Ala Tyr His Cys20 25 30Pro Leu Leu Pro Arg Ile Ser Ser Val Trp Phe Pro Leu Arg Pro Thr35 40 45Gln Ala Arg Leu Ala Met Thr Asp Thr Ala Tyr His Gln Leu Ile Ala50 55 60Asp Phe Gly Arg Leu Ile Gly Ile Asp Ser Leu Asn Pro Gly Ala Gly65 70 75 80Gly Leu Cys Gln Leu Ile Phe Glu Pro Cys Ala Pro Val Phe Ile Ala85 90 95Pro Val His Ala Arg Thr Glu Ile Met Ile Ser Cys Val Leu Gly Thr100 105 110Ala Asp Ala Ala Asn Pro Ala Ser Met ala Arg Ala Asn Phe Met Gln115 120 125Ala Gly Ser Gly Val Val Ala Cys Ile Gly Gly Asp Gly Leu Phe Tyr130 135 140Leu Gln Gln Ala Ile Pro Leu Ser Arg Ala Thr Pro Ala Ile Leu Leu145 150 155 160Asp His Cys Glu Arg Leu Leu Gln Glu Ala Ser Arg Trp Arg Val Gly165 170 175Asp His Asp Gly Cys Ala Thr Ser Ala Pro Asn Ile Ala Ala Leu Thr180 185 190Arg Gly Val195<210>45<211>369<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(369)<400>45atg atg ccg cat acc cta ccc tcg ccc agc ctt cag gta cgc gaa ctt48Met Met Pro His Thr Leu Pro Ser Pro Ser Leu Gln Val Arg Glu Leu1 5 10 15ctg caa ttg ctt gcg cac cac tac cag ttg cag cgc caa tgg agc aag96Leu Gln Leu Leu Ala His His Tyr Gln Leu Gln Arg Gln Trp Ser Lys20 25 30acg gtt gcc ctg ctg gcg gcc ctg gat gcc ctg gac gcg atc gac agc144Thr Val Ala Leu Leu Ala Ala Leu Asp Ala Leu Asp Ala Ile Asp Ser35 40 45cag tcc ctg ctg gcc ctg gcg ctg ggc tat ctg cac cag ggc gaa ccg192Gln Ser Leu Leu Ala Leu Ala Leu Gly Tyr Leu His Gln Gly Glu Pro50 55 60cgc atg gcc ttg gtc acg ctg gac aag cgc gca ctg cgc gcc aca ccc240Arg Met ala Leu Val Thr Leu Asp Lys Arg Ala Leu Arg Ala Thr Pro65 70 75 80gat gcc gcc ggc cat ctg gtc cgg gcg cag gcc atg cag gcg ctg aac288Asp Ala Ala Gly His Leu Val Arg Ala Gln Ala Met Gln Ala Leu Asn85 90 95cga ccc gac gac gcc cgc cag gcc atg cgt gac tat atg gcg cta cgg336Arg Pro Asp Asp Ala Arg Gln Ala Met Arg Asp Tyr Met ala Leu Arg100 105 110gcg gcg tct tgc gcg agc gcg acc ccg cca tga369Ala Ala Ser Cys Ala Ser Ala Thr Pro Pro *115 120<210>46<211>122<212>PRT<213>百日咳博德特氏菌<400>46Met Met Pro His Thr Leu Pro Ser Pro Ser Leu Gln Val Arg Glu Leu15 10 15Leu Gln Leu Leu Ala His His Tyr Gln Leu Gln Arg Gln Trp Ser Lys20 25 30Thr Val Ala Leu Leu Ala Ala Leu Asp Ala Leu Asp Ala Ile Asp Ser35 40 45Gln Ser Leu Leu Ala Leu Ala Leu Gly Tyr Leu His Gln Gly Glu Pro50 55 60Arg Met ala Leu Val Thr Leu Asp Lys Arg Ala Leu Arg Ala Thr Pro65 70 75 80Asp Ala Ala Gly His Leu Val Arg Ala Gln Ala Met Gln Ala Leu Asn85 90 95Arg Pro Asp Asp Ala Arg Gln Ala Met Arg Asp Tyr Met ala Leu Arg100 105 110Ala Ala Ser Cys Ala Ser Ala Thr Pro Pro115 120<210>47<211>411<212>DNA<213>百日咳博德特氏菌<220>
<22l>CDS<222>(1)...(411)<400>47atg tcc agc gcc gta ccc ggc atg cat gcc atg cac ctc ggc ctg gag48Met Ser Ser Ala Val Pro Gly Met His Ala Met His Leu Gly Leu Glu1 5 10 15cgc ggc gtc gac cac atc gtg cgc ggt ccc cgc tgc gag ccc gcc ccc96Arg Gly Val Asp His Ile Val Arg Gly Pro Arg Cys Glu Pro Ala Pro20 25 30acc ctg cca ccc gag cgc tgg ctc gaa ccg ccc gcc acc ggc gcg gtc144Thr Leu Pro Pro Glu Arg Trp Leu Glu Pro Pro Ala Thr Gly Ala Val35 40 45gat cat ctg aaa gcc ctg ctc gta cgc ccg gac ctg agc gcg atg ctc192Asp His Leu Lys Ala Leu Leu Val Arg Pro Asp Leu Ser Ala Met Leu50 55 60gac gag tcg gcg cgg cct cgc ctg acg gat ggc gca tta ttc cag ccc240Asp Glu Ser Ala Arg Pro Arg Leu Thr Asp Gly Ala Leu Phe Gln Pro65 70 75 80gcg cag ttc gag cgc gcc ctg gcc cag gcg cgc gac gaa ctg tcc cgg288Ala Gln Phe Glu Arg Ala Leu Ala Gln Ala Arg Asp Glu Leu Ser Arg85 90 95gcc atg gaa ctg cat gcc ggc aac acc gcg cca gcc tta agc cgc gcc336Ala Met Glu Leu His Ala Gly Asn Thr Ala Pro Ala Leu Ser Arg Ala100 105 110ttg cac gta ctc aac gag gcc gga aag ctg cgc gac ctg gct gcc atg384Leu His Val Leu Asn Glu Ala Gly Lys Leu Arg Asp Leu Ala Ala Met115 120 125tat cgc agc gcg ctc tac cag gga tga411Tyr Arg Ser Ala Leu Tyr Gln Gly *130 135<210>48<211>136<212>PRT<213>百日咳博德特氏菌<400>48Met Ser Ser Ala Val Pro Gly Met His Ala Met His Leu Gly Leu Glu1 5 10 15Arg Gly Val Asp His Ile Val Arg Gly Pro Arg Cys Glu Pro Ala Pro20 25 30Thr Leu Pro Pro Glu Arg Trp Leu Glu Pro Pro Ala Thr Gly Ala Val35 40 45Asp His Leu Lys Ala Leu Leu Val Arg Pro Asp Leu Ser Ala Met Leu50 55 60Asp Glu Ser Ala Arg Pro Arg Leu Thr Asp Gly Ala Leu Phe Gln Pro65 70 75 80Ala Gln Phe Glu Arg Ala Leu Ala Gln Ala Arg Asp Glu Leu Ser Arg
85 90 95Ala Met Glu Leu His Ala Gly Asn Thr Ala Pro Ala Leu Ser Arg Ala100 105 110Leu His Val Leu Asn Glu Ala Gly Lys Leu Arg Asp Leu Ala Ala Met115 120 125Tyr Arg Ser Ala Leu Tyr Gln Gly130 135<210>49<211>378<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(378)<400>49atg aat act gcc gat agg gcg ctg cat cag ttc ggc cag gat atc ggc48Met Asn Thr Ala Asp Arg Ala Leu His Gln Phe Gly Gln Asp Ile Gly1 5 10 15atc gag ggc ctg gca ttc ggg ccg tcc gga tcg gcg tcg ctg gcg ctg96Ile Glu Gly Leu Ala Phe Gly Pro Ser Gly Ser Ala Ser Leu Ala Leu20 25 30tcc aac ggg cgc cgc ctg ggc gtc gaa tgc gtc gcc ggc gcg gcc ctg144Ser Asn Gly Arg Arg Leu Gly Val Glu Cys Val Ala Gly Ala Ala Leu35 40 45gtc cac ctg gcc cag cgg gtc gag cgc gac gcc gcc tcc gtg ttg ctg192Val His Leu Ala Gln Arg Val Glu Arg Asp Ala Ala Ser Val Leu Leu50 55 60gcg gca tgg aaa cgg gcc cat ggg cag cgc gga agc gcc gca tcc atc240Ala Ala Trp Lys Arg Ala His Gly Gln Arg Gly Ser Ala Ala Ser Ile65 70 75 80cag acg tca ctc tgg tcg gag ggc agc gag gac tgg atc gtc gcg cag288Gln Thr Ser Leu Trp Ser Glu Gly Ser Glu Asp Trp Ile Val Ala Gln85 90 95aca cga ctg ccc gaa cgc tcg ctc gac gca gcg gcg ttg cgc ctg gcg336Thr Arg Leu Pro Glu Arg Ser Leu Asp Ala Ala Ala Leu Arg Leu Ala100 105 110gtg ctg ggc ctg acg aac tgg ctc gac cgc ctg gag gcg tga378Val Leu Gly Leu Thr Asn Trp Leu Asp Arg Leu Glu Ala *115 120 125
<210>50<211>125<212>PRT<213>百日咳博德特氏菌<400>50Met Asn Thr Ala Asp Arg Ala Leu His Gln Phe Gly Gln Asp Ile Gly1 5 10 15Ile Glu Gly Leu Ala Phe Gly Pro Ser Gly Ser Ala Ser Leu Ala Leu20 25 30Ser Asn Gly Arg Arg Leu Gly Val Glu Cys Val Ala Gly Ala Ala Leu35 40 45Val His Leu Ala Gln Arg Val Glu Arg Asp Ala Ala Ser Val Leu Leu50 55 60Ala Ala Trp Lys Arg Ala His Gly Gln Arg Gly Ser Ala Ala Ser Ile65 70 75 80Gln Thr Ser Leu Trp Ser Glu Gly Ser Glu Asp Trp Ile Val Ala Gln85 90 95Thr Arg Leu Pro Glu Arg Ser Leu Asp Ala Ala Ala Leu Arg Leu Ala100 105 110Val Leu Gly Leu Thr Asn Trp Leu Asp Arg Leu Glu Ala115 120 125<210>51<211>783<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(783)<400>51atg ggg agt cct cgg aga agg aac cat ttg cct act ggt gca gtg agt48Met Gly Ser Pro Arg Arg Arg Asn His Leu Pro Thr Gly Ala Val Ser1 5 10 15gtc gcg cgc gcg gtc atg gtt ccc gga aac ggg cgc gat att ggg caa96Val Ala Arg Ala Val Met Val Pro Gly Asn Gly Arg Asp Ile Gly Gln20 25 30ttc gca gcc tgg aac ttg ccg cgg gcg cag ggt tac tca gca tgc gtc144Phe Ala Ala Trp Asn Leu Pro Arg Ala Gln Gly Tyr Ser Ala Cys Val35 40 45ttt caa ctc gaa gga gct ctc atg agc att gat ctc gga gtt tca ctc192Phe Gln Leu Glu Gly Ala Leu Met Ser Ile Asp Leu Gly Val Ser Leu50 55 60acg tcg cag gcc ggc ggc ctg caa ggc atc gac ctc aag agc atg gat240Thr Ser Gln Ala Gly Gly Leu Gln Gly Ile Asp Leu Lys Ser Met Asp65 70 75 80atc cag act ctc atg gtg tat gtg cag ggt cgt cgc gcc gaa ctc ctc 288Ile Gln Thr Leu Met Val Tyr Val Gln Gly Arg Arg Ala Glu Leu Leu85 90 95acg gct caa atg cag acc cag gcc gaa gtg gtg cag aag gcc aat gaa 336Thr Ala Gln Met Gln Thr Gln Ala Glu Val Val Gln Lys Ala Asn Glu100 105 110cgc atg gcg cag ctc aac gag gtc ctg tcc gcg ctg tcc cgg gcc aag 384Arg Met ala Gln Leu Asn Glu Val Leu Ser Ala Leu Ser Arg Ala Lys115 120 125gcc gag ttt ccg ccc aat ccg aag ccg ggc gac acc atc ccg ggc tgg 432Ala Glu Phe Pro Pro Asn Pro Lys Pro Gly Asp Thr Ile Pro Gly Trp130 135 140gac agc cag aag atc agc cgg atc gag gtt cct ctc aat gat gcg ctg 480Asp Ser Gln Lys Ile Ser Arg Ile Glu Val Pro Leu Asn Asp Ala Leu145150 155 160cgt gcc gcc ggc ctg acg ggc atg ttc gaa gcg cgc gat ggc cgg gtg 528Arg Ala Ala Gly Leu Thr Gly Met Phe Glu Ala Arg Asp Gly Arg Val165 170 175acc ggc ccc gac ggc cgg ggt acg cag gtc gtg aac ggc acg ggc gtc 576Thr Gly Pro Asp Gly Arg Gly Thr Gln Val Val Asn Gly Thr Gly Val180 185 190atg gcc ggt tcc acg acc tat aag gaa ctc gaa agt gcc tac acc acc 624Met ala Gly Ser Thr Thr Tyr Lys Glu Leu Glu Ser Ala Tyr Thr Thr195 200 205gta aag ggg atg ctg gat acg gcg tcc aat acg caa cag atg gac atg 672Val Lys Gly Met Leu Asp Thr Ala Ser Asn Thr Gln Gln Met Asp Met210 215 220atc agg ctg cag gcc gcc agc aac aag cgc aac gag gct ttc gag gtc 720Ile Arg Leu Gln Ala Ala Ser Asn Lys Arg Asn Glu Ala Phe Glu Val225 230 235 240atg acc aac acc gag aag cgg cgc agc gac ttg aac agc tcc atc acc 768Met Thr Asn Thr Glu Lys Arg Arg Ser Asp Leu Asn Ser Ser Ile Thr245 250 255agc aac atg cgc taa 783Ser Asn Met Arg *260
