识别凝血酶阴性的葡萄球菌和金黄色葡萄球菌的表面蛋白的交叉反应性单克隆抗体和多...的制作方法

专利名称：识别凝血酶阴性的葡萄球菌和金黄色葡萄球菌的表面蛋白的交叉反应性单克隆抗体和多 ...的制作方法
技术领域：
总的来说，本发明涉及来自金黄色葡萄球菌的表面蛋白质及其活性的区域例如它们的A功能域，它具有凝固酶阴性的葡萄球菌例如表皮葡萄球菌和溶血葡萄球菌上的同源蛋白质，以及识别上述的蛋白质的抗体，具体地说，涉及分离的单克隆和多克隆抗体，它们识别来自金黄色葡萄球菌和凝固酶阴性的葡萄球菌的特定的蛋白质，以及抗金黄色葡萄球菌和凝固酶阴性的葡萄球菌的交叉反应，因此可用于疫苗，以及用于预防或治疗各种各样的由葡萄球菌的细菌引起的感染的方法。
背景技术：
对于大多数引起动物和人类的传染病的微生物，在宿主中成功地形成集落是一个必需的过程。微生物的粘附是最后导致疾病的一系列事件的第一个重要步骤。借助细菌表面的特定的adhesins，通过粘附到宿主组织或血清限定的移植入的生物材料，例如导管，人工的关节，血管的移植片，病原微生物在宿主中形成集落。MSCRAMMs(识别粘附的基质分子的微生物表面成分)是细胞表面adhesins的一个家族，它们识别和特定地结合到宿主的细胞外的基质的不同的组分。一旦细菌成功地粘附并且在宿主组织中形成集落，他们的生理机能被急剧改变并且分泌受损害的成分例如毒素和水解蛋白质的酶。此外，粘连的细菌常常产生生物膜并且很快地变得对大多数的抗生素的杀戮效果更具抗性。
金黄色葡萄球菌产生从皮肤上的损害例如伤口感染，脓疱病，疖到包括肺炎，脓毒性的关节炎，脓毒病，心内膜炎的威胁生命的状况以及生物材料相关的传染病的范围的感染谱。已知金黄色葡萄球菌表达不同的MSCRAMMs的全部技能，它们独立地或者协调地促进微生物的粘附到特定的宿主组织组分。另外，葡萄球菌细菌的另一个类型被识别为凝固酶阴性的细菌，包括例如已知同样地表示MSCRAMMs的表皮葡萄球菌和溶血葡萄球菌，也对各种各样的细菌的感染和有关的疾病负有责任。关于这一点，通常MSCRAMMs由抗体，多克隆和单克隆抗体，对免疫学的攻击提供出色的靶。
然而，由于抗体本性非常特异的，就不同类型的葡萄球菌而言，例如一方面金黄色葡萄球菌(凝固酶阳性)和另一方面表皮葡萄球菌和溶血葡萄球菌(凝固酶-阴性的)，研制显示不同的类型的细菌之间年的交叉反应性的抗体，仍然是值得注意的问题。这样的交叉的反应的抗体是特别令人想望的，因为他们具有使人类和动物患者致免疫的潜在性和提供抵抗由两种类型的葡萄球菌的细菌，即凝固酶阳性的细菌例如金黄色葡萄球菌和凝固酶阴性的细菌，例如表皮葡萄球菌和溶血葡萄球菌引起的感染潜在性。因此这样的抗体在预防或治疗各种各样的由葡萄球菌的细菌引起的感染中非常地有效。
发明概述因此，本发明的目的之一提供识别来自凝固酶阳性的细菌例如金黄色葡萄球菌的MSCRAMMs以及来自凝固酶阴性的细菌，例如表皮葡萄球菌和溶血葡萄球菌的MSCRAMMs的单克隆抗体。
本发明的还有一个目的是识别和分离来自葡萄球菌的细菌的MSCRAMMs，以及他们的活性的区域例如A区域，该区域可用于产生与凝固酶阳性的和凝固酶阴性的葡萄球菌发生交叉反应的单克隆和多克隆抗体。
本发明的另一个目的提供可以识别来自凝固酶阴性的葡萄球菌的表面蛋白质例如DgsK蛋白质的A区域，同时识别来自金黄色葡萄球菌的表面蛋白质例如SasA蛋白质的分离的抗体。
本发明的又一方面是利用本发明的分离的蛋白质，A区域和抗体生产用于治疗或预防葡萄球菌的感染的疫苗，以及提供了其中将本发明的疫苗和抗体可用于预防或治疗葡萄球菌的感染的方法。
这些和其他的目的通过本发明实现，所述的发明包括识别和分离一种类型的葡萄球菌的细菌例如凝固酶阴性的或凝固酶阳性的葡萄球菌的表面蛋白质，它导致产生识别两种类型的葡萄球菌的表面蛋白质的交叉的反应性的抗体，并且因此将抗体用于治疗或预防各种各样葡萄球菌的感染的疫苗和方法。本发明还涉及从这些表面蛋白质产生多克隆和单克隆抗体，以及它们用于预防或治疗葡萄球菌的感染。
对本领域内技术人员而言，根据阅读本说明书和/或在此处记载的参考文献，这些具体的实施方案和在公开的发明的精神和范围之内的其他的供替代的选择和改变是显而易见的，所有引用的文献都被并入本文。
附图简述附

图1是本发明的in silico预测的蛋白质的一级的结构图。
附图2显示本发明的开放读框表达的N-末端His标记的纯化的重组的蛋白质的考马斯亮蓝染色凝胶。
附图3A-3C显示分别用抗-KesK抗体(附图3A)，抗-KnkA抗体(附图3B)和抗DsqA抗体(附图3C)探测时金黄色葡萄球菌的细胞壁提取物的Western印迹。
附图4A-4B显示表达金黄色葡萄球菌的MSCRAMM称为KnkA(附图4A)和KesK(附图4B)的乳酸乳球菌的点印迹和Western免疫印迹。
附图5A-5D代表用康复的人血清体内检测表达本发明重组LPXTG蛋白质，包括RrKn和RrKN2(附图5A)，Kesk1和Kesk2A(附图5B)，KnkA(附图5C)和DsqA2(附图5D)的探测情况。
附图6显示证实抗金黄色葡萄球菌SasA的兔多克隆抗体与从表皮葡萄球菌HB细胞表面释放的蛋白质以及来自从表皮葡萄球菌克隆的DsgK的重组A-区域发生交叉反应的Western印迹分析结果。
优选的实施方案的详细描述根据本发明，提供了来自凝固酶阳性的葡萄球菌的细菌例如金黄色葡萄球菌以及凝固酶阴性的葡萄球菌的细菌例如表皮葡萄球菌和溶血葡萄球菌，的特定的表面蛋白质，包括其活性片段例如这些蛋白质或可以产生识别完整的蛋白质的抗体的其他的表位区域的A区域。根据本发明，基于MSCRAMMs(识别粘附基质分子的微生物表面成分)的一些共有的特征实施候选肽序列和蛋白质的鉴定和分离，在大多数情况下，这些特征共价地固定到细胞壁肽葡聚糖。这些表面蛋白质具有下列的共有的特征可用于确定和分离本发明的序列，即(i)Sec-依赖性分泌必需的N-末端信号肽(长度大约40个残基)，(ii)一个富含脯氨酸和甘氨酸残基或由丝氨酸和门冬氨酸盐二肽重复组成点的跨壁功能域；(iii)蛋白质共价固定到肽葡聚糖中的五个甘氨酸交联桥必需的LPXTG基序，(iv)一个疏水的跨膜功能区，跟随其后的(v)几个带正电荷的残基。
根据本发明，按照以上所述的特性，通过开发金黄色葡萄球菌的完整的基因组，识别了至少八个新的开放读框编码的蛋白质，具有分泌和固定象征MSCRAMMs的基序(即的携带N-末端信号肽和跟随疏水的功能区之后的C-末端LPXTG基序和一个带正电的尾部)。表1阐明所识别的蛋白质的一览表，包括在金黄色葡萄球菌基因组中的分布，蛋白质的大小和C-末端细胞壁排序序列。
表1
缩写eMRSA-16；N，8325；C，COL；S，MSSA；J，N315，M，Mu50。
8个蛋白质中的6个存在于已经被测序的所有六个葡萄球菌的基因组中，剩余的2个存在于5/6的这些基因组中。
根据本发明，获得上述的蛋白质的氨基酸和编码它的核酸序列，如下所述Ekes MRSA-SEQ ID NO1(DNA序列)；EkeS～MRSA-SEQ ID NO2(蛋白质序列)；DsqA(8325)-SEQ IDNO3(DNA序列)；DsqA(8325)-SEQ ID NO4(蛋白质序列)；KesK1(8325)-SEQ ID NO5(DNA序列)；KesK1(8325)-SEQ IDNO6(蛋白质序列)；KrkN2(8325)--SEQ ID NO7(DNA序列)；KrkN2(8325)-SEQ ID NO8(蛋白质序列)；KrkN(8325)-SEQ IDNO9(DNA序列)；KrkN(8325)-SEQ ID NO10(蛋白质序列)；RkaS(COL)-SEQ ID NO11(DNA序列)；RkaS(COL)-SEQ IDNO12(蛋白质序列)；RrkN(8325)-SEQ ID NO13(DNA序列)；RrkN(8325)-SEQ ID NO14(蛋白质序列)；KnkA(8325)-SEQ IDNO15(DNA序列)；KnkA(8325)-SEQII7 NO16(蛋白质序列)。
根据本发明，可以从上述的蛋白质或他们的活性区域例如A区域产生分离的抗体以便能够识别上述的蛋白质和/或上述的功能区。这些抗体可以是单克隆的或多克隆的。如果需要多克隆抗体，可以本领域内熟知的许多常规方法的任何一种制备。一种典型的方法是，可以将需要的表面蛋白质或其活性的区域注射到合适的宿主动物，例如，小鼠或兔子，在合适的时间期间之后，可以从该宿主动物分离和回收想要的抗体。对于单克隆抗体，根据本发明，可以许多合适的方法包括例如熟知的Kohler和Milstein，Nature 256495-497(1975)方法，本领域内已知的任何其他方法，例如美国专利Nos.6,331,415；5,981,216；5,807,715；和4,816,567；欧洲专利申请519,596；和PCT公开WO 00/71585公开的方法制备，所有这些专利文献都通过在此引述而合并于本文。这些方法包括其制备如以本领域内熟知的方式进行嵌合，人性化，或人的单克隆抗体。仍然进一步地，可以从一个单链，例如轻或重链制备，以及还可以从保持完整的抗体的结合特性(例如，特异性和/或亲合力)的抗体的活性的片段制备单克隆抗体。活性的片段是指具有与完全抗体相同的结合特异性的抗体片段，所述完全抗体结合到来自不同的类型的葡萄球菌细菌(即凝固酶阴性的，或凝固酶阳性的)的特定的表面蛋白质或其同系物。本文术语″抗体″包括所述的片段。另外，也可以利用本发明的单克隆或多克隆抗体制备抗血清，可以本领域内熟知的许多合适方式制备。
如上所述，可以本领域内熟知的许多合适的方式制备本发明的抗分离的表面蛋白质和/或其活性的区域的抗体，例如上述所述的完全建立的Kohler和Milstein方法，可用于产生单克隆抗体。例如，在这样的一种方法的预备的步骤中使用，可以在长时间的时期用本发明的纯化的重组MSCRAMM或其活性的部分长期给小鼠每一周腹膜内注射一次，随后测试从免疫接种的老鼠获得的血液以测定与纯化的蛋白质的反应性。在识别老鼠与该蛋白质的反应性之后，将从小鼠脾脏分离的淋巴细胞与小鼠脊髓细胞融合以产生抗本发明的表面蛋白质的抗体阳性的杂交瘤，然后分离和培养，随后纯化和使之形成同位型。
为了产生本发明的单克隆抗体，优选的是利用本发明的重组制备的MSCRAMM′s来制备，利用本领域内熟知的许多标准方法可以制备这些重组子并且分离。例如，一种这样的方法使用了大肠杆菌表达载体pQE-30作为克隆和表达重组蛋白质和肽的表达载体。在一个优选的方法中，利用PCR，从上面描述的序列扩增识别为DgsK或SasA的表面蛋白质的A区域，再克隆到大肠杆菌表达载体PQE-30(Qiagen)，以允许表达一种含有六个组氨酸残基的重组融合蛋白质。
随后将该载体转化到大肠杆菌ATCC55151，在15-升发酵罐中生长到光密度(OD600)为0.7，用0.2mM异丙基-1β-D半乳糖苷(IPTG)诱导4小时。利用AGTechnologies中空纤维组件(孔大小0.45um)收集细胞，将细胞浆在-80C冷冻。利用2次穿过French Press，1100psi，将细胞在1×PBS(10mL缓冲液/1克的细胞浆)裂解。将裂解的细胞17,000rpm旋转30分钟以除去细胞碎片。将上清液经过装备了0.1M NiCI2的5-毫升Hi俘获Cheating(Pharmacia)柱。在上样之后，用5个柱体积的10mM Tris，pH 8.0，100mM NaCI(缓冲液A)洗涤柱子。利用超过30个柱体积的0-100％梯度的10mM Tris，pH 8.0，100mM NaCI，200mM咪唑(Buffer B)洗脱蛋白质。SdrGN1N2N3或SdrGN2N3在-13％Buffer B(～26mM咪唑)被洗脱。监测280nm处的吸光率。在1×PBS中透析含有SdrGN1 N2N3或SdrGN2N3的馏分。
接著，然后将各个蛋白质经过一个内毒素除去方案。在该方案中所使用的缓冲液通过一个5-毫升Mono-Q琼脂糖(Pharmacia)柱而没有内毒素。将蛋白质平衡地分到4×15毫升试管中。各个试管的体积用缓冲液A加到9毫升。将1毫升的10％Triton X-114加入到各个试管中，在4℃旋转保温1小时。将试管置于37℃水浴中以使相位分离。将试管以2,000rpm旋转10分钟，收集各个试管的上面的含水的相，用去污剂反复提取。将2次提取的含水相合并，传递过带有0.1M NiCl2的5-mL IDAcheating(Sigma)柱，以除去剩余的洗涤剂。用9个柱体积的Buffer A洗涤该柱，然后用3个柱体积的Buffer B洗脱该蛋白质。将洗脱液通过5-毫升的Detoxigel(Sigma)柱，收集流出物，再上样到柱。收集第二次过柱的流出物，在1×PBS中透析。分析纯化的产物的浓度，纯度和内毒素水平，然后给小鼠给药。
在该优选的方法，以下面的方式从上面识别的重组蛋白质可以制备本发明的单克隆抗体。在该方法中，将大肠杆菌表达的和纯化的重组SasA和DsgK蛋白质用于制备一组鼠的单克隆抗体，而将小鼠血清用作为多克隆抗体的来源。简单地说，一组Balb/C或SJL小鼠接收了一系列皮下的免疫接种1-10mg溶液蛋白质或与下面表2描述的佐剂混合的蛋白质。
表2免疫接种方案RIMMS注射天数数量(微克)途径佐剂#1 0 5 皮下FCA/RIBI#2 2 1 皮下FCA/RIBI#3 4 1 皮下FCA/RIBI#4 7 1 皮下FCA/RIBI#5 9 1 皮下FCA/RIBI常规注射天数数量(微克)途径佐剂原始0 5 皮下FCA加强#1 14 1 腹膜内 RIBI加强#2 28 1 腹膜内 RIBI加强#3 42 1 腹膜内 RIBI在杀死时(RIMMS)或加强后几天收集(常规的)血清，在ELISA测定中测定抗MSCRAMM蛋白质或对完整的细胞(表皮葡萄球菌和金黄色葡萄球菌)的效价。在最后的加强之后3天，去除脾脏或淋巴结，挑开成单细胞悬浮液，收集淋巴细胞。然后将淋巴细胞与P3X63Ag8.653骨髓瘤细胞系(ATCC & CRL-1580)融合。根据Current Protocols in Immunology(第2章，第2单元)中的生产单克隆抗体的方案进行细胞融合，随后涂覆平板和喂养，所述文献都通过在此引述而合并于本文。
然后利用标准的ELISA分析方法，从融合产生的任何克隆中筛选特异的抗-SasA抗体的产生。扩增阳性的克隆，进一步通过流式血细胞计数测试完整的细菌的细胞结合测定中的活性以及Biacore分析中SasA的结合。在整个Biacore分析中，流速保持在10毫升/分钟不变。在注射SasA或DgsK之前，借助于RAM-Fc结合，将测试抗体吸附到芯片。在时间0，将浓度为30mg/ml的SasA或DgsK注射过芯片3分钟，随后解离2分钟。对该阶段的分析测定Mab/SasA和DgsK相互作用的相对关系和解离常数。
接著，对上面提到的抗体与完整细菌的结合情况进行测试。在该测试中，用兔子IgG(50mg/ml)封闭之后，收集，洗涤细菌的样品金黄色葡萄球菌Newman，金黄色葡萄球菌60，金黄色葡萄球菌397(Sal6)，金黄色葡萄球菌Wood，金黄色葡萄球菌8325-4，甲氧苯青霉素抗性的金黄色葡萄球菌MRSA 16，表皮葡萄球菌ATCC 35984，表皮葡萄球菌HB，表皮葡萄球菌CN-899和溶血葡萄球菌ATCC 43253，用浓度为2pg/ml的Mab或仅用PBS(对照)保温。在用抗体保温之后，将用作为检测抗体的山羊-F(ab′)2-抗-小鼠-F(ab′)2-FITC用于孵育细菌的细胞。在抗体标记之后，用FACS管径流式细胞计数器抽吸细菌的细胞以分析荧光射出(刺激488，发射570)。对于各个细菌的菌株，收集和检测10,000个样品。这些资料显示抗金黄色SasA的抗体能够识别凝固酶阴性的葡萄球菌的表面的同源的蛋白质。该资料支持Western印迹分析，表明抗金黄色葡萄球菌SasA的兔子多克隆抗体与从表皮葡萄球菌HB细胞表面释放的蛋白质以及来自于从表皮葡萄球菌克隆的DsgK的重组A-区域发生交叉反应(参见附图6和下表3)。
表3多克隆血清反应性
虽然利用重组形式的本发明的表面蛋白质生产抗体是优选的，但是该抗体可以从这些蛋白质天然分离的和纯化形式或其活性区域例如A区域制备抗体，以类似于上面所述的方式利用这些蛋白质或活性的区域可以产生单克隆抗体或多克隆抗体以获得这样的抗体。仍然其他的常规的方法，利用重组或天然的纯化蛋白质或其活性区域，可用于产生本发明的抗体，正如本领域内技术人员可以识别的。
正如本领域内技术人员可以认识到的，本发明的抗体也可以配制成可用于人或动物患者给药的合适的药学的组合物，以便治疗或预防葡萄球菌的细菌引起的感染。含有本发明的抗体的药物组合物，或其有效的片段，可以与本领域内常用的合适的药物介质，赋形剂或载体结合配制，包括例如盐水，葡萄糖，水，甘油，乙醇，其他的治疗化合物和其结合使用。本领域内技术人员应该认识到，所使用的特定的介质，赋形剂或载体将依据患者和患者的状况而改变，如本领域内技术人员将会认识到的各种各样的给药方式适用于本发明的组合物。在本申请中公开的以药物组合物给药的合适的方式包括，但不限于，局部的，口部的，肛门的，阴道的，静脉内的，腹膜内的，肌内的，皮下的，鼻内的和真皮内的给药。
对于局部的给药，将组合物配制成软膏，乳膏，凝胶，洗液，滴剂(例如眼药水和耳朵滴剂)，或溶液(例如漱口剂)。可以将组合物埋植于伤口或外科的敷料，缝合线和气溶胶。该组合物可以含有常规的添加剂。例如防腐剂，溶剂以促进渗透和润滑药。局部的制剂也可以含有常规的载体例如乳膏或油膏基底，乙醇或油酰醇。另外，通常也可以将抗体组合物，和涉及其他MSCRAMM的组合物，疫苗，方法和应用的其他的信息用于本发明，涉及前面提到的MSCRAMM和它们的活性的区域和其抗体，这些其它的MSCRAMM公开于例如美国专利5,175,096；5,320,951；5,416,021；5,440,014；5,571,514；5,652,217；5,707,702；5,789,549；5,840,846；5,980,908；6,086,895；6,008,341；6,177,084；5,851,794和6,288,214；所有这些专利都通过在此引述而合并于本文。
本发明的抗体组合物也可以与其量可有效地加强致免疫的应答的合适的佐剂一起给药。例如，合适的佐剂可以包括广泛用于人的明矾(磷酸铝或氢氧化铝)，和其他的佐剂例如皂甙和其纯化的组分Quil A，弗氏完全佐剂，RIBBI佐剂，和其它用于研究和兽医应用的佐剂，还有其他以化学方法定义的制剂例如muramyl二肽，一磷酰脂类A，磷脂轭合物例如Goodman-Snitkoff等人在J.Immunol.147410-415(1991)描述的，该文献通过在此引述而合并于本文，蛋白脂质体内的偶合物的形成胶囊如Miller等人，J.Exp.Med.1761739-1744(1992)，该文献通过在此引述而合并于本文，脂类载体中蛋白质形成胶囊如NovasomeTM脂类载体(Micro Vescular System，Inc.，Nashua，NH)也可以使用。
在任何情况下，本发明的识别蛋白质或如上文提出的它们的活性的区域的抗体组合物可用于预防或治疗葡萄球菌的感染的方法，可用于抑制葡萄球菌的细菌与宿主组织和/或细胞的结合。根据本发明，提供了预防或治疗葡萄球菌的感染的方法，包括以有效量的抗体给本文提出的表面蛋白或其活性的亚区域以便治疗或预防葡萄球菌的感染。另外，这些单克隆抗体可用于损害葡萄球菌的细菌与宿主细胞的结合。
因此，根据本发明，将本发明的抗体以如上所述的任何常规的方法(例如，局部的，非肠道的，肌内的，等)给药，因此当将有效量的抗体组合物给人或动物患者给药时，提供了一种治疗或预防人类或动物患者的葡萄球菌的感染的非常地有用的方法。有效量是指足以阻止细菌粘合，抑制葡萄球菌的细菌结合到宿主细胞从而可用于治疗或阻止葡萄球菌的感染的使用水平，例如抗体效价的水平。如本领域内普通技术人员认识到的，有效地治疗或预防葡萄球菌的感染所必需的抗体效价的水平将依据患者的特性和健康状况，和/或预存在的葡萄球菌的感染的严重程度。
除了用于治疗或预防葡萄球菌的感染的方法中，本发明的抗体也可用于葡萄球菌的蛋白质的特定的检测或作为研究工具。本文所述的术语″抗体″包括单克隆，多克隆，嵌合，单链，双特异性，simianize，和人性化的或灵长目化的抗体以及Fab片段，例如保持抗体与上面详细说明的表面蛋白质的抗体的结合特异性的那些片段，包括Fab免疫球蛋白表达文库的产物。因此，本发明涉及单链的使用例如抗体的可变的重和轻链。这些类型的抗体或抗体片段的制备是本领域内技术人员熟知的。在本发明中，抗表面蛋白质或上面所述的它们的活性的区域的抗体可以制备，分离和/或纯化，然后用于治疗或免受葡萄球菌的感染。
上面描述的任何抗体可直接用可检测的标签进行标记以识别葡萄球菌细菌和对其定性。免疫测定中使用的标记物通常是本领域内技术人员已知的，并且包括酶，放射性同位素，荧光，发光的和产色的物质，包括有色的颗粒例如胶态的金或胶乳珠子。合适的免疫测定包括酶联免疫吸附(ELISA)。
或者，通过与具有免疫球蛋白亲合力的标记的物质可以间接标记该抗体。该抗体可以与第二物质偶合，用已标记的第三物质进行检测，该第三物质与偶合到抗体的第二物质具有亲和力。例如，可以将抗体结合到生物素，利用标记的抗生物素蛋白或链生抗生物素蛋白可以检测抗体-生物素偶合物。类似地，可以将该抗体偶合到半肝素，利用标记的抗半肝素抗体检测抗体-半肝素偶合物。这些和其他标记抗体和分析偶合物的其他方法是本领域内技术人员熟知的。
根据本发明，还提供了被设计为治疗或保护其抵抗葡萄球菌感染的主动或被动免疫接种疫苗，可以利用许多本领域内熟知的常规疫苗制备方法从以上提出的表面蛋白质或活性区域制备这些疫苗。对于典型的疫苗，将致免疫量的合适的表面蛋白或其活性片段与合适的药物学可接受载体，媒介物或赋形剂结合，合适的是可以以有效地免疫接种人或动物患者的疫苗的量给药。本领域技术人员认识到致免疫量是指能够在人或动物患者中产生免疫应答的蛋白质或活性片段或其亚片段的量。
除了借助于将免疫量的本发明的蛋白质或活性片段导入或给药产生的抗体的活性疫苗以外，也可以将本发明的分离的抗体或活性片段用于研制抗钩环感染的被动免疫接种的疫苗。在这样的情况下，可以将如上所述的抗体组合物，即有效量的抗体和药物学可接受的载体，媒介物或赋形剂以适当方式给药到人或动物患者。
因此，本发明的蛋白质或其活性片段可用作为活性疫苗，本发明的抗体可用作为被动疫苗，可用于提供合适的抗体以治疗或予防葡萄球菌感染。本领域内技术人员将会认识到，可以将一秒包装以适用于许多方式给药，例如非肠道的(即肌内的，透皮的或皮下的)给药或鼻咽的(即鼻内的)给药。一种这样的方式是将疫苗通过肌内注射到例如三角肌，但是，给药的特定方式将依赖于待治疗的细菌感染的特性和患者的状况。优选的是将该疫苗与药物学可接受的载体，媒介物或赋形剂结合以有助于给药，通常该载体是有或没有防腐剂的水，或缓冲盐水。该疫苗可以是在给药时重悬浮的冻干形式或溶液形式。
另外，在某些情况下，如必要可以将本发明的抗体进行修饰，以对给药患者的致免疫性降低，例如，如果该患者是人。通过将杂交瘤衍生的抗体的complimentarity表位移植到如Jones等人，Nature 321522-525(1986)或Tempest等人/Biotechnology 9266-273(1991)描述的人单克隆抗体而将抗体“人性化”或通过将免疫球蛋白可变区中的表面接触的鼠骨架残基改变为模拟同源的人骨架配体如由Padlan，Molecular Imm.28489-498(1991)描述而进行″胶合″，这些文献都通过在此引述而合并于本文。甚至于进一步地，如果需要，本发明的单克隆抗体可以与合适的抗生素结合给药以进一步加强所述组合物抗击细菌感染的能力。
除了治疗人或动物患者以外，本发明的组合物也可以用于停止或予防医学装置或其他生物材料例如移植片的感染。待用抗体，蛋白质和活性片段包被的医学装置或聚合物生物材料包括，但不限于钩环，缝合线，复位心脏瓣膜，心脏的协助装置，硬的和软的接触透镜，眼内的透镜植入片(前的室或后的室)，其他的植入片例如角膜的镶嵌物，kerato-修补物，血管的斯滕特氏印模，epikeratophalia装置，绿内障分流术，视网膜的钩环，巩膜的扣子，牙齿的修补物，thyroplastic装置，laryngoplastic装置，血管的移植片，软的和硬的组织修补物，包括但不限于，泵，电的装置包括刺激器和记录器，听觉的修复术，起搏器，人工的喉，牙齿的植入片，乳房的植入片，阴茎的植入片，cranio/脸的腱，人工的关节，腱，纽带，menisci，和磁盘，人工的骨骼，人工器官包括人工胰腺，人工心脏，人工的肢，和心脏瓣膜，斯滕特氏印模，金属丝，导向装置丝，静脉内的和中央的静脉的导管，激光和气球血管成形术装置，血管的和心脏装置(管子，导管，气球)，室的协助，血透析组合物，血氧合器，尿道的/输尿管的/泌尿的装置(Foley导管，斯滕特氏印模，管和气球)，导气管导管(气管内的和气管造口术管和胶管管头)，肠内的喂养管(包括nasogastric，胃内的和空肠的管)，伤口排水管，用于引流体腔的管例如胸膜的，腹膜的，颅侧的和心包的cavities，血袋，测试管，血收集管，vacutainers，注射器，针，吸移管，吸量管尖端，和血造管。
本领域内技术人员将会明白，本文所用的术语″被包被″或″包被″是指将抗体或活性片段或其衍生的药物组合物应用到装置的表面，优选的是与葡萄球菌细菌感染接触的外表面。该装置的表面不必完全被蛋白质，抗体或活性片段所覆盖。
本发明抗体给药的优选剂量是有效地抑制或治疗葡萄球菌感染的量，易于认识到该量将大大依赖于感染的特性和患者的状况。如上所述，用于本发明的抗体或药物制剂的“有效量”是指无毒但足以产生需要的予防或治疗效果的剂量。正如上文指出的，所需要的抗体或特定试剂的精确量将随患者而变化，取决于患者的种类，年龄，和总的状态，待治疗的状况的严重程度，所使用的特定的载体或赋形剂和给药方式等等。因此，特定抗体组合物的“有效量″将基于特定的环境而变化，普通技术人员利用唯一的常规试验可以测定各种应用情况下合适的有效量。可以将该剂量调节到适应于以该组合物给药的个体，并且随患者的年龄，重量，代谢而变化。该组合物还可以含有稳定剂或药物学可接受的防腐剂，例如硫汞撒(乙基(2-mercaptobenzoate-S)汞钠盐)(Sigma Chemical Company，St.Louis，MO)。
当与合适的标记物或其他合适的可检测的生物分子或化学物质一起使用时，本文描述单克隆抗体可用于体内或体外诊断葡萄球菌感染或检测葡萄球菌细菌的目的。通过这样的抗体的使用也有助于试验室研究。各种类型的标记物和将标记物偶合到本发明的抗体的方法是本领域内技术人员熟知的，例如下面列出的一种方法。
例如，可以将抗体偶合到(直接或通过螯合剂)放射性标记物例如，但不限于32P，3H，14C，35S，125I，或131I。通过例如闪烁计数，γ射线分光光度计放射自显影的方法可以检测标记物。也可以使用生物发光标记物，例如萤火虫荧光素衍生物。采用常规方法生物发光物质可以共价结合到该蛋白质，当酶例如荧光酶催化引起生物发光的与ATP的反应以散发光子时，可以检测标记物蛋白质。也可以将荧光团用于标记蛋白质。荧光团的例子包括荧光和衍生物，藻红蛋白，allo-藻蓝蛋白，藻蓝蛋白，硷性蕊香红，和德克萨斯红。通常荧光团可以通过荧光检测器进行检测。
通过对如上所述的抗体进行标记和根据本领域内技术人员熟知的方法例如免疫荧光显微镜，利用如Warren等人(Mol.Cell.Biol.，71326-1337，1987)描述的程序，对标记物进行检测可以测定细胞中配体的定位。
如上所述，本发明的单克隆抗体，或活性部分或其片段特别适用于干扰负责感染的葡萄球菌病原体和哺乳动物宿主之间的起始物理相互作用，这种对物理相互作用的干扰可用于治疗患者和in-dwelling医学装置予防或降低细菌感染以使它们在使用时更安全。
在本发明的另一个实施例中，提供了一种可用于分离和识别葡萄球菌细菌感染的试剂盒，包括在单个容器中的合适形式，例如冻干的本发明的抗体，然后通过加入怀疑含有葡萄球菌细菌的含水的样品而变成活性。这样的试剂盒通常包括用于储存合适形式抗体和合适的免疫检测试剂的合适的容器，所述的免疫检测试剂允许识别结合到本发明的表面蛋白质或抗体的复合物。通常，这些试剂盒含有本发明的抗体和识别当来自患者的样品导入到该抗体时识别结合的抗体的工具。例如，合适的免疫检测试剂可以包括合适的可检测的信号或标记物，例如生物素或产生可检测的颜色的酶等，可以连接到抗体或用于其他合适的方式中标记物以便当抗体结合到抗原时提供可检测的结果。
简单地说，识别和结合到本发明的表面蛋白质的本发明的抗体或其活性片段可用于治疗各种各样的人和动物患者的葡萄球菌感染，以及用于医疗或其他in-dwelling装置。根据本发明，由于这些蛋白质的特性，以及它们含有与其他类型的葡萄球菌细菌的蛋白质共有的表位，即来自凝血酶阴性葡萄球菌的蛋白质将产生识别来自金黄色葡萄球菌的同源蛋白的抗体或者相反，本发明的抗体将显示交叉反应性和应该有效地抗击广谱的葡萄球菌感染。因此，本发明提供了用于治疗或抵抗广谱葡萄球菌感染的方法和改善的方法中的组合物。
实施例提供下面的实施例，举例说明本发明的优选的实施方式的各个方面。本领域内技术人员应该认识到实施例中公开的技术允许代表发明人发现的技术以实施本发明，因此可以认为构成了实施本发明的优选的方式。但是，本领域内技术人员应该认识到，在本发明公开的基础上，可以在公开的特定的实施方式中进行许多改变，仍然可获得类似的结构而不脱离本发明的精神和范围。
实施例1.金黄色葡萄球菌的MSCRAMM′s的分离和测序已知金黄色葡萄球菌表达一类表面结合的蛋白质，所述的蛋白质通过允许细菌避免宿主防御和通过作为adhesins对致病性起重要作用。已知这些蛋白质是MSCRAMMs(识别粘附的基质的分子的微生物表面成分)，在大多数情况下，可以共价结合到细胞壁肽葡聚糖。它们有几个共同的特征(i)Sec-依赖性分秘所需要的N-末端信号肽(长度约40个残基)，(ii)富含脯氨酸和甘氨酸残基或由丝氨酸和天冬酰胺二肽重复区组成的跨膜区，(iii)该蛋白质与肽葡聚糖中的五甘氨酸桥键共价结合需要的LPXTG基序，(iv)疏水的跨膜区，以及其后的(v)几个带正电荷的残基。
通过开发金黄色葡萄球菌的整个基因组序列，鉴别8个编码具有分秘和锚基基序指示MSCRAMMs的蛋白质的新的开放读框(即携带N-末端的信号肽和C-末端LPXTG基序，随后跟随一个疏水的功能域和一个带正电荷的尾部)。下面的表说明了所识别的蛋白质名录，包括他们在基因组中的分布，他们的蛋白质大小，和挑选C-末端细胞壁的序列。
缩写eMRSA-16；N，8325；C，COL；S，MSSA；J，N315，M，Mu50。
8个蛋白质中的6个存在于已经被测序的所有六个葡萄球菌的基因组中，剩余的2个存在于5/6的这些基因组中。
下面是DNA和蛋白质序列的名录Ekes MRSA(SEQ ID NO1)acaacacagcagagaatagacaaccaggaggaaaacgaaatgaatttgttaaagaaaaataaatatagtattagaaaatataaagtagggatattctctactttaatcgggacagttttattactttcaaacccaaatggtgcacaagctttaactacggatcataatgtgcaaggtggttcaaatcaagcattacctggcaactcacaaaatacaaatgccgatactaatcgagacatagtaaatgattcgcaaaatactcctaatgcacatgcaacagacaatacatcaacaaatcaagcattgactaatcatcaaaacgttgatgtggcaaatcaagtcgggcctgctccaatacagcctagcgcgtcgcctgcgcaaaataataataattctaatgctaattcaacagcaacagagccagcggcgaatacaaataataatttagcatcaaataacaatacattaaacgtgcctaataatacagataacaatgattcagcgcgtcatctgactttaaaagaaattcaagaagatgttcgtcattcgtctgataagccagagttagttgcgattgctgaagaagcatctaatagaccgaaaaagagaagcagacgtgctgcgccaacagatcctaatgcaacaccagcagatccaacggctacaccagcagatccaacggcaggaaatggtagtgcaccagttgcaattacagcgccatacacgccaacaactgatcccaatgccaataatataggacaaaatgcacctaacgaagtgctttcatttgatgataacaacattagaccaagtacgaaccgttctgtgcctacagtaactgttgttgataatttaccaggctacacactgattaatggtggtaaagtaggggtgtttagtcatgcaatggtaagaacgagcatgtttgattcaggagatgccaagaactatcaagcgcaaggcaatgtaattgcattgggtcgtattagaggaaatgatacaaatgatcatggcgattttaatggtatcgagaaaacattaacagtaaatccgaattctgaattaatctttgaatttaatactatgactactaaaaactatcaaggtatgacaaatttaatcattaaaaatgctgataacgatactgttattggtgaaaaagtagttgcttatggtccgatttggcgcttattaaaagtacctgaaaatgttagtcatctaaaaattcaatttgtacctaaaaatgacgcaataacagatgcacgtggtatttatcaattacgagatggatataaatactatgactttgtagactcaatcggtcttcattctgggtcacatgtctatgttgaaagacgtacaatggagccaacagcaacaaataataaagaatttacagttacaacgtcattaaagaataatggtaactttggcgcttcattcaatacagatgattttgtatataaaattcaattacctgaaggt
gttgaatatgtaaataattcattgactaaagattttcctagcggtaattcaggtgttgatattaatgatatgaatgtgacgtatgacgcagcaaatcgaattattacaattaaaagtactggtggaggtacagggaattcgccggcacgactaatgcctgataaaatattggatttgaagtataagctacgtgtgaacaatgtgccaacaccaagaacagtaacatttaacgatacattaacgtataaaacatattcacaagattttattaattcacctgctgaaagtcatactgtaagtacaaatccatatacaattgatatcatcatgaataaagacgcattgcaagccgaagtcgatagacgaattcaacaagcggattatacatttgcatcattagatatttttaatgatcttaaaagacgcgcacaaacaattttagatgaaaaccgtaacaatgtacctttaaacaaaagagtttctcaagcagatatcgattcattagcaaatcagatgcaacatacgttaattcgcagtgttgacgctgaaaatgccgttaatagaaaagttgatgacatggaagatttagttaaccaaaatgatgaactgacagatgaagaaaaacaagcagcgattcaagtcatcgaggaacataaaaatgaaattattgggaatattggtgaccaaacgactgatgatggcgttactagaattaaagatcaaggtatacagactttaagtggagacactgcaacaccagttgttaaaccaaatgctaaacaagctatacgtgataaagcagcgaaacaaagagaaattatcaatcacacgccagatgctactcaagatgaaattcaagatgcattaaatcaattaacaacggatgaaacagatgctattgataatgttacgaatgctactaccaatgctgatgttgaaacagctaaaaataatggtattaatacaattggtgcagttgcgccacaagtgacacacaaacaagctgcaagagatgcaattaatcaagcgacagcaacgaaacgacaacaaataaatagcaatagagaagcaacacaagaagagaaaaatgcagcattgaatgaattaacgcaagccacgaaccacgcattagaacaaatcaatcaagcgacaaccaatgatgatgtagatactgccaaaggtgatggtctgaatgccattaatcctattgcgcctgtaactgttgtcaagcaagcagcaagagatgccgtatcacatgatgcacaacagcatatcgcagagatcaatgcaaatcctgatgcgactcaagaagaaagacaagcagcaatagagaaagtaaatgctgctgtagctgttgcgaatactaatatattaaatgctaataccaatgctgatgttgagcaagtaaagacaaatgcaattcaaggtatacaagccattgaaccagctacaaaggttaaaacagatgctaaaaacgctattgatcaaagtgcggaaacgcaacataatgcgatatttaataataatgatgcgaccttagaagagcaacaagcagcacaacaattgcttgatcaagctgtagccacagcgaagcaaaatattaatgcagcagatacgaatcaagaagttgcacaagcaaaagatcagggcacacaaaatatagttgtgattcaaccggcaacacaagttaaaacggatgcacgcaatgctgtaaatgaaaaagcgcgagaggcgataacaaatatcaatgctacacctggcgcgactcgagaagagaaacaagaagccgataaatcgtgtcaatacacttaaaaatagagcattaaatgatattggtgtgacgtctactactgcgatggtcaatagtattagagacgatgcagtcaatcaaatcggtgcagttcaaccgcatgtaacgaagaaacaaactgctacaggtgtattaacggacttagcaactgcaaaaaaacaagaaattaatcaaaatacaaatgcaaccactgaagaaaagcaagtagcattaaatcaagtagaccaagatttagcaacggcaattaataatataaatcaagctgatactaatgcagaagtagatcaagcacaacaattaggtacaaaagcaattaatgcgattcagccaaatattgtaaaaaaacctgcagcattagcacaaaccaatcagcattatagtgctaaattagttgaaatcaatgctacaccagatgcaacagatgatgagaaaaatgctgcgatcaatactttaaatcaagacagacaacaagctattgaaagtattaaacaagcaaatacaaatgcggaagtagaccaagctgcgacagtggcagagaataatatcgatgctgttcaagttgacgttgtaaaaaaacaagcagcgcgagataaaatcactgctgaagtagcgaagcgtattgaagcggttaaacaaacacctaatgcaactgacgaagaaaagcaggctgcagttaatcaaatcaatcaacttaaagatcaagcgtttaatcaaattaatcaaaaccaaacaaatgatcaggtagacgcaactacaaatcaagcgattaatgctatagataatgttgaagctgaagtagtaattaaaccaaaggcaattgcagatattgaaaaagctgttaaagaaaagcaacagcaaattgataatagtcttgattcaacagataatgagaaagaagttgctttacaagcattagctaaagaaaaagaaaaagcacttgcagctattgaccaagctcaaacgaatagtcaggtgaatcaagcggcaacaaatggtgtatcagcgattaaaattattcaacctgaaacaaaaattaaaccagcagcacgtgaaaaaatcaatcaaaaagcgaatgaattacgtgcgcaaattaatcaagataaagaagcgacagcagaagaaagacaagcggcgttagataaaatcaatgatttagttgctaaagctatgacaaatatcacgaatgatagaacaaatcagcaagttaatgactcaacaaatcaagcgcttgacgacattgcattagtgacgcctgaccatattgttagagcagctgctagagatgcagttaagcaacaatatgaagctaaaaagcacgaaattgagcaagcggaacatg cgactgatgaagaaaaacaagttgctttaaatcaattagcgaataatgaaaaacgtgcattacaaaacattaatcaagcaatagcgaataatgatgtgaaacgtgttgaatcaaatggtattgctacgttaaaaggcgtagaaccgcacattgtggttaaacctgaagctcaagaagccataaaagcgagcgcagataaccaagtagaatctataaaagatacaccacatgctacgacagatgaattagatgaagcaaaccaacaaataaacgacacacttaaacaaggtcaacaagatatagacaatacgacacaagatgcagctgtcaatgatgttagaaaccaaacgattaaggcaatcgaacaaattaaaccgaaagttagacgcaaacgtgcagcgttggataacattgatgaaagtaataataatcaactcgatgcaatacgaaatacgctagatacaacgcaagatgaacgaaatgttgctattgctgcgttaaataaaattgttaatgcaattaaaaatgatattgcacaaaacaaaacgaatgcagaagtggatcaaactgaggctgatggtaacaacaacatcaaagtgattttacctaaagttcaagttaaaccagcagcgcgtcaatctgtcagcgcaaaagctgaag
ctcaaaatgcacttattgatcaaagtgatttatctaccgaagaagaaagattagctgctaaacatttagtagaacaagcacttaatcaagctattgatcagatcaatcacgcagataagactgcgcaagttaatcaaaatagtatcgatgctcaaaatattatttcaaaaattaaaccagcgacaacagttaaagcaacagcattacaacaaattcaaaatatcgctacaaataaaattaatttaattaaagcaaataacgaagcgacagatgaagaacaaaatgctgcaatagtacaagttgaaaaagagttaattaaagctaaacaacaaattgctggtgcagtgactaatgctgatgtggcatatttattgcatgatgggaaaaacgaaattcgtgaaatcgaacctgttattaataaaaaagcaactgcgcgagaacaattaacaacattattcaacgataagaaacaagcaattgaagcgaatgttcaagcaacagtagaagaaagaaatagtattttagcacagttacaaaacatttatgacactgctattggacaaattgatcaagatcgtagcaatgcacaagttgataaaacagcaacattaaatctacaaacaatacatgatttagacgtacatcctattaaaaagccagatgctgaaaaaacgattaatgatgatcttgcacgtgttacacatttagtgcaaaattatcgaaaagtaagtgatcgtaataaggctgatgcattaaaagctataactgcattaaaattacaaatggatgaagaattaaaaacagcacgcactaatgctgatgttgatgcagttttaaaacgatttaatgttgcattaggcgatatagaagcagtaattactgaaaaagaaaatagcttactgcgcattgataacattgctcaacaaacatatgcgaaattcaaagcgatcgcaacaccagaacaattagctaaagtaaaagcattaattgatcaatatgttgcagatggcaatagaatggttgatgaagatgcgacattaaatgacatcaaaaaagatacgcaactcattattgatgaaattttagcaattaaattacctgctgaagtgataaaagcgtcaccaaaagtggggcaacctgctccaaaagtttgtacgcctattaaaaaagaagataaacaagaagtgcgaaaagttgtaaaagaacttccaaatactggttctgaagaaatggatttaccattaaaagaattagcactaattacaggcgcagcattattagctagaagacgttctaaaaaagaaaaagaatcataaEkeS_MRSA(SEQ ID NO2)MNLLKKNKYSIRKYKVGIFSTLIGTVLLLSNPNGAQALTTDHNVQGGSNQALPGNSQNTNADTNRDIVNDSQNTPNAHATDNTSTNQALTNHQNVDVANQVGPAPIQPSASPAQNNNNSNANSTATEPAANTNNNLASNNNTLNVPNNTDNNDSARHLTLKEIQEDVRHSSDKPELVAIAEEASNRPKKRSRRAAPTDPNATPADPTATPADPTAGNGSAPVAITAPYTPTTDPNANNIGQNAPNEVLSFDDNNIRPSTNRSVPTVTVVDNLPGYTLINGGKVGVFSHAMVRTSMFDSGDAKNYQAQGNVIALGRIRGNDTNDHGDFNGIEKTLTVNPNSELIFEFNTMTTKNYQGMTNLIIKNADNDTVIGEKVVAYGPIWRLLKVPENVSHLKIQFVPKNDAITDARGIYQLRDGYKYYDFVDSIGLHSGSHVYVERRTMEPTATNNKEFTVTTSLKNNGNFGASFNTDDFVYKIQLPEGVEYVNNSLTKDFPSGNSGVDINDMNVTYDAANRIITIKSTGGGTGNSPARLMPDKILDLKYKLRVNNVPTPRTVTFNDTLTYKTYSQDFINSPAESHTVSTNPYTIDIIMNKDALQAEVDRRIQQADYTFASLDIFNDLKRRAQTILDENRNNVPLNKRVSQADIDSLANQMQHTLIRSVDAENAVNRKVDDMEDLVNQNDELTDEEKQAAIQVIEEHKNEIIGNIGDQTTDDGVTRIKDQGIQTLSGDTATPVVKPNAKQAIRDKAAKQREIINHTPDATQDEIQDALNQLTTDETDAIDNVTNATTNADVETAKNNGINTIGAVAPQVTHKQAARDAINQATATKRQQINSNREATQEEKNAALNELTQATNHALEQINQATTNDDVDTAKGDGLNAINPIAPVTVVKQAARDAVSHDAQQHIAEINANPDATQEERQAAIEKVYAAVAVANTNILNANTNADVEQVKTNAIQGIQAIEPATKVKTDAKNAIDQSAETQHNAIFNNNDATLEEQQAAQQLLDQAVATAKQNINAADTNQEVAQAKDQGTQNIVVIQPATQVKTDARNAVNEKAREAITNINATPGATREEKQEAINRVNTLKNRALNDIGVTSTTAMVNSIRDDAVNQIGAVQPHVTKKQTATGVLTDLATAKKQEINQNTNATTEEKQVALNQVDQDLATAINNINQADTNAEVDQAQQLGTKAINAIQPNIVKKPAALAQTNQHYSAKLVEINATPDATDDEKNAAINTLNQDRQQAIESIKQANTNAEVDQAATVAENNIDAVQVDVVKKQAARDKITAEVAKRIEAVKQTPNATDEEKQAAVNQINQLKDQAFNQINQNQTNDQVDATTNQAINAIDNVEAEVVIKPKAIADIEKAVKEKQQQIDNSLDSTDNEKEVALQALAKEKEKALAAIDQAQTNSQVNQAATNGVSAIKIIQPETKIKPAAREKINQKANELRAQINQDKEATAEERQAALDKINDLVAKAMTNITNDRTNQQVNDSTNQALDDIALVTPDHIVRAAARDAVKQQYEAKKHEIEQAEHATDEEKQVALNQLANNEKRALQNINQAIANNDVKRVESNGIA
TLKGVEPHIVVKPEAQEAIKASADNQVESIKDTPHATTDELDEANQQINDTLKQGQQDIDNTTQDAAVNDVRNQTIKAIEQIKPKVRRKRAALDNIDESNNNQLDAIRNTLDTTQDERNVAIAALNKIVNAIKNDIAQNKTNAEVDQTEADGNNNIKVILPKVQVKPAARQSVSAKAEAQNALIDQSDLSTEEERLAAKHLVEQALNQAIDQINHADKTAQVNQNSIDAQNIISKIKPATTVKATALQQIQNIATNKINLIKANNEATDEEQNAAIVQVEKELIKAKQQIAGAVTNADVAYLLHDGKNEIREIEPVINKKATAREQLTTLFNDKKQAIEANVQATVEERNSILAQLQNIYDTAIGQIDQDRSNAQVDKTATLNLQTIHDLDVHPIKKPDAEKTINDDLARVTHLVQNYRKVSDRNKADALKAITALKLQMDEELKTARTNADVDAVLKRFNVALGDIEAVITEKENSLLRIDNIAQQTYAKFKAIATPEQLAKVKALIDQYVADGNRMVDEDATLNDIKKDTQLIIDEILAIKLPAEVIKASPKVGQPAPKVCTPIKKEDKQEVRKVVKELPNTGSEEMDLPLKELALITGAALLARRRSKKEKESDsqA(8325)(SEQ ID NO3)tctaatgaatgtaaagataatacaaggagttattacatgagtaaaagacagaaagcatttcatgacagcttagcaaacgaaaaaacaagagtaagactttataaatctggaaaaaattgggtaaaatccggaattaaagaaatagaaatgttcaaaattatggggctaccatttattagtcatagtttagtgagtcaagataatcaaagcattagtaaaaaaatgacgggatacggactgaaaactacggcggttattggtggtgcattcacggtaaatatgttgcatgaccagcaagcttttgcggcttctgatgcaccattaacttctgaattaaacacacaaagtgaaacagtaggtaatcaaaactcaacgacaatcgaagcatcaacatcaacagccgattccacaagtgtaacgaaaaatagtagttcggtacaaacatcaaatagtgacacagtctcaagtgaaaagtctgaaaaggtcacttcgacaactaatagtacaagcaatcaacaagagaaattgacatctacatcagaatcaacatcctcaaagaatactacatcaagttctgatactaaatctgtagcttcaacttcaagtacagaacaaccaattaatacatcaacaaatcaaagtactgcatcaaataacacttcacaaagcacaacgccatcttcggtcaacttaaacaaaactagcacaacgtcaactagcaccgcaccagtaaaacttcgaactttcagtcgcttagctatgtcaacatttgcgtcagcagcgacgacaaccgcagtaactgctaatacaattacagttaataaagataacttaaaacaatatatgacaacgtcaggtaatgctacctatgatcaaagtaccggtattgtgacgttaacacaggatgcatacagccaaaaaggtgctattacattaggaacacgtattgactctaataagagttttcatttttctggaaaagtaaatttaggtaacaaatatgaagggcatggaaatggtggagatggtatcggttttgccttttcaccaggtgtattaggtgaaacagggttaaacggtgccgcagtaggtattggtggcttaagtaacgcatttggcttcaaattggatacgtatcacaatacatctaaaccaaattcagctgcaaaggcgaatgctgacccatctaatgtagctggtggaggtgcgtttggtgcatttgtaacaacagatagttatggtgttgcgacaacgtatacatcaagttcaacagctgataatgctgcgaagttaaatgttcaacctacaaataacacgttccaagattttgatattaactataatggtgatacaaaggttatgactgtcaaatatgcaggtcaaacatggacacgtaatatttcagattggattgcgaaaagtggtacgaccaacttttcattatcaatgacagcctcaacaggtggcgcgacaaatttacaacaagtacaatttggaacattcgaatatacagagtctgctgttacacaagtgagatacgttgatgtaacaacaggtaaagatattattccaccaaaaacatattcaggaaatgttgatcaagtcgtgacaatcgataatcagcaatctgcattgactgctaaaggatataactacacgtccgtcgatagttcatatgcgtcaacttataatgatacaaataaaactgtaaaaatgacgaatgctggacaatcagtgacatattattttactgatgtaaaagcaccaactgtaactgtaggcaatcaaaccatagaagtgggtaaaacaatgaatcctattgtattgactacaacggataatggtactgggactgtgacaaatacagttacaggattaccaagcggattaagttacgatagtgcaacgaattcaatcattgggacaccaacaaaaattggtcaatcaacagtgacagttgtgtctactgaccaagcaaataacaaatcgacgacaacttttacaataaatgttgtggatacgacagcaccaacagtgacaccaataggagatcaatcatcagaagtgtattcaccaatatccccgattaaaattgctacgcaag ataacagtggaaatgcggtgacgaatacagtgactggattgccatccggactaacatttgatagtacaaataatactattagtggtacaccaacaaacattggtacaagtactatatcaatcgtttctacagatgcgagcggtaacaaaacgacgacaacttttaaatatgaagtaacaagaaatagcatgagtgattccgtatcaacatcaggaagtacacaacaatctcaaagtgtgtcaacaagtaaagctgactcacaaagtgcatcaacgagtacatcaggatcgattgtggtatctacatcagctagtacctcgaaatcgacaagtgtaagcctatctgattctgtgagtgcatctaagtcattaagcacatctgaaagtaatagtgtatcaagctcaacaagcacaagtttagtgaattcacaaagtgtatcatcaagcatgtcggattcagctagtaaatcaacatcattaagcgattctatttcaaactctagcagtactgaaaaatccgaaagtctatcaacaagtacatctgattcattgcgtacatcaacatcactcagtgactcattaagtatgagtacatcaggaagcttgtctaagtcacaaagcttatcaacgagtatatcagggtcgtctagtacatcagcatcattaagtgacagtacatcgaatgcaattagtacatcaacatcattgagcgagtcagctagcacctcggactctatcagtatttcaaatagcatagccaactctcaaagtgcgtcaacaa