<210>52<211>260<212>PRT<213>百日咳博德特氏菌<400>52Met Gly Ser Pro Arg Arg Arg Asn His Leu Pro Thr Gly Ala Val Ser1 5 10 15Val Ala Arg Ala Val Met Val Pro Gly Asn Gly Arg Asp Ile Gly Gln20 25 30Phe Ala Ala Trp Asn Leu Pro Arg Ala Gln Gly Tyr Ser Ala Cys Val35 40 45Phe Gln Leu Glu Gly Ala Leu Met Ser Ile Asp Leu Gly Val Ser Leu50 55 60Thr Ser Gln Ala Gly Gly Leu Gln Gly Ile Asp Leu Lys Ser Met Asp65 70 75 80Ile Gln Thr Leu Met Val Tyr Val Gln Gly Arg Arg Ala Glu Leu Leu85 90 95Thr Ala Gln Met Gln Thr Gln Ala Glu Val Val Gln Lys Ala Asn Glu100 105 110Arg Met ala Gln Leu Asn Glu Val Leu Ser Ala Leu Ser Arg Ala Lys115 120 125Ala Glu Phe Pro Pro Asn Pro Lys Pro Gly Asp Thr Ile Pro Gly Trp130 135 140Asp Ser Gln Lys Ile Ser Arg Ile Glu Val Pro Leu Asn Asp Ala Leu145 150 155 160Arg Ala Ala Gly Leu Thr Gly Met Phe Glu Ala Arg Asp Gly Arg Val165 170 175Thr Gly Pro Asp Gly Arg Gly Thr Gln Val Val Asn Gly Thr Gly Val180 185 190Met ala Gly Ser Thr Thr Tyr Lys Glu Leu Glu Ser Ala Tyr Thr Thr195 200 205Val Lys Gly Met Leu Asp Thr Ala Ser Asn Thr Gln Gln Met Asp Met210 215 220Ile Arg Leu Gln Ala Ala Ser Asn Lys Arg Asn Glu Ala Phe Glu Val225 230 235 240Met Thr Asn Thr Glu Lys Arg Arg Ser Asp Leu Asn Ser Ser Ile Thr245 250 255Ser Asn Met Arg260<210>53<211>276<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(276)<400>53atg cag gag caa ggc atc caa tcc atc atg cgc gcc gcg gaa gag ctg48Met Gln Glu Gln Gly Ile Gln Ser Ile Met Arg Ala Ala Glu Glu Leu1 5 10 15gtc gag cag acc cgc cag gcg ttg tac agc gtc gac gag atc tac gcc96Val Glu Gln Thr Arg Gln Ala Leu Tyr Ser Val Asp Glu Ile Tyr Ala20 25 30cac gtt ggc gtc gac ccc gct cgc ctg cgc aat ctg gcg gtc gag cag144His Val Gly Val Asp Pro Ala Arg Leu Arg Asn Leu Ala Val Glu Gln35 40 45gcc agg ata gag gcc gag gcc cag gcg gcg ttc cgt gat gac ctc gcg192Ala Arg Ile Glu Ala Glu Ala Gln Ala Ala Phe Arg Asp Asp Leu Ala50 55 60gac atc gag cgc gag gcg gcg cgc gtc aag gcg gcc tgc acc gat gcg240Asp Ile Glu Arg Glu Ala Ala Arg Val Lys Ala Ala Cys Thr Asp Ala65 70 75 80ccg cag gcc cgc agg gtg ctt cac aac cac gtc tga276Pro Gln Ala Arg Arg Val Leu His Asn His Val *85 90<210>54<211>91<212>PRT<213>百日咳博德特氏菌<400>54Met Gln Glu Gln Gly Ile Gln Ser Ile Met Arg Ala Ala Glu Glu Leu1 5 10 15Val Glu Gln Thr Arg Gln Ala Leu Tyr Ser Val Asp Glu Ile Tyr Ala20 25 30His Val Gly Val Asp Pro Ala Arg Leu Arg Asn Leu Ala Val Glu Gln35 40 45Ala Arg Ile Glu Ala Glu Ala Gln Ala Ala Phe Arg Asp Asp Leu Ala50 55 60Asp Ile Glu Arg Glu Ala Ala Arg Val Lys Ala Ala Cys Thr Asp Ala65 70 75 80Pro Gln Ala Arg Arg Val Leu His Asn His Val85 90<210>55<211>942<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS
<222>(1)...(942)<400>55atg tct gtt tct ccg act tcg ccc ggc tct ttc ggg gcc ggc cct gtc 48Met Ser Val Ser Pro Thr Ser Pro Gly Ser Phe Gly Ala Gly Pro Val1 5 10 15ttt gac tcc gaa ttg cag gcc ccg gcc ccg tcg gcg cag cgt cgc ggc 96Phe Asp Ser Glu Leu Gln Ala Pro Ala Pro Ser Ala Gln Arg Arg Gly20 25 30ggt gcg gcg cct gtg ccg ccg ccc gtc gat cgg cgc ggc gtc gag ccg 144Gly Ala Ala Pro Val Pro Pro Pro Val Asp Arg Arg Gly Val Glu Pro35 40 45gga gat ccc acg ctg ggc atg ctg ccc gcg cca gat ttg ctc gcg ggg 192Gly Asp Pro Thr Leu Gly Met Leu Pro Ala Pro Asp Leu Leu Ala Gly50 55 60ggc gcc gtc agc cgc acc cgc gcg gcg ctc gac gat ctg gac gcc gca 240Gly Ala Val Ser Arg Thr Arg Ala Ala Leu Asp Asp Leu Asp Ala Ala65 70 75 80cgg ctc ggt gaa gac atc tac gcc ttg atg gcg gtg ttg caa cag gcc 288Arg Leu Gly Glu Asp Ile Tyr Ala Leu Met ala Val Leu Gln Gln Ala85 90 95agt cag cag atg cgg gac gcc gcc cgt atc gct cgt gat gcc gag gct 336Ser Gln Gln Met Arg Asp Ala Ala Arg Ile Ala Arg Asp Ala Glu Ala100 105 110acg cgg caa acg cag gct ctc ggc gat gcg gcc agc cag atg cgc cag 384Thr Arg Gln Thr Gln Ala Leu Gly Asp Ala Ala Ser Gln Met Arg Gln115 120 125gcg gcg agc gag cgc atg gcc gga gcg atc gtg gcg ggc gcc atg cag 432Ala Ala Ser Glu Arg Met ala Gly Ala Ile Val Ala Gly Ala Met Gln130 135 140ata gcg ggt ggt ttc gtg cag ctg ggg gcg ggc ctg gca gcg ggt ttg 480Ile Ala Gly Gly Phe Val Gln Leu Gly Ala Gly Leu Ala Ala Gly Leu145 150 155 160cag gcc atg ggt ggc gct gct gcg caa gcc aag ggc gcc gca ttc tcc 528Gln Ala Met Gly Gly Ala Ala Ala Gln Ala Lys Gly Ala Ala Phe Ser165 170 175gag cag gcc tcg aca agc cgc aag gtg gcg gcc ggc ttg cac gat gcc 576Glu Gln Ala Ser Thr Ser Arg Lys Val Ala Ala Gly Leu His Asp Ala180 185 190ccc gag ctg cag gca acg gtg cag gcc cgc gca acc cag ctc gaa gcg624Pro Glu Leu Gln Ala Thr Val Gln Ala Arg Ala Thr Gln Leu Glu Ala195 200 205caa gcg gcc tcg ttt ggt gcg gac gcg gct cgt tcg tcg gca aag tcg672Gln Ala Ala Ser Phe Gly Ala Asp Ala Ala Arg Ser Ser Ala Lys Ser210 215 220cag cgc gta tcg agc gtt gcc cag gcc ggc gcc gca gcg gcc ggc ggt720Gln Arg Val Ser Ser Val Ala Gln Ala Gly Ala Ala Ala Ala Gly Gly225 230 235 240atc ggc ggc ctg acc agc gcc gcc cag gaa cgc cgc gcc gcc gag cac768Ile Gly Gly Leu Thr Ser Ala Ala Gln Glu Arg Arg Ala Ala Glu His245 250 255gag gcc agg cgc gcg gag ctg gac gtc gaa gcg aag gtg cat gaa acg816Glu Ala Arg Arg Ala Glu Leu Asp Val Glu Ala Lys Val His Glu Thr260 265 270gcc tcg cgg cgg gcc gac gaa gcc atg cag cag atg ctc gac atc atc864Ala Ser Arg Arg Ala Asp Glu Ala Met Gln Gln Met Leu Asp Ile Ile275 280 285cgc ggc atc agg gaa aag ctg gcc ggg atg gag cag tcc cgc agc gag912Arg Gly Ile Arg Glu Lys Leu Ala Gly Met Glu Gln Ser Arg Ser Glu290 295 300acc gcc cgt agc gtg gcc cgc aat atc tga942Thr Ala Arg Ser Val Ala Arg Asn Ile *305 310<210>56<211>313<212>PRT<213>百日咳博德特氏菌<400>56Met Ser Val Ser Pro Thr Ser Pro Gly Ser Phe Gly Ala Gly Pro Val1 5 10 15Phe Asp Ser Glu Leu Gln Ala Pro Ala Pro Ser Ala Gln Arg Arg Gly20 25 30Gly Ala Ala Pro Val Pro Pro Pro Val Asp Arg Arg Gly Val Glu Pro35 40 45Gly Asp Pro Thr Leu Gly Met Leu Pro Ala Pro Asp Leu Leu Ala Gly50 55 60Gly Ala Val Ser Arg Thr Arg Ala Ala Leu Asp Asp Leu Asp Ala Ala65 70 75 80Arg Leu Gly Glu Asp Ile Tyr Ala Leu Met ala Val Leu Gln Gln Ala85 90 95Ser Gln Gln Met Arg Asp Ala Ala Arg Ile Ala Arg Asp Ala Glu Ala100 105 110Thr Arg Gln Thr Gln Ala Leu Gly Asp Ala Ala Ser Gln Met Arg Gln115 120 125Ala Ala Ser Glu Arg Met ala Gly Ala Ile Val Ala Gly Ala Met Gln130 135 140Ile Ala Gly Gly Phe Val Gln Leu Gly Ala Gly Leu Ala Ala Gly Leu145 150 155 160Gln Ala Met Gly Gly Ala Ala Ala Gln Ala Lys Gly Ala Ala Phe Ser165 170 175Glu Gln Ala Ser Thr Ser Arg Lys Val Ala Ala Gly Leu His Asp Ala180 185 190Pro Glu Leu Gln Ala Thr Val Gln Ala Arg Ala Thr Gln Leu Glu Ala195 200 205Gln Ala Ala Ser Phe Gly Ala Asp Ala Ala Arg Ser Ser Ala Lys Ser210 215 220Gln Arg Val Ser Ser Val Ala Gln Ala Gly Ala Ala Ala Ala Gly Gly225 230 235 240Ile Gly Gly Leu Thr Ser Ala Ala Gln Glu Arg Arg Ala Ala Glu His245 250 255Glu Ala Arg Arg Ala Glu Leu Asp Val Glu Ala Lys Val His Glu Thr260 265 270Ala Ser Arg Arg Ala Asp Glu Ala Met Gln Gln Met Leu Asp Ile Ile275 280 285Arg Gly Ile Arg Glu Lys Leu Ala Gly Met Glu Gln Ser Arg Ser Glu290 295 300Thr Ala Arg Ser Val Ala Arg Asn Ile305 310<210>57<211>1203<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1203)<400>57atg acc gtc atg agt acg acc ata tcc aca gcc ccg agc ggc gcc gcg48Met Thr Val Met Ser Thr Thr Ile Ser Thr Ala Pro Ser Gly Ala Ala1 5 10 15ctt gcg ccg tct cgc ata gat atg cgg gcg ccg gag ccc ggg agt gcc96Leu Ala Pro Ser Arg Ile Asp Met Arg Ala Pro Glu Pro Gly Ser Ala20 25 30ggc gaa ggc gcc ggt atc ctg gcg ccg gtg acg acg ctg gct ctg gcg144Gly Glu Gly Ala Gly Ile Leu Ala Pro Val Thr Thr Leu Ala Leu Ala35 40 45gcg ggc cgg ccg gct ttg cca gcg tca ccg tcg ctg cgc acc gcg ccc 192Ala Gly Arg Pro Ala Leu Pro Ala Ser Pro Ser Leu Arg Thr Ala Pro50 55 60gtc ctg gat ccg cca gtg cgc gat ctc agc ccc gcc gac ttg gcc gac 240Val Leu Asp Pro Pro Val Arg Asp Leu Ser Pro Ala Asp Leu Ala Asp65 70 75 80ctg ctg cgc gtc ttg cga tcc agg gcg gtg gac ggg cag ttg gcc acg 288Leu Leu Arg Val Leu Arg Ser Arg Ala Val Asp Gly Gln Leu Ala Thr85 90 95gcg cgc gag aac ctg cag gat gcg caa gtc aag gcg aag cag aac acc 336Ala Arg Glu Asn Leu Gln Asp Ala Gln Val Lys Ala Lys Gln Asn Thr100 105 110cag gcc cag ctc gac aag ctg gac gca tgg ttt cgg aag gct gag gac 384Gln Ala Gln Leu Asp Lys Leu Asp Ala Trp Phe Arg Lys Ala Glu Asp115 120 125gcc gag agc aag ggc tgg ctg agc aag gtg ttc ggc tgg atc ggg aag 432Ala Glu Ser Lys Gly Trp Leu Ser Lys Val Phe Gly Trp Ile Gly Lys130 135 140gtg ctg gcg gtc gtg gca tcg gcc ctg gct gtg ggc ttt gct gcc gtc 480Val Leu Ala Val Val Ala Ser Ala Leu Ala Val Gly Phe Ala Ala Val145 150 155 160gcc agc gtg gtc acc ggc gcg gcg gcc acg ccc atg ctg gtg ctc agc 528Ala Ser Val Val Thr Gly Ala Ala Ala Thr Pro Met Leu Val Leu Ser165 170 175ggc atg gca ttg gtc agc gcc gtg aca tcg ctg gcc gac cag ata tcg 576Gly Met ala Leu Val Ser Ala Val Thr Ser Leu Ala Asp Gln Ile Ser180 185 190cga gag gcg gga ggg ccg cct atc agc ctg ggc ggg ttt ctc tcc ggg 624Arg Glu Ala Gly Gly Pro Pro Ile Ser Leu Gly Gly Phe Leu Ser Gly195 200 205ctg gcc gga cgt ctg ctg aca gcg ttg ggg gtg gat cag tcg cag gcc 672Leu Ala Gly Arg Leu Leu Thr Ala Leu Gly Val Asp Gln Ser Gln Ala210 215 220gac caa att gcc aag atc gtc gcc ggc ctg gcc gtg ccc gcc gtc ttg 720Asp Gln Ile Ala Lys Ile Val Ala Gly Leu Ala Val Pro Ala Val Leu225 230 235 240ctg atc gaa ccc cag atg ctg ggc gaa atg gcc gaa ggc gtg gcc agg 768Leu Ile Glu Pro Gln Met Leu Gly Glu Met ala Glu Gly Val Ala Arg245 250 255ctg gcg ggc gcc ggc gat gcc acc gcg gga tac ata gcc atg gcg atg816Leu Ala Gly Ala Gly Asp Ala Thr Ala Gly Tyr Ile Ala Met ala Met260 265 270tcc atc gtg gcg gcg atc gcg gtc gcc gcg atc aat gcc gcc ggt acg864Ser Ile Val Ala Ala Ile Ala Val Ala Ala Ile Asn Ala Ala Gly Thr275 280 285gcc ggc gcg ggc agc gcc tcg gcg atc agg ggt gcc tgg gat cgg gcc912Ala Gly Ala Gly Ser Ala Ser Ala Ile Arg Gly Ala Trp Asp Arg Ala290 295 300gcc gcg gta gcc acc cag gtc ctt cag ggg ggt acg gca gtg gcg caa960Ala Ala Val Ala Thr Gln Val Leu Gln Gly Gly Thr Ala Val Ala Gln305 310 315 320ggc ggc gtc ggc gtg tcg atg gca gtc gat cgc aaa cag gcc gat ctc1008Gly Gly Val Gly Val Ser Met ala Val Asp Arg Lys Gln Ala Asp Leu325 330 335ctg gtc gcc gac aag gcg gat ctg gcg gcg agc ctg aca aaa ctg cgg1056Leu Val Ala Asp Lys Ala Asp Leu Ala Ala Ser Leu Thr Lys Leu Arg340 345 350gcg gcc atg gag cgt gag gcg gac gat atc aag aag atc ctg gct caa1104Ala Ala Met Glu Arg Glu Ala Asp Asp Ile Lys Lys Ile Leu Ala Gln355 360 365ttc gac gcg gcc tat cac atg atc gcg cag atg atc agc gac atg gcg1152Phe Asp Ala Ala Tyr His Met Ile Ala Gln Met Ile Ser Asp Met ala370 375 380agc acg cac agc cag gtc agc gcc aac ctc gga cgg cgc cag gcg gtg1200Ser Thr His Ser Gln Val Ser Ala Asn Leu Gly Arg Arg Gln Ala Val385 390 395 400tag1203<210>58<211>400<212>PRT<213>百日咳博德特氏菌<400>58Met Thr Val Met Ser Thr Thr Ile Ser Thr Ala Pro Ser Gly Ala Ala1 5 10 15Leu Ala Pro Ser Arg Ile Asp Met Arg Ala Pro Glu Pro Gly Ser Ala20 25 30Gly Glu Gly Ala Gly Ile Leu Ala Pro Val Thr Thr Leu Ala Leu Ala35 40 45Ala Gly Arg Pro Ala Leu Pro Ala Ser Pro Ser Leu Arg Thr Ala Pro50 55 60Val Leu Asp Pro Pro Val Arg Asp Leu Ser Pro Ala Asp Leu Ala Asp65 70 75 80Leu Leu Arg Val Leu Arg Ser Arg Ala Val Asp Gly Gln Leu Ala Thr85 90 95Ala Arg Glu Asn Leu Gln Asp Ala Gln Val Lys Ala Lys Gln Asn Thr100 105 110Gln Ala Gln Leu Asp Lys Leu Asp Ala Trp Phe Arg Lys Ala Glu Asp115 120 125Ala Glu Ser Lys Gly Trp Leu Ser Lys Val Phe Gly Trp Ile Gly Lys130 135 140Val Leu Ala Val Val Ala Ser Ala Leu Ala Val Gly Phe Ala Ala Val145 150 155 160Ala Ser Val Val Thr Gly Ala Ala Ala Thr Pro Met Leu Val Leu Ser165 170 175Gly Met ala Leu Val Ser Ala Val Thr Ser Leu Ala Asp Gln Ile Ser180 185 190Arg Glu Ala Gly Gly Pro Pro Ile Ser Leu Gly Gly Phe Leu Ser Gly195 200 205Leu Ala Gly Arg Leu Leu Thr Ala Leu Gly Val Asp Gln Ser Gln Ala210 215 220Asp Gln Ile Ala Lys Ile Val Ala Gly Leu Ala Val Pro Ala Val Leu225 230 235 240Leu Ile Glu Pro Gln Met Leu Gly Glu Met ala Glu Gly Val Ala Arg245 250 255Leu Ala Gly Ala Gly Asp Ala Thr Ala Gly Tyr Ile Ala Met ala Met260 265 270Ser Ile Val Ala Ala Ile Ala Val Ala Ala Ile Asn Ala Ala Gly Thr275 280 285Ala Gly Ala Gly Ser Ala Ser Ala Ile Arg Gly Ala Trp Asp Arg Ala290 295 300Ala Ala Val Ala Thr Gln Val Leu Gln Gly Gly Thr Ala Val Ala Gln305 310 315 320Gly Gly Val Gly Val Ser Met ala Val Asp Arg Lys Gln Ala Asp Leu325 330 335Leu Val Ala Asp Lys Ala Asp Leu Ala Ala Ser Leu Thr Lys Leu Arg340 345 350Ala Ala Met Glu Arg Glu Ala Asp Asp Ile Lys Lys Ile Leu Ala Gln355 360 365Phe Asp Ala Ala Tyr His Met Ile Ala Gln Met Ile Ser Asp Met ala370 375 380Ser Thr His Ser Gln Val Ser Ala Asn Leu Gly Arg Arg Gln Ala Val385 390 395 400<210>59<211>462<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(462)<400>59atg act gtt cat gac gac gcg gcc gcg gcg ctg cgc gcc cgg ctg gat48Met Thr Val His Asp Asp Ala Ala Ala Ala Leu Arg Ala Arg Leu Asp1 5 10 15gcg ttg ccg ggc agc cgg cgc ctg aca gcc gag caa ttg gaa gtg att96Ala Leu Pro Gly Ser Arg Arg Leu Thr Ala Glu Gln Leu Glu Val Ile20 25 30tac gcg atg gcg tat gcg cac gtc gcc agg tgc gag tac ggc aag gcg144Tyr Ala Met ala Tyr Ala His Val Ala Arg Cys Glu Tyr Gly Lys Ala35 40 45ctg ccc att ttc gcc ttc ctc gcg cag tac ggc ccc acg cgc aag cac192Leu Pro Ile Phe Ala Phe Leu Ala Gln Tyr Gly Pro Thr Arg Lys His50 55 60tac tgg gcc ggc ctg gcg cta tgc ctg cag aag acc gac cgt ccc gac240Tyr Trp Ala Gly Leu Ala Leu Cys Leu Gln Lys Thr Asp Arg Pro Asp65 70 75 80gag gcg cgc aat atc tat gcg ttg atc ctc acg ctc tat cca gat tcc288Glu Ala Arg Asn Ile Tyr Ala Leu Ile Leu Thr Leu Tyr Pro Asp Ser85 90 95gcg gat gcc gtg ttg cgc acg gcc gag tgc gag ctg gcg ttg ggt gag336Ala Asp Ala Val Leu Arg Thr Ala Glu Cys Glu Leu Ala Leu Gly Glu100 105 110aac gaa cgg gcg cag gcg gcc ctg ttc ggc gca atc gcc atc gat gca384Asn Glu Arg Ala Gln Ala Ala Leu Phe Gly Ala Ile Ala Ile Asp Ala115 120 125gaa agt ggg cag cca ggt ccg gtc tcg cac cgt gcg cgc gct ttg ctc432Glu Ser Gly Gln Pro Gly Pro Val Ser His Arg Ala Arg Ala Leu Leu130 135 140gat ctt att tca gtt tca cat ccg gag taa462Asp Leu Ile Ser Val Ser His Pro Glu *145 150<210>60<211>153<212>PRT<213>百日咳博德特氏菌<400>60Met Thr Val His Asp Asp Ala Ala Ala Ala Leu Arg Ala Arg Leu Asp1 5 10 15Ala Leu Pro Gly Ser Arg Arg Leu Thr Ala Glu Gln Leu Glu Val Ile20 25 30Tyr Ala Met ala Tyr Ala His Val Ala Arg Cys Glu Tyr Gly Lys Ala35 40 45Leu Pro Ile Phe Ala Phe Leu Ala Gln Tyr Gly Pro Thr Arg Lys His50 55 60Tyr Trp Ala Gly Leu Ala Leu Cys Leu Gln Lys Thr Asp Arg Pro Asp65 70 75 80Glu Ala Arg Asn Ile Tyr Ala Leu Ile Leu Thr Leu Tyr Pro Asp Ser85 90 95Ala Asp Ala Val Leu Arg Thr Ala Glu Cys Glu Leu Ala Leu Gly Glu100 105 110Asn Glu Arg Ala Gln Ala Ala Leu Phe Gly Ala Ile Ala Ile Asp Ala115 120 125Glu Ser Gly Gln Pro Gly Pro Val Ser His Arg Ala Arg Ala Leu Leu130 135 140Asp Leu Ile Ser Val Ser His Pro Glu145 150<210>61<211>522<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(522)<400>6latg cac tca gac tca ggt tca gat tca ggc tca gac tca ggc tca ggc48Met His Ser Asp Ser Gly Ser Asp Ser Gly Ser Asp Ser Gly Ser Gly1 5 10 15tca ccc atg gtc tcg tcg ata cat cca tcg gaa ccg ata cag ccg atg96Ser Pro Met Val Ser Ser Ile His Pro Ser Glu Pro Ile Gln Pro Met20 25 30gag cat gtg ctc gag gag gcc gac gcc cgc ctg ctt acc gaa gtg ggt144Glu His Val Leu Glu Glu Ala Asp Ala Arg Leu Leu Thr Glu Val Gly35 40 45ttt ctg gcg gcg gcc gtc agc gat ctg acg cgc gcg gac gcc att ttc192Phe Leu Ala Ala Ala Val Ser Asp Leu Thr Arg Ala Asp Ala Ile Phe50 55 60aat gca ttg caa cgt gta cgg ccg ggc cgg acg cat ccc tgc atc ggc240Asn Ala Leu Gln Arg Val Arg Pro Gly Arg Thr His Pro Cys Ile Gly65 70 75 80ctg gcg gtc gcc cgc atg aac gcc ggg ctg ccc gac gaa gcc gcc gag288Leu Ala Val Ala Arg Met Asn Ala Gly Leu Pro Asp Glu Ala Ala Glu85 90 95atc ctg gcg aat ttc cag ccg gca cag ccg gag gac cgc tcg gaa ctg336Ile Leu Ala Asn Phe Gln Pro Ala Gln Pro Glu Asp Arg Ser Glu Leu100 105 110gac gcc tgg tgc ggg ttc gct ctg ttg ctg gct ggc cgc tcg gac gag384Asp Ala Trp Cys Gly Phe Ala Leu Leu Leu Ala Gly Arg Ser Asp Glu115 120 125gcg cgc cgc atg ctg cag cga gcc atc gat gcg ggt ggc gag gcg gca432Ala Arg Arg Met Leu Gln Arg Ala Ile Asp Ala Gly Gly Glu Ala Ala130 135 140agg ctg gcg cag gtc gtg ttg gac agc gga ccc gcc atg atg cgg ccc480Arg Leu Ala Gln Val Val Leu Asp Ser Gly Pro Ala Met Met Arg Pro145 150 155 160gcg ccg ttg cag tcc gag cca tta cct gga gct cct gga tga522Ala Pro Leu Gln Ser Glu Pro Leu Pro Gly Ala Pro Gly *165 170<210>62<211>173<212>PRT<213>百日咳博德特氏菌<400>62Met His Ser Asp Ser Gly Ser Asp Ser Gly Ser Asp Ser Gly Ser Gly1 5 10 15Ser Pro Met Val Ser Ser Ile His Pro Ser Glu Pro Ile Gln Pro Met20 25 30Glu His Val Leu Glu Glu Ala Asp Ala Arg Leu Leu Thr Glu Val Gly35 40 45PhezLeu Ala Ala Ala Val Ser Asp Leu Thr Arg Ala Asp Ala Ile Phe50 55 60Asn Ala Leu Gln Arg Val Arg Pro Gly Arg Thr His Pro Cys Ile Gly65 70 75 80Leu Ala Val Ala Arg Met Asn Ala Gly Leu Pro Asp Glu Ala Ala Glu85 90 95Ile Leu Ala Asn Phe Gln Pro Ala Gln Pro Glu Asp Arg Ser Glu Leu100 105 110Asp Ala Trp Cys Gly Phe Ala Leu Leu Leu Ala Gly Arg Ser Asp Glu115 120 125Ala Arg Arg Met Leu Gln Arg Ala Ile Asp Ala Gly Gly Glu Ala Alal30 135 140Arg Leu Ala Gln Val Val Leu Asp Ser Gly Pro Ala Met Met Arg Pro145 150 155 160Ala Pro Leu Gln Ser Glu Pro Leu Pro Gly Ala Pro Gly165 170<210>63<211>180<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(180)<400>63atg gtt gcg cga ggt cct tgc gca cca cct ggt gta gtc ccc ata cgt48Met Val Ala Arg Gly Pro Cys Ala Pro Pro Gly Val Val Pro Ile Arg1 5 10 15cgc gtc gta agt ttt acg gaa gac gtt cag cgc gtt gta tct gaa tgc96Arg Val Val Ser Phe Thr Glu Asp Val Gln Arg Val Val Ser Glu Cys20 25 30tcc ggt agc gac cgc gat ccg aca tta gtt tcg gaa gtt aac aac tgc144Ser Gly Ser Asp Arg Asp Pro Thr Leu Val Ser Glu Val Asn Asn Cys35 40 45cgg ata aaa ccg gca gtc agt atc ttg gtg acg tga180Arg Ile Lys Pro Ala Val Ser Ile Leu Val Thr *50 55<210>64<211>59<212>PRT<213>百日咳博德特氏菌<400>64Met Val Ala Arg Gly Pro Cys Ala Pro Pro Gly Val Val Pro Ile Arg1 5 10 15Arg Val Val Ser Phe Thr Glu Asp Val Gln Arg Val Val Ser Glu Cys20 25 30Ser Gly Ser Asp Arg Asp Pro Thr Leu Val Ser Glu Val Asn Asn Cys35 40 45Arg Ile Lys Pro Ala Val Ser Ile Leu Val Thr50 55<210>65<211>975<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(975)<400>65atg cgg ttt cga gca ggt tat agc cgt tat caa gcc cgc tca ggc cat48Met Arg Phe Arg Ala Gly Tyr Ser Arg Tyr Gln Ala Arg Ser Gly His1 5 10 15ggg gac cgc cca ccc ccc gca cag gcg cgc gtc cag acg gta ctc ctg96Gly Asp Arg Pro Pro Pro Ala Gln Ala Arg Val Gln Thr Val Leu Leu20 25 30cac gga ctc tcc gcg ttg acg gcg caa gtt gcg cag cgc ttc gaa atg144His Gly Leu Ser Ala Leu Thr Ala Gln Val Ala Gln Arg Phe Glu Met35 40 45gcg cgc cac cgg atg gct ggc ccc ggt cgc acg aca ggc cac cac cat192Ala Arg His Arg Met ala Gly Pro Gly Arg Thr Thr Gly His His His50 55 60ttc cag ctc gag gcc cag cgt atg gcc gac act ttg cgc agc gtt caa240Phe Gln Leu Glu Ala Gln Arg Met ala Asp Thr Leu Arg Ser Val Gln65 70 75 80ggc gag cct cgg tgg ccg gac ggg agc gag gcc tgc atg ccg tcg ggt288Gly Glu Pro Arg Trp Pro Asp Gly Ser Glu Ala Cys Met Pro Ser Gly85 90 95ttg tca tgc cgg cat gga acc gaa gag ccg aaa gcg tca cac agt gca336Leu Ser Cys Arg His Gly Thr Glu Glu Pro Lys Ala Ser His Ser Ala100 105 110tat tcc atg ttc ccg ctc cgt aga acg cga tac acc caa gga ttc gag384Tyr Ser Met Phe Pro Leu Arg Arg Thr Arg Tyr Thr Gln Gly Phe Glu115 120 125aca acg gcc cac cgc atg aac ttc cag atc cca ccc gct tta cct gct432Thr Thr Ala His Arg Met Asn Phe Gln Ile Pro Pro Ala Leu Pro Ala130 135 140ttg gag ctt gat gtc ttt gcg cgc gcc gcc agc caa gga gag acc cta480Leu Glu Leu Asp Val Phe Ala Arg Ala Ala Ser Gln Gly Glu Thr Leu145 150 155 160tat gtc acc aaa gca ggc gag cag ttc cag gtc atc gca tcc ggc acg528Tyr Val Thr Lys Ala Gly Glu Gln Phe Gln Val Ile Ala Ser Gly Thr165 170 175acg ccg tca ggg cgc aac gta tcc tgg gtc gcc acc gac gag gac acg576Thr Pro Ser Gly Arg Asn Val Ser Trp Val Ala Thr Asp Glu Asp Thr180 185 190ctt gtc atg ttt tcc agc gcg ctg gcg ctg gcc tac ggc acg gga atc624Leu Val Met Phe Ser Ser Ala Leu Ala Leu Ala Tyr Gly Thr Gly Ile195 200 205gcc cgc gcc gtc gcc aag gag ctc gat ctg cac gcg gtc ccg acg aca672Ala Arg Ala Val Ala Lys Glu Leu Asp Leu His Ala Val Pro Thr Thr210 215 220tcg ctg tcg gcg cgc gtc gtc acg cga gcg gtc gac atg gcg gaa acc720Ser Leu Ser Ala Arg Val Val Thr Arg Ala Val Asp Met ala Glu Thr225 230 235 240tca cgc cac gcc ctg cag ggc gtg gat ttc ctt acc ttc ctg tcc tgg768Ser Arg His Ala Leu Gln Gly Val Asp Phe Leu Thr Phe Leu Ser Trp245 250 255tcg gcc cgc gcc gac gcc gcc ggc ttc cga cag gtc tgt cac gac acc816Ser Ala Arg Ala Asp Ala Ala Gly Phe Arg Gln Val Cys His Asp Thr260 265 270ggt gtc tct ccc gat cag ata tcc gga acg ttg cgc gcc acg atc gac864Gly Val Ser Pro Asp Gln Ile Ser Gly Thr Leu Arg Ala Thr Ile Asp275 280 285gaa agc atg cag cag cgc ttc gca tcc gcc gca caa tca ggt aag gcg912Glu Ser Met Gln Gln Arg Phe Ala Ser Ala Ala Gln Ser Gly Lys Ala290 295 300ccg gta tcc gcc cat acg gcg caa gaa tgg ttg cgc gag gtc ctt gcg960Pro Val Ser Ala His Thr Ala Gln Glu Trp Leu Arg Glu Val Leu Ala305 310 315 320cac cac ctg gtg tag975His His Leu Val *<210>66<211>324<212>PRT<213>百日咳博德特氏菌<400>66Met Arg Phe Arg Ala Gly Tyr Ser Arg Tyr Gln Ala Arg Ser Gly His1 5 10 15Gly Asp Arg Pro Pro Pro Ala Gln Ala Arg Val Gln Thr Val Leu Leu20 25 30His Gly Leu Ser Ala Leu Thr Ala Gln Val Ala Gln Arg Phe Glu Met35 40 45Ala Arg His Arg Met ala Gly Pro Gly Arg Thr Thr Gly His His His