gcaaatcagattcacaaagtacatcaatatcattaagtacaagtgattcaaaatcgatgagtacatcagaatcattgagcgattcgacgagcacaagtggttctgtttctggatcactaagcatagcagcatcacaaagtgtctcaacaagtacatcagactcgatgagtacttcagagatagtaagtgactctatcagtacaagtgggtcattatctgcatcagacagtaaatcaatgtccgtaagtagttcaatgagcacgtctcagtcaggtagtacatcagaatcattaagtgattcacaaagtacatctgattctgatagtaagtcattatcacaaagtactagtcaatcaggttcaacaagtacatcaacgtcgacaagtgcttcagtacgtacttcggaatcacaaagtacgtctggttcaatgagtgcaagtcaatccgattcaatgagcatatcaacgtcgtttagtgattcaacgagtgatagcaaatcagcatcaactgcatcaagtgaatcaatatcacaaagtgcttctacgagcacatctggttcggtaagtacttcgacatcgttaagtacaagtaattcagaacgtacatcaacatctatgagtgattccacaagcttaagtacatcagagtctgattcaataagtgaatcaacgtcaacgagcgactctataagtgaagcaatatctgcttcagagagcacgtttatatcattaagtgaatcaaatagtactagcgattcagaatcacaaagtgcatctgcctttttaagtgaatcattaagtgaaagtacgtctgaatcaacatcagagtcagtgagtagttcgacaagtgagagtacgtcattatcagacagtacatcagaatctggtagcacatcaacatcattaagtaattcaacaagtggtagtacgtccatttcaacatcgacaagtatcagtgaatcaacgtcaacgtttaagagcgagagtgtttcaacatcactgagtatgtcaacgagtacaagtttgtctgactctacaagtttgtcaacatcattaagtgattccacaagtgatagtaagtctgattcattaagtacatcaatgtcgacaagtgattcaatcagtacaagtaaatctgattccattagtacatccacatcattaagtggttctacaagtgaaagtgaatccgactcaacatcatcaagtgaaagtaaatccgattcaacatcaatgagcataagtatgtctcaatcaacatcaggaagtacaagtacgtcaacgagtacaagtttgtctgactcaacgagtacatcattgtcactaagtgcctcaatgaatcaaagcggagtagactcaaactcagcaagccaaagtgcctcaaactcaacaagtacaagcacgagcgaatccgattcacaaagcacatcatcatatacaagtcagtcaacaagccaaagtgaatccacatcgacatcaacgtcactaagcgattcaacaagtatatctaaaagtacgagtcaatcaggttcggtaagcacatcagcgtcattaagtggttcagagagtgaatctgattcacaaagtatctcaacaagtgcaagtgagtcaacatcagaaagtgcgtcaacatcactcagtgactcaacaagtacaagtaactcaggatcagcaagtacgtcaacatcgctcagtaactcagcaagcgcaagtgaatccgatttgtcgtcaacatctttaagtgattcaacatctgcgtcaatgcaaagcagtgaatccgattcacaaagcacatcagcatcattaagtgattcgctaagtacatcaacttcaaaccgcatgtcgaccattgcaagtttatctacatcggtaagtacatcagagtctggctcaacatcagaaagtacaagtgaatccgattcaacatcaacatcattaagcgattcacaaagcacatcaagaagtacaagtgcatcaggatcagcaagtacatcaacatcaacaagtgactctcgtagtacatcagcttcaactagtacttcgatgcgtacaagtactagtgattcacaaagtatgtcgctttcgacaagtacatcaacaagtatgagtgattcaacgtcattatctgatagtgttagtgattcaacatcagactcaacaagtgcgagtacatctggttcgatgagtgtgtctatatcgttaagtgattcgacaagtacatcaacatcggctagtgaagtaatgagcgcaagcatatctgattcacaaagtatgtcagaatctgtaaatgattcagaaagtgtaagtgaatctaattctgaaagtgactctaaatcgatgagtggctcaacaagtgtcagtgattctggctcattgagcgtctcaacgtcattaagaaaatcagaaagtgtaagcgagtcaagttcattgagttgctcacaatcgatgagcgattcagtaagcacaagcgattcgtcatcattaagtgtatcgacgtcactaagaagttcagaaagcgtgagtgaatctgattcattaagtgattcaaaatcaacaagtggttcgacttcaacaagtacatctggttcattgagtacctcaacatcattaagtggttcagaaagcgtaagcgagtctacctcgctaagtgattcaatatcaatgagtgattctactagtacaagtgactccgactcattaagtggatcaatatctttaagtggttccacaagtcttagcacttcggattcattaagtgattcaaaatcattgagtagctcgcaaagtatgagtggatcagaatcaacgtcaacaagtgtgagcgattcgcagtcaagctcaacaagtaatagtcaatttgactctatgagcatcagtgcatcagaaagcgactcaatgtctacaagtgattcgtctagcatcagtggatcaaattcaacgagtacatcactttcaacatctgactcaatgagcggaagcgtatcagtttcaacatcgacaagtttaagtgactcaatatcaggttcaacaagtgtaagtgactcgagctcaacaagcacatctacatcattaagtgattcaatgtcacaaagccagtcaacaagtacaagtgcatctggttccttaagtacatcgatatcaacatcaatgtcaatgagtgctagtacatcgtcatcacaaagcacatcggtgtcgacatcattatcaacatcagacagtatcagtgattctacttcaataagtatcagtggttcacaaagtacagtagaatcagaatctacaagtgattcaacttctatcagtgactcagaatcattgagtacatcagattcagactcgacatcgacaagtacatcggactcaacaagtggttcaacttcaacaagcatatctgaatcattaagtacgtctggttcaggttcaacgagcgtatctgactcaacatcaatgagtgaatctaattcatcgagtgtttcaatgtcacaagacaaatccgactcaacatcaattagtgactcagaatcagtgtcaacaagcacatcaacgtcattgagcacatccgattcgacaagcacatccgaatcactgagtacatctatgtctggttcacaaagcatttctgactcaacatcaacaagtatgtccggctcaacaagtacatctgaatctaactcaatgcatccgtcagactcaatgagtatgcatcatactcacagcacgagcacatctcgcttatcaagtgaagcaacaacgagcacgagtgaatctcagtctacattaagtgcaacatctgaagtgactaaacataatggcacaccagcacaaagtgaaaaaagattgccagatacaggtgactcaataaaacaaaatggattactaggtggcgttatgacattattagttggtttaggtttaatgaagagaaagaaaaagaaagatgaaaatgatcaagatgattctcaagcataa
DsqA(8325)(SEQ ID NO4)SNECKDNTRSYYMSKRQKAFHDSLANEKTRVRLYKSGKNWVKSGIKEIEMFKIMGLPFISHSLVSQDNQSISKKMTGYGLKTTAVIGGAFTVNMLHDQQAFAASDAPLTSELNTQSETVGNQNSTTIEASTSTADSTSVTKNSSSVQTSNSDTVSSEKSEKVTSTTNSTSNQQEKLTSTSESTSSKNTTSSSDTKSVASTSSTEQPINTSTNQSTASNNTSQSTTPSSVNLNKTSTTSTSTAPVKLRTFSRLAMSTFASAATTTAVTANTITVNKDNLKQYMTTSGNATYDQSTGIVTLTQDAYSQKGAITLGTRIDSNKSFHFSGKVNLGNKYEGHGNGGDGIGFAFSPGVLGETGLNGAAVGIGGLSNAFGFKLDTYHNTSKPNSAAKANADPSNVAGGGAFGAFVTTDSYGVATTYTSSSTADNAAKLNVQPTNNTFQDFDINYNGDTKVMTVKYAGQTWTRNISDWIAKSGTTNFSLSMTASTGGATNLQQVQFGTFEYTESAVTQVRYVDVTTGKDIIPPKTYSGNVDQVVTIDNQQSALTAKGYNYTSVDSSYASTYNDTNKTVKMTNAGQSVTYYFTDVKAPTVTVGNQTIEVGKTMNPIVLTTTDNGTGTVTNTVTGLPSGLSYDSATNSIIGTPTKIGQSTVTVVSTDQANNKSTTTFTINVVDTTAPTVTPIGDQSSEVYSPISPIKIATQDNSGNAVTNTVTGLPSGLTFDSTNNTISGTPTNIGTSTISIVSTDASGNKTTTTFKYEVTRNSMSDSVSTSGSTQQSQSVSTSKADSQSASTSTSGSIVVSTSASTSKSTSVSLSDSVSASKSLSTSESNSVSSSTSTSLVNSQSVSSSMSDSASKSTSLSDSISNSSSTEKSESLSTSTSDSLRTSTSLSDSLSMSTSGSLSKSQSLSTSISGSSSTSASLSDSTSNAISTSTSLSESASTSDSISISNSIANSQSASTSKSDSQSTSISLSTSDSKSMSTSESLSDSTSTSGSVSGSLSIAASQSVSTSTSDSMSTSEIVSDSISTSGSLSASDSKSMSVSSSMSTSQSGSTSESLSDSQSTSDSDSKSLSQSTSQSGSTSTSTSTSASVRTSESQSTSGSMSASQSDSMSISTSFSDSTSDSKSASTASSESISQSASTSTSGSVSTSTSLSTSNSERTSTSMSDSTSLSTSESDSISESTSTSDSISEAISASESTFISLSESNSTSDSESQSASAFLSESLSESTSESTSESVSSSTSESTSLSDSTSESGSTSTSLSNSTSGSTSISTSTSISESTSTFKSESVSTSLSMSTSTSLSDSTSLSTSLSDSTSDSKSDSLSTSMSTSDSISTSKSDSISTSTSLSGSTSESESDSTSSSESKSDSTSMSISMSQSTSGSTSTSTSTSLSDSTSTSLSLSASMNQSGVDSNSASQSASNSTSTSTSESDSQSTSSYTSQSTSQSESTSTSTSLSDSTSISKSTSQSGSVSTSASLSGSESESDSQSISTSASESTSESASTSLSDSTSTSNSGSASTSTSLSNSASASESDLSSTSLSDSTSASMQSSESDSQSTSASLSDSLSTSTSNRMSTIASLSTSVSTSESGSTSESTSESDSTSTSLSDSQSTSRSTSASGSASTSTSTSDSRSTSASTSTSMRTSTSDSQSMSLSTSTSTSMSDSTSLSDSVSDSTSDSTSASTSGSMSVSISLSDSTSTSTSASEVMSASISDSQSMSESVNDSESVSESNSESDSKSMSGSTSVSDSGSLSVSTSLRKSESVSESSSLSCSQSMSDSVSTSDSSSLSVSTSLRSSESVSESDSLSDSKSTSGSTSTSTSGSLSTSTSLSGSESVSESTSLSDSISMSDSTSTSDSDSLSGSISLSGSTSLSTSDSLSDSKSLSSSQSMSGSESTSTSVSDSQSSSTSNSQFDSMSISASESDSMSTSDSSSISGSNSTSTSLSTSDSMSGSVSVSTSTSLSDSISGSTSVSDSSSTSTSTSLSDSMSQSQSTSTSASGSLSTSISTSMSMSASTSSSQSTSVSTSLSTSDSISDSTSISISGSQSTVESESTSDSTSISDSESLSTSDSDSTSTSTSDSTSGSTSTSISESLSTSGSGSTSVSDSTSMSESNSSSVSMSQDKSDSTSISDSESVSTSTSTSLSTSDSTSTSESLSTSMSGSQSISDSTSTSMSGSTSTSESNSMHPSDSMSMHHTHSTSTSRLSSEATTSTSESQSTLSATSEVTKHNGTPAQSEKRLPDTGDSIKQNGLLGGVMTLLVGLGLMKRKKKKDENDQDDSQA
KesK1(8325)(SEQ ID NO5)ttattatcaattaaatataatcttataggagttgttaacaacatgaacaaacatcacccaaaattaaggtctttctattctattagaaaatcaactctaggcgttgcatcggtcattgtcagtacactatttttaattacttctcaacatcaagcacaagcagcagaaaatacaaatacttcagataaaatctcggaaaatcaaaataataatgcaactacaactcagccacctaaggatacaaatcaaacacaacctgctacgcaaccagcaaacactgcgaaaaactatcctgcagcggatgaatcacttaaagatgcaattaaagatcctgcattagaaaataaagaacatgatataggtccaagagaacaagtcaatttccagttattagataaaaacaatgaaacgcagtactatcactttttcagcatcaaagatccagcagatgtgtattacactaaaaagaaagcagaagttgaattagacatcaatactgcttcaacatggaagaagtttgaagtctatgaaaacaatcaaaaattgccagtgagacttgtatcatatagtcctgtaccagaagaccatgcctatattcgattcccagtttcagatggcacacaagaattgaaaattgtttcttcgactcaaattgatgatggagaagaaacaaattatgattatactaaattagtatttgctaaacctatttataacgatccttcacttgtaaaatcagatacaaatgatgcagtagtaacgaatgatcaatcaagttcagtcgcaagtaatcaaacaaacacgaatacatctaatcaaaatatatcaacgatcaacaatgctaataatcaaccgcaggcaacgaccaatatgagtcaacctgcacaaccaaaatcgtcaacgaatgcagatcaagcgtcaagccaaccagctcatgaaacaaattctaatggtaatactaacgataaaacgaatgagtcaagtaatcagtcggatgttaatcaacagtatccaccagcagatgaatcactacaagatgcaattaaaaacccggctatcatcgataaagaacatacagctgataattggcgaccaattgattttcaaatgaaaaatgataaaggtgaaagacagttctatcattatgctagtactgttgaaccagcaactgtcatttttacaaaaacaggaccaataattgaattaggtttaaagacagcttcaacatggaagaaatttgaagtttatgaaggtgacaaaaagttaccagtcgaattagtatcatatgattctgataaagattatgcctatattcgtttcccagtatctaatggtacgagagaagttaaaattgtgtcatctattgaatatggtgagaacatccatgaagactatgattatacgctaatggtctttgcacagcctattactaataacccagacgactatgtggatgaagaaacatacaatttacaaaaattattagctccgtatcacaaagctaaaacgttagaaagacaagtttatgaattagaaaaattacaagagaaattgccagaaaaatataaggcggaatataaaaagaaattagatcaaactagagtagagttagctgatcaagttaaatcagcagtgacggaatttgaaaatgttacacctacaaatgatcaattaacagatttacaagaagcgcattttgttgtttttgaaagtgaagaaaatagtgagtcagttatggacggctttgttgaacatccattctatacagcaactttaaatggtcaaaaatatgtagtgatgaaaacaaaggatgacagttactggaaagatttaattgtagaaggtaaacgtgtcactactgtttctaaagatcctaaaaataattctagaacgctgattttcccatatatacctgacaaagcagtttacaatgcgattgttaaagtcgttgtggcaaacattggttatgaaggtcaatatcatgtcagaattataaatcaggatatcaatacaaaagatgatgatacatcacaaaataacacgagtgaaccgctaaatgtacaaacaggacaagaaggtaaggttgctgatacagatgtagctgaaaatagcagcactgcaacaaatcctaaagatgcgtctgataaagcagatgtgatagaaccagagtctgacgtggttaaagatgctgataataatattgataaagatgtgcaacatgatgttgatcatttatccgatatgtcggataataatcacttcgataaatatgatttaaaagaaatggatactcaaattgccaaagatactgatagaaatgtggataaagatgccgataatagcgttggtatgtcatctaatgtcgatactgataaagactctaataaaaataaagacaaagtcatacagctgaatcatattgccgataaaaataatcatactggaaaagcagcaaagcttgacgtagtgaaacaaaattataataatacagacaaagttactgacaaaaaaacaactgaacatctgccgagtgatattcataaaactgtagataaaacagtgaaaacaaaagaaaaagccggcacaccatcgaaagaaaacaaacttagtcaatctaaaatgctaccaaaaactggagaaacaacttcaagccaatcatggtggggcttatatgcgttattaggtatgttagctttattcattcctaaattcagaaaagaatctaaataa
KesK1(8325)(SEQ ID NO6)LLSIKYNLIGVVNNMNKHHPKLRSFYSIRKSTLGVASVIVSTLFLITSQHQAQAAENTNTSDKISENQNNNATTTQPPKDTNQTQPATQPANTAKNYPAADESLKDAIKDPALENKEHDIGPREQVNFQLLDKNNETQYYHFFSIKDPADVYYTKKKAEVELDINTASTWKKFEVYENNQKLPVRLVSYSPVPEDHAYIRFPVSDGTQELKIVSSTQIDDGEETNYDYTKLVFAKPIYNDPSLVKSDTNDAVVTNDQSSSVASNQTNTNTSNQNISTINNANNQPQATTNMSQPAQPKSSTNADQASSQPAHETNSNGNTNDKTNESSNQSDVNQQYPPADESLQDAIKNPAIIDKEHTADNWRPIDFQMKNDKGERQFYHYASTVEPATVIFTKTGPIIELGLKTASTWKKFEVYEGDKKLPVELVSYDSDKDYAYIRFPVSNGTREVKIVSSIEYGENIHEDYDYTLMVFAQPITNNPDDYVDEETYNLQKLLAPYHKAKTLERQVYELEKLQEKLPEKYKAEYKKKLDQTRVELADQVKSAVTEFENVTPTNDQLTDLQEAHFVVFESEENSESVMDGFVEHPFYTATLNGQKYVVMKTKDDSYWKDLIVEGKRVTTVSKDPKNNSRTLIFPYIPDKAVYNAIVKVVVANIGYEGQYHVRIINQDINTKDDDTSQNNTSEPLNVQTGQEGKVADTDVAENSSTATNPKDASDKADVIEPESDVVKDADNNIDKDVQHDVDHLSDMSDNNHFDKYDLKEMDTQIAKDTDRNVDKDADNSVGMSSNVDTDKDSNKNKDKVIQLNHIADKNNHTGKAAKLDVVKQNYNNTDKVTDKKTTEHLPSDIHKTVDKTVKTKEKAGTPSKENKLSQSKMLPKTGETTSSQSWWGLYALLGMLALFIPKFRKESKKrkN2(8325)(SEQ ID NO7)gaggaaaacaacatgacaaaacattatttaaacagtaagtatcaatcagaacaacgttcatcagctatgaaaaagattacaatgggtacagcatctatcattttaggttcccttgtatacataggcgcagacagccaacaagtcaatgcggcaacagaagctacgaacgcaactaataatcaaagcacacaagtttctcaagcaacatcacaaccaattaatttccaagtgcaaaaagatggctcttcagagaagtcacacatggatgactatatgcaacaccctggtaaagtaattaaacaaaataataaatattatttccaaaccgtgttaaacaatgcatcattctggaaagaatacaaattttacaatgcaaacaatcaagaattagcaacaactgttgttaacgataataaaaaagcggatactagaacaatcaatgttgcagttgaacctggatataagagcttaactactaaagtacatattgtcgtgccacaaattaattacaatcatagatatactacgcatttggaatttgaaaaagcaattcctacattagctgacgcagcaaaaccaaacaatgttaaaccggttcaaccaaaaccagctcaacctaaaacacctactgagcaaactaaaccagttcaacctaaagttgaaaaagttaaacctactgtaactacaacaagcaaagttgaagacaatcactctactaaagttgtaagtactgacacaacaaaagatcaaactaaaacacaaactgctcatacagttaaaacagcacaaactgctcaagaacaaaataaagttcaaacacctgttaaagatgttgcaacagcgaaatctgaaagcaacaatcaagctgtaagtgataataaatcacaacaaactaacaaagttacaaaacataacgaaacgcctaaacaagcatctaaagctaaagaattaccaaaaactggtttaacttcagttgataactttattagcacagttgccttcgcaacacttgcccttttaggttcattatctttattacttttcaaaagaaaagaatctaaataa
KrkN2(8325)(SEQ ID NO8)EENNMTKHYLNSKYQSEQRSSAMKKITMGTASIILGSLVYIGADSQQVNAATEATNATNNQSTQVSQATSQPINFQVQKDGSSEKSHMDDYMQHPGKVIKQNNKYYFQTVLNNASFWKEYKFYNANNQELATTVVNDNKKADTRTINVAVEPGYKSLTTKVHIVVPQINYNHRYTTHLEFEKAIPTLADAAKPNNVKPVQPKPAQPKTPTEQTKPVQPKVEKVKPTVTTTSKVEDNHSTKVVSTDTTKDQTKTQTAHTVKTAQTAQEQNKVQTPVKDVATAKSESNNQAVSDNKSQQTNKVTKHNETPKQASKAKELPKTGLTSVDNFISTVAFATLALLGSLSLLLFKRKESKKrkN(8325)(SEQ ID NO9)tatacaattaggagttgtttctacaacatgaacaaacagcaaaaagaatttaaatcattttattcaattagaaagtcatcactaggcgttgcatctgtagcaattagtacacttttattattaatgtcaaatggcgaagcacaagcagcagctgaagaaacaggtggtacaaatacagaagcacaaccaaaaactgaagcagttgcaagtccaacaacaacatctgaaaaagctccagaaactaaaccagtagctaatgctgtctcagtatctaataaagaagttgaggcccctacttctgaaacaaaagaagctaaagaagttaaagaagttaaagcccctaaggaaacaaaagaagttaaaccagcagcaaaagccactaacaatacatatcctattttgaatcaggaacttagagaagcgattaaaaaccctgcaataaaagacaaagatcatagcgcaccaaactctcgtccaattgattttgaaatgaaaaagaaagatggaactcaacagttttatcattatgcaagttctgttaaacctgctagagttattttcactgattcaaaaccagaaattgaattaggattacaatcaggtcaattttggagaaaatttgaagtttatgaaggtgacaaaaagttgccaattaaattagtatcatacgatactgttaaagattatgcttacattcgcttctctgtatcaaacggaacaaaagctgttaaaattgttagttcaacacacttcaataacaaagaagaaaaatacgattacacattaatggaattcgcacaaccaatttataacagtgcagataaattcaaaactgaagaagattataaagctgaaaaattattagcgccatataaaaaagcgaaaacactagaaagacaagtttatgaattaaataaaattcaagataaacttcctgaaaaattaaaggctgagtacaagaagaaattagaggatacaaagaaagctttagatgagcaagtgaaatcagctattactgaattccaaaatgtacaaccaacaaatgaaaaaatgactgatttacaagatacaaaatatgttgtttatgaaagtgttgagaataacgaatctatgatggatacttttgttaaacaccctattaaaacaggtatgcttaacggcaaaaaatatatggtcatggaaactactaatgacgattactggaaagatttcatggttgaaggtcaacgtgttagaactataagcaaagatgctaaaaataatactagaacaattattttcccatatgttgaaggtaaaactctatatgatgctatcgttaaagttcacgtaaaaacgattgattatgatggacaataccatgtcagaatcgttgataaagaagcatttacaaaagccaataccgataaatctaacaaaaaagaacaacaagataactcagctaagaaggaagctactccagctacgcctagcaaaccaacaccatcacctgttgaaaaagaatcacaaaaacaagacagccaaaaagatgacaataaacaattaccaagtgttgaaaaagaaaatgacgcatctagtgagtcaggtaaagacaaaacgcctgctacaaaaccaactaaaggtgaagtagaatcaagtagtacaactccaactaaggtagtatctacgactcaaaatgttgcaaaaccaacaactgcttcatcaaaaacaacaaaagatgttgttcaaacttcagcaggttctagcgaagcaaaagatagtgctccattacaaaaagcaaacattaaaaacacaaatgatggacacactcaaagccaaaacaataaaaatacacaagaaaataaagcaaaatcattaccacaaactggtgaagaatcaaataaagatatgacattaccattaatggcattattagctttaagtagcatcgttgcattcgtattacctagaaaacgtaaaaactaa