50 55 60Phe Gln Leu Glu Ala Gln Arg Met ala Asp Thr Leu Arg Ser Val Gln65 70 75 80Gly Glu Pro Arg Trp Pro Asp Gly Ser Glu Ala Cys Met Pro Ser Gly85 90 95Leu Ser Cys Arg His Gly Thr Glu Glu Pro Lys Ala Ser His Ser Ala100 105 110Tyr Ser Met Phe Pro Leu Arg Arg Thr Arg Tyr Thr Gln Gly Phe Glu115 120 125Thr Thr Ala His Arg Met Asn Phe Gln Ile Pro Pro Ala Leu Pro Ala130 135 140Leu Glu Leu Asp Val Phe Ala Arg Ala Ala Ser Gln Gly Glu Thr Leu145 150 155 160Tyr Val Thr Lys Ala Gly Glu Gln Phe Gln Val Ile Ala Ser Gly Thr165 170 175Thr Pro Ser Gly Arg Asn Val Ser Trp Val Ala Thr Asp Glu Asp Thr180 185 190Leu Val Met Phe Ser Ser Ala Leu Ala Leu Ala Tyr Gly Thr Gly Ile195 200 205Ala Arg Ala Val Ala Lys Glu Leu Asp Leu His Ala Val Pro Thr Thr210 215 220Ser Leu Ser Ala Arg Val Val Thr Arg Ala Val Asp Met ala Glu Thr225 230 235 240Ser Arg His Ala Leu Gln Gly Val Asp Phe Leu Thr Phe Leu Ser Trp245 250 255Ser Ala Arg Ala Asp Ala Ala Gly Phe Arg Gln Val Cys His Asp Thr260 265 270Gly Val Ser Pro Asp Gln Ile Ser Gly Thr Leu Arg Ala Thr Ile Asp275 280 285Glu Ser Met Gln Gln Arg Phe Ala Ser Ala Ala Gln Ser Gly Lys Ala290 295 300Pro Val Ser Ala His Thr Ala Gln Glu Trp Leu Arg Glu Val Leu Ala305 310 315 320His His Leu Val<210>67<211>1146<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1146)<400>67atg ctg atc aac gcg gcc gag cac ccc gcc gcc agc ctg gat gcc gac48Met Leu Ile Asn Ala Ala Glu His Pro Ala Ala Ser Leu Asp Ala Asp1 5 10 15tgg tac cgg cga gtg cgg gtg ccg cgg ccc atc tac gag gaa ctc gtc96Trp Tyr Arg Arg Val Arg Val Pro Arg Pro Ile Tyr Glu Glu Leu Val20 25 30ggc cag cga ggc tgg ctg cac cgg atc ggg ata gac gcc aag gca cag 144Gly Gln Arg Gly Trp Leu His Arg Ile Gly Ile Asp Ala Lys Ala Gln35 40 45aac agc ccc tgc acg tcg gtt ccc gtg gcc atc gcc gcg cgc tgc ctg 192Asn Ser Pro Cys Thr Ser Val Pro Val Ala Ile Ala Ala Arg Cys Leu50 55 60aac gtc gtg ctg gcg ctg gct ccc gcg cag atc gcc atg ttc gcc aac 240Asn Val Val Leu Ala Leu Ala Pro Ala Gln Ile Ala Met Phe Ala Asn65 70 75 80agc ccg ctg gag gca ggg cgg gtg acc ggt ctc aag gaa aac cgc ctg 288Ser Pro Leu Glu Ala Gly Arg Val Thr Gly Leu Lys Glu Asn Arg Leu85 90 95acc ctg tgg ccg cgc atg ttc cga ggc gcg cgc tac ctg ggc gac gac 336Thr Leu Trp Pro Arg Met Phe Arg Gly Ala Arg Tyr Leu Gly Asp Asp100 105 110ctg ctg cat cgc ctg cct gca agg ccg ttt cgc gat ctc ggc gat tat 384Leu Leu His Arg Leu Pro Ala Arg Pro Phe Arg Asp Leu Gly Asp Tyr115 120 125ttc cgc tgg atg ttc ggc gga ttg acc gcc agc cgg gcg cta ccg ccg 432Phe Arg Trp Met Phe Gly Gly Leu Thr Ala Ser Arg Ala Leu Pro Pro130 135 140ggc gac gct tgc gac tac aag aac gcc gat gtg gcc tgc ctg gtg gga 480Gly Asp Ala Cys Asp Tyr Lys Asn Ala Asp Val Ala Cys Leu Val Gly145 150 155 160gcc cct tcg ctg gca gag ttc ctg tat gcg ggc gcg tgg tcc gcg cga 528Ala Pro Ser Leu Ala Glu Phe Leu Tyr Ala Gly Ala Trp Ser Ala Arg165 170 175aac ctg aat gat ggc ggt tcc gtg cgt ctg gcc gcg cgc agc gaa cat 576Asn Leu Asn Asp Gly Gly Ser Val Arg Leu Ala Ala Arg Ser Glu His180 185 190ttc gtc tat tcg cag ttc gcg cag ttc ctg gac gcg cgt tgg cgc tac 624Phe Val Tyr Ser Gln Phe Ala Gln Phe Leu Asp Ala Arg Trp Arg Tyr195200 205agg atg ccg att gtc ccc gcc ttg ccg gcg ctg ttg cga gcc tgg gac 672Arg Met Pro Ile Val Pro Ala Leu Pro Ala Leu Leu Arg Ala Trp Asp210 215 220agg cag ggc ggc ctg gaa gcg ctg ttc gag cag gcc ggc gcg caa ggc720Arg Gln Gly Gly Leu Glu Ala Leu Phe Glu Gln Ala Gly Ala Gln Gly225 230 235 240tac atc gag ggg cgc gcg ccg ggc gcg gta ttt gcc gat gcc gac ttg768Tyr Ile Glu Gly Arg Ala Pro Gly Ala Val Phe Ala Asp Ala Asp Leu245 250 255ctg agc tca gcc ggc gat gca gtc gcg gcc agt gcg ccg atg gcg gcg816Leu Ser Ser Ala Gly Asp Ala Val Ala Ala Ser Ala Pro Met ala Ala260265 270tcg gcg ctg caa ttg ggg ctg ttg cgc aat ctg cac gac gcc gag gcc864Ser Ala Leu Gln Leu Gly Leu Leu Arg Asn Leu His Asp Ala Glu Ala275 280 285ctg gtg agg cga tgg ggc tgg ctg cgc ttg cgt gcg ttg cgc gat cgg912Leu Val Arg Arg Trp Gly Trp Leu Arg Leu Arg Ala Leu Arg Asp Arg290 295 300gcc atc gct ttg gcg ttg gac gat gcg cag gtg cgc tgc ctt tgc caa960Ala Ile Ala Leu Ala Leu Asp Asp Ala Gln Val Arg Cys Leu Cys Gln305 310 315 320cag gtc gtg gcg gta gcc gaa ggc ggg ctg gcc ggc gac gag cag caa1008Gln Val Val Ala Val Ala Glu Gly Gly Leu Ala Gly Asp Glu Gln Gln325 330 335tgg ctc gat tat gtg cgt tac gtg gtg gaa acc ggc gag acc gcc gcg1056Trp Leu Asp Tyr Val Arg Tyr Val Val Glu Thr Gly Glu Thr Ala Ala340 345 350gac cgc atg ctg cgc ttg tgg cgc cag gcg cgc ggc acg cct gag atg1104Asp Arg Met Leu Arg Leu Trp Arg Gln Ala Arg Gly Thr Pro Glu Met355 360 365cgc cgc gca cag gcg tgc cgg cag cgc gcg gtg ctg tcc tag1146Arg Arg Ala Gln Ala Cys Arg Gln Arg Ala Val Leu Ser *370 375 380<210>68<211>381<212>PRT<213>百日咳博德特氏菌<400>68Met Leu Ile Asn Ala Ala Glu His Pro Ala Ala Ser Leu Asp Ala Asp1 5 10 15Trp Tyr Arg Arg Val Arg Val Pro Arg Pro Ile Tyr Glu Glu Leu Val20 25 30Gly Gln Arg Gly Trp Leu His Arg Ile Gly Ile Asp Ala Lys Ala Gln35 40 45Asn Ser Pro Cys Thr Ser Val Pro Val Ala Ile Ala Ala Arg Cys Leu50 55 60Asn Val Val Leu Ala Leu Ala Pro Ala Gln Ile Ala Met Phe Ala Asn65 70 75 80Ser Pro Leu Glu Ala Gly Arg Val Thr Gly Leu Lys Glu Asn Arg Leu85 90 95Thr Leu Trp Pro Arg Met Phe Arg Gly Ala Arg Tyr Leu Gly Asp Asp100 105 110Leu Leu His Arg Leu Pro Ala Arg Pro Phe Arg Asp Leu Gly Asp Tyr115 120 125Phe Arg Trp Met Phe Gly Gly Leu Thr Ala Ser Arg Ala Leu Pro Pro130 135 140Gly Asp Ala Cys Asp Tyr Lys Asn Ala Asp Val Ala Cys Leu Val Gly145 150 155 160Ala Pro Ser Leu Ala Glu Phe Leu Tyr Ala Gly Ala Trp Ser Ala Arg165 170 175Asn Leu Asn Asp Gly Gly Ser Val Arg Leu Ala Ala Arg Ser Glu His180 185 190Phe Val Tyr Ser Gln Phe Ala Gln Phe Leu Asp Ala Arg Trp Arg Tyr195 200 205Arg Met Pro Ile Val Pro Ala Leu Pro Ala Leu Leu Arg Ala Trp Asp210 215 220Arg Gln Gly Gly Leu Glu Ala Leu Phe Glu Gln Ala Gly Ala Gln Gly225 230 235 240Tyr Ile Glu Gly Arg Ala Pro Gly Ala Val Phe Ala Asp Ala Asp Leu245 250 255Leu Ser Ser Ala Gly Asp Ala Val Ala Ala Ser Ala Pro Met ala Ala260 265 270Ser Ala Leu Gln Leu Gly Leu Leu Arg Asn Leu His Asp Ala Glu Ala275 280 285Leu Val Arg Arg Trp Gly Trp Leu Arg Leu Arg Ala Leu Arg Asp Arg290 295 300Ala Ile Ala Leu Ala Leu Asp Asp Ala Gln Val Arg Cys Leu Cys Gln305 310 315 320Gln Val Val Ala Val Ala Glu Gly Gly Leu Ala Gly Asp Glu Gln Gln325 330 335Trp Leu Asp Tyr Val Arg Tyr Val Val Glu Thr Gly Glu Thr Ala Ala340 345 350Asp Arg Met Leu Arg Leu Trp Arg Gln Ala Arg Gly Thr Pro Glu Met355 360 365Arg Arg Ala Gln Ala Cys Arg Gln Arg Ala Val Leu Ser370 375 380<210>69<211>1233<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1233)<400>69atg acc ctt cgc ccc gga ata ctc gcc ccg ctc gcc ctg ctt ctt ggc48Met Thr Leu Arg Pro Gly Ile Leu Ala Pro Leu Ala Leu Leu Leu Gly1 5 10 15ctc gcg tgg ccc atc gcg gcg gca tcc agc cct cct cct gtc gcg gcc96Leu Ala Trp Pro Ile Ala Ala Ala Ser Ser Pro Pro Pro Val Ala Ala20 25 30ctg agc tcc ggc gtc gcc ctt acc tcg ccc cgc ctt ccg cct ccc tcc144Leu Ser Ser Gly Val Ala Leu Thr Ser Pro Arg Leu Pro Pro Pro Ser35 40 45cat aca tcc ggc cgg aaa tgg cgc atc ggt tat gtg ggt agc ggc gag192His Thr Ser Gly Arg Lys Trp Arg Ile Gly Tyr Val Gly Ser Gly Glu50 55 60tac gag gag tat ccg cgc acg ctc tac gcg atc gcg cgc gca ttg caa240Tyr Glu Glu Tyr Pro Arg Thr Leu Tyr Ala Ile Ala Arg Ala Leu Gln65 70 75 80caa ctc gga tgg ctg cgt atc gac gac atg ccc gag ata acc gat atg288Gln Leu Gly Trp Leu Arg Ile Asp Asp Met Pro Glu Ile Thr Asp Met85 90 95cga aag gcc tgg ctt tac ctg gcc acg cat gcc cgc agc aac tac atc336Arg Lys Ala Trp Leu Tyr Leu Ala Thr His Ala Arg Ser Asn Tyr Ile100 105 110gag ttc gtg ccc gat gcg tgg tgg cag ccc ggc aac ttc gac acc gcc384Glu Phe Val Pro Asp Ala Trp Trp Gln Pro Gly Asn Phe Asp Thr Ala115 120 125ttg cgg cct gcc gtg cgc gaa gcc gtt gcg gca cgc ctg cat ggc gcc432Leu Arg Pro Ala Val Arg Glu Ala Val Ala Ala Arg Leu His Gly Ala130 135 140aag gac atc gac ctg atc atc gcc atg ggt acc tgg gct gga cag gac480Lys Asp Ile Asp Leu Ile Ile Ala Met Gly Thr Trp Ala Gly Gln Asp145 150 155 160atg gtc gaa ctg ggc acg ccg gta ccc acc gtg gtc gtc tcg tcg acc528Met Val Glu Leu Gly Thr Pro Val Pro Thr Val Val Val Ser Ser Thr165 170 175gac ccg ata agc gcc cgg atc ata ccc agt gcg gcc gac agc ggc cag576Asp Pro Ile Ser Ala Arg Ile Ile Pro Ser Ala Ala Asp Ser Gly Gln180 185 190gac aac ctg cat gcc cgg gta cag ccc