KrkN(8325)(SEQ ID NO10)YTIRSCFYNMNKQQKEFKSFYSIRKSSLGVASVAISTLLLLMSNGEAQAAAEETGGTNTEAQPKTEAVASPTTTSEKAPETKPVANAVSVSNKEVEAPTSETKEAKEVKEVKAPKETKEVKPAAKATNNTYPILNQELREAIKNPAIKDKDHSAPNSRPIDFEMKKKDGTQQFYHYASSVKPARVIFTDSKPEIELGLQSGQFWRKFEVYEGDKKLPIKLVSYDTVKDYAYIRFSVSNGTKAVKIVSSTHFNNKEEKYDYTLMEFAQPIYNSADKFKTEEDYKAEKLLAPYKKAKTLERQVYELNKIQDKLPEKLKAEYKKKLEDTKKALDEQVKSAITEFQNVQPTNEKMTDLQDTKYWYESVENNESMMDTFVKHPIKTGMLNGKKYMVMETTNDDYWKDFMVEGQRVRTISKDAKNNTRTIIFPYVEGKTLYDAIVKVHVKTIDYDGQYHVRIVDKEAFTKANTDKSNKKEQQDNSAKKEATPATPSKPTPSPVEKESQKQDSQKDDNKQLPSVEKENDASSESGKDKTPATKPTKGEVESSSTTPTKVVSTTQNVAKPTTASSKTTKDVVQTSAGSSEAKDSAPLQKANIKNTNDGHTQSQNNKNTQENKAKSLPQTGEESNKDMTLPLMALLALSSIVAFVLPRKRKNRkaS(COL)(SEQ ID NO11)tttataaataatttacataaaatcaatcattttaatataaggattatgataatatattggtgtatgacagttaatggagggaacgaaatgaaagctttattacttaaaacaagtgtatggctcgttttgctttttagtgtaatgggattatggcaagtctcgaacgcggctgagcagcatacaccaatgaaagcacatgcagtaacaacgatagacaaagcaacaacagataagcaacaagtaccgccaacaaaggaagcggctcatcattctggcaaagaagcggcaaccaacgtatcagcatcagcgcagggaacagctgatgatacaaacagcaaagtaacatccaacgcaccatctaacaaaccatctacagtagtttcaacaaaagtaaacgaaacacgcgacgtagatacacaacaagcctcaacacaaaaaccaactcacacagcaacgttcaaattatcaaatgctaaaacagcatcactttcaccacgaatgtttgctgctaatgcaccacaaacaacaacacataaaatattacatacaaatgatatccatggccgactagccgaagaaaaagggcgtgtcatcggtatggctaaattaaaaacagtaaaagaacaagaaaagcctgatttaatgttagacgcaggagacgccttccaaggtttaccactttcaaaccagtctaaaggtgaagaaatggctaaagcaatgaatgcagtaggttatgatgctatggcagtcggtaaccatgaatttgactttggatacgatcagttgaaaaagttagagggtatgttagacttcccgatgctaagtactaacgtttataaagatggaaaacgcgcgtttaagccttcaacgattgtaacaaaaaatggtattcgttatggaattattggtgtaacgacaccagaaacaaagacgaaaacaagacctgaaggcattaaaggcgttgaatttagagatccattacaaagtgtgacagcggaaatgatgcgtatttataaagacgtagatacatttgttgttatatcacatttaggaattgatccttcaacacaagaaacatggcgtggtgattacttagtgaaacaattaagtcaaaatccacaattgaagaaacgtattacagttattgatggtcattcacatacagtacttcaaaatggtcaaatttataacaatgatgcattggcacaaacaggtacagcacttgcgaatatcggtaagattacatttaattatcgcaatggagaggtatcgaatattaaaccgtcattgattaatgttaaagacgttgaaaatgtaacaccgaacaaagcattagctgaacaaattaatcaagctgatcaaacatttagagcacaaactgcagaggtaattattccaaacaataccattgatttcaaaggagaaagagatgacgttagaacgcgtgaaacaaatttaggaaacgcgattgcagatgctatggaagcgtatggcgttaagaatttctctaaaaagactgactttgccgtgacaaatggtggaggtattcgtgcctctatcgcaaaaggtaaggtgacacgctatgatttaatctcagtattaccatttggaaatacgattgcgcaaattgatgtaaaaggttcagacgtctggacggctttcgaacatagtttaggcgcaccaacaacacaaaaggacggtaagacagtgttaacagcgaatggcggtttactacatatctctgattcaatccgtgtttactatgatataaataaaccgtctggcaaacgaattaatgctattcaaattttaaataaagagacaggtaagtttgaaaatattgatttaaaacgtgtatatcacgtaacgatgaatgacttcacagcatcaggtggcgacggatatagtatgttcggtggtcctagagaagaaggtatttcattagatcaagtactagcaagttatttaaaaacagctaacttagctaagtatgatacgacagaaccacaacgtatgttattaggtaaaccagcagtaagtgaacaaccagctaaaggacaacaaggtagcaaaggtagtaagtctggtaaagatacacaaccaattggtgacgacaaagtgatggatccagcgaaaaaaccagctccaggtaaagttgtattgttgctagcgcatagaggaactgttagtagcggtacagaaggttctggtcgcacaatagaaggagctactgtatcaagcaagagtgggaaacaattggctagaatgtcagtgcctaaaggtagcgcgcatgagaaacagttaccaaaaactggaactaatcaaagttcaagcccagaagcgatgtttgtattattagcaggtataggtttaatcgcgactgtacgacgtagaaaagctagctaa
RkaS(COL)(SEQ ID NO12)FINNLHKINHFNIRIMIIYWCMTVNGGNEMKALLLKTSVWLVLLFSVMGLWQVSNAAEQHTPMKAHAVTTIDKATTDKQQVPPTKEAAHHSGKEAATNVSASAQGTADDTNSKVTSNAPSNKPSTVVSTKVNETRDVDTQQASTQKPTHTATFKLSNAKTASLSPRMFAANAPQTTTHKILHTNDIHGRLAEEKGRVIGMAKLKTVKEQEKPDLMLDAGDAFQGLPLSNQSKGEEMAKAMNAVGYDAMAVGNHEFDFGYDQLKKLEGMLDFPMLSTNVYKDGKRAFKPSTIVTKNGIRYGIIGVTTPETKTKTRPEGIKGVEFRDPLQSVTAEMMRIYKDVDTFVVISHLGIDPSTQETWRGDYLVKQLSQNPQLKKRITVIDGHSHTVLQNGQIYNNDALAQTGTALANIGKITFNYRNGEVSNIKPSLINVKDVENVTPNKALAEQINQADQTFRAQTAEVIIPNNTIDFKGERDDVRTRETNLGNAIADAMEAYGVKNFSKKTDFAVTNGGGIRASIAKGKVTRYDLISVLPFGNTIAQIDVKGSDVWTAFEHSLGAPTTQKDGKTVLTANGGLLHISDSIRVYYDINKPSGKRINAIQILNKETGKFENIDLKRVYHVTMNDFTASGGDGYSMFGGPREEGISLDQVLASYLKTANLAKYDTTEPQRMLLGKPAVSEQPAKGQQGSKGSKSGKDTQPIGDDKVMDPAKKPAPGKVVLLLAHRGTVSSGTEGSGRTIEGATVSSKSGKQLARMSVPKGSAHEKQLPKTGTNQSSSPEAMFVLLAGIGLIATVRRRKASRrkN(8325)(SEQ ID NO13)agtggaaaatatggaaaaaggagtatgcaaatgagagataagaaaggaccggtaaataaaagagtagattttctatcaaataaattgaataaatattcaataagaaaatttacagttggaacagcatctattttaattggctcactaatgtatttgggaactcaacaagaggcagaagcagctgaaaacaatattgagaatccaactacattaaaagataatgtccaatcaaaagaagtgaagattgaagaagtaacaaacaaagacactgcaccacagggtgtagaagctaaatctgaagtaacttcaaacaaagacacaatcgaacatgaaccatcagtaaaagctgaagatatatcaaaaaaggaggatacaccaaaagaagtagctgatgttgctgaagttcagccgaaatcgtcagtcactcataacgcagagacacctaaggttagaaaagctcgttctgttgatgaaggctcttttgatattacaagagattctaaaaatgtagttgaatctaccccaattacaattcaaggtaaagaacattttgaaggttacggaagtgttgatatacaaaaaaaaccaacagatttaggggtatcagaggtaaccaggtttaatgttggtaatgaaagtaatggtttgataggagctttacaattaaaaaataaaatagattttagtaaggatttcaattttaaagttagagtggcaaataaccatcaatcaaataccacaggtgctgatggttgggggttcttatttagtaaaggaaatgcagaagaatatttaactaatggtggaatccttggggataaaggtctggtaaattcaggcggatttaaaattgatactggatacatttatacaagttccatggacaaaactgaaaagcaagctggacaaggttatagaggatacggagcttttgtgaaaaatgacagttctggtaattcacaaatggttggagaaaatattgataaatcaaaaactaattttttaaactatgcggacaattcaactaatacatcagatggaaagtttcatgggcaacgtttaaatgatgtcatcttaacttatgttgcttcaactggtaaaatgagagcagaatatgctggtaaaacttgggagacttcaataacagatttaggtttatctaaaaatcaggcatataatttcttaattacatctagtcaaagatggggccttaatcaagggataaatgcaaatggctggatgagaactgacttgaaaggttcagagtttacttttacaccagaagcgccaaaaacaataacagaattagaaaaaaaagttgaagagattccattcaagaaagaacgtaaatttaatccggatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacaccaacactaaaaaatccattaactggagtaattattagtaaaggtgaaccaaaagaagagattacaaaagatccgattaatgaattaacagaatacggacctgaaacaatagcgccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaggaagttccaggtaaaccaggaattaagaatccagaaacaggagacgtagttagaccgccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagagattccattcgagaaagaacgtaaatttaatcctgatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacgccaacactaaaaaatccattaactggagaaattattagtaaaggtgaatcgaaagaagaaatcacaaaagatccgattaatgaattaacagaatacggaccagaaacgataacaccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaggaagttccaggtaaaccaggaattaagaatccagaaacaggagatgtagttagaccaccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagagattccattcgagaaagaacgtaaatttaatcctgatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacaccaacactaaaaaatc
cattaactggagtaattattagtaaaggtgaaccaaaagaagaaatcacaaaagatccgattaatgaattaacagaatacggaccagaaacgataacaccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaagaagttccaggtaaaccaggaattaagaatccagaaacaggagacgtagttagaccaccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagagattccattcaagaaagaacgtaaatttaatccggatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacgccaacactaaaaaatccattaactggagaaattattagtaaaggtgaatcgaaagaagaaatcacaaaagatccgattaatgaattaacagaatacggaccagaaacgataacaccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaggaagttccaggtaaaccaggaattaagaatccagaaacaggagatgtagttagaccaccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagagattccattcgagaaagaacgtaaatttaatcctgatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacgccaacactaaaaaatccattaactggagaaattattagtaaaggtgaatcgaaagaagaaatcacaaaagatccgattaatgaattaacagaatacggaccagaaacgataacaccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaggaagttccaggtaaaccaggaattaagaatccagaaacaggagacgtagttagaccaccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagaaattccattcaagaaagaacgtaaatttaatcctgatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacgccaacactaaaaaatccattaactggagaaattattagtaaaggtgaatcgaaagaagaaatcacaaaagatccgattaatgaattaacagaatacggaccagaaacgataacaccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaggaagttccaggtaaaccaggaattaagaatccagaaacaggagatgtagttagaccaccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagaaattccattcgagaaagaacgtaaatttaatcctgatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacgccaacactaaaaaatccattaactggagaaattattagtaaaggtgaatcgaaagaagaaatcacaaaagatccgattaatgaattaacagaatacggaccagaaacgataacaccaggtcatcgagacgaatttgatccgaagttaccaacaggagagaaagaggaagttccaggtaaaccaggaattaagaatccagaaacaggagatgtagttagaccaccggtcgatagcgtaacaaaatatggacctgtaaaaggagactcgattgtagaaaaagaagaaattccattcgagaaagaacgtaaatttaatcctgatttagcaccagggacagaaaaagtaacaagagaaggacaaaaaggtgagaagacaataacgacgccaacactaaaaaatccattaactggagaaattattagtaaaggtgaatcgaaagaagaaatcacaaaagatccagttaatgaattaacagaattcggtggcgagaaaataccgcaaggtcataaagatatctttgatccaaacttaccaacagatcaaacggaaaaagtaccaggtaaaccaggaatcaagaatccagacacaggaaaagtgatcgaagagccagtggatgatgtgattaaacacggaccaaaaacgggtacaccagaaacaaaaacagtagagataccgtttgaaacaaaacgtgagtttaatccaaaattacaacctggtgaagagcgagtgaaacaagaaggacaaccaggaagtaagacaatcacaacaccaatcacagtgaacccattaacaggtgaaaaagttggcgagggtcaaccaacagaagagatcacaaaacaaccagtagataagattgtagagttcggtggagagaaaccaaaagatccaaaaggacctgaaaacccagagaagccgagcagaccaactcatccaagtggcccagtaaatcctaacaatccaggattatcgaaagacagagcaaaaccaaatggcccagttcattcaatggataaaaatgataaagttaaaaaatctaaaattgctaaagaatcagtagctaatcaagagaaaaaacgagcagaattaccaaaaacaggtttagaaagcacgcaaaaaggtttgatctttagtagtataattggaattgctggattaatgttattgg ctcgtagaagaaagaattaaRrkN(8325)(SEQ ID NO14)SGKYGKRSMQMRDKKGPVNKRVDFLSNKLNKYSIRKFTVGTASILIGSLMYLGTQQEAEAAENNIENPTTLKDNVQSKEVKIEEVTNKDTAPQGVEAKSEVTSNKDTIEHEPSVKAEDISKKEDTPKEVADVAEVQPKSSVTHNAETPKVRKARSVDEGSFDITRDSKNVVESTPITIQGKEHFEGYGSVDIQKKPTDLGVSEVTRFNVGNESNGLIGALQLKNKIDFSKDFNFKVRVANNHQSNTTGADGWGFLFSKGNAEEYLTNGGILGDKGLVNSGGFKIDTGYIYTSSMDKTEKQAGQGYRGYGAFVKNDSSGNSQMVGENIDKSKT
NFLNYADNSTNTSDGKFHGQRLNDVILTYVASTGKMRAEYAGKTVVETSITDLGLSKNQAYNFLITSSQRWGLNQGINANGWMRTDLKGSEFTFTPEAPKTITELEKKVEEIPFKKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGVIISKGEPKEEITKDPINELTEYGPETIAPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPINELTEYGPETITPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGVIISKGEPKEEITKDPINELTEYGPETITPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFKKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPINELTEYGPETITPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPINELTEYGPETITPGH RDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFKKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPINELTEYGPETITPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPINELTEYGPETITPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPVNELTEFGGEKIPQGHKDIFDPNLPTDQTEKVPGKPGIKNPDTGKVIEEPVDDVIKHGPKTGTPETKTVEIPFETKREFNPKLQPGEERVKQEGQPGSKTITTPITVNPLTGEKVGEGQPTEEITKQPVDKIVEFGGEKPKDPKGPENPEKPSRPTHPSGPVNPNNPGLSKDRAKPNGPVHSMDKNDKVKKSKIAKESVANQEKKRAELPKTGLESTQKGLIFSSIIGIAGLMLLARRRKNKnkA(8325)(SEQ ID NO15)ggaaggagtatgttgatggctaaatatcgagggaaaccgtttcaattatatgtaaagttatcgtgttcgacaatgatggcgacaagtatcattttaacgaatatcttgccgtacgatgcccaagctgcatctgaaaaggatactgaaattacaaaagagatattatctaagcaagatttattagacaaagttgacaaggcaattcgtcaaattgagcaattaaaacagttatcggcttcatctaaagaacattataaagcacaactaaatgaagcgaaaacagcatcgcaaatagatgaaatcataaaacgagctaatgagttggatagcaaagacaataaaagttctcacactgaaatgaacggtcaaagtgatatagacagtaaattagatcaattgcttaaagatttaaatgaggtttcttcaaatgttgataggggtcaacaaagtggcgaggacgatcttaatgcaatgaaaaatgatatgtcacaaacggctacaacaaaacatggagaaaaagatgataaaaatgatgaagcaatggtaaataaggcgttagaagacctagaccatttgaatcagcaaatacacaaatcgaaagatgcatcgaaagatacatcggaagatccagcagtgtctacaacagataataatcatgaagtagctaaaacgccaaataatgatggttctggacatgttgtgttaaataaattcctttcaaatgaagagaatcaaagccatagtaatcgactcactgataaattacaaggaagcgataaaattaatcatgctatgattgaaaaattagctaaaagtaatgcctcaacgcaacattacacatatcataaactgaatacgttacaatctttagatcaacgtattgcaaatacgcaacttcctaaaaatcaaaaatcagacttaatgagcgaagtaaataagacgaaagagcgtataaaaagtcaacgaaatattattttggaagaacttgcacgtactgatgataaaaagtatgctacacaaagcattttagaaagtatatttaataaagacgaggcagttaaaattctaaaagatatacgtgttgatggtaaaacagatcaacaaattgcagatcaaattactcgtcatattgatcaattatctctgacaacgagtgatgatttattaacgtcattgattgatcaatcacaagataagtcgctattgatttctcaaattttacaaacgaaattaggaaaagctgaagcagataaattggctaaagattggacgaataaaggattatcaaatcgccaaatcgttgaccaattgaagaaacattttgcatcaactggcgacacgtcttcagatgatatattaaaagcaattttgaataatgccaaagataaaaaacaagcaattgaaacgattttagcaacacgtatagaaagacaaaaggcaaaattactggcagatttaattactaaaata
gaaacagatcaaaataaaatttttaatttagttaaatcggcattgaatggtaaagcggatgatttattgaatttacaaaagagactcaatcaaacgaaaaaagatatagattatattttatcaccaatagtaaatcgtccaagtttactagatcgattgaataaaaatgggaaaacgacagatttaaataagttagcaaatttaatgaatcaaggatcagatttattagacagtattccagatatacccacaccaaagccagaaaagacgttaacacttggtaaaggtaatggattgttaagtggattattaaatgctgatggtaatgtatctttgcctaaagcgggggaaacgataaaagaacattggttgccgatatctgtaattgttggtgcaatgggtgtactaatgatttggttatcacgacgcaataagttgaaaaataaagcataaKnkA(8325)(SEQ ID NO16)GRSMLMAKYRGKPFQLYVKLSCSTMMATSIILTNILPYDAQAASEKDTEITKEILSKQDLLDKVDKAIRQIEQLKQLSASSKEHYKAQLNEAKTASQIDEIIKRANELDSKDNKSSHTEMNGQSDIDSKLDQLLKDLNEVSSNVDRGQQSGEDDLNAMKNDMSQTATTKHGEKDDKNDEAMVNKALEDLDHLNQQIHKSKDASKDTSEDPAVSTTDNNHEVAKTPNNDGSGHVVLNKFLSNEENQSHSNRLTDKLQGSDKINHAMIEKLAKSNASTQHYTYHKLNTLQSLDQRIANTQLPKNQKSDLMSEVNKTKERIKSQRNIILEELARTDDKKYATQSILESIFNKDEAVKILKDIRVDGKTDQQIADQITRHIDQLSLTTSDDLLTSLIDQSQDKSLLISQILQTKLGKAEADKLAKDWTNKGLSNRQIVDQLKKHFASTGDTSSDDILKAILNNAKDKKQAIETILATRIERQKAKLLADLITKIETDQNKIFNLVKSALNGKADDLLNLQKRLNQTKKDIDYILSPIVNRPSLLDRLNKNGKTTDLNKLANLMNQGSDLLDSIPDIPTPKPEKTLTLGKGNGLLSGLLNADGNVSLPKAGETIKEHWLPISVIVGAMGVLMIWLSRRNKLKNKA一级结构分析将生物信息方法(bioinformatic approach)用于一级结构和功能预测(附图1)。蛋白质RrkN和DsqA具有与先前描述的MSCRAMMs相似的结构组织。RrkN的结构类似于分别为金黄色葡萄球菌和表皮葡萄球菌的Pls/Aap蛋白质。它在N-末端含有200个残基的区域显示与Pis和Aap有40％相同。该蛋白质的C-末端突出地由128个残基重复区域组成，菌株与菌株之间重复区的数量是变化的。这些重复也存在于Pis和Aap。推定的sar同源区和fnbpA和fnbpB正好位于基因组上RrkN的上游。
DsqA的结构的组织相似于蛋白质的Sdr家族。它含有典型的A区域，其后跟随TYYFTDVK基序，该基序类似于在所有Sdr蛋白质中存在的保守的TYTFTVYVD基序。该基序的功能至今没有被测定。2个88个残基的重复区域居留在该蛋白质的中心，其后跟随类似于在所Sdr蛋白质中存在的SD-重复基序的C-末端SX-重复基序。该重复区的大小随菌株变化。在基因组上DsqA邻接secY和secA。DsqA同系物(＞90％同一性)也存在于表皮葡萄球菌。
KnkA的序列中不含有重复区域。次级的结构预测分析显示该蛋白质主要地由α-螺旋。
RkaS的序列中不含有重复区域。BLAST分析显示它类似于5′核苷酸酶UDP-糖水解酶。编码RkaS的基因正好地位于fromoryx的上游，mec元件的插入位点。
KesK在该蛋白质的N-末端含有二个140个残基的重复区域，有38％相同。水疗法绘制图分析(Kyte和Doolittle，1982)显示在该蛋白质的中心点有一个大的亲水的区域(残基500-560)。
EkeS在该蛋白质的N-末端含有二个300个残基的重复区域，有38％相同。Blast分析显示该蛋白质的N-末端(残基1-1268，携带两种重复)与FmtB有49％等同，所述的FmtBan是具有17个串联的重复的LPXTG蛋白质。FmtB被推测间接参与甲氧苯青霉素抗性，因为fmt8的灭活去除了甲氧苯青霉素抗性。这似乎是由于通过增加细胞壁前体葡萄糖胺-1-磷酸盐的产生可以减轻对细胞壁组成的影响如甲氧苯青霉素敏感性(Komatsuzawa等人，2000)。
KrkN和KrkN2在基因组上互相邻接。
表达分析由于与蛋白质数据库之序列没有同源性，不能预知这些蛋白质中每一个的推定的功能，因此以分子的近似值为依据。利用Qiagen pQE-30表达系统，四个开放读框的独特的区域可以在大肠杆菌中表达为重组his-标记的融合蛋白质。附图2表示纯化的N-末端his-标记的融合蛋白的考马斯亮蓝染色的SDS-PAGE凝胶。将重组蛋白质RrkN1，DsqA2，KesK1和KnkA用于在兔子中产生抗体。金黄色葡萄球菌细胞壁提取物的Western印迹分析显示KesK，KnkA和DsqA被表达并且细胞壁相关(附图3)。菌株eMRSA-16代表aknkA-阴性的菌株，因为它缺乏knkA基因。65kDa的免疫反应谱带与菌株8325-4的指数和静止的时期细胞的细胞壁提取物起反应(附图3，B)。菌株eMRSA-16中没有该谱带暗示它代表了knkA的基因产物。
在指数和静止期的培养物中，利用抗-KesK抗体进行的菌株8325-4的细胞壁成分的Western免疫印迹识别了150kDa免疫反应谱带。表达质粒(pKS80)上全长KesK的表达获得的从乳酸乳球菌的细胞壁成分释放的类似大小的免疫反应蛋白质暗示该150kDa谱带代表kesK基因产物(数据未显示)。需要金黄色葡萄球菌的kesK敲出突变体来证实细胞壁释放的KesK蛋白质的大小。