gac cac tac cag cgg cag atc 624Asp Asn Leu His Ala Arg Val Gln Pro Asp His Tyr Gln Arg Gln Ile195 200 205cag ctg ctc cat gac atc gtg ccg ttc aag acg ctt gga ctg gtc tac 672Gln Leu Leu His Asp Ile Val Pro Phe Lys Thr Leu Gly Leu Val Tyr210 215 220gaa gac acc gaa gca ggt cgc acc tac gca gcc atc gat aag gtc gcc 720Glu Asp Thr Glu Ala Gly Arg Thr Tyr Ala Ala Ile Asp Lys Val Ala225 230 235 240gca cta atg ccg gca ttg gat ttc tcc gtc aag cgt tgc gac gca cgc 768Ala Leu Met Pro Ala Leu Asp Phe Ser Val Lys Arg Cys Asp Ala Arg245 250 255gcg acc ggc atc ccc atc gcc acg gca acc cag aac gtt ctg gct tgc 816Ala Thr Gly Ile Pro Ile Ala Thr Ala Thr Gln Asn Val Leu Ala Cys260 265 270tac cag aag ctg tcg agc gaa gtc gac gcc ttt tac gtc acc gag cac 864Tyr Gln Lys Leu Ser Ser Glu Val Asp Ala Phe Tyr Val Thr Glu His275 280 285cgg ggc atc acc tcg acg tcc gtc aag cag ctc gcc gcg ctg ctg cgc 912Arg Gly Ile Thr Ser Thr Ser Val Lys Gln Leu Ala Ala Leu Leu Arg290 295 300gcc gcc cgc gtg ccg agt ttc tcg atg caa ggc tcc gac gag gtc aag 960Ala Ala Arg Val Pro Ser Phe Ser Met Gln Gly Ser Asp Glu Val Lys305 310 315 320gcc ggc ctg ttg atg agc ctg gcc aag gcg gac tac tcc agc gta ggc 1008Ala Gly Leu Leu Met Ser Leu Ala Lys Ala Asp Tyr Ser Ser Val Gly325 330 335atg ttc cac gcc cag acc att gcc cgc att ttc aat ggg gaa aag ccg 1056Met Phe His Ala Gln Thr Ile Ala Arg Ile Phe Asn Gly Glu Lys Pro340 345 350cgc agc atc agc cag gtc tgg aat gcc ccc gcc aag ata gcc atc aat 1104Arg Ser Ile Ser Gln Val Trp Asn Ala Pro Ala Lys Ile Ala Ile Asn355 360 365ctg gaa acg gcg cgg cgc atc ggc ttc gac cca ccg gtg gat att ctg 1152Leu Glu Thr Ala Arg Arg Ile Gly Phe Asp Pro Pro Val Asp Ile Leu370 375 380ctg gcg gcc gac gag gtg tac gaa gcg gag cac tga cag gcc tgg cca 1200Leu Ala Ala Asp Glu Val Tyr Glu Ala Glu His * Gln Ala Trp Pro385 390 395acg aga cct ggc aag gaa tgt gcc gga tcc tag1233Thr Arg Pro Gly Lys Glu Cys Ala Gly Ser *400 405<210>70<211>409<212>PRT<213>百日咳博德特氏菌<400>70Met Thr Leu Arg Pro Gly Ile Leu Ala Pro Leu Ala Leu Leu Leu Gly1 5 10 15Leu Ala Trp Pro Ile Ala Ala Ala Ser Ser Pro Pro Pro Val Ala Ala20 25 30Leu Ser Ser Gly Val Ala Leu Thr Ser Pro Arg Leu Pro Pro Pro Ser35 40 45His Thr Ser Gly Arg Lys Trp Arg Ile Gly Tyr Val Gly Ser Gly Glu50 55 60Tyr Glu Glu Tyr Pro Arg Thr Leu Tyr Ala Ile Ala Arg Ala Leu Gln65 70 75 80Gln Leu Gly Trp Leu Arg Ile Asp Asp Met Pro Glu Ile Thr Asp Met85 90 95Arg Lys Ala Trp Leu Tyr Leu Ala Thr His Ala Arg Ser Asn Tyr Ile100 105 110Glu Phe Val Pro Asp Ala Trp Trp Gln Pro Gly Asn Phe Asp Thr Ala115 120 125Leu Arg Pro Ala Val Arg Glu Ala Val Ala Ala Arg Leu His Gly Ala130 135 140Lys Asp Ile Asp Leu Ile Ile Ala Met Gly Thr Trp Ala Gly Gln Asp145 150 155 160Met Val Glu Leu Gly Thr Pro Val Pro Thr Val Val Val Ser Ser Thr165 170 175Asp Pro Ile Ser Ala Arg Ile Ile Pro Ser Ala Ala Asp Ser Gly Gln180 185 190Asp Asn Leu His Ala Arg Val Gln Pro Asp His Tyr Gln Arg Gln Ile195 200 205Gln Leu Leu His Asp Ile Val Pro Phe Lys Thr Leu Gly Leu Val Tyr210 215 220Glu Asp Thr Glu Ala Gly Arg Thr Tyr Ala Ala Ile Asp Lys Val Ala225 230 235 240Ala Leu Met Pro Ala Leu Asp Phe Ser Val Lys Arg Cys Asp Ala Arg245 250 255Ala Thr Gly Ile Pro Ile Ala Thr Ala Thr Gln Asn Val Leu Ala Cys260 265 270Tyr Gln Lys Leu Ser Ser Glu Val Asp Ala Phe Tyr Val Thr Glu His275 280 285Arg Gly Ile Thr Ser Thr Ser Val Lys Gln Leu Ala Ala Leu Leu Arg290 295 300Ala Ala Arg Val Pro Ser Phe Ser Met Gln Gly Ser Asp Glu Val Lys305 310 315 320Ala Gly Leu Leu Met Ser Leu Ala Lys Ala Asp Tyr Ser Ser Val Gly325 330 335Met Phe His Ala Gln Thr Ile Ala Arg Ile Phe Asn Gly Glu Lys Pro340 345 350Arg Ser Ile Ser Gln Val Trp Asn Ala Pro Ala Lys Ile Ala Ile Asn355 360 365Leu Glu Thr Ala Arg Arg Ile Gly Phe Asp Pro Pro Val Asp Ile Leu370 375 380Leu Ala Ala Asp Glu Val Tyr Glu Ala Glu His Gln Ala Trp Pro Thr385 390 395 400Arg Pro Gly Lys Glu Cys Ala Gly Ser405<210>71<211>645<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(645)<400>71atg gag cag ctg gat ctg ccg ctg gta gtg gtc ggg ctg tac ccg ggc48Met Glu Gln Leu Asp Leu Pro Leu Val Val Val Gly Leu Tyr Pro Gly1 5 10 15atg cag gtt gtc ctg gcc gct gtc ggc cgc act ggg tat gat ccg ggc96Met Gln Val Val Leu Ala Ala Val Gly Arg Thr Gly Tyr Asp Pro Gly20 25 30gct tat cgg gtc ggt cga cga gac gac cac ggt ggg tac cgg cgt gcc144Ala Tyr Arg Val Gly Arg Arg Asp Asp His Gly Gly Tyr Arg Arg Ala35 40 45cag ttc gac cat gtc ctg tcc agc cca ggt acc cat ggc gat gat cag192Gln Phe Asp His Val Leu Ser Ser Pro Gly Thr His Gly Asp Asp Gln50 55 60gtc gat gtc ctt ggc gcc atg cag gcg tgc cgc aac ggc ttc gcg cac240Val Asp Val Leu Gly Ala Met Gln Ala Cys Arg Asn Gly Phe Ala His65 70 75 80ggc agg ccg caa ggc ggt gtc gaa gtt gcc ggg ctg cca cca cgc atc288Gly Arg Pro Gln Gly Gly Val Glu Val Ala Gly Leu Pro Pro Arg Ile85 90 95ggg cac gaa ctc gat gta gtt gct gcg ggc atg cgt ggc cag gta aag336Gly His Glu Leu Asp Val Val Ala Ala Gly Met Arg Gly Gln Val Lys
100 105 110cca ggc ctt tcg cat atc ggt tat ctc ggg cat gtc gtc gat acg cag384Pro Gly Leu Ser His Ile Gly Tyr Leu Gly His Val Val Asp Thr Gln115 120 125cca tcc gag ttg ttg caa tgc gcg cgc gat cgc gta gag cgt gcg cgg432Pro Ser Glu Leu Leu Gln Cys Ala Arg Asp Arg Val Glu Arg Ala Arg130 135 140ata ctc ctc gta ctc gcc gct acc cac ata acc gat gcg cca ttt ccg480Ile Leu Leu Val Leu Ala Ala Thr His Ile Thr Asp Ala Pro Phe Pro145 150 155 160gcc gga tgt atg gga ggg agg cgg aag gcg ggg cga ggt aag ggc gac528Ala Gly Cys Met Gly Gly Arg Arg Lys Ala Gly Arg Gly Lys Gly Asp165 170 175gcc gga gct cag ggc cgc gac agg agg agg gct gga tgc cgc cgc gat576Ala Gly Ala Gln Gly Arg Asp Arg Arg Arg Ala Gly Cys Arg Arg Asp180 185 190ggg cca cgc gag gcc aag aag cag ggc gag cgg ggc gag tat tcc ggg624Gly Pro Arg Glu Ala Lys Lys Gln Gly Glu Arg Gly Glu Tyr Ser Gly195 200 205gcg aag ggt cat ggg cga tga645Ala Lys Gly His Gly Arg *210<210>72<211>214<212>PRT<213>百日咳博德特氏菌<400>72Met Glu Gln Leu Asp Leu Pro Leu Val Val Val Gly Leu Tyr Pro Gly1 5 10 15Met Gln Val Val Leu Ala Ala Val Gly Arg Thr Gly Tyr Asp Pro Gly20 25 30Ala Tyr Arg Val Gly Arg Arg Asp Asp His Gly Gly Tyr Arg Arg Ala35 40 45Gln Phe Asp His Val Leu Ser Ser Pro Gly Thr His Gly Asp Asp Gln50 55 60Val Asp Val Leu Gly Ala Met Gln Ala Cys Arg Asn Gly Phe Ala His65 70 75 80Gly Arg Pro Gln Gly Gly Val Glu Val Ala Gly Leu Pro Pro Arg Ile85 90 95Gly His Glu Leu Asp Val Val Ala Ala Gly Met Arg Gly Gln Val Lys100 105 110Pro Gly Leu Ser His Ile Gly Tyr Leu Gly His Val Val Asp Thr Gln115 120 125Pro Ser Glu Leu Leu Gln Cys Ala Arg Asp Arg Val Glu Arg Ala Arg130 135 140Ile Leu Leu Val Leu Ala Ala Thr His Ile Thr Asp Ala Pro Phe Pro145 150 155 160Ala Gly Cys Met Gly Gly Arg Arg Lys Ala Gly Arg Gly Lys Gly Asp165 170 175Ala Gly Ala Gln Gly Arg Asp Arg Arg Arg Ala Gly Cys Arg Arg Asp180 185 190Gly Pro Arg Glu Ala Lys Lys Gln Gly Glu Arg Gly Glu Tyr Ser Gly195 200 205Ala Lys Gly His Gly Arg210<210>73<211>1314<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1314)<400>73atg tcc aat acc tat ttc ccg cgc tgg cgg ctg gcc gac gac acc gtg48Met Ser Asn Thr Tyr Phe Pro Arg Trp Arg Leu Ala Asp Asp Thr Val1 5 10 15ccg ggc gcg gtc arc gcg ccc gac gaa cgc ctg tcc tgg ccc aag aac96Pro Gly Ala Val Ile Ala Pro Asp Glu Arg Leu Ser Trp Pro Lys Asn20 25 30atc gcc atg ggg gcc cag cac gtg gtc gcc atg ttc ggt tcc acc gtg144Ile Ala Met Gly Ala Gln His Val Val Ala Met Phe Gly Ser Thr Val35 40 45ctg gcg ccg ctg ctg atg ggt ttc gac ccc aat gtg gcg atc ctc atg192Leu Ala Pro Leu Leu Met Gly Phe Asp Pro Asn Val Ala Ile Leu Met50 55 60tcc ggc atc ggc acg ctg atc ttc ttc ctg ttc gtc ggc ggc cgg gtg240Ser Gly Ile Gly Thr Leu Ile Phe Phe Leu Phe Val Gly Gly Arg Val65 70 75 80ccc agc tac ctg ggc tcc agc ttc gcc ttc atc ggc ggg gtg gtg gcg288Pro Ser Tyr Leu Gly Ser Ser Phe Ala Phe Ile Gly Gly Val Val Ala85 90 95gtc acc ggc tat gtg gcg ccc ggc gcc aac gcc aat atc ggc gtg gcg336Val Thr Gly Tyr Val Ala Pro Gly Ala Asn Ala Asn Ile Gly Val Ala
100 105 110ctc ggc gcg atc atc gcc tgt ggc ctg gtg tac gcg ctg atc ggc ctg 384Leu Gly Ala Ile Ile Ala Cys Gly Leu Val Tyr Ala Leu Ile Gly Leu115 120 125gtc gta tgg gcg gcc agc gcg cgc ggc aac ggg gcg cgc tgg atc gag 432Val Val Trp Ala Ala Ser Ala Arg Gly Asn Gly Ala Arg Trp Ile Glu130 135 140gcc atg atg ccg ccg gtc gtc acg ggc gcg gtg gtg gcg gtg atc ggc 480Ala Met Met Pro Pro Val Val Thr Gly Ala Val Val Ala Val Ile Gly145 150 155 160ctg aac ctg gcc ccg atc gcc gcc aag ggc gcc atg ggt tcg tcc ggc 528Leu Asn Leu Ala Pro Ile Ala Ala Lys Gly Ala Met Gly Ser Ser Gly165 170 175ttc gag gcc agc atg gcg ttg atg acc atc ctg tgc gtg ggc ggc atc 576Phe Glu Ala Ser Met ala Leu Met Thr Ile Leu Cys Val Gly Gly Ile180 185 190gcc gtc tac acg cgc ggc atg gtg cag cgg ctg ctg atc ctg gtc ggc 624Ala Val Tyr Thr Arg Gly Met Val Gln Arg Leu Leu Ile Leu Val Gly195 200 205ctg gtg ctg gcc tgc gtc atc tac gcg gtc tgc gcc aac ggc ctg ggg 672Leu Val Leu Ala Cys Val Ile Tyr Ala Val Cys Ala Asn Gly Leu Gly210 215 220ctg ggc gcg ccc atg gac ttc gcc aag gtg gcc gcc gcg ccg tgg ttc 720Leu Gly Ala Pro Met Asp Phe Ala Lys Val Ala Ala Ala Pro Trp Phe225 230 235 240ggc ctg ccc agc ttc gcc gcg ccg gtg ttc gag ccg cag gcc atg ggc 768Gly Leu Pro Ser Phe Ala Ala Pro Val Phe Glu Pro Gln Ala Met Gly245 250 255ctg atc gtg ccg gtg gcc atc atc ctg gtg gcc gag aac ctg ggc cac 816Leu Ile Val Pro Val Ala Ile Ile Leu Val Ala Glu Asn Leu Gly His260 265 270gtg aag gcg gtc gcc gcc atg acc gga cag gac ctg gac cgc tac gtg 864Val Lys Ala Val Ala Ala Met Thr Gly Gln Asp Leu Asp Arg Tyr Val275 280 285ggc cgc gcc ttc gtg ggc gac ggc gtg gcg acc atg gtt tcc ggc