利用抗-DsqA抗体，金黄色葡萄球菌菌株MSSA和eMRSA-16的细胞壁成分的Western免疫印迹结果识别了一个130kDa免疫反应谱带。在静止期细胞中表达水平更高。
在乳酸乳球菌中异源表达以前已经将金黄色葡萄球菌的的表面蛋白质在乳酸乳球菌(L.lactis)中的异源表达用作为研究蛋白质功能的工具(Sinha等人，2000)。在该研究中，也将该替代系统用于在乳酸乳球菌表面表达各个in silico-预知的MSCRAMMs以查出功能。已经将KesK和KnkA克隆到乳酸乳球菌，并且通过点印迹显示是在表面表达的(附图4)。阴性的对照中没有观察到交叉反应(pKS80质粒没有插入物)，显示这是一个特异反应。通过用溶菌酶和mutanolysin消化产生了携带pKS-KnkA和pKS-KesK的乳酸乳球菌的细胞壁和原生质体成分，并且用于分别利用抗-KnkA和抗-KesK的抗体进行的Western印迹研究。与在金黄色葡萄球菌中观察到的不同，在乳酸乳球菌细胞壁成分中没有检测到KnkA，但是发现与原生质体成分相关。KnkA的固定用基序不同于一致LPXTG序列，其中含有一个丙氨酸残基而不是苏氨酸(即LPKAG)(表1)。最近已经公开了金黄色葡萄球菌含有二个sortase基因，srtA和srtB(Pallen，2001)。由第二个sortase基因加工LPXTG基序的变异形式是可能的，乳酸乳球菌中没有该基因。这也可以解释在原生质体成分中观察到的KnkA蛋白质大小的轻微的增加，而细胞壁分类拣选信号没有被裂解。
在乳酸乳球菌的细胞壁成分中探测到KesK，但是迁移的分子量小于从金黄色葡萄球菌细胞壁释放的KesK蛋白质。在乳酸乳球菌表面表达的大多数MSCRAMMs被证明在细胞壁提取程序中发生蛋白水解(Louise O′Brien，个人通讯)。因此，从乳酸乳球菌表面释放KesK蛋白质表示截短形式的KesK是可能的。已经证明用溶菌酶和mutanolysin缩短消化时间以限制蛋白水解的程度。
in silico-预测的MSCRAMMs体内表达在ELISA分析中对来自金黄色葡萄球菌感染痊愈的33个患者的康复的人-时期血清进行测试，以检测其识别纯化的N-末端his-标记的融合蛋白质的能力。将从儿童和健康的供血者采掘的血清用作为阴性的对照。将阳性的反应其值作为等同于或高于阴性的对照的二倍。附图5A-5D阐明在27-42％患者中识别了所有的蛋白质，这暗示这些蛋白质在体内表达并且在宿主感染期间是致免疫的。
引证文献Komatsuzawa，H.，Ohta，K.，Sugai，M.，Fujiwara，T.，Glanzmann，P.，Berger-Bachi，B.，Suginaka，H.(2000)编码细胞壁相关的蛋白质的fmt B基因的Tn551-介导的插入失活消除了金黄色葡萄球菌的甲氧苯青霉素抗性。J.Antimicrob.Chemother.45421-31。
Sinha，B.，Francois，P.，Que，Y.A.，Hussain，M.，Heilmann，C.，Moreillon，P.，Lew，D.，Krause，K.H.，Peters，G.，Herrmann，M.(2000)异源表达的金黄色葡萄球菌粘连蛋白结合蛋白质足以侵入宿主细胞Infect.Immun.686871-6878。
Pallen，M.J.，Lam，A.C.，Antonio，M.，Dunbar，K.(2000)sortases的阻碍-一种丰富的反应物？Trends.Microbiol.997-101。
实施例2从金黄色葡萄球菌和从凝固酶阴性的葡萄球菌分离交叉反应的蛋白质并且进行测序最近已经证明表皮葡萄球菌含有结构上与金黄色葡萄球菌MSCRAMM蛋白质相关的表面蛋白质(US09/386,962)。从金黄色葡萄球菌中分离一个蛋白质特别有利，因为它与表皮葡萄球菌紧密同源。该蛋白质被称为DsqA或SasA(金黄色葡萄球菌)和DgsK(表皮葡萄球菌)。它们的特征是包括一个大约500个氨基酸残基的典型的″A″区域，其后跟随约有40％相同有，各为88个残基的二个B重复区，和一个独特的SXSX二肽重复区，该区域的长度随菌株而变化。金黄色葡萄球菌的DsqA/SasA的A区域内含有一个180残基的区域，该区域与金黄色葡萄球菌蛋白质的RrkN，Pls和表皮葡萄球菌蛋白质Aap的A区域内的一个相似的大小的区域有40％一致性。DsqA/SasA和DgsK蛋白质的A区域在氨基酸水平上有46％的一致性，BB重复区是50％一致性。所包括的主动和被动免疫接种策略；疫苗，识别金黄色葡萄球菌和凝固酶阴性的葡萄球菌的蛋白质的多克隆抗体和单克隆抗体是本发明的主题。
与凝固酶阴性的葡萄球菌和金黄色葡萄球菌发生交叉反应的抗体的特异性例子凝固酶阴性的葡萄球菌DgsK A-区域氨基酸序列(SEQID NO17)ASETPITSEISSNSETVANQNSTTIKNSQKETVNSTSLESNHSNSTNKQMSSEVTNTAQSSEKAGISQQSSETSNQSSKLNTYASTDHVESTTINNDNTAQQDQNKSSNVTSKSTQSNTSSSEKNISSNLTQSIETKATDSLATSEARTSTNQISNLTSTSTSNQSSPTSFANLRTFSRFTVLNTMAAPTTTSTTTTSSLTSNSVWNKDNFNEHMNLSGSATYDPKTGIATLTPDAYSQKGAISLNTRLDSNRSFRFIGKVNLGNRYEGYSPDGVAGGDGIGFAFSPGPLGQIGKEGAAVGIGGLNNAFGFKLDTYHNTSTPRSDAKAKADPRNVGGGGAFGAFVSTDRNGMATTEESTAAKLNVQPTDNSFQDFVIDYNGDTKVMTVTYAGQTFTRNLTDWIKNSGGTTFSLSMTASTGGAKNLQQVQFGTFEYTESAVAKVRYVDANTGKDIIPPPKTIAGEVDGTVNIDKQLNNFKNLGYSYVGTDALKAPNYTETSGTPTLKLTNSSQTVIYKFKDVQ
金黄色葡萄球菌SasA A-区域氨基酸序列(SEQ ID NO18)ASDAPLTSELNTQSETVGNQNSTTIEASTSTADSTSVTKNSSSVQTSNSDTVSSEKSEKVTSTTNSTSNQQEKLTSTSESTSSKNTTSSSDTKSVASTSSTEQPINTSTNQSTASNNTSQSTTPSSVNLNKTSTTSTSTAPVKLRTFSRLAMSTFASAATTTAVTANTITVNKDNLKQYMTTSGNATYDQSTGIVTLTQDAYSQKGAITLGTRIDSNKSFHFSGKVNLGNKYEGHGNGGDGIGFAFSPGVLGETGLNGAAVGIGGLSNAFGFKLDTYHNTSKPNSAAKANADPSNVAGGGAFGAFVTTDSYGVATTYTSSSTADNAAKLNVQPTNNTFQDFDINYNGDTKVMTVKYAGQTWTRNISDWIAKSGTTNFSLSMTASTGGATNLQQVQFGTFEYTESAVTQVRYVDVTTGKDIIPPKTYSGNVDQVVTIDNQQSALTAKGYNYTSVDSSYASTYNDTNKTVKMTNAGQSVTYYFTDWAap蛋白质的完整序列和它的编码DNA(指示有A区域存在)显示如下表皮葡萄球菌Aap蛋白质(加下划线的为A-区域)(SEQ IDNO19)MGKRRQGPINKKVDFLPNKLNKYSIRKFTVGTASILLGSTLIFGSSSHEAKAAEEKQVDPITQANQNDSSERSLENTNQPTVNNEAPQMSSTLQAEEGSNAEAPQSEPTKAEEGGNAEAAQSEPTKAEEGGNAEAPQSEPTKAEEGGNAEAAQSEPTKTEEGSNVKAAQSEPTKAEEGSNAEAPQSEPTKTEEGSNAKAAQSEPTKAEEGGNAEAAQSEPTKTEEGSNAEAPQSEPTKAEEGGNAEAPQSEPTKTEEGGNAEAPNVPTIKANSDNDTQTQFSEAPTRNDLARKEDIPAVSKNEELQSSQPNTDSKIEPTTSEPVNLNYSSPFMSLLSMPADSSSNNTKNTIDIPPTTVKGRDNYDFYGRVDIESNPTDLNATNLTRYNYGQPPGTTTAGAVQFKNQVSFDKDFDFNIRVANNRQSNTTGADGWGFMFSKKDGDDFLKNGGILREKGTPSAAGFRIDTGYYNNDPLDKIQKQAGQGYRGYGTFVK
NDSQGNTSKVGSGTPSTDFLNYADNTTNDLDGKFHGQKLNNVNLKYNASNQTFTATYAGKTWTATLSELGLSPTDSYNFLVTSSQYGNGNSGTYASGVMRADLDGATLTYTPKAVDGDPIISTKEIPFNKKREFDPNLAPGTEKVVQKGEPGIETTTTPTYVNPNTGEKVGEGEPTEKITKQPVDEIVHYGGEEIKPGHKDEFDPNAPKGSQTTQPGKPGVKNPDTGEVVTPPVDDVTKYGPVDGDPITSTEEIPFDKKREFNPDLKPGEERVKQKGEPGTKTITTPTTKNPLTGEKVGEGEPTEKITKQPVDEITEYGGEEIKPGHKDEFDPNAPKGSQEDVPGKPGVKNPGTGEVVTPPVDDVTKYGPVDGDPITSTEEIPFDKKREFNPDLKPGEERVKQKGEPGTKTITTPTTKNPLTGEKVGEGEPTEKITKQPVDEIVHYGGEQIPQGHKDEFDPNAPVDSKTEVPGKPGVKNPDTGEVVTPPVDDVTKYGPVDGDSITSTEEIPFDKKREFDPNLAPGTEKVVQKGEPGTKTITTPTTKNPLTGEKVGEGKSTEKVTKQPVDEIVEYGPTKAEPGKPAEPGKPAEPGKPAEPGTPAEPGKPAEPGTPAEPGKPAEPGKPAEPGKPAEPGKPAEPGTPAEPGTPAEPGKPAEPGTPAEPGKPAEPGTPAEPGKPAESGKPVEPGTPAQSGAPEQPNRSMHSTDNKNQLPDTGENRQANEGTLVGSLLAIVGSLFIFGRRKKGNEK表皮葡萄球菌aap DNA(SEQ ID NO20)atgggcaaac gtagacaagg tcctattaat aaaaaagtggattttttacc taacaaatta aacaagtatt ctataagaaa attcactgtt ggtacggcctcaatattact tggttcgaca cttatttttg gaagtagtag ccatgaagcg aaagctgcagaagaaaaaca agttgatcca attacacaag ctaatcaaaa tgatagtagt gaaagatcacttgaaaacac aaatcaacct actgtaaaca atgaagcacc acagatgtct tctacattgcaagcagaaga aggaagcaat gcagaagcac ctcaatctga gccaacgaag gcagaagaaggaggcaatgc agaagcagct caatctgagc caacgaaggc agaagaagga ggcaatgcagaagcacctca atctgagcca acgaaggcag aagaaggagg caatgcagaa gcagctcaatctgagccaac gaagacagaa gaaggaagca acgtaaaagc agctcaatct gagccaacgaaggcagaaga aggaagcaat gcagaagcac ctcaatctga gccaacgaag acagaagaaggaagcaacgc aaaagcagct caatctgagc caacgaaggc agaagaagga ggcaatgcagaagcagctca atctgagcca acgaagacag aagaaggaag caatgcagaa gcacctcaatctgagccaac gaaggcagaa gaaggaggca atgcagaagc acctcaatct gagccaacgaagacagaaga aggaggcaat gcagaagcac cgaatgttcc aactatcaaa gctaattcag
ataatgatac acaaacacaa ttttcagaag cccctacaag aaatgaccta gctagaaaagaagatatccc tgctgtttct aaaaacgagg aattacaatc atcacaacca aacactgacagtaaaataga acctacaact tcagaacctg tgaatttaaa ttatagttct ccgtttatgtccttattaag catgcctgct gatagttcat ccaataacac taaaaataca atagatataccgccaactac ggttaaaggt agagataatt acgattttta cggtagagta gatatcgaaagtaatcctac agatttaaat gcgacaaatt taacgagata taattatgga cagccacctggtacaacaac agctggtgca gttcaattta aaaatcaagt tagttttgat aaagatttcgactttaacat tagagtagca aacaatcgtc aaagtaatac aactggtgca gatggttggggctttatgtt cagcaagaaa gatggggatg atttcctaaa aaacggtggt atcttacgtgaaaaaggtac acctagtgca gctggtttca gaattgatac aggatattat aataacgatccattagataa aatacagaaa caagctggtc aaggctatag agggtatggg acatttgttaaaaatgactc ccaaggtaat acttctaaag taggatcagg tactccatca acagattttcttaactacgc agataatact actaatgatt tagatggtaa attccatggt caaaaattaaataatgttaa tttgaaatat aatgcttcaa atcaaacttt tacagctact tatgctggtaaaacttggac ggctacgtta tctgaattag gattgagtcc aactgatagt tacaattttttagttacatc aagtcaatat ggaaatggta atagtggtac atacgcaagt ggcgttatgagagctgattt agatggtgca acattgacat acactcctaa agcagtcgat ggagatccaattatatcaac taaggaaata ccatttaata agaaacgtga atttgatcca aacttagccccaggtacaga aaaagtagtc caaaaaggtg aaccaggaat tgaaacaaca acaacaccaacttatgtcaa tcctaataca ggagaaaaag ttggcgaagg tgaaccaaca gaaaaaataacaaaacaacc agtggatgaa atcgttcatt atggtggcga agaaatcaag ccaggccataaggatgaatt tgatccaaat gcaccgaaag gtagtcaaac aacgcaacca ggtaagccgggggttaaaaa tcctgataca ggcgaagtag ttactccacc tgtggatgat gtgacaaaatatggtccagt tgatggagat ccgatcacgt caacggaaga aattccattc gacaagaaacgtgaattcaa tcctgattta aaaccaggtg aagagcgtgt taaacaaaaa ggtgaaccaggaacaaaaac aattacaaca ccaacaacta agaacccatt aacaggggaa aaagttggcgaaggtgaacc aacagaaaaa ataacaaaac aaccagtaga tgaaatcaca gaatatggtggcgaagaaat caagccaggc cataaggatg aatttgatcc aaatgcaccg aaaggtagccaagaggacgt tccaggtaaa ccaggagtta aaaaccctgg aacaggcgaa gtagtcacac
caccagtgga tgatgtgaca aaatatggtc cagttgatgg agatccgatc acgtcaacggaagaaattcc attcgacaag aaacgtgaat tcaatcctga tttaaaacca ggtgaagagcgcgttaaaca gaaaggtgaa ccaggaacaa aaacaattac aacgccaaca actaagaacccattaacagg agaaaaagtt ggcgaaggtg aaccaacaga aaaaataaca aaacaaccagtggatgagat tgttcattat ggtggtgaac aaataccaca aggtcataaa gatgaatttgatccaaatgc acctgtagat agtaaaactg aagttccagg taaaccagga gttaaaaatcctgatacagg tgaagttgtt accccaccag tggatgatgt gacaaaatat ggtccagttgatggagattc gattacgtca acggaagaaa ttccgtttga taaaaaacgc gaatttgatccaaacttagc gccaggtaca gagaaagtcg ttcaaaaagg tgaaccagga acaaaaacaattacaacgcc aacaactaag aacccattaa caggagaaaa agttggcgaa ggtaaatcaacagaaaaagt cactaaacaa cctgttgacg aaattgttga gtatggtcca acaaaagcagaaccaggtaa accagcggaa ccaggtaaac cagcggaacc aggtaaacca gcggaaccaggtacgccagc agaaccaggt aaaccagcgg aaccaggtac gccagcagaa ccaggtaaaccagcggaacc aggtaaacca gcggaaccag gtaaaccagc ggaaccaggt aaaccagcggaaccaggtac gccagcagaa ccaggtacgc cagcagaacc aggtaaacca gcggaaccaggtacgccagc agaaccaggt aaaccagcgg aaccaggtac gccagcagaa ccaggtaaaccagcggaatc aggtaaacca gtggaaccag gtacgccagc acaatcaggt gcaccagaacaaccaaatag atcaatgcat tcaacagata ataaaaatca attacctgat acaggtgaaaatcgtcaagc taatgaggga actttagtcg gatctctatt agcaattgtc ggatcattgttcatatttgg tcgtcgtaaa aaaggtaatg aaaaataatt tcatataaaa actttctgccattaa从A-区域到表皮葡萄球菌Aap(氨基酸55-600)(SEQ IDNO21)55EKQVDPITQANQNDSSERSLENTNQPTVNNEAPQMSSTLQAEEGSNAEAPQSEPTKAEEGGNAEAAQSEPTKAEEGGNAEAPQSEPTKAEEGGNAEAAQSEPTKTEEGSNVKAAQSEPTKAEEGSNAEAPQSEPTKTEEGSNAKAAQSEPTKAEEGGNAEAAQSEPTKTEEGSNAEAPQSEPTKAEEGGNAEAPQSEPTKTEEGGNAEAPNVPTIKANSDNDTQTQFSEAPTRNDLARKEDIPAVSKNEELQSSQPNTDSKIEPTTSEPVNLNYSSPFMSLLSMPADSSSNNTKNTIDIPPTTVKGRDNYDFYGRVDIESNPTDLNAT
NLTRYNYGQPPGTTTAGAVQFKNQVSFDKDFDFNIRVANNRQSNTTGADGWGFMFSKKDGDDFLKNGGILREKGTPSAAGFRIDTGYYNNDPLDKIQKQAGQGYRGYGTFVKNDSQGNTSKVGSGTPSTDFLNYADNTTNDLDGKFHGQKLNNVNLKYNASNQTFTATYAGKTWTATLSELGLSPTDSYNFLVTSSQYGNGNSGTYASGVMRADLDGA600蛋白质的生产和纯化利用PCR，从上面描述的序列扩增识别为DgsK或SasA的表面蛋白质的A区域，再克隆到大肠杆菌表达载体PQE-30(Qiagen)，以允许表达一种含有六个组氨酸残基的重组融合蛋白质。随后将该载体转化到大肠杆菌ATCC 55151，在15-升发酵罐中生长到光密度(OD600)为0.7，用0.2mM异丙基-1β-D半乳糖苷(IPTG)诱导4小时。利用AG Technologies中空纤维组件(孔大小0.45um)收集细胞，将细胞浆在-80C冷冻。利用2次穿过French Press，1100psi，将细胞在1×PBS(10mL缓冲液/1克的细胞浆)裂解。将裂解的细胞17,000rpm旋转30分钟以除去细胞碎片。将上清液经过装备了0.1M NiCI2的5-mL Hi俘获Cheating(Pharmacia)柱。在上样之后，用5个柱体积的10mM Tris，pH 8.0，100mM NaCI(缓冲液A)洗涤柱子。利用超过30个柱体积的0-100％梯度的10mM Tris，pH 8.0，100mM NaCI，200mM咪唑(Buffer B)洗脱蛋白质。SdrGN1N2N3或SdrGN2N3在～13％Buffer B(～26mM咪唑)被洗脱。监测280nm处的吸光率。在1×PBS中透析含有SdrGN1N2N3或SdrGN2N3的馏分。
然后将各个蛋白质经过一个内毒素除去方案。在该方案中所使用的缓冲液通过一个5-mL Mono-Q琼脂糖(Pharmacia)柱而没有内毒素。将蛋白质平衡地分到4×15毫升试管中。各个试管的体积用缓冲液A加到9毫升。将1毫升的10％Triton X-114加入到各个试管中，在4℃旋转保温1小时。将试管置于37℃水浴中以使相位分离。将试管以2,000rpm旋转10分钟，收集各个试管的上面的含水的相，用去污剂反复提取。将2次提取的含水相合并，传递过带有0.1M NiCl2的5-毫升IDA cheating(Sigma)柱，以除去剩余的洗涤剂。用9个柱体积的Buffer A洗涤该柱，然后用3个柱体积的Buffer B洗脱该蛋白质。将洗脱液通过5-毫升的Detoxigel(Sigma)柱，收集流出物，再上样到柱。收集第二次过柱的流出物，在1×PBS中透析。分析纯化的产物的浓度，纯度和内毒素水平，然后给小鼠给药。
单克隆抗体的生产将大肠杆菌表达的和纯化的重组SasA和DsgK蛋白质用于制备一组鼠的单克隆抗体，而将小鼠血清用作为多克隆抗体的来源。简单地说，一组Balb/C或SJL小鼠接收了一系列皮下的免疫接种1-10mg溶液蛋白质或与下面表描述的佐剂混合的蛋白质。
免疫接种方案RIMMS注射天数数量(微克)途径佐剂#1 0 5 皮下FCA/RIBI#2 2 1 皮下FCA/RIBI#3 4 1 皮下FCA/RIBI#4 7 1 皮下FCA/RIBI#5 9 1 皮下FCA/RIBI
常规注射 天数数量(微克)途径佐剂原始 0 5 皮下FCA加强#114 1 腹膜内 RIBI加强#228 1 腹膜内 RIBI加强#342 1 腹膜内 RIBI在杀死时(RIMMS)或加强后几天收集(常规的)血清，在ELISA测定中测定抗MSCRAMM蛋白质或对完整的细胞(表皮葡萄球菌和金黄色葡萄球菌)的效价。在最后的加强之后3天，去除脾脏或淋巴结，挑开成单细胞悬浮液，收集淋巴细胞。然后将淋巴细胞与P3X63Ag8.653骨髓瘤细胞系(ATCC &CRL-1580)融合。根据Current Protocols in Immunology(第2章，第2单元)中的生产单克隆抗体的方案进行细胞融合，随后涂覆平板和喂养。
然后利用标准的ELISA分析方法，从融合产生的任何克隆中筛选特异的抗-SasA抗体的产生。扩增阳性的克隆，进一步通过流式血细胞计数测试完整的细菌的细胞结合测定中的活性以及Biacore分析中SasA的结合。
Biacore分析在整个分析中，流速保持在10毫升/分钟不变。在注射SasA或DgsK之前，借助于RAM-Fc结合，将测试抗体吸附到芯片。在时间0，将浓度为30mg/ml的SasA或DgsK注射过芯片3分钟，随后解离2分钟。该阶段的分析测定了Mab/SasA或DgsK相互作用的相对关系和分离动力学。
与完整细菌的结合用兔子IgG(50mg/ml)封闭之后，收集，洗涤细菌的样品金黄色葡萄球菌Newman，金黄色葡萄球菌60，金黄色葡萄球菌397(Sal6)，金黄色葡萄球菌Wood，金黄色葡萄球菌8325-4，甲氧苯青霉素抗性的金黄色葡萄球菌MRSA 16，表皮葡萄球菌ATCC 35984，表皮葡萄球菌HB，表皮葡萄球菌CN-899和溶血葡萄球菌ATCC 43253，用浓度为2pg/ml的Mab或仅用PBS(对照)保温。在用抗体保温之后，将用作为检测抗体的山羊-F(ab′)2-抗-小鼠-F(ab′)2-FITC用于孵育细菌的细胞。在抗体标记之后，用FACS caliber流式细胞计数器抽吸细菌的细胞以分析荧光射出(刺激488，发射570)。对于各个细菌的菌株，收集和检测10,000个样品。这些资料显示抗金黄色SasA的抗体能够识别凝固酶阴性的葡萄球菌的表面的同源的蛋白质。该资料支持Western印迹分析，表明抗金黄色葡萄球菌SasA的兔子多克隆抗体与从表皮葡萄球菌HB细胞表面释放的蛋白质以及来自于从表皮葡萄球菌克隆的DsgK的重组A-区域发生交叉反应(参见附图6和下表)。
多克隆血清反应性
序列表<110>蒂莫西·J·福斯特等人<120>识别凝血酶阴性的葡萄球菌和金黄色葡萄球菌的表面蛋白的交叉反应性单克隆抗体和多克隆抗体<130>P07263US01/BAS<140>US 10/172,502<141>2002-06-17<150>US 60/298,098<151>2001-06-15<160>29<170>Patent In version 3.1<210>1<211>6609<212>DNA<213>Staphylococcus epidermidis<400>1acaacacagc agagaataga caaccaggag gaaaacgaaa tgaatttgtt aaagaaaaat 60aaatatagta ttagaaaata taaagtaggg atattctcta ctttaatcgg gacagtttta120ttactttcaa acccaaatgg tgcacaagct ttaactacgg atcataatgt gcaaggtggt180tcaaatcaag cattacctgg caactcacaa aatacaaatg ccgatactaa tcgagacata240gtaaatgatt cgcaaaatac tcctaatgca catgcaacag acaatacatc aacaaatcaa300gcattgacta atcatcaaaa cgttgatgtg gcaaatcaag tcgggcctgc tccaatacag360cctagcgcgt cgcctgcgca aaataataat aattctaatg ctaattcaac agcaacagag420ccagcggcga atacaaataa taatttagca tcaaataaca atacattaaa cgtgcctaat480aatacagata acaatgattc agcgcgtcat ctgactttaa aagaaattca agaagatgtt540cgtcattcgt ctgataagcc agagttagtt gcgattgctg aagaagcatc taatagaccg600aaaaagagaa gcagacgtgc tgcgccaaca gatcctaatg caacaccagc agatccaacg660gctacaccag cagatccaac ggcaggaaat ggtagtgcac cagttgcaat tacagcgcca720tacacgccaa caactgatcc caatgccaat aatataggac aaaatgcacc taacgaagtg780ctttcatttg atgataacaa cattagacca agtacgaacc gttctgtgcc tacagtaact840gttgttgata atttaccagg ctacacactg attaatggtg gtaaagtagg ggtgtttagt900
catgcaatgg taagaacgag catgtttgat tcaggagatg ccaagaacta tcaagcgcaa960ggcaatgtaa ttgcattggg tcgtattaga ggaaatgata caaatgatca tggcgatttt 1020aatggtatcg agaaaacatt aacagtaaat ccgaattctg aattaatctt tgaatttaat 1080actatgacta ctaaaaacta tcaaggtatg acaaatttaa tcattaaaaa tgctgataac 1140gatactgtta ttggtgaaaa agtagttgct tatggtccga tttggcgctt attaaaagta 1200cctgaaaatg ttagtcatct aaaaattcaa tttgtaccta aaaatgacgc aataacagat 1260gcacgtggta tttatcaatt acgagatgga tataaatact atgactttgt agactcaatc 1320ggtcttcatt ctgggtcaca tgtctatgtt gaaagacgta caatggagcc aacagcaaca 1380aataataaag aatttacagt tacaacgtca ttaaagaata atggtaactt tggcgcttca 1440ttcaatacag atgattttgt atataaaatt caattacctg aaggtgttga atatgtaaat 1500aattcattga ctaaagattt tcctagcggt aattcaggtg ttgatattaa tgatatgaat 1560gtgacgtatg acgcagcaaa tcgaattatt acaattaaaa gtactggtgg aggtacaggg 1620aattcgccgg cacgactaat gcctgataaa atattggatt tgaagtataa gctacgtgtg 1680aacaatgtgc caacaccaag aacagtaaca tttaacgata cattaacgta taaaacatat 1740tcacaagatt ttattaattc acctgctgaa agtcatactg taagtacaaa tccatataca 1800attgatatca tcatgaataa agacgcattg caagccgaag tcgatagacg aattcaacaa 1860gcggattata catttgcatc attagatatt tttaatgatc ttaaaagacg cgcacaaaca 1920attttagatg aaaaccgtaa caatgtacct ttaaacaaaa gagtttctca agcagatatc 1980gattcattag caaatcagat gcaacatacg ttaattcgca gtgttgacgc tgaaaatgcc 2040gttaatagaa aagttgatga catggaagat ttagttaacc aaaatgatga actgacagat 2100gaagaaaaac aagcagcgat tcaagtcatc gaggaacata aaaatgaaat tattgggaat 2160attggtgacc aaacgactga tgatggcgtt actagaatta aagatcaagg tatacagact 2220ttaagtggag acactgcaac accagttgtt aaaccaaatg ctaaacaagc tatacgtgat 2280aaagcagcga aacaaagaga aattatcaat cacacgccag atgctactca agatgaaatt 2340caagatgcat taaatcaatt aacaacggat gaaacagatg ctattgataa tgttacgaat 2400gctactacca atgctgatgt tgaaacagct aaaaataatg gtattaatac aattggtgca 2460gttgcgccac aagtgacaca caaacaagct gcaagagatg caattaatca agcgacagca 2520acgaaacgac aacaaataaa tagcaataga gaagcaacac aagaagagaa aaatgcagca 2580
ttgaatgaat taacgcaagc cacgaaccac gcattagaac aaatcaatca agcgacaacc 2640aatgatgatg tagatactgc caaaggtgat ggtctgaatg ccattaatcc tattgcgcct 2700gtaactgttg tcaagcaagc agcaagagat gccgtatcac atgatgcaca acagcatatc 2760gcagagatca atgcaaatcc tgatgcgact caagaagaaa gacaagcagc aatagagaaa 2820gtaaatgctg ctgtagctgt tgcgaatact aatatattaa atgctaatac caatgctgat 2880gttgagcaag taaagacaaa tgcaattcaa ggtatacaag ccattgaacc agctacaaag 2940gttaaaacag atgctaaaaa cgctattgat caaagtgcgg aaacgcaaca taatgcgata 3000tttaataata atgatgcgac cttagaagag caacaagcag cacaacaatt gcttgatcaa 3060gctgtagcca cagcgaagca aaatattaat gcagcagata cgaatcaaga agttgcacaa 3120gcaaaagatc agggcacaca aaatatagtt gtgattcaac cggcaacaca agttaaaacg 3180gatgcacgca atgctgtaaa tgaaaaagcg cgagaggcga taacaaatat caatgctaca 3240cctggcgcga ctcgagaaga gaaacaagaa gcgataaatc gtgtcaatac acttaaaaat 3300agagcattaa atgatattgg tgtgacgtct actactgcga tggtcaatag tattagagac 3360gatgcagtca atcaaatcgg tgcagttcaa ccgcatgtaa cgaagaaaca aactgctaca 3420ggtgtattaa cggacttagc aactgcaaaa aaacaagaaa ttaatcaaaa tacaaatgca 3480accactgaag aaaagcaagt agcattaaat caagtagacc aagatttagc aacggcaatt 3540aataatataa atcaagctga tactaatgca gaagtagatc aagcacaaca attaggtaca 3600aaagcaatta atgcgattca gccaaatatt gtaaaaaaac ctgcagcatt agcacaaacc 3660aatcagcatt atagtgctaa attagttgaa atcaatgcta caccagatgc aacagatgat 3720gagaaaaatg ctgcgatcaa tactttaaat caagacagac aacaagctat tgaaagtatt 3780aaacaagcaa atacaaatgc ggaagtagac caagctgcga cagtggcaga gaataatatc 3840gatgctgttc aagttgacgt tgtaaaaaaa caagcagcgc gagataaaat cactgctgaa 3900gtagcgaagc gtattgaagc ggttaaacaa acacctaatg caactgacga agaaaagcag 3960gctgcagtta atcaaatcaa tcaacttaaa gatcaagcgt ttaatcaaat taatcaaaac 4020caaacaaatg atcaggtaga cgcaactaca aatcaagcga ttaatgctat agataatgtt 4080gaagctgaag tagtaattaa accaaaggca attgcagata ttgaaaaagc tgttaaagaa 4140aagcaacagc aaattgataa tagtcttgat tcaacagata atgagaaaga agttgcttta 4200caagcattag ctaaagaaaa agaaaaagca cttgcagcta ttgaccaagc tcaaacgaat 4260
agtcaggtga atcaagcggc aacaaatggt gtatcagcga ttaaaattat tcaacctgaa 4320acaaaaatta aaccagcagc acgtgaaaaa atcaatcaaa aagcgaatga attacgtgcg 4380caaattaatc aagataaaga agcgacagca gaagaaagac aagcggcgtt agataaaatc 4440aatgatttag ttgctaaagc tatgacaaat atcacgaatg atagaacaaa tcagcaagtt 4500aatgactcaa caaatcaagc gcttgacgac attgcattag tgacgcctga ccatattgtt 4560agagcagctg ctagagatgc agttaagcaa caatatgaag ctaaaaagca cgaaattgag 4620caagcggaac atgcgactga tgaagaaaaa caagttgctt taaatcaatt agcgaataat 4680gaaaaacgtg cattacaaaa cattaatcaa gcaatagcga ataatgatgt gaaacgtgtt 4740gaatcaaatg gtattgctac gttaaaaggc gtagaaccgc acattgtggt taaacctgaa 4800gctcaagaag ccataaaagc gagcgcagat aaccaagtag aatctataaa agatacacca 4860catgctacga cagatgaatt agatgaagca aaccaacaaa taaacgacac acttaaacaa 4920ggtcaacaag atatagacaa tacgacacaa gatgcagctg tcaatgatgt tagaaaccaa 4980acgattaagg caatcgaaca aattaaaccg aaagttagac gcaaacgtgc agcgttggat 5040aacattgatg aaagtaataa taatcaactc gatgcaatac gaaatacgct agatacaacg 5100caagatgaac gaaatgttgc tattgctgcg ttaaataaaa ttgttaatgc aattaaaaat 5160gatattgcac aaaacaaaac gaatgcagaa gtggatcaaa ctgaggctga tggtaacaac 5220aacatcaaag tgattttacc taaagttcaa gttaaaccag cagcgcgtca atctgtcagc 5280gcaaaagctg aagctcaaaa tgcacttatt gatcaaagtg atttatctac cgaagaagaa 5340agattagctg ctaaacattt agtagaacaa gcacttaatc aagctattga tcagatcaat 5400cacgcagata agactgcgca agttaatcaa aatagtatcg atgctcaaaa tattatttca 5460aaaattaaac cagcgacaac agttaaagca acagcattac aacaaattca aaatatcgct 5520acaaataaaa ttaatttaat taaagcaaat aacgaagcga cagatgaaga acaaaatgct 5580gcaatagtac aagttgaaaa agagttaatt aaagctaaac aacaaattgc tggtgcagtg 5640actaatgctg atgtggcata tttattgcat gatgggaaaa acgaaattcg tgaaatcgaa 5700cctgttatta ataaaaaagc aactgcgcga gaacaattaa caacattatt caacgataag 5760aaacaagcaa ttgaagcgaa tgttcaagca acagtagaag aaagaaatag tattttagca 5820cagttacaaa acatttatga cactgctatt ggacaaattg atcaagatcg tagcaatgca 5880caagttgata aaacagcaac attaaatcta caaacaatac atgatttaga cgtacatcct 5940
attaaaaagc cagatgctga aaaaacgatt aatgatgatc ttgcacgtgt tacacattta 6000gtgcaaaatt atcgaaaagt aagtgatcgt aataaggctg atgcattaaa agctataact 6060gcattaaaat tacaaatgga tgaagaatta aaaacagcac gcactaatgc tgatgttgat 6120gcagttttaa aacgatttaa tgttgcatta ggcgatatag aagcagtaat tactgaaaaa 6180gaaaatagct tactgcgcat tgataacatt gctcaacaaa catatgcgaa attcaaagcg 6240atcgcaacac cagaacaatt agctaaagta aaagcattaa ttgatcaata tgttgcagat 6300ggcaatagaa tggttgatga agatgcgaca ttaaatgaca tcaaaaaaga tacgcaactc 6360attattgatg aaattttagc aattaattta cctgctgaag tgataaaagc gtcaccaaaa 6420gtggggcaac ctgctccaaa agtttgtacg cctattaaaa aagaagataa acaagaagtg 6480cgaaaagttg taaaagaact tccaaatact ggttctgaag aaatggattt accattaaaa 6540gaattagcac taattacagg cgcagcatta ttagctagaa gacgttctaa aaaagaaaaa 6600gaatcataa 6609<210>2<211>2189<212>PRT<213>Staphylococcus epidermidis<400>2Met Asn Leu Leu Lys Lys Asn Lys Tyr Ser Ile Arg Lys Tyr Lys Val1 5 1015Gly Ile Phe Ser Thr Leu Ile Gly Thr Val Leu Leu Leu Ser Asn Pro20 2530Asn Gly Ala Gln Ala Leu Thr Thr Asp His Asn Val Gln Gly Gly Ser35 40 45Asn Gln Ala Leu Pro Gly Asn Ser Gln Asn Thr Asn Ala Asp Thr Asn50 55 60Arg Asp Ile Val Asn Asp Ser Gln Asn Thr Pro Asn Ala His Ala Thr65 70 75 80Asp Asn Thr Ser Thr Asn Gln Ala Leu Thr Asn His Gln Asn Val Asp85 9095
Val Ala Asn Gln Val Gly Pro Ala Pro Ile Gln Pro Ser Ala Ser Pro100 105 110Ala Gln Asn Asn Asn Asn Ser Asn Ala Asn Ser Thr Ala Thr Glu Pro115120 125Ala Ala Asn Thr Asn Asn Asn Leu Ala Ser Asn Asn Asn Thr Leu Asn130135 140Val Pro Asn Asn Thr Asp Asn Asn Asp Ser Ala Arg His Leu Thr Leu145150155 160Lys Glu Ile Gln Glu Asp Val Arg His Ser Ser Asp Lys Pro Glu Leu165 170 175Val Ala Ile Ala Glu Glu Ala Ser Asn Arg Pro Lys Lys Arg Ser Arg180185 190Arg Ala Ala Pro Thr Asp Pro Asn Ala Thr Pro Ala Asp Pro Thr Ala195 200 205Thr Pro Ala Asp Pro Thr Ala Gly Asn Gly Ser Ala Pro Val Ala Ile210215 220Thr Ala Pro Tyr Thr Pro Thr Thr Asp Pro Asn Ala Asn Asn Ile Gly225 230235 240Gln Asn Ala Pro Asn Glu Val Leu Ser Phe Asp Asp Asn Asn Ile Arg245 250255Pro Ser Thr Asn Arg Ser Val Pro Thr Val Thr Val Val Asp Asn Leu260 265 270Pro Gly Tyr Thr Leu Ile Asn Gly Gly Lys Val Gly Val Phe Ser His275 280 285Ala Met Val Arg Thr Ser Met Phe Asp Ser Gly Asp Ala Lys Asn Tyr290295300Gln Ala Gln Gly Asn Val Ile Ala Leu Gly Arg Ile Arg Gly Asn Asp305 310 315320
Thr Asn Asp His Gly Asp Phe Asn Gly Ile Glu Lys Thr Leu Thr Val325 330335Asn Pro Asn Ser Glu Leu Ile Phe Glu Phe Asn Thr Met Thr Thr Lys340 345350Asn Tyr Gln Gly Met Thr Asn Leu Ile Ile Lys Asn Ala Asp Asn Asp355 360 365Thr Val Ile Gly Glu Lys Val Val Ala Tyr Gly Pro Ile Trp Arg Leu370 375 380Leu Lys Val Pro Glu Asn Val Ser His Leu Lys Ile Gln Phe Val Pro385 390 395 400Lys Asn Asp Ala Ile Thr Asp Ala Arg Gly Ile Tyr Gln Leu Arg Asp405 410 415Gly Tyr Lys Tyr Tyr Asp Phe Val Asp Ser Ile Gly Leu His Ser Gly420 425430Ser His Val Tyr Val Glu Arg Arg Thr Met Glu Pro Thr Ala Thr Asn435 440445Asn Lys Glu Phe Thr Val Thr Thr Ser Leu Lys Asn Asn Gly Asn Phe450 455 460Gly Ala Ser Phe Asn Thr Asp Asp Phe Val Tyr Lys Ile Gln Leu Pro465 470475 480Glu Gly Val Glu Tyr Val Asn Asn Ser Leu Thr Lys Asp Phe Pro Ser485490495Gly Asn Ser Gly Val Asp Ile Asn Asp Met Asn Val Thr Tyr Asp Ala500 505 510Ala Asn Arg Ile Ile Thr Ile Lys Ser Thr Gly Gly Gly Thr Gly Asn515520 525Ser Pro Ala Arg Leu Met Pro Asp Lys Ile Leu Asp Leu Lys Tyr Lys530 535540
Leu Arg Val Asn Asn Val Pro Thr Pro Arg Thr Val Thr Phe Asn Asp545550555 560Thr Leu Thr Tyr Lys Thr Tyr Ser Gln Asp Phe Ile Asn Ser Pro Ala565 570575Glu Ser His Thr Val Ser Thr Asn Pro Tyr Thr Ile Asp Ile Ile Met580 585 590Asn Lys Asp Ala Leu Gln Ala Glu Val Asp Arg Arg Ile Gln Gln Ala595 600 605Asp Tyr Thr Phe Ala Ser Leu Asp Ile Phe Asn Asp Leu Lys Arg Arg610 615 620Ala Gln Thr Ile Leu Asp Glu Asn Arg Asn Asn Val Pro Leu Asn Lys625 630635 640Arg Val Ser Gln Ala Asp Ile Asp Ser Leu Ala Asn Gln Met Gln His645 650 655Thr Leu Ile Arg Ser Val Asp Ala Glu Asn Ala Val Asn Arg Lys Val660 665 670Asp Asp Met Glu Asp Leu Val Asn Gln Asn Asp Glu Leu Thr Asp Glu675 680 685Glu Lys Gln Ala Ala Ile Gln Val Ile Glu Glu His Lys Asn Glu Ile690 695 700Ile Gly Asn Ile Gly Asp Gln Thr Thr Asp Asp Gly Val Thr Arg Ile705 710 715 720Lys Asp Gln Gly Ile Gln Thr Leu Ser Gly Asp Thr Ala Thr Pro Val725730735Val Lys Pro Asn Ala Lys Gln Ala Ile Arg Asp Lys Ala Ala Lys Gln740 745 750Arg Glu Ile Ile Asn His Thr Pro Asp Ala Thr Gln Asp Glu Ile Gln755 760 765
Asp Ala Leu Asn Gln Leu Thr Thr Asp Glu Thr Asp Ala Ile Asp Asn770775780Val Thr Asn Ala Thr Thr Asn Ala Asp Val Glu Thr Ala Lys Asn Asn785 790795 800Gly Ile Asn Thr Ile Gly Ala Val Ala Pro Gln Val Thr His Lys Gln805 810 815Ala Ala Arg Asp Ala Ile Asn Gln Ala Thr Ala Thr Lys Arg Gln Gln820 825830Ile Asn Ser Asn Arg Glu Ala Thr Gln Glu Glu Lys Asn Ala Ala Leu835 840 845Asn Glu Leu Thr Gln Ala Thr Asn His Ala Leu Glu Gln Ile Asn Gln850 855860Ala Thr Thr Asn Asp Asp Val Asp Thr Ala Lys Gly Asp Gly Leu Asn865870 875 880Ala Ile Asn Pro Ile Ala Pro Val Thr Val Val Lys Gln Ala Ala Arg885 890 895Asp Ala Val Ser His Asp Ala Gln Gln His Ile Ala Glu Ile Asn Ala900 905 910Asn Pro Asp Ala Thr Gln Glu Glu Arg Gln Ala Ala Ile Glu Lys Val915 920 925Tyr Ala Ala Val Ala Val Ala Asn Thr Asn Ile Leu Asn Ala Asn Thr930 935 940Asn Ala Asp Val Glu Gln Val Lys Thr Asn Ala Ile Gln Gly Ile Gln945950 955960Ala Ile Glu Pro Ala Thr Lys Val Lys Thr Asp Ala Lys Asn Ala Ile965 970 975Asp Gln Ser Ala Glu Thr Gln His Asn Ala Ile Phe Asn Asn Asn Asp980 985 990
Ala Thr Leu Glu Glu Gln Gln Ala Ala Gln Gln Leu Leu Asp Gln Ala995 1000 1005Val Ala Thr Ala Lys Gln Asn Ile Asn Ala Ala Asp Thr Asn Gln1010 1015 1020Glu Val Ala Gln Ala Lys Asp Gln Gly Thr Gln Asn Ile Val Val1025 1030 1035Ile Gln Pro Ala Thr Gln Val Lys Thr Asp Ala Arg Asn Ala Val1040 1045 1050Asn Glu Lys Ala Arg Glu Ala Ile Thr Asn Ile Asn Ala Thr Pro1055 1060 1065Gly Ala Thr Arg Glu Glu Lys Gln Glu Ala Ile Asn Arg Val Asn1070 1075 1080Thr Leu Lys Asn Arg Ala Leu Asn Asp Ile Gly Val Thr Ser Thr1085 1090 1095Thr Ala Met Val Asn Ser Ile Arg Asp Asp Ala Val Asn Gln Ile1100 1105 1110Gly Ala Val Gln Pro His Val Thr Lys Lys Gln Thr Ala Thr Gly1115 1120 1125Val Leu Thr Asp Leu Ala Thr Ala Lys Lys Gln Glu Ile Asn Gln1130 1135 1140Asn Thr Asn Ala Thr Thr Glu Glu Lys Gln Val Ala Leu Asn Gln1145 1150 1155Val Asp Gln Asp Leu Ala Thr Ala Ile Asn Asn Ile Asn Gln Ala1160 1165 1170Asp Thr Asn Ala Glu Val Asp Gln Ala Gln Gln Leu Gly Thr Lys1175 1180 1185Ala Ile Asn Ala Ile Gln Pro Asn Ile Val Lys Lys Pro Ala Ala1190 1195 1200
Leu Ala Gln Thr Asn Gln His Tyr Ser Ala Lys Leu Val Glu Ile1205 1210 1215Asn Ala Thr Pro Asp Ala Thr Asp Asp Glu Lys Asn Ala Ala Ile1220 1225 1230Asn Thr Leu Asn Gln Asp Arg Gln Gln Ala Ile Glu Ser Ile Lys1235 1240 1245Gln Ala Asn Thr Asn Ala Glu Val Asp Gln Ala Ala Thr Val Ala1250 1255 1260Glu Asn Asn Ile Asp Ala Val Gln Val Asp Val Val Lys Lys Gln1265 1270 1275Ala Ala Arg Asp Lys Ile Thr Ala Glu Val Ala Lys Arg Ile Glu1280 1285 1290Ala Val Lys Gln Thr Pro Asn Ala Thr Asp Glu Glu Lys Gln Ala1295 1300 1305Ala Val Asn Gln Ile Asn Gln Leu Lys Asp Gln Ala Phe Asn Gln1310 1315 1320Ile Asn Gln Asn Gln Thr Asn Asp Gln Val Asp Ala Thr Thr Asn1325 1330 1335Gln Ala Ile Asn Ala Ile Asp Asn Val Glu Ala Glu Val Val Ile1340 1345 1350Lys Pro Lys Ala Ile Ala Asp Ile Glu Lys Ala Val Lys Glu Lys1355 1360 1365Gln Gln Gln Ile Asp Asn Ser Leu Asp Ser Thr Asp Asn Glu Lys1370 1375 1380Glu Val Ala Leu Gln Ala Leu Ala Lys Glu Lys Glu Lys Ala Leu1385 1390 1395Ala Ala Ile Asp Gln Ala Gln Thr Asn Ser Gln Val Asn Gln Ala1400 1405 1410