gcc 912Gly Arg Ala Phe Val Gly Asp Gly Val Ala Thr Met Val Ser Gly Ala290 295 300gtc ggc ggc acc ggg gtg acc acc tac gcc gag aat atc ggc gtg atg 960Val Gly Gly Thr Gly Val Thr Thr Tyr Ala Glu Asn Ile Gly Val Met305 310 315 320gcc gtg acg cgc atc tat tcc acg ctg gtg ttc gtg gtg gcg gcc gtg1008Ala Val Thr Arg Ile Tyr Ser Thr Leu Val Phe Val Val Ala Ala Val325 330 335atc gcg ctg gtg ctg ggg ttc tcg ccc aag ttc ggc gcg ctg atc cag1056Ile Ala Leu Val Leu Gly Phe Ser Pro Lys Phe Gly Ala Leu Ile Gln340 345 350acc atc ccc ggc ccc gtg ctg ggg ggc atg tcg gtc gtg gtg ttc ggc1104Thr Ile Pro Gly Pro Val Leu Gly Gly Met Ser Val Val Val Phe Gly355 360 365ctg atc gcc atc gcc ggc gcg cgc atc tgg gtg gtc aac cag gtc gat1152Leu Ile Ala Ile Ala Gly Ala Arg Ile Trp Val Val Asn Gln Val Asp370 375 380ttc agc gac aac cgc aat ctg atc gtg gcc gcc gtg acc ctg gtg ctg1200Phe Ser Asp Asn Arg Asn Leu Ile Val Ala Ala Val Thr Leu Val Leu385 390 395 400ggg gcg ggc gac ttc agc gtc aag ctg ggc gat ttc tcg atg aac ggc1248Gly Ala Gly Asp Phe Ser Val Lys Leu Gly Asp Phe Ser Met Asn Gly405 410 415atc ggc acc gcc acg ttc ggc gcc atc atc ctg tac gcc ctg ctg ggc1296Ile Gly Thr Ala Thr Phe Gly Ala Ile Ile Leu Tyr Ala Leu Leu Gly420 425 430ctg gcg cgt cgc cgc tga1314Leu Ala Arg Arg Arg *435<210>74<211>437<212>PRT<213>百日咳博德特氏菌<400>74Met Ser Asn Thr Tyr Phe Pro Arg Trp Arg Leu Ala Asp Asp Thr Val1 5 10 15Pro Gly Ala Val Ile Ala Pro Asp Glu Arg Leu Ser Trp Pro Lys Asn20 25 30Ile Ala Met Gly Ala Gln His Val Val Ala Met Phe Gly Ser Thr Val35 40 45Leu Ala Pro Leu Leu Met Gly Phe Asp Pro Asn Val Ala Ile Leu Met50 55 60Ser Gly Ile Gly Thr Leu Ile Phe Phe Leu Phe Val Gly Gly Arg Val65 70 75 80Pro Ser Tyr Leu Gly Ser Ser Phe Ala Phe Ile Gly Gly Val Val Ala85 90 95Val Thr Gly Tyr Val Ala Pro Gly Ala Asn Ala Asn Ile Gly Val Ala100 105 110Leu Gly Ala Ile Ile Ala Cys Gly Leu Val Tyr Ala Leu Ile Gly Leu115 120 125Val Val Trp Ala Ala Ser Ala Arg Gly Asn Gly Ala Arg Trp Ile Glu130 135 40Ala Met Met Pro Pro Val Val Thr Gly Ala Val Val Ala Val Ile Gly145 150 155 160Leu Asn Leu Ala Pro Ile Ala Ala Lys Gly Ala Met Gly Ser Ser Gly165 170 175Phe Glu Ala Ser Met ala Leu Met Thr Ile Leu Cys Val Gly Gly Ile180 185 190Ala Val Tyr Thr Arg Gly Met Val Gln Arg Leu Leu Ile Leu Val Gly195 200 205Leu Val Leu Ala Cys Val Ile Tyr Ala Val Cys Ala Asn Gly Leu Gly210 215 220Leu Gly Ala Pro Met Asp Phe Ala Lys Val Ala Ala Ala Pro Trp Phe225 230 235 240Gly Leu Pro Ser Phe Ala Ala Pro Val Phe Glu Pro Gln Ala Met Gly245 250 255Leu Ile Val Pro Val Ala Ile Ile Leu Val Ala Glu Asn Leu Gly His260 265 270Val Lys Ala Val Ala Ala Met Thr Gly Gln Asp Leu Asp Arg Tyr Val275 280 285Gly Arg Ala Phe Val Gly Asp Gly Val Ala Thr Met Val Ser Gly Ala290 295 300Val Gly Gly Thr Gly Val Thr Thr Tyr Ala Glu Asn Ile Gly Val Met305 310 315 320Ala Val Thr Arg Ile Tyr Ser Thr Leu Val Phe Val Val Ala Ala Val325 330 335Ile Ala Leu Val Leu Gly Phe Ser Pro Lys Phe Gly Ala Leu Ile Gln340 345 350Thr Ile Pro Gly Pro Val Leu Gly Gly Met Ser Val Val Val Phe Gly355 360 365Leu Ile Ala Ile Ala Gly Ala Arg Ile Trp Val Val Asn Gln Val Asp370 375 380Phe Ser Asp Asn Arg Asn Leu Ile Val Ala Ala Val Thr Leu Val Leu385 390 395 400Gly Ala Gly Asp Phe Ser Val Lys Leu Gly Asp Phe Ser Met Asn Gly405 410 415Ile Gly Thr Ala Thr Phe Gly Ala Ile Ile Leu Tyr Ala Leu Leu Gly420 425 430Leu Ala Arg Arg Arg435<210>75<211>1536<212>DNA
<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1536)<400>75atg ctt gaa agg atc aag gtc cgc acc gcg atg gtg gcg gta ttc gcg48Met Leu Glu Arg Ile Lys Val Arg Thr Ala Met Val Ala Val Phe Ala1 5 10 15tgc ttc ctg gcg gtg ctg atg ctg tcg ggc gcc ctg acg tgg cgc aac96Cys Phe Leu Ala Val Leu Met Leu Ser Gly Ala Leu Thr Trp Arg Asn20 25 30gcg ggc agg agc gcc gcc gag atc gag ggg ctg aac cag gtc gcc gtc144Ala Gly Arg Ser Ala Ala Glu Ile Glu Gly Leu Asn Gln Val Ala Val35 40 45aac cag gtc gac ccg ctg ttc gag gcc agc ggc gcg gcg cag cgc cag192Asn Gln Val Asp Pro Leu Phe Glu Ala Ser Gly Ala Ala Gln Arg Gln50 55 60gcg gcc acg caa ttc cag cgc tac gtg gac gtg ccc aag gag ccg gcc240Ala Ala Thr Gln Phe Gln Arg Tyr Val Asp Val Pro Lys Glu Pro Ala65 70 75 80gcg gcc gag ctg gcc gcg acc ctg cag acg cgc tgg cgc gcc tac cag288Ala Ala Glu Leu Ala Ala Thr Leu Gln Thr Arg Trp Arg Ala Tyr Gln85 90 95tcg gtg ctg gac gag ctg gcc gcc gcc gtc gac gcc ggc cag gcc gag336Ser Val Leu Asp Glu Leu Ala Ala Ala Val Asp Ala Gly Gln Ala Glu100 105 110ccc gcc ctg gcc gcc atg cat cgc gcg cag cag gcc gaa cat gca ttc384Pro Ala Leu Ala Ala Met His Arg Ala Gln Gln Ala Glu His Ala Phe115 120 125cag cgc gac atg gaa gcc ttt ctg gcc agg gta cag gcg cac agc gac432Gln Arg Asp Met Glu Ala Phe Leu Ala Arg Val Gln Ala His Ser Asp130 135 140gaa gtg cgc agc ggc gcc gag gac acc cat gtc gtg gcc cgc tgg agc480Glu Val Arg Ser Gly Ala Glu Asp Thr His Val Val Ala Arg Trp Ser145 150 155 160gcc atc gcg ctg acc acg ctg ggc gtg ctg ctg acc ctg gcc ggc tgg528Ala Ile Ala Leu Thr Thr Leu Gly Val Leu Leu Thr Leu Ala Gly Trp165 170 175ctg ttc gtg cgc cgc gcg gtg ctg cgc ccc ttg ctg gag gcc ggc cat 576Leu Phe Val Arg Arg Ala Val Leu Arg Pro Leu Leu Glu Ala Gly His180 185 190cat ttc gac cgc atc gcc gac ggc gac ctc acc gcg cgc atc gag gtg 624His Phe Asp Arg Ile Ala Asp Gly Asp Leu Thr Ala Arg Ile Glu Val195 200 205cgc tcg gcc aat gaa atc ggc gcg ctg ttc gcg gcg ctc aag cgc atg 672Arg Ser Ala Asn Glu Ile Gly Ala Leu Phe Ala Ala Leu Lys Arg Met210 215 220cag gaa ggc ctg acg cgc acc atc gcc gtc atg cgg cgc ggc gtc gac 720Gln Glu Gly Leu Thr Arg Thr Ile Ala Val Met Arg Arg Gly Val Asp225 230 235 240gaa atc aac gtc ggc gcg gcc gag atc tcg gcc ggc aac gcc aac ctg 768Glu Ile Asn Val Gly Ala Ala Glu Ile Ser Ala Gly Asn Ala Asn Leu245 250 255tcc agc cgc acg gag gag cag gcc gcc gcc ctg gaa gag acc gcg gcc 816Ser Ser Arg Thr Glu Glu Gln Ala Ala Ala Leu Glu Glu Thr Ala Ala260 265 270acc atg gag gaa ctg gcc acc acg gtc aag cag aac gcc gac aat gcc 864Thr Met Glu Glu Leu Ala Thr Thr Val Lys Gln Asn Ala Asp Asn Ala275 280 285gcg cag gcc aat cag ctg gcc gcc gtc agc atg cag gtg gcg cag cgc 912Ala Gln Ala Asn Gln Leu Ala Ala Val Ser Met Gln Val Ala Gln Arg290 295 300ggc ggc gag tcg gtc gcg cag gtg gtg cag acc atg cac ggc atc tcc 960Gly Gly Glu Ser Val Ala Gln Val Val Gln Thr Met His Gly Ile Ser305 310 315 320gcg agc tcg cgc cag atc gcc gac atc gtc acc gtg atc gac ggc atc 1008Ala Ser Ser Arg Gln Ile Ala Asp Ile Val Thr Val Ile Asp Gly Ile325 330 335gcc ttc cag acc aat atc ctg gcg ctg aac gcc gcg gtc gag gcg gcg 1056Ala Phe Gln Thr Asn Ile Leu Ala Leu Asn Ala Ala Val Glu Ala Ala340 345 350cgc gcc ggc gaa cag ggc aag ggc ttc gcg gtg gtg gcg ggc gag gtg 1104Arg Ala Gly Glu Gln Gly Lys Gly Phe Ala Val Val Ala Gly Glu Val355 360 365cgc agc ctg gcc cag cgc gcc gcg cag gcg gcc aag gag atc aag gcc 1152Arg Ser Leu Ala Gln Arg Ala Ala Gln Ala Ala Lys Glu Ile Lys Ala370 375 380ctg atc gag agc tcg gtg gcg acg gtg cgc gcc ggc tcg caa cag gtc1200Leu Ile Glu Ser Ser Val Ala Thr Val Arg Ala Gly Ser Gln Gln Val385 390 395 400gcc agc gcc ggc ggc acc atg gac gag gtg gtg gcc tcg gta cag cgc1248Ala Ser Ala Gly Gly Thr Met Asp Glu Val Val Ala Ser Val Gln Arg405 410 415gtg gcc gac atc atg ggg gag atc tcg gcc gcc tcg gcc cag cag gcc1296Val Ala Asp Ile Met Gly Glu Ile Ser Ala Ala Ser Ala Gln Gln Ala420 425 430agc ggc atc gac cag gtc agc ctg gcg att tcg caa atg gac gaa acc1344Ser Gly Ile Asp Gln Val Ser Leu Ala Ile Ser Gln Met Asp Glu Thr435 440 445acc cag cag aat gcc gcg ctg gtc gaa cag gcc gcg gcg gcg gcc acg1392Thr Gln Gln Asn Ala Ala Leu Val Glu Gln Ala Ala Ala Ala Ala Thr450 455 460gcc atg gaa gaa cag gcc cgc cac ctg gcg gcc gcg gcg gcg gtc ttc1440Ala Met Glu Glu Gln Ala Arg His Leu Ala Ala Ala Ala Ala Val Phe465 470 475 480agg acg cag ggc ggc gcc atc atc gac gtc gcc gcc gcg ccg ctg gcc1488Arg Thr Gln Gly Gly Ala Ile Ile Asp Val Ala Ala Ala Pro Leu Ala485 490 495ggg ccg gcg ggc ggc cat gcc gcc ctg ccg ccg gcc gcg gcc cac tga1536Gly Pro Ala Gly Gly His Ala Ala Leu Pro Pro Ala Ala Ala His *500 505 510<210>76<211>511<212>PRT<213>百日咳博德特氏菌<400>76Met Leu Glu Arg Ile Lys Val Arg Thr Ala Met Val Ala Val Phe Ala1 5 10 15Cys Phe Leu Ala Val Leu Met Leu Ser Gly Ala Leu Thr Trp Arg Asn20 25 30Ala Gly Arg Ser Ala Ala Glu Ile Glu Gly Leu Asn Gln Val Ala Val35 40 45Asn Gln Val Asp Pro Leu Phe Glu Ala Ser Gly Ala Ala Gln Arg Gln50 55 60Ala Ala Thr Gln Phe Gln Arg Tyr Val Asp Val Pro Lys Glu Pro Ala65 70 75 80Ala Ala Glu Leu Ala Ala Thr Leu Gln Thr Arg Trp Arg Ala Tyr Gln