Ala Thr Asn Gly Val Ser Ala Ile Lys Ile Ile Gln Pro Glu Thr1415 14201425Lys Ile Lys Pro Ala Ala Arg Glu Lys Ile Asn Gln Lys Ala Asn1430 14351440Glu Leu Arg Ala Gln Ile Asn Gln Asp Lys Glu Ala Thr Ala Glu1445 14501455Glu Arg Gln Ala Ala Leu Asp Lys Ile Asn Asp Leu Val Ala Lys1460 14651470Ala Met Thr Asn Ile Thr Asn Asp Arg Thr Asn Gln Gln Val Asn1475 14801485Asp Ser Thr Asn Gln Ala Leu Asp Asp Ile Ala Leu Val Thr Pro1490 14951500Asp His Ile Val Arg Ala Ala Ala Arg Asp Ala Val Lys Gln Gln1505 15101515Tyr Glu Ala Lys Lys His Glu Ile Glu Gln Ala Glu His Ala Thr1520 15251530Asp Glu Glu Lys Gln Val Ala Leu Asn Gln Leu Ala Asn Asn Glu1535 15401545Lys Arg Ala Leu Gln Asn Ile Asn Gln Ala Ile Ala Asn Asn Asp1550 15551560Val Lys Arg Val Glu Ser Asn Gly Ile Ala Thr Leu Lys Gly Val1565 15701575Glu Pro His Ile Val Val Lys Pro Glu Ala Gln Glu Ala Ile Lys1580 15851590Ala Ser Ala Asp Asn Gln Val Glu Ser Ile Lys Asp Thr Pro His1595 16001605Ala Thr Thr Asp Glu Leu Asp Glu Ala Asn Gln Gln Ile Asn Asp1610 16151620
Thr Leu Lys Gln Gly Gln Gln Asp Ile Asp Asn Thr Thr Gln Asp1625 1630 1635Ala Ala Val Asn Asp Val Arg Asn Gln Thr Ile Lys Ala Ile Glu1640 1645 1650Gln Ile Lys Pro Lys Val Arg Arg Lys Arg Ala Ala Leu Asp Asn1655 1660 1665Ile Asp Glu Ser Asn Asn Asn Gln Leu Asp Ala Ile Arg Asn Thr1670 1675 1680Leu Asp Thr Thr Gln Asp Glu Arg Asn Val Ala Ile Ala Ala Leu1685 1690 1695Asn Lys Ile Val Asn Ala Ile Lys Asn Asp Ile Ala Gln Asn Lys1700 1705 1710Thr Asn Ala Glu Val Asp Gln Thr Glu Ala Asp Gly Asn Asn Asn1715 1720 1725Ile Lys Val Ile Leu Pro Lys Val Gln Val Lys Pro Ala Ala Arg1730 1735 1740Gln Ser Val Ser Ala Lys Ala Glu Ala Gln Asn Ala Leu Ile Asp1745 1750 1755Gln Ser Asp Leu Ser Thr Glu Glu Glu Arg Leu Ala Ala Lys His1760 1765 1770Leu Val Glu Gln Ala Leu Asn Gln Ala Ile Asp Gln Ile Asn His1775 1780 1785Ala Asp Lys Thr Ala Gln Val Asn Gln Asn Ser Ile Asp Ala Gln1790 1795 1800Asn Ile Ile Ser Lys Ile Lys Pro Ala Thr Thr Val Lys Ala Thr1805 1810 1815Ala Leu Gln Gln Ile Gln Asn Ile Ala Thr Asn Lys Ile Asn Leu1820 1825 1830
Ile Lys Ala Asn Asn Glu Ala Thr Asp Glu Glu Gln Asn Ala Ala1835 1840 1845Ile Val Gln Val Glu Lys Glu Leu Ile Lys Ala Lys Gln Gln Ile1850 1855 1860Ala Gly Ala Val Thr Asn Ala Asp Val Ala Tyr Leu Leu His Asp1865 1870 1875Gly Lys Asn Glu Ile Arg Glu Ile Glu Pro Val Ile Asn Lys Lys1880 1885 1890Ala Thr Ala Arg Glu Gln Leu Thr Thr Leu Phe Asn Asp Lys Lys1895 1900 1905Gln Ala Ile Glu Ala Asn Val Gln Ala Thr Val Glu Glu Arg Asn1910 1915 1920Ser Ile Leu Ala Gln Leu Gln Asn Ile Tyr Asp Thr Ala Ile Gly1925 1930 1935Gln Ile Asp Gln Asp Arg Ser Asn Ala Gln Val Asp Lys Thr Ala1940 1945 1950Thr Leu Asn Leu Gln Thr Ile His Asp Leu Asp Val His Pro Ile1955 1960 1965Lys Lys Pro Asp Ala Glu Lys Thr Ile Asn Asp Asp Leu Ala Arg1970 1975 1980Val Thr His Leu Val Gln Asn Tyr Arg Lys Val Ser Asp Arg Asn1985 1990 1995Lys Ala Asp Ala Leu Lys Ala Ile Thr Ala Leu Lys Leu Gln Met2000 2005 2010Asp Glu Glu Leu Lys Thr Ala Arg Thr Asn Ala Asp Val Asp Ala2015 2020 2025Val Leu Lys Arg Phe Asn Val Ala Leu Gly Asp Ile Glu Ala Val2030 2035 2040
Ile Thr Glu Lys Glu Asn Ser Leu Leu Arg Ile Asp Asn Ile Ala2045 20502055Gln Gln Thr Tyr Ala Lys Phe Lys Ala Ile Ala Thr Pro Glu Gln2060 20652070Leu Ala Lys Val Lys Ala Leu Ile Asp Gln Tyr Val Ala Asp Gly2075 20802085Asn Arg Met Val Asp Glu Asp Ala Thr Leu Asn Asp Ile Lys Lys2090 20952100Asp Thr Gln Leu Ile Ile Asp Glu Ile Leu Ala Ile Lys Leu Pro2105 21102115Ala Glu Val Ile Lys Ala Ser Pro Lys Val Gly Gln Pro Ala Pro2120 21252130Lys Val Cys Thr Pro Ile Lys Lys Glu Asp Lys Gln Glu Val Arg2135 21402145Lys Val Val Lys Glu Leu Pro Asn Thr Gly Ser Glu Glu Met Asp2150 21552160Leu Pro Leu Lys Glu Leu Ala Leu Ile Thr Gly Ala Ala Leu Leu2165 21702175Ala Arg Arg Arg Ser Lys Lys Glu Lys Glu Ser2180 2185<210>3<211>6852<212>DNA<213>Staphylococcus epidermidis<400>3tctaatgaat gtaaagataa tacaaggagt tattacatga gtaaaagaca gaaagcattt 60catgacagct tagcaaacga aaaaacaaga gtaagacttt ataaatctgg aaaaaattgg120gtaaaatccg gaattaaaga aatagaaatg ttcaaaatta tggggctacc atttattagt180catagtttag tgagtcaaga taatcaaagc attagtaaaa aaatgacggg atacggactg240aaaactacgg cggttattgg tggtgcattc acggtaaata tgttgcatga ccagcaagct300
tttgcggctt ctgatgcacc attaacttct gaattaaaca cacaaagtga aacagtaggt360aatcaaaact caacgacaat cgaagcatca acatcaacag ccgattccac aagtgtaacg420aaaaatagta gttcggtaca aacatcaaat agtgacacag tctcaagtga aaagtctgaa480aaggtcactt cgacaactaa tagtacaagc aatcaacaag agaaattgac atctacatca540gaatcaacat cctcaaagaa tactacatca agttctgata ctaaatctgt agcttcaact600tcaagtacag aacaaccaat taatacatca acaaatcaaa gtactgcatc aaataacact660tcacaaagca caacgccatc ttcggtcaac ttaaacaaaa ctagcacaac gtcaactagc720accgcaccag taaaacttcg aactttcagt cgcttagcta tgtcaacatt tgcgtcagca780gcgacgacaa ccgcagtaac tgctaataca attacagtta ataaagataa cttaaaacaa840tatatgacaa cgtcaggtaa tgctacctat gatcaaagta ccggtattgt gacgttaaca900caggatgcat acagccaaaa aggtgctatt acattaggaa cacgtattga ctctaataag960agttttcatt tttctggaaa agtaaattta ggtaacaaat atgaagggca tggaaatggt 1020ggagatggta tcggttttgc cttttcacca ggtgtattag gtgaaacagg gttaaacggt 1080gccgcagtag gtattggtgg cttaagtaac gcatttggct tcaaattgga tacgtatcac 1140aatacatcta aaccaaattc agctgcaaag gcgaatgctg acccatctaa tgtagctggt 1200ggaggtgcgt ttggtgcatt tgtaacaaca gatagttatg gtgttgcgac aacgtataca 1260tcaagttcaa cagctgataa tgctgcgaag ttaaatgttc aacctacaaa taacacgttc 1320caagattttg atattaacta taatggtgat acaaaggtta tgactgtcaa atatgcaggt 1380caaacatgga cacgtaatat ttcagattgg attgcgaaaa gtggtacgac caacttttca 1440ttatcaatga cagcctcaac aggtggcgcg acaaatttac aacaagtaca atttggaaca 1500ttcgaatata cagagtctgc tgttacacaa gtgagatacg ttgatgtaac aacaggtaaa 1560gatattattc caccaaaaac atattcagga aatgttgatc aagtcgtgac aatcgataat 1620cagcaatctg cattgactgc taaaggatat aactacacgt ccgtcgatag ttcatatgcg 1680tcaacttata atgatacaaa taaaactgta aaaatgacga atgctggaca atcagtgaca 1740tattatttta ctgatgtaaa agcaccaact gtaactgtag gcaatcaaac catagaagtg 1800ggtaaaacaa tgaatcctat tgtattgact acaacggata atggtactgg gactgtgaca 1860aatacagtta caggattacc aagcggatta agttacgata gtgcaacgaa ttcaatcatt 1920gggacaccaa caaaaattgg tcaatcaaca gtgacagttg tgtctactga ccaagcaaat 1980
aacaaatcga cgacaacttt tacaataaat gttgtggata cgacagcacc aacagtgaca 2040ccaataggag atcaatcatc agaagtgtat tcaccaatat ccccgattaa aattgctacg 2100caagataaca gtggaaatgc ggtgacgaat acagtgactg gattgccatc cggactaaca 2160tttgatagta caaataatac tattagtggt acaccaacaa acattggtac aagtactata 2220tcaatcgttt ctacagatgc gagcggtaac aaaacgacga caacttttaa atatgaagta 2280acaagaaata gcatgagtga ttccgtatca acatcaggaa gtacacaaca atctcaaagt 2340gtgtcaacaa gtaaagctga ctcacaaagt gcatcaacga gtacatcagg atcgattgtg 2400gtatctacat cagctagtac ctcgaaatcg acaagtgtaa gcctatctga ttctgtgagt 2460gcatctaagt cattaagcac atctgaaagt aatagtgtat caagctcaac aagcacaagt 2520ttagtgaatt cacaaagtgt atcatcaagc atgtcggatt cagctagtaa atcaacatca 2580ttaagcgatt ctatttcaaa ctctagcagt actgaaaaat ccgaaagtct atcaacaagt 2640acatctgatt cattgcgtac atcaacatca ctcagtgact cattaagtat gagtacatca 2700ggaagcttgt ctaagtcaca aagcttatca acgagtatat cagggtcgtc tagtacatca 2760gcatcattaa gtgacagtac atcgaatgca attagtacat caacatcatt gagcgagtca 2820gctagcacct cggactctat cagtatttca aatagcatag ccaactctca aagtgcgtca 2880acaagcaaat cagattcaca aagtacatca atatcattaa gtacaagtga ttcaaaatcg 2940atgagtacat cagaatcatt gagcgattcg acgagcacaa gtggttctgt ttctggatca 3000ctaagcatag cagcatcaca aagtgtctca acaagtacat cagactcgat gagtacttca 3060gagatagtaa gtgactctat cagtacaagt gggtcattat ctgcatcaga cagtaaatca 3120atgtccgtaa gtagttcaat gagcacgtct cagtcaggta gtacatcaga atcattaagt 3180gattcacaaa gtacatctga ttctgatagt aagtcattat cacaaagtac tagtcaatca 3240ggttcaacaa gtacatcaac gtcgacaagt gcttcagtac gtacttcgga atcacaaagt 3300acgtctggtt caatgagtgc aagtcaatcc gattcaatga gcatatcaac gtcgtttagt 3360gattcaacga gtgatagcaa atcagcatca actgcatcaa gtgaatcaat atcacaaagt 3420gcttctacga gcacatctgg ttcggtaagt acttcgacat cgttaagtac aagtaattca 3480gaacgtacat caacatctat gagtgattcc acaagcttaa gtacatcaga gtctgattca 3540ataagtgaat caacgtcaac gagcgactct ataagtgaag caatatctgc ttcagagagc 3600acgtttatat cattaagtga atcaaatagt actagcgatt cagaatcaca aagtgcatct 3660
gcctttttaa gtgaatcatt aagtgaaagt acgtctgaat caacatcaga gtcagtgagt 3720agttcgacaa gtgagagtac gtcattatca gacagtacat cagaatctgg tagcacatca 3780acatcattaa gtaattcaac aagtggtagt acgtccattt caacatcgac aagtatcagt 3840gaatcaacgt caacgtttaa gagcgagagt gtttcaacat cactgagtat gtcaacgagt 3900acaagtttgt ctgactctac aagtttgtca acatcattaa gtgattccac aagtgatagt 3960aagtctgatt cattaagtac atcaatgtcg acaagtgatt caatcagtac aagtaaatct 4020gattccatta gtacatccac atcattaagt ggttctacaa gtgaaagtga atccgactca 4080acatcatcaa gtgaaagtaa atccgattca acatcaatga gcataagtat gtctcaatca 4140acatcaggaa gtacaagtac gtcaacgagt acaagtttgt ctgactcaac gagtacatca 4200ttgtcactaa gtgcctcaat gaatcaaagc ggagtagact caaactcagc aagccaaagt 4260gcctcaaact caacaagtac aagcacgagc gaatccgatt cacaaagcac atcatcatat 4320acaagtcagt caacaagcca aagtgaatcc acatcgacat caacgtcact aagcgattca 4380acaagtatat ctaaaagtac gagtcaatca ggttcggtaa gcacatcagc gtcattaagt 4440ggttcagaga gtgaatctga ttcacaaagt atctcaacaa gtgcaagtga gtcaacatca 4500gaaagtgcgt caacatcact cagtgactca acaagtacaa gtaactcagg atcagcaagt 4560acgtcaacat cgctcagtaa ctcagcaagc gcaagtgaat ccgatttgtc gtcaacatct 4620ttaagtgatt caacatctgc gtcaatgcaa agcagtgaat ccgattcaca aagcacatca 4680gcatcattaa gtgattcgct aagtacatca acttcaaacc gcatgtcgac cattgcaagt 4740ttatctacat cggtaagtac atcagagtct ggctcaacat cagaaagtac aagtgaatcc 4800gattcaacat caacatcatt aagcgattca caaagcacat caagaagtac aagtgcatca 4860ggatcagcaa gtacatcaac atcaacaagt gactctcgta gtacatcagc ttcaactagt 4920acttcgatgc gtacaagtac tagtgattca caaagtatgt cgctttcgac aagtacatca 4980acaagtatga gtgattcaac gtcattatct gatagtgtta gtgattcaac atcagactca 5040acaagtgcga gtacatctgg ttcgatgagt gtgtctatat cgttaagtga ttcgacaagt 5100acatcaacat cggctagtga agtaatgagc gcaagcatat ctgattcaca aagtatgtca 5160gaatctgtaa atgattcaga aagtgtaagt gaatctaatt ctgaaagtga ctctaaatcg 5220atgagtggct caacaagtgt cagtgattct ggctcattga gcgtctcaac gtcattaaga 5280aaatcagaaa gtgtaagcga gtcaagttca ttgagttgct cacaatcgat gagcgattca 5340
gtaagcacaa gcgattcgtc atcattaagt gtatcgacgt cactaagaag ttcagaaagc 5400gtgagtgaat ctgattcatt aagtgattca aaatcaacaa gtggttcgac ttcaacaagt 5460acatctggtt cattgagtac ctcaacatca ttaagtggtt cagaaagcgt aagcgagtct 5520acctcgctaa gtgattcaat atcaatgagt gattctacta gtacaagtga ctccgactca 5580ttaagtggat caatatcttt aagtggttcc acaagtctta gcacttcgga ttcattaagt 5640gattcaaaat cattgagtag ctcgcaaagt atgagtggat cagaatcaac gtcaacaagt 5700gtgagcgatt cgcagtcaag ctcaacaagt aatagtcaat ttgactctat gagcatcagt 5760gcatcagaaa gcgactcaat gtctacaagt gattcgtcta gcatcagtgg atcaaattca 5820acgagtacat cactttcaac atctgactca atgagcggaa gcgtatcagt ttcaacatcg 5880acaagtttaa gtgactcaat atcaggttca acaagtgtaa gtgactcgag ctcaacaagc 5940acatctacat cattaagtga ttcaatgtca caaagccagt caacaagtac aagtgcatct 6000ggttccttaa gtacatcgat atcaacatca atgtcaatga gtgctagtac atcgtcatca 6060caaagcacat cggtgtcgac atcattatca acatcagaca gtatcagtga ttctacttca 6120ataagtatca gtggttcaca aagtacagta gaatcagaat ctacaagtga ttcaacttct 6180atcagtgact cagaatcatt gagtacatca gattcagact cgacatcgac aagtacatcg 6240gactcaacaa gtggttcaac ttcaacaagc atatctgaat cattaagtac gtctggttca 6300ggttcaacga gcgtatctga ctcaacatca atgagtgaat ctaattcatc gagtgtttca 6360atgtcacaag acaaatccga ctcaacatca attagtgact cagaatcagt gtcaacaagc 6420acatcaacgt cattgagcac atccgattcg acaagcacat ccgaatcact gagtacatct 6480atgtctggtt cacaaagcat ttctgactca acatcaacaa gtatgtccgg ctcaacaagt 6540acatctgaat ctaactcaat gcatccgtca gactcaatga gtatgcatca tactcacagc 6600acgagcacat ctcgcttatc aagtgaagca acaacgagca cgagtgaatc tcagtctaca 6660ttaagtgcaa catctgaagt gactaaacat aatggcacac cagcacaaag tgaaaaaaga 6720ttgccagata caggtgactc aataaaacaa aatggattac taggtggcgt tatgacatta 6780ttagttggtt taggtttaat gaagagaaag aaaaagaaag atgaaaatga tcaagatgat 6840tctcaagcat aa 6852<210>4<211>2283<212>PRT
<213>Staphylococcus epidermidis<400>4Ser Asn Glu Cys Lys Asp Asn Thr Arg Ser Tyr Tyr Met Ser Lys Arg1 5 10 15Gln Lys Ala Phe His Asp Ser Leu Ala Asn Glu Lys Thr Arg Val Arg20 25 30Leu Tyr Lys Ser Gly Lys Asn Trp Val Lys Ser Gly Ile Lys Glu Ile35 40 45Glu Met Phe Lys Ile Met Gly Leu Pro Phe Ile Ser His Ser Leu Val50 55 60Ser Gln Asp Asn Gln Ser Ile Ser Lys Lys Met Thr Gly Tyr Gly Leu65 70 75 80Lys Thr Thr Ala Val Ile Gly Gly Ala Phe Thr Val Asn Met Leu His85 90 95Asp Gln Gln Ala Phe Ala Ala Ser Asp Ala Pro Leu Thr Ser Glu Leu100 105 110Asn Thr Gln Ser Glu Thr Val Gly Asn Gln Asn Ser Thr Thr Ile Glu115 120125Ala Ser Thr Ser Thr Ala Asp Ser Thr Ser Val Thr Lys Asn Ser Ser130 135 140Ser Val Gln Thr Ser Asn Ser Asp Thr Val Ser Ser Glu Lys Ser Glu145 150 155 160Lys Val Thr Ser Thr Thr Asn Ser Thr Ser Asn Gln Gln Glu Lys Leu165170175Thr Ser Thr Ser Glu Ser Thr Ser Ser Lys Asn Thr Thr Ser Ser Ser180 185 190Asp Thr Lys Ser Val Ala Ser Thr Ser Ser Thr Glu Gln Pro Ile Asn195 200 205
Thr Ser Thr Asn Gln Ser Thr Ala Ser Asn Asn Thr Ser Gln Ser Thr210215 220Thr Pro Ser Ser Val Asn Leu Asn Lys Thr Ser Thr Thr Ser Thr Ser225 230 235 240Thr Ala Pro Val Lys Leu Arg Thr Phe Ser Arg Leu Ala Met Ser Thr245250255Phe Ala Ser Ala Ala Thr Thr Thr Ala Val Thr Ala Asn Thr Ile Thr260 265 270Val Asn Lys Asp Asn Leu Lys Gln Tyr Met Thr Thr Ser Gly Asn Ala275 280 285Thr Tyr Asp Gln Ser Thr Gly Ile Val Thr Leu Thr Gln Asp Ala Tyr290 295 300Ser Gln Lys Gly Ala Ile Thr Leu Gly Thr Arg Ile Asp Ser Asn Lys305 310 315 320Ser Phe His Phe Ser Gly Lys Val Asn Leu Gly Asn Lys Tyr Glu Gly325330 335His Gly Asn Gly Gly Asp Gly Ile Gly Phe Ala Phe Ser Pro Gly Val340 345 350Leu Gly Glu Thr Gly Leu Asn Gly Ala Ala Val Gly Ile Gly Gly Leu355360 365Ser Asn Ala Phe Gly Phe Lys Leu Asp Thr Tyr His Asn Thr Ser Lys370375 380Pro Asn Ser Ala Ala Lys Ala Asn Ala Asp Pro Ser Asn Val Ala Gly385 390 395 400Gly Gly Ala Phe Gly Ala Phe Val Thr Thr Asp Ser Tyr Gly Val Ala405 410 415Thr Thr Tyr Thr Ser Ser Ser Thr Ala Asp Asn Ala Ala Lys Leu Asn420 425430