85 90 95Ser Val Leu Asp Glu Leu Ala Ala Ala Val Asp Ala Gly Gln Ala Glu100 105 110Pro Ala Leu Ala Ala Met His Arg Ala Gln Gln Ala Glu His Ala Phe115 120 125Gln Arg Asp Met Glu Ala Phe Leu Ala Arg Val Gln Ala His Ser Asp130 135 140Glu Val Arg Ser Gly Ala Glu Asp Thr His Val Val Ala Arg Trp Ser145 150 155 160Ala Ile Ala Leu Thr Thr Leu Gly Val Leu Leu Thr Leu Ala Gly Trp165 170 175Leu Phe Val Arg Arg Ala Val Leu Arg Pro Leu Leu Glu Ala Gly His180 185 190His Phe Asp Arg Ile Ala Asp Gly Asp Leu Thr Ala Arg Ile Glu Val195 200 205Arg Ser Ala Asn Glu Ile Gly Ala Leu Phe Ala Ala Leu Lys Arg Met210 215 220Gln Glu Gly Leu Thr Arg Thr Ile Ala Val Met Arg Arg Gly Val Asp225 230 235 240Glu Ile Asn Val Gly Ala Ala Glu Ile Ser Ala Gly Asn Ala Asn Leu245 250 255Ser Ser Arg Thr Glu Glu Gln Ala Ala Ala Leu Glu Glu Thr Ala Ala260 265 270Thr Met Glu Glu Leu Ala Thr Thr Val Lys Gln Asn Ala Asp Asn Ala275 280285Ala Gln Ala Asn Gln Leu Ala Ala Val Ser Met Gln Val Ala Gln Arg290 295 300Gly Gly Glu Ser Val Ala Gln Val Val Gln Thr Met His Gly Ile Ser305 310 315 320Ala Ser Ser Arg Gln Ile Ala Asp Ile Val Thr Val Ile Asp Gly Ile325 330 335Ala Phe Gln Thr Asn Ile Leu Ala Leu Asn Ala Ala Val Glu Ala Ala340 345 350Arg Ala Gly Glu Gln Gly Lys Gly Phe Ala Val Val Ala Gly Glu Val355360 365Arg Ser Leu Ala Gln Arg Ala Ala Gln Ala Ala Lys Glu Ile Lys Ala370375 380Leu Ile Glu Ser Ser Val Ala Thr Val Arg Ala Gly Ser Gln Gln Val385 390 395 400Ala Ser Ala Gly Gly Thr Met Asp Glu Val Val Ala Ser Val Gln Arg405 410 415Val Ala Asp Ile Met Gly Glu Ile Ser Ala Ala Ser Ala Gln Gln Ala420425 430Ser Gly Ile Asp Gln Val Ser Leu Ala Ile Ser Gln Met Asp Glu Thr435 440 445Thr Gln Gln Asn Ala Ala Leu Val Glu Gln Ala Ala Ala Ala Ala Thr450 455 460Ala Met Glu Glu Gln Ala Arg His Leu Ala Ala Ala Ala Ala Val Phe465 470 475 480Arg Thr Gln Gly Gly Ala Ile Ile Asp Val Ala Ala Ala Pro Leu Ala485 490 495Gly Pro Ala Gly Gly His Ala Ala Leu Pro Pro Ala Ala Ala His500 505 510<210>77<211>477<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(477)<400>77atg tct gcc att cct ttg acc gtg cgc ggg gcc gag cgc ttg cag caa48Met Ser Ala Ile Pro Leu Thr Val Arg Gly Ala Glu Arg Leu Gln Gln1 5 10 15gaa ctg cat cgg ctt aag acc gtt gag cgt cct gcg gtg atc agc gcc96Glu Leu His Arg Leu Lys Thr Val Glu Arg Pro Ala Val Ile Ser Ala20 25 30att gcg gag gcg cgt gcg cag ggt gat ttg tcg gaa aat gcc gag tac144Ile Ala Glu Ala Arg Ala Gln Gly Asp Leu Ser Glu Asn Ala Glu Tyr35 40 45gac gcc gcc cgc gaa cgc cag ggc ttc atc gaa ggc cgg atc tcc gaa192Asp Ala Ala Arg Glu Arg Gln Gly Phe Ile Glu Gly Arg Ile Ser Glu50 55 60ctc gag ggc acg ctt tcg aac gcg cac ctc atc gat cca acg gcg ctc240Leu Glu Gly Thr Leu Ser Asn Ala His Leu Ile Asp Pro Thr Ala Leu65 70 75 80gac gcc gaa ggc cgt gcc gtg ttc ggc gcg acc gtg gaa atc gaa gac288Asp Ala Glu Gly Arg Ala Val Phe Gly Ala Thr Val Glu Ile Glu Asp85 90 95ctc gac tcg ggc gac cgc ctg acc tac cag atc gtg ggc gac gtc gaa336Leu Asp Ser Gly Asp Arg Leu Thr Tyr Gln Ile Val Gly Asp Val Glu100 105 110gcc gac atc aag tcc aac ctg att tcg gtc tcc agc ccg gtg gcc cgc384Ala Asp Ile Lys Ser Asn Leu Ile Ser Val Ser Ser Pro Val Ala Arg115 120 125gcc ctg atc ggc aaa tcc gag ggc gat gtg gtc gaa gtg aag gtg ccg432Ala Leu Ile Gly Lys Ser Glu Gly Asp Val Val Glu Val Lys Val Pro130 135 140gct ggc gtg cgc gag tac gaa gtc atc ggt gtg cgt tat ctc tga477Ala Gly Val Arg Glu Tyr Glu Val Ile Gly Val Arg Tyr Leu *145 150 155<210>78<211>158<212>PRT<213>百日咳博德特氏菌<400>78Met Ser Ala Ile Pro Leu Thr Val Arg Gly Ala Glu Arg Leu Gln Gln1 5 10 15Glu Leu His Arg Leu Lys Thr Val Glu Arg Pro Ala Val Ile Ser Ala20 25 30Ile Ala Glu Ala Arg Ala Gln Gly Asp Leu Ser Glu Asn Ala Glu Tyr35 40 45Asp Ala Ala Arg Glu Arg Gln Gly Phe Ile Glu Gly Arg Ile Ser Glu50 55 60Leu Glu Gly Thr Leu Ser Asn Ala His Leu Ile Asp Pro Thr Ala Leu65 70 75 80Asp Ala Glu Gly Arg Ala Val Phe Gly Ala Thr Val Glu Ile Glu Asp85 90 95Leu Asp Ser Gly Asp Arg Leu Thr Tyr Gln Ile Val Gly Asp Val Glu100 105 110Ala Asp Ile Lys Ser Asn Leu Ile Ser Val Ser Ser Pro Val Ala Arg115 120 125Ala Leu Ile Gly Lys Ser Glu Gly Asp Val Val Glu Val Lys Val Pro130 135 140Ala Gly Val Arg Glu Tyr Glu Val Ile Gly Val Arg Tyr Leu145 150 155<210>79<211>951<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(951)<400>79atg aac acc cat aag cat gcc cga ttg acc ttc cta cgt cga ctc gaa48Met Asn Thr His Lys His Ala Arg Leu Thr Phe Leu Arg Arg Leu Glu1 5 10 15atg gtc cag caa ttg atc gcc cat caa gtt tgt gtg cct gaa gcg gcc96Met Val Gln Gln Leu Ile Ala His Gln Val Cys Val Pro Glu Ala Ala20 25 30cgc gcc tat ggg gtc acc gcg ccg act gtg cgc aaa tgg ctg ggc cgc144Arg Ala Tyr Gly Val Thr Ala Pro Thr Val Arg Lys Trp Leu Gly Arg35 40 45ttc ctg gct cag ggc cag gcg ggc ttg gcc gat gcg tcc tcg cgc ccg 192Phe Leu Ala Gln Gly Gln Ala Gly Leu Ala Asp Ala Ser Ser Arg Pro50 55 60acg gtc tcg ccc cga gcg att gcg ccg gcc aag gcg ctg gct atc gtg 240Thr Val Ser Pro Arg Ala Ile Ala Pro Ala Lys Ala Leu Ala Ile Val65 70 75 80gag ctg cgc cgc aag cgg ctg acc caa gcg cgc atc gcc cag gcg ctg 288Glu Leu Arg Arg Lys Arg Leu Thr Gln Ala Arg Ile Ala Gln Ala Leu85 90 95ggc gtg tca gcc agc acc gtc agc cgc gtc ctg gcc cgc gcc ggt ctg 336Gly Val Ser Ala Ser Thr Val Ser Arg Val Leu Ala Arg Ala Gly Leu100 105 110tcg cac ctg gcc gac ctg gag ccg gcc gag ccg gtg gtg cgc tac gag 384Ser His Leu Ala Asp Leu Glu Pro Ala Glu Pro Val Val Arg Tyr Glu115 120 125cat cag gcc ccc ggc gat ctg ctg cac atc gac atc aag aag ctg gga 432His Gln Ala Pro Gly Asp Leu Leu His Ile Asp Ile Lys Lys Leu Gly130 135 140cgt atc cag cgc cct ggc cac cgg gtc acg ggc aac cga cgc gat acc 480Arg Ile Gln Arg Pro Gly His Arg Val Thr Gly Asn Arg Arg Asp Thr145 150 155 160gtt gag ggg gcc ggc tgg gac ttc gtc ttc gtg gcc atc gat gac cac 528Val Glu Gly Ala Gly Trp Asp Phe Val Phe Val Ala Ile Asp Asp His165 170 175gcc cgc gtg gcc ttc acc gac atc cac ccc gac gag cgc ttc ccc agc 576Ala Arg Val Ala Phe Thr Asp Ile His Pro Asp Glu Arg Phe Pro Ser180 185 190gcc gtc cag ttc ctc aag gac gca gtg gcc tac tac cag cgc ctg ggc 624Ala Val Gln Phe Leu Lys Asp Ala Val Ala Tyr Tyr Gln Arg Leu Gly195 200 205gtg acc atc cag cgc ttg ctc acc gac aat ggc tcg gcc ttt cgc agc 672Val Thr Ile Gln Arg Leu Leu Thr Asp Asn Gly Ser Ala Phe Arg Ser210 215 220cgc gcc ttc gcc gcg ctg tgc cat gag ctg ggc atc aag cac cgc ttt 720Arg Ala Phe Ala Ala Leu Cys His Glu Leu Gly Ile Lys His Arg Phe225 230 235 240acc cga cct tac cgc cca cag acc aat ggc aag gcc gaa cgc ttc atc 768Thr Arg Pro Tyr Arg Pro Gln Thr Asn Gly Lys Ala Glu Arg Phe Ile
245 250 255cag tcg gcc ttg cgt gag tgg gct tac gct cac acc tac cag aac tcc816Gln Ser Ala Leu Arg Glu Trp Ala Tyr Ala His Thr Tyr Gln Asn Ser260 265 270caa cac cga gcc gat gcc atg aaa tcc tgg cta cac cac tac aac tgg864Gln His Arg Ala Asp Ala Met Lys Ser Trp Leu His His Tyr Asn Trp275 280 285cat cga ccc cac caa ggc atc ggg cgc gct gta ccc atc tcc aga ctc912His Arg Pro His Gln Gly Ile Gly Arg Ala Val Pro Ile Ser Arg Leu290 295 300aac ctg gac gaa tac aac cta ttg aca gtt cac acc tag951Asn Leu Asp Glu Tyr Asn Leu Leu Thr Val His Thr *305 310 315<210>80<211>316<212>PRT<213>百日咳博德特氏菌<400>80Met Asn Thr His Lys His Ala Arg Leu Thr Phe Leu Arg Arg Leu Glu1 5 10 15Met Val Gln Gln Leu Ile Ala His Gln Val Cys Val Pro Glu Ala Ala20 25 30Arg Ala Tyr Gly Val Thr Ala Pro Thr Val Arg Lys Trp Leu Gly Arg35 40 45Phe Leu Ala Gln Gly Gln Ala Gly Leu Ala Asp Ala Ser Ser Arg Pro50 55 60Thr Val Ser Pro Arg Ala Ile Ala Pro Ala Lys Ala Leu Ala Ile Val65 70 75 80Glu Leu Arg Arg Lys Arg Leu Thr Gln Ala Arg Ile Ala Gln Ala Leu85 90 95Gly Val Ser Ala Ser Thr Val Ser Arg Val Leu Ala Arg Ala Gly Leu100 105 110Ser His Leu Ala Asp Leu Glu Pro Ala Glu Pro Val Val Arg Tyr Glu115 120 125His Gln Ala Pro Gly Asp Leu Leu His Ile Asp Ile Lys Lys Leu Gly130 135 140Arg Ile Gln Arg Pro Gly His Arg Val Thr Gly Asn Arg Arg Asp Thr145 150 155 160Val Glu Gly Ala Gly Trp Asp Phe Val Phe Val Ala Ile Asp Asp His165 170 175Ala Arg Val Ala Phe Thr Asp Ile His Pro Asp Glu Arg Phe Pro Ser180 185 190Ala Val Gln Phe Leu Lys Asp Ala Val Ala Tyr Tyr Gln Arg Leu Gly195 200 205Val Thr Ile Gln Arg Leu Leu Thr Asp Asn Gly Ser Ala Phe Arg Ser210 215 220Arg Ala Phe Ala Ala Leu Cys His Glu Leu Gly Ile Lys His Arg Phe225 230 235 240Thr Arg Pro Tyr Arg Pro Gln Thr Asn Gly Lys Ala Glu Arg Phe Ile245 250 255Gln Ser Ala Leu Arg Glu Trp Ala Tyr Ala His Thr Tyr Gln Asn Ser260 265 270Gln His Arg Ala Asp Ala Met Lys Ser Trp Leu His His Tyr Asn Trp275 280 285His Arg Pro His Gln Gly Ile Gly Arg Ala Val Pro Ile Ser Arg Leu290 295 300Asn Leu Asp Glu Tyr Asn Leu Leu Thr Val His Thr305 310 315<210>81<211>1851<212>DNA<213>百日咳博德特氏菌<220>
<221>CDS<222>(1)...(1851)<400>81atg gac ttg gta gtt cgt gac acc gac acg cgc tgg tcc acg ctg ctc48Met Asp Leu Val Val Arg Asp Thr Asp Thr Arg Trp Ser Thr Leu Leu1 5 10 15gac gac aag atc cgc acc atc cgg gaa agc cgc agg caa ctg atc caa96Asp Asp Lys Ile Arg Thr Ile Arg Glu Ser Arg Arg Gln Leu Ile Gln20 25 30ctc agc gcg gtc gtg aca tcg gtg ctg aac gcc tat gcc gcg cag gcc144Leu Ser Ala Val Val Thr Ser Val Leu Asn Ala Tyr Ala Ala Gln Ala35 40 45gag cgc gga cac gtc act acc ggc gcc gcc aag ggc atg gcg cgt gtc192Glu Arg Gly His Val Thr Thr Gly Ala Ala Lys Gly Met ala Arg Val50 55 60tgg ctg aac cat ctc gac ctg gga ccg cgc cgc gtc gcc ttc gcc tat240Trp Leu Asn His Leu Asp Leu Gly Pro Arg Arg Val Ala Phe Ala Tyr65 70 75 80gac gcg gaa ggc acc gtg ctg gcc agc acc aac ccc cgg atg atc gac288Asp Ala Glu Gly Thr Val Leu Ala Ser Thr Asn Pro Arg Met Ile Asp85 90 95cgg gac ctc tcc ggg atc cgc gac ttc aag ggc cgg ccg ctc gcc gcc336Arg Asp Leu Ser Gly Ile Arg Asp Phe Lys Gly Arg Pro Leu Ala Ala
100 105 110gcc atg tac gag gaa agc cgc aac gac ggt cgc ggc ttc gcc atc tac 384Ala Met Tyr Glu Glu Ser Arg Asn Asp Gly Arg Gly Phe Ala Ile Tyr115 120 125ccg tct ccc ctg gac gag tcc gcc cag atg cga cac gcc tat ttc gtg 432Pro Ser Pro Leu Asp Glu Ser Ala Gln Met Arg His Ala Tyr Phe Val130 135 140tac ttc ccc gcg tgg aag tgg gtt ctc gcc atc tcc gat agc tca caa 480Tyr Phe Pro Ala Trp Lys Trp Val Leu Ala Ile Ser Asp Ser Ser Gln145 150 155 160gcc atc atc gac aag gtt gcc gcc cag aaa gcc aac atg att gcc gcg 528Ala Ile Ile Asp Lys Val Ala Ala Gln Lys Ala Asn Met Ile Ala Ala165 170 175ata gac cgg aac ctg tcg gag ctg cgg ctc agc cgc cat ggt ttc gtg 576Ile Asp Arg Asn Leu Ser Glu Leu Arg Leu Ser Arg His Gly Phe Val180 185 190ttc gtg gtt gcg gac gat ggc acg gtg atc gtg ccg cca ccc cca tcg 624Phe Val Val Ala Asp Asp Gly Thr Val Ile Val Pro Pro Pro Pro Ser195 200 205gcc gcc cgg ctg ctg gac tcg aca gac gtc gaa tcg gga cgg gta ttg 672Ala Ala Arg Leu Leu Asp Ser Thr Asp Val Glu Ser Gly Arg Val Leu210 215 220cat tcg atg ctt gcc gaa atc tcg tct acc cgc ggc ctg acg ttg cgc 720His Ser Met Leu Ala Glu Ile Ser Ser Thr Arg Gly Leu Thr Leu Arg225 230 235 240ttt acc aac ggc gaa agc gcc tgg cag atc gac gcc ctg cga tac aag 768Phe Thr Asn Gly Glu Ser Ala Trp Gln Ile Asp Ala Leu Arg Tyr Lys245 250 255ccg ctg cat tgg acc atc atc ggt gtc gtt ccc gag ccg gac ctg acc 816Pro Leu His Trp Thr Ile Ile Gly Val Val Pro Glu Pro Asp Leu Thr260 265 270gac ccg gca cag aat ctg gtg cgc cgg cag gca ctg atc ttc gcc gcc 864Asp Pro Ala Gln Asn Leu Val Arg Arg Gln Ala Leu Ile Phe Ala Ala275 280 285acc ttg ctg gcc ggg ctg atg ctg gca tgg gtg gtg gcg gtg cgc atc 912Thr Leu Leu Ala Gly Leu Met Leu Ala Trp Val Val Ala Val Arg Ile290 295 300gcc cgg ccg ttg gcg caa ctg agc aac tac gct cgc cag ctt ccc acc 960Ala Arg Pro Leu Ala Gln Leu Ser Asn Tyr Ala Arg Gln Leu Pro Thr305 310 315 320cag gac ctc acc gag ccg atc cgg gtt ccg ccg tcg gtg gca tgc ctg 1008Gln Asp Leu Thr Glu Pro Ile Arg Val Pro Pro Ser Val Ala Cys Leu325 330 335ccg cgc cgg cgg cgc gac gaa gtc gga cag ctc gcc gaa tcg ttc ctg 1056Pro Arg Arg Arg Arg Asp Glu Val Gly Gln Leu Ala Glu Ser Phe Leu340 345 350ttc atg aac gaa cag ctg cac cac aat gtg cgg gcc ctg atg gcg cag 1104Phe Met Asn Glu Gln Leu His His Asn Val Arg Ala Leu Met ala Gln355 360 365ata tcg aac cgc gaa cgc ctc gaa agc gaa ttg agc atc gcc cgc tcc 1152Ile Ser Asn Arg Glu Arg Leu Glu Ser Glu Leu Ser Ile Ala Arg Ser370 375 380atc caa ctt ggc ctg ctt ccc cag ccg ttg ccc gat gcg gcc acg cgc 1200Ile Gln Leu Gly Leu Leu Pro Gln Pro Leu Pro Asp Ala Ala Thr Arg385 390 395 400ggc agc cag ttg cgt gcc gtc atg tac ccg gcc cgg gag gtc ggt ggg 1248Gly Ser Gln Leu Arg Ala Val Met Tyr Pro Ala Arg Glu Val Gly Gly405 410 415gat ttc tac gac tac ttc gtg ctg gca gac ggg cgt ctg tgc ttt gcc 1296Asp Phe Tyr Asp Tyr Phe Val Leu Ala Asp Gly Arg Leu Cys Phe Ala420 425 430atc ggc gac gta tcc gga aaa ggc gtg ccc gcg gcc ctg ttc atg gcc 1344Ile Gly Asp Val Ser Gly Lys Gly Val Pro Ala Ala Leu Phe Met ala435 440 445atc gtc agg acc ttg ata cgc agc gtg gcg gaa gaa gag cac gac ccg 1392Ile Val Arg Thr Leu Ile Arg Ser Val Ala Glu Glu Glu His Asp Pro450 455 460ggc gcc atc gcc acc aag gtg aac cac cgt ctg gcc gag aac aac ccc 1440Gly Ala Ile Ala Thr Lys Val Asn His Arg Leu Ala Glu Asn Asn Pro465 470 475 480aag ctg atg ttt gtc acc ttg ctg ata ggc gtc ttc acc ccg gaa aca 1488Lys Leu Met Phe Val Thr Leu Leu Ile Gly Val Phe Thr Pro Glu Thr485 490 495ggc gcc ctg gcc tgg gtc aac gcc ggc cac ccg ccg ccg ctg ctc atc 1536Gly Ala Leu Ala Trp Val Asn Ala Gly His Pro Pro Pro Leu Leu Ile500 505 510gac gaa cgt ggc gag gtc cgc ctg ctt caa gga agc agc ggc gcg gcc1584Asp Glu Arg Gly Glu Val Arg Leu Leu Gln Gly Ser Ser Gly Ala Ala515 520 525tgc ggc gtg ctg gac aac gag gcg tat tcc acc ctg agc acc acc ttg1632Cys Gly Val Leu Asp Asn Glu Ala Tyr Ser Thr Leu Ser Thr Thr Leu530 535 540ccg aac ggc acc tcg ctg gtc gcg ttt acc gac ggc gtc acc gaa gcc1680Pro Asn Gly Thr Ser Leu Val Ala Phe Thr Asp Gly Val Thr Glu Ala545 550 555 560atc cac ggc ggc tgc gcc cag tatggt ctg ccg cgg ctg gtc gcc ctg1728Ile His Gly Gly Cys Ala Gln Tyr Gly Leu Pro Arg Leu Val Ala Leu565 570 575atg cag ggc gcg ccg cac gca gcg gcc gaa ctc atc gag cac att ctg1776Met Gln Gly Ala Pro His Ala Ala Ala Glu Leu Ile Glu His Ile Leu580 585 590cac gac cta cgc gaa ttc gcc gcc gat tcc gaa caa tcc gac gat ctc1824His Asp Leu Arg Glu Phe Ala Ala Asp Ser Glu Gln Ser Asp Asp Leu595 600 605acc atc atc gcc att cat cgc cca tga1851Thr Ile Ile Ala Ile His Arg Pro *610 615<210>82<211>616<212>PRT<213>百日咳博德特氏菌<400>82Met Asp Leu Val Val Arg Asp Thr Asp Thr Arg Trp Ser Thr Leu Leu1 5 10 15Asp Asp Lys Ile Arg Thr Ile Arg Glu Ser Arg Arg Gln Leu Ile Gln20 25 30Leu Ser Ala Val Val Thr Ser Val Leu Asn Ala Tyr Ala Ala Gln Ala35 40 45Glu Arg Gly His Val Thr Thr Gly Ala Ala Lys Gly Met ala Arg Val50 55 60Trp Leu Asn His Leu Asp Leu Gly Pro Arg Arg Val Ala Phe Ala Tyr65 70 75 80Asp Ala Glu Gly Thr Val Leu Ala Ser Thr Asn Pro Arg Met Ile Asp85 90 95Arg Asp Leu Ser Gly Ile Arg Asp Phe Lys Gly Arg Pro Leu Ala Ala100 105 110Ala Met Tyr Glu Glu Ser Arg Asn Asp Gly Arg Gly Phe Ala Ile Tyr115 120 125Pro Ser Pro Leu Asp Glu Ser Ala Gln Met Arg His Ala Tyr Phe Val130 135 140Tyr Phe Pro Ala Trp Lys Trp Val Leu Ala Ile Ser Asp Ser Ser Gln145 150 155 160Ala Ile Ile Asp Lys Val Ala Ala Gln Lys Ala Asn Met Ile Ala Ala165170 175Ile Asp Arg Asn Leu Ser Glu Leu Arg Leu Ser Arg His Gly Phe Val180 185 190Phe Val Val Ala Asp Asp Gly Thr Val Ile Val Pro Pro Pro Pro Ser195 200 205Ala Ala Arg Leu Leu Asp Ser Thr Asp Val Glu Ser Gly Arg Val Leu210 215 220His Ser Met Leu Ala Glu Ile Ser Ser Thr Arg Gly Leu Thr Leu Arg225 230 235 240Phe Thr Asn Gly Glu Ser Ala Trp Gln Ile Asp Ala Leu Arg Tyr Lys245 250 255Pro Leu His Trp Thr Ile Ile Gly Val Val Pro Glu Pro Asp Leu Thr260 265 270Asp Pro Ala Gln Asn Leu Val Arg Arg Gln Ala Leu Ile Phe Ala Ala275 280 285Thr Leu Leu Ala Gly Leu Met Leu Ala Trp Val Val Ala Val Arg Ile290 295 300Ala Arg Pro Leu Ala Gln Leu Ser Asn Tyr Ala Arg Gln Leu Pro Thr305 310 315 320Gln Asp Leu Thr Glu Pro Ile Arg Val Pro Pro Ser Val Ala Cys Leu325330 335Pro Arg Arg Arg Arg Asp Glu Val Gly Gln Leu Ala Glu Ser Phe Leu340 345 350Phe Met Asn Glu Gln Leu His His Asn Val Arg Ala Leu Met ala Gln355 360 365Ile Ser Asn Arg Glu Arg Leu Glu Ser Glu Leu Ser Ile Ala Arg Ser370 375 380Ile Gln Leu Gly Leu Leu Pro Gln Pro Leu Pro Asp Ala Ala Thr Arg385 390 395 400Gly Ser Gln Leu Arg Ala Val Met Tyr Pro Ala Arg Glu Val Gly Gly405 410 415Asp Phe Tyr Asp Tyr Phe Val Leu Ala Asp Gly Arg Leu Cys Phe Ala420 425 430Ile Gly Asp Val Ser Gly Lys Gly Val Pro Ala Ala Leu Phe Met ala435 440 445Ile Val Arg Thr Leu Ile Arg Ser Val Ala Glu Glu Glu His Asp Pro450 455 460Gly Ala Ile Ala Thr Lys Val Asn His Arg Leu Ala Glu Asn Asn Pro465 470 475 480Lys Leu Met Phe Val Thr Leu Leu Ile Gly Val Phe Thr Pro Glu Thr485 490 495Gly Ala Leu Ala Trp Val Asn Ala Gly His Pro Pro Pro Leu Leu Ile500 505 510Asp Glu Arg Gly Glu Val Arg Leu Leu Gln Gly Ser Ser Gly Ala Ala515 520 525Cys Gly Val Leu Asp Asn Glu Ala Tyr Ser Thr Leu Ser Thr Thr Leu
530 535 540Pro Asn Gly Thr Ser Leu Val Ala Phe Thr Asp Gly Val Thr Glu Ala545 550 555 560Ile His Gly Gly Cys Ala Gln Tyr Gly Leu Pro Arg Leu Val Ala Leu565 570575Met Gln Gly Ala Pro His Ala Ala Ala Glu Leu Ile Glu His Ile Leu580 585 590His Asp Leu Arg Glu Phe Ala Ala Asp Ser Glu Gln Ser Asp Asp Leu595 600 605Thr Ile Ile Ala Ile His Arg Pro610 61权利要求
1.一种分离的多肽，它包含与选自SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70和72的氨基酸序列具有至少75％同一性的氨基酸序列。
2.权利要求1中要求保护的多肽，它包含选自SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70和72的氨基酸序列。
3.一种SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70或72的分离的多肽。
4.一种分离的多肽，它包含权利要求1-3中任一项要求保护的多肽的片段。
5.权利要求4的多肽，其中所述片段是免疫原性的。
6.一种分离的多核苷酸或与所述分离的多核苷酸互补的一种核苷酸序列，所述分离的多核苷酸包含编码分别在SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70或72的全长范围内与SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70或72的氨基酸序列具有至少75％同一性的多肽的核苷酸序列。
7.一种分离的多核苷酸或与所述分离的多核苷酸互补的一种核苷酸序列，所述分离的多核苷酸包含与编码SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70或72的多肽的核苷酸序列具有至少75％同一性的核苷酸序列。
8.一种分离的多核苷酸或与所述分离的多核苷酸互补的一种核苷酸序列，所述分离的多核苷酸包含分别在SEQ ID NO41、43、45、47、49、51、53、55、57、59、61、63、65、67、69或71的全长范围内与SEQ ID NO41、43、45、47、49、51、53、55、57、59、61、63、65、67、69或71的核苷酸序列具有至少75％同一性的核苷酸序列。
9.权利要求6-8中任一项要求保护的分离的多核苷酸，其中所述同一性是与SEQ ID NO41、43、45、47、49、51、53、55、57、59、61、63、65、67、69或71至少95％的同一性。
10.一种分离的多核苷酸，它包含编码SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70或72的多肽的核苷酸序列。
11.一种分离的多核苷酸，它包含SEQ ID NO41、43、45、47、49、51、53、55、57、59、61、63、65、67、69或71的多核苷酸。
12.一种分离的多核苷酸，它包含编码SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70或72的多肽的核苷酸序列，所述核苷酸序列可通过在严格杂交条件下用具有SEQ IDNO41、43、45、47、49、51、53、55、57、59、61、63、65、67、69或71的序列或其片段的标记探针筛选合适的文库而获得。
13.一种表达载体或一种重组活的微生物，它包含一种按照权利要求6-12中任一项的分离的多核苷酸。
14.一种包含权利要求13的表达载体的宿主细胞、或所述宿主细胞的表达一种分离的多肽的一种亚细胞部分或膜，所述分离的多肽包含与选自42、44、46、48、50、52、54、56、58、60、62、64、66、68、70和72的氨基酸序列具有至少75％同一性的氨基酸序列。
15.一种产生多肽的方法，所述多肽包含与选自SEQ ID NO42、44、46、48、50、52、54、56、58、60、62、64、66、68、70和72的氨基酸序列具有至少75％同一性的氨基酸序列，所述方法包括在足以产生所述多肽的条件下培养权利要求14的宿主细胞，并从所述培养基中回收所述多肽。
16.一种表达权利要求6-12中任一项的多核苷酸的方法，所述方法包括用包含至少其中一种所述多核苷酸的表达载体转化宿主细胞，并在足以表达所述多核苷酸的任一种的条件下培养所述宿主细胞。
17.一种疫苗组合物，它包含有效量的权利要求1-5中任一项的多肽和一种药学上可接受的载体。
18.权利要求17的疫苗组合物，其中所述多肽具有选自SEQ IDNO42、46、48、50、52、54、56、58、60和62的氨基酸序列。
19.一种疫苗组合物，它包含有效量的权利要求6-12中任一项的多核苷酸和一种药学上可接受的载体。
20.按照权利要求17-19中任一项的疫苗组合物，其中所述组合物包含至少一种其它百日咳博德特氏菌抗原。
21.一种对于权利要求1-5中任一项要求保护的多肽或片段具有免疫专一性的抗体。
22.一种诊断百日咳博德特氏菌感染的方法，所述方法包括鉴定在来自怀疑患这样一种感染的动物的生物学样品中存在的、权利要求1-5中任一项要求保护的多肽、或对所述多肽具有免疫专一性的抗体。
23.包含免疫有效量的权利要求1-5中任一项要求保护的多肽的组合物的用途，用于制备用来在动物中产生免疫应答的药物。
24.包含免疫有效量的权利要求6-12中任一项要求保护的多核苷酸的组合物的用途，用于制备用来在动物中产生免疫应答的药物。
25.一种可用于治疗患有百日咳博德特氏菌疾病的人的治疗组合物，所述组合物包含至少一种针对权利要求1-5的多肽的抗体和一种合适的药用载体。
26.一种用于诊断人类百日咳博德特氏菌细菌感染的试剂盒，所述试剂盒包含一种权利要求3-18的多核苷酸或一种权利要求19的多肽。
27.一种从来自细菌致病性菌株的含有III型分泌系统的致病性岛鉴定毒力基因的方法，所述方法包括设计与耶尔森氏菌属LcrD多肽特有的高度保守区互补的简并PCR引物；扩增含有存在于所述细菌致病性菌株中的lcrD样基因的引物之间(包括其DNA序列)的DNA序列的多核苷酸；对所述lcrD样基因进行测序；测定所述DNA序列是否与lcrD样基因的毒力相关家族或lcrD样基因的鞭毛相关家族更同源；和如果是一种毒力相关成员，则对所述整个致病性岛测序，并且鉴定在该序列内的基因。
28.一种确定特定菌株是否含有涉及致病性的III型分泌系统的方法，所述方法包括设计与耶尔森氏菌属LcrD多肽特有的高度保守区互补的简并PCR引物；扩增含有所述引物之间(包括其DNA序列)的DNA序列的多核苷酸，以确定所述细菌菌株中的任何lcrD样基因的存在；如果成功地扩增，则对所述lcrD样基因进行测序；和确定所述DNA序列是否与lcrD样基因的毒力相关家族或lcrD样基因的鞭毛相关家族更同源。
全文摘要
本发明涉及用于检测含有一种III型分泌系统的细菌致病菌株的一种基本方法。更具体地说,本发明涉及适用于病原体百日咳博德特氏菌的方法。此外,本发明涉及在这些区内新鉴定的多核苷酸、由它们编码的毒力多肽,并涉及这类多核苷酸和多肽的应用,还涉及它们的产生。更具体地说,本发明的多核苷酸和多肽涉及与百日咳博德特氏菌的III型分泌系统相关的毒力效应蛋白,所述多核苷酸和多肽特别适用于疫苗目的。
文档编号A61K48/00GK1371389SQ99816279
公开日2002年9月25日 申请日期1999年12月21日 优先权日1998年12月21日
发明者A·波伦, A·福康尼尔, E·格德弗罗伊德 申请人:布鲁塞尔大学