Val Gln Pro Thr Asn Asn Thr Phe Gln Asp Phe Asp Ile Asn Tyr Asn435440 445Gly Asp Thr Lys Val Met Thr Val Lys Tyr Ala Gly Gln Thr Trp Thr450455 460Arg Asn Ile Ser Asp Trp Ile Ala Lys Ser Gly Thr Thr Asn Phe Ser465 470 475 480Leu Ser Met Thr Ala Ser Thr Gly Gly Ala Thr Asn Leu Gln Gln Val485 490 495Gln Phe Gly Thr Phe Glu Tyr Thr Glu Ser Ala Val Thr Gln Val Arg500 505 510Tyr Val Asp Val Thr Thr Gly Lys Asp Ile Ile Pro Pro Lys Thr Tyr515 520 525Ser Gly Asn Val Asp Gln Val Val Thr Ile Asp Asn Gln Gln Ser Ala530535 540Leu Thr Ala Lys Gly Tyr Asn Tyr Thr Ser Val Asp Ser Ser Tyr Ala545 550 555 560Ser Thr Tyr Asn Asp Thr Asn Lys Thr Val Lys Met Thr Asn Ala Gly565570 575Gln Ser Val Thr Tyr Tyr Phe Thr Asp Val Lys Ala Pro Thr Val Thr580 585 590Val Gly Asn Gln Thr Ile Glu Val Gly Lys Thr Met Asn Pro Ile Val595 600605Leu Thr Thr Thr Asp Asn Gly Thr Gly Thr Val Thr Asn Thr Val Thr610 615 620Gly Leu Pro Ser Gly Leu Ser Tyr Asp Ser Ala Thr Asn Ser Ile Ile625 630 635 640Gly Thr Pro Thr Lys Ile Gly Gln Ser Thr Val Thr Val Val Ser Thr645 650 655
Asp Gln Ala Asn Asn Lys Ser Thr Thr Thr Phe Thr Ile Asn Val Val660 665 670Asp Thr Thr Ala Pro Thr Val Thr Pro Ile Gly Asp Gln Ser Ser Glu675 680 685Val Tyr Ser Pro Ile Ser Pro Ile Lys Ile Ala Thr Gln Asp Asn Ser690695 700Gly Asn Ala Val Thr Asn Thr Val Thr Gly Leu Pro Ser Gly Leu Thr705 710 715 720Phe Asp Ser Thr Asn Asn Thr Ile Ser Gly Thr Pro Thr Asn IIe Gly725 730 735Thr Ser Thr Ile Ser Ile Val Ser Thr Asp Ala Ser Gly Asn Lys Thr740 745750Thr Thr Thr Phe Lys Tyr Glu Val Thr Arg Asn Ser Met Ser Asp Ser755 760 765Val Ser Thr Ser Gly Ser Thr Gln Gln Ser Gln Ser Val Ser Thr Ser770 775 780Lys Ala Asp Ser Gln Ser Ala Ser Thr Ser Thr Ser Gly Ser Ile Val785 790 795 800Val Ser Thr Ser Ala Ser Thr Ser Lys Ser Thr Ser Val Ser Leu Ser805 810815Asp Ser Val Ser Ala Ser Lys Ser Leu Ser Thr Ser Glu Ser Asn Ser820 825 830Val Ser Ser Ser Thr Ser Thr Ser Leu Val Asn Ser Gln Ser Val Ser835 840 845Ser Ser Met Ser Asp Ser Ala Ser Lys Ser Thr Ser Leu Ser Asp Ser850 855 860Ile Ser Asn Ser Ser Ser Thr Glu Lys Ser Glu Ser Leu Ser Thr Ser865 870 875 880
Thr Ser Asp Ser Leu Arg Thr Ser Thr Ser Leu Ser Asp Ser Leu Ser885 890 895Met Ser Thr Ser Gly Ser Leu Ser Lys Ser Gln Ser Leu Ser Thr Ser900 905 910Ile Ser Gly Ser Ser Ser Thr Ser Ala Ser Leu Ser Asp Ser Thr Ser915 920 925Asn Ala Ile Ser Thr Ser Thr Ser Leu Ser Glu Ser Ala Ser Thr Ser930 935 940Asp Ser Ile Ser Ile Ser Asn Ser Ile Ala Asn Ser Gln Ser Ala Ser945 950 955 960Thr Ser Lys Ser Asp Ser Gln Ser Thr Ser Ile Ser Leu Ser Thr Ser965 970 975Asp Ser Lys Ser Met Ser Thr Ser Glu Ser Leu Ser Asp Ser Thr Ser980 985 990Thr Ser Gly Ser Val Ser Gly Ser Leu Ser Ile Ala Ala Ser Gln Ser9951000 1005Val Ser Thr Ser Thr Ser Asp Ser Met Ser Thr Ser Glu Ile Val1010 1015 1020Ser Asp Ser Ile Ser Thr Ser Gly Ser Leu Ser Ala Ser Asp Ser1025 1030 1035Lys Ser Met Ser Val Ser Ser Ser Met Ser Thr Ser Gln Ser Gly1040 1045 1050Ser Thr Ser Glu Ser Leu Ser Asp Ser Gln Ser Thr Ser Asp Ser1055 1060 1065Asp Ser Lys Ser Leu Ser Gln Ser Thr Ser Gln Ser Gly Ser Thr1070 1075 1080Ser Thr Ser Thr Ser Thr Ser Ala Ser Val Arg Thr Ser Glu Ser1085 10901095
Gln Ser Thr Ser Gly Ser Met Ser Ala Ser Gln Ser Asp Ser Met1100 11051110Ser Ile Ser Thr Ser Phe Ser Asp Ser Thr Ser Asp Ser Lys Ser1115 11201125Ala Ser Thr Ala Ser Ser Glu Ser Ile Ser Gln Ser Ala Ser Thr1130 11351140Ser Thr Ser Gly Ser Val Ser Thr Ser Thr Ser Leu Ser Thr Ser1145 11501155Asn Ser Glu Arg Thr Ser Thr Ser Met Ser Asp Ser Thr Ser Leu1160 11651170Ser Thr Ser Glu Ser Asp Ser Ile Ser Glu Ser Thr Ser Thr Ser1175 11801185Asp Ser Ile Ser Glu Ala Ile Ser Ala Ser Glu Ser Thr Phe Ile1190 11951200Ser Leu Ser Glu Ser Asn Ser Thr Ser Asp Ser Glu Ser Gln Ser1205 12101215Ala Ser Ala Phe Leu Ser Glu Ser Leu Ser Glu Ser Thr Ser Glu1220 12251230Ser Thr Ser Glu Ser Val Ser Ser Ser Thr Ser Glu Ser Thr Ser1235 12401245Leu Ser Asp Ser Thr Ser Glu Ser Gly Ser Thr Ser Thr Ser Leu1250 12551260Ser Asn Ser Thr Ser Gly Ser Thr Ser Ile Ser Thr Ser Thr Ser1265 12701275Ile Ser Glu Ser Thr Ser Thr Phe Lys Ser Glu Ser Val Ser Thr1280 12851290Ser Leu Ser Met Ser Thr Ser Thr Ser Leu Ser Asp Ser Thr Ser1295 13001305
Leu Ser Thr Ser Leu Ser Asp Ser Thr Ser Asp Ser Lys Ser Asp1310 13151320Ser Leu Ser Thr Ser Met Ser Thr Ser Asp Ser Ile Ser Thr Ser1325 13301335Lys Ser Asp Ser Ile Ser Thr Ser Thr Ser Leu Ser Gly Ser Thr1340 13451350Ser Glu Ser Glu Ser Asp Ser Thr Ser Ser Ser Glu Ser Lys Ser1355 13601365Asp Ser Thr Ser Met Ser Ile Ser Met Ser Gln Ser Thr Ser Gly1370 13751380Ser Thr Ser Thr Ser Thr Ser Thr Ser Leu Ser Asp Ser Thr Ser1385 13901395Thr Ser Leu Ser Leu Ser Ala Ser Met Asn Gln Ser Gly Val Asp1400 14051410Ser Asn Ser Ala Ser Gln Ser Ala Ser Asn Ser Thr Ser Thr Ser1415 14201425Thr Ser Glu Ser Asp Ser Gln Ser Thr Ser Ser Tyr Thr Ser Gln1430 14351440Ser Thr Ser Gln Ser Glu Ser Thr Ser Thr Ser Thr Ser Leu Ser1445 14501455Asp Ser Thr Ser Ile Ser Lys Ser Thr Ser Gln Ser Gly Ser Val1460 14651470Ser Thr Ser Ala Ser Leu Ser Gly Ser Glu Ser Glu Ser Asp Ser1475 14801485Gln Ser Ile Ser Thr Ser Ala Ser Glu Ser Thr Ser Glu Ser Ala1490 14951500Ser Thr Ser Leu Ser Asp Ser Thr Ser Thr Ser Asn Ser Gly Ser1505 15101515
Ala Ser Thr Ser Thr Ser Leu Ser Asn Ser Ala Ser Ala Ser Glu1520 1525 1530Ser Asp Leu Ser Ser Thr Ser Leu Ser Asp Ser Thr Ser Ala Ser1535 1540 1545Met Gln Ser Ser Glu Ser Asp Ser Gln Ser Thr Ser Ala Ser Leu1550 1555 1560Ser Asp Ser Leu Ser Thr Ser Thr Ser Asn Arg Met Ser Thr Ile1565 1570 1575Ala Ser Leu Ser Thr Ser Val Ser Thr Ser Glu Ser Gly Ser Thr1580 1585 1590Ser Glu Ser Thr Ser Glu Ser Asp Ser Thr Ser Thr Ser Leu Ser1595 1600 1605Asp Ser Gln Ser Thr Ser Arg Ser Thr Ser Ala Ser Gly Ser Ala1610 1615 1620Ser Thr Ser Thr Ser Thr Ser Asp Ser Arg Ser Thr Ser Ala Ser1625 1630 1635Thr Ser Thr Ser Met Arg Thr Ser Thr Ser Asp Ser Gln Ser Met1640 1645 1650Ser Leu Ser Thr Ser Thr Ser Thr Ser Met Ser Asp Ser Thr Ser1655 1660 1665Leu Ser Asp Ser Val Ser Asp Ser Thr Ser Asp Ser Thr Ser Ala1670 1675 1680Ser Thr Ser Gly Ser Met Ser Val Ser Ile Ser Leu Ser Asp Ser1685 1690 1695Thr Ser Thr Ser Thr Ser Ala Ser Glu Val Met Ser Ala Ser Ile1700 1705 1710Ser Asp Ser Gln Ser Met Ser Glu Ser Val Asn Asp Ser Glu Ser1715 1720 1725
Val Ser Glu Ser Asn Ser Glu Ser Asp Ser Lys Ser Met Ser Gly1730 17351740Ser Thr Ser Val Ser Asp Ser Gly Ser Leu Ser Val Ser Thr Ser1745 17501755Leu Arg Lys Ser Glu Ser Val Ser Glu Ser Ser Ser Leu Ser Cys1760 17651770Ser Gln Ser Met Ser Asp Ser Val Ser Thr Ser Asp Ser Ser Ser1775 17801785Leu Ser Val Ser Thr Ser Leu Arg Ser Ser Glu Ser Val Ser Glu1790 17951800Ser Asp Ser Leu Ser Asp Ser Lys Ser Thr Ser Gly Ser Thr Ser1805 18101815Thr Ser Thr Ser Gly Ser Leu Ser Thr Ser Thr Ser Leu Ser Gly1820 18251830Ser Glu Ser Val Ser Glu Ser Thr Ser Leu Ser Asp Ser Ile Ser1835 18401845Met Ser Asp Ser Thr Ser Thr Ser Asp Ser Asp Ser Leu Ser Gly1850 18551860Ser Ile Ser Leu Ser Gly Ser Thr Ser Leu Ser Thr Ser Asp Ser1865 1870 1875Leu Ser Asp Ser Lys Ser Leu Ser Ser Ser Gln Ser Met Ser Gly1880 18851890Ser Glu Ser Thr Ser Thr Ser Val Ser Asp Ser Gln Ser Ser Ser1895 19001905Thr Ser Asn Ser Gln Phe Asp Ser Met Ser Ile Ser Ala Ser Glu1910 19151920Ser Asp Ser Met Ser Thr Ser Asp Ser Ser Ser Ile Ser Gly Ser1925 19301935
Asn Ser Thr Ser Thr Ser Leu Ser Thr Ser Asp Ser Met Ser Gly1940 1945 1950Ser Val Ser Val Ser Thr Ser Thr Ser Leu Ser Asp Ser Ile Ser1955 1960 1965Gly Ser Thr Ser Val Ser Asp Ser Ser Ser Thr Ser Thr Ser Thr1970 1975 1980Ser Leu Ser Asp Ser Met Ser Gln Ser Gln Ser Th Ser Thr Ser1985 1990 1995Ala Ser Gly Ser Leu Ser Thr Ser Ile Ser Thr Ser Met Ser Met2000 2005 2010Ser Ala Ser Thr Ser Ser Ser Gln Ser Thr Ser Val Ser Thr Ser2015 2020 2025Leu Ser Thr Ser Asp Ser Ile Ser Asp Ser Thr Ser Ile Ser Ile2030 2035 2040Ser Gly Ser Gln Ser Thr Val Glu Ser Glu Ser Thr Ser Asp Ser2045 20502055Thr Ser Ile Ser Asp Ser Glu Ser Leu Ser Thr Ser Asp Ser Asp2060 20652070Ser Thr Ser Thr Ser Thr Ser Asp Ser Thr Ser Gly Ser Thr Ser2075 20802085Thr Ser Ile Ser Glu Ser Leu Ser Thr Ser Gly Ser Gly Ser Thr2090 20952100Ser Val Ser Asp Ser Thr Ser Met Ser Glu Ser Asn Ser Ser Ser2105 2110 2115Val Ser Met Ser Gln Asp Lys Ser Asp Ser Thr Ser Ile Ser Asp2120 2125 2130Ser Glu Ser Val Ser Thr Ser Thr Ser Thr Ser Leu Ser Thr Ser2135 2140 2145
Asp Ser Thr Ser Thr Ser Glu Ser Leu Ser Thr Ser Met Ser Gly2150 2155 2160Ser Gln Ser Ile Ser Asp Ser Thr Ser Thr Ser Met Ser Gly Ser2165 2170 2175Thr Ser Thr Ser Glu Ser Asn Ser Met His Pro Ser Asp Ser Met2180 2185 2190Ser Met His His Thr His Ser Thr Ser Thr Ser Arg Leu Ser Ser2195 2200 2205Glu Ala Thr Thr Ser Thr Ser Glu Ser Gln Ser Thr Leu Ser Ala2210 2215 2220Thr Ser Glu Val Thr Lys His Asn Gly Thr Pro Ala Gln Ser Glu2225 2230 2235Lys Arg Leu Pro Asp Thr Gly Asp Ser Ile Lys Gln Asn Gly Leu2240 2245 2250Leu Gly Gly Val Met Thr Leu Leu Val Gly Leu Gly Leu Met Lys2255 22602265Arg Lys Lys Lys Lys Asp Glu Asn Asp Gln Asp Asp Ser Gln Ala2270 22752280<210>5<211>2730<212>DNA<213>Staphylococcus epidermidis<400>5ttattatcaa ttaaatataa tcttatagga gttgttaaca acatgaacaa acatcaccca 60aaattaaggt ctttctattc tattagaaaa tcaactctag gcgttgcatc ggtcattgtc120agtacactat ttttaattac ttctcaacat caagcacaag cagcagaaaa tacaaatact180tcagataaaa tctcggaaaa tcaaaataat aatgcaacta caactcagcc acctaaggat240acaaatcaaa cacaacctgc tacgcaacca gcaaacactg cgaaaaacta tcctgcagcg300gatgaatcac ttaaagatgc aattaaagat cctgcattag aaaataaaga acatgatata360ggtccaagag aacaagtcaa tttccagtta ttagataaaa acaatgaaac gcagtactat420
cactttttca gcatcaaaga tccagcagat gtgtattaca ctaaaaagaa agcagaagtt480gaattagaca tcaatactgc ttcaacatgg aagaagtttg aagtctatga aaacaatcaa540aaattgccag tgagacttgt atcatatagt cctgtaccag aagaccatgc ctatattcga600ttcccagttt cagatggcac acaagaattg aaaattgttt cttcgactca aattgatgat660ggagaagaaa caaattatga ttatactaaa ttagtatttg ctaaacctat ttataacgat720ccttcacttg taaaatcaga tacaaatgat gcagtagtaa cgaatgatca atcaagttca780gtcgcaagta atcaaacaaa cacgaataca tctaatcaaa atatatcaac gatcaacaat840gctaataatc aaccgcaggc aacgaccaat atgagtcaac ctgcacaacc aaaatcgtca900acgaatgcag atcaagcgtc aagccaacca gctcatgaaa caaattctaa tggtaatact960aacgataaaa cgaatgagtc aagtaatcag tcggatgtta atcaacagta tccaccagca 1020gatgaatcac tacaagatgc aattaaaaac ccggctatca tcgataaaga acatacagct 1080gataattggc gaccaattga ttttcaaatg aaaaatgata aaggtgaaag acagttctat 1140cattatgcta gtactgttga accagcaact gtcattttta caaaaacagg accaataatt 1200gaattaggtt taaagacagc ttcaacatgg aagaaatttg aagtttatga aggtgacaaa 1260aagttaccag tcgaattagt atcatatgat tctgataaag attatgccta tattcgtttc 1320ccagtatcta atggtacgag agaagttaaa attgtgtcat ctattgaata tggtgagaac 1380atccatgaag actatgatta tacgctaatg gtctttgcac agcctattac taataaccca 1440gacgactatg tggatgaaga aacatacaat ttacaaaaat tattagctcc gtatcacaaa 1500gctaaaacgt tagaaagaca agtttatgaa ttagaaaaat tacaagagaa attgccagaa 1560aaatataagg cggaatataa aaagaaatta gatcaaacta gagtagagtt agctgatcaa 1620gttaaatcag cagtgacgga atttgaaaat gttacaccta caaatgatca attaacagat 1680ttacaagaag cgcattttgt tgtttttgaa agtgaagaaa atagtgagtc agttatggac 1740ggctttgttg aacatccatt ctatacagca actttaaatg gtcaaaa ta tgtagtgatg 1800aaaacaaagg atgacagtta ctggaaagat ttaattgtag aaggtaaacg tgtcactact 1860gtttctaaag atcctaaaaa taattctaga acgctgattt tcccatatat acctgacaaa 1920gcagtttaca atgcgattgt taaagtcgtt gtggcaaaca ttggttatga aggtcaatat 1980catgtcagaa ttataaatca ggatatcaat acaaaagatg atgatacatc acaaaataac 2040acgagtgaac cgctaaatgt acaaacagga caagaaggta aggttgctga tacagatgta 2100
gctgaaaata gcagcactgc aacaaatcct aaagatgcgt ctgataaagc agatgtgata 2160gaaccagagt ctgacgtggt taaagatgct gataataata ttgataaaga tgtgcaacat 2220gatgttgatc atttatccga tatgtcggat aataatcact tcgataaata tgatttaaaa 2280gaaatggata ctcaaattgc caaagatact gatagaaatg tggataaaga tgccgataat 2340agcgttggta tgtcatctaa tgtcgatact gataaagact ctaataaaaa taaagacaaa 2400gtcatacagc tgaatcatat tgccgataaa aataatcata ctggaaaagc agcaaagctt 2460gacgtagtga aacaaaatta taataataca gacaaagtta ctgacaaaaa aacaactgaa 2520catctgccga gtgatattca taaaactgta gataaaacag tgaaaacaaa agaaaaagcc 2580ggcacaccat cgaaagaaaa caaacttagt caatctaaaa tgctaccaaa aactggagaa 2640acaacttcaa gccaatcatg gtggggctta tatgcgttat taggtatgtt agctttattc 2700attcctaaat tcagaaaaga atctaaataa2730<210>6<211>909<212>PRT<213>Staphylococcus epidermidis<400>6Leu Leu Ser Ile Lys Tyr Asn Leu Ile Gly Val Val Asn Asn Met Asn1 510 15Lys His His Pro Lys Leu Arg Ser Phe Tyr Ser Ile Arg Lys Ser Thr20 25 30Leu Gly Val Ala Ser Val Ile Val Ser Thr Leu Phe Leu Ile Thr Ser3540 45Gln His Gln Ala Gln Ala Ala Glu Asn Thr Asn Thr Ser Asp Lys Ile50 55 60Ser Glu Asn Gln Asn Asn Asn Ala Thr Thr Thr Gln Pro Pro Lys Asp65 70 7580Thr Asn Gln Thr Gln Pro Ala Thr Gln Pro Ala Asn Thr Ala Lys Asn85 90 95Tyr Pro Ala Ala Asp Glu Ser Leu Lys Asp Ala Ile Lys Asp Pro Ala100105110
Leu Glu Asn Lys Glu His Asp Ile Gly Pro Arg Glu Gln Val Asn Phe115 120125Gln Leu Leu Asp Lys Asn Asn Glu Thr Gln Tyr Tyr His Phe Phe Ser130 135 140Ile Lys Asp Pro Ala Asp Val Tyr Tyr Thr Lys Lys Lys Ala Glu Val145 150 155 160Glu Leu Asp Ile Asn Thr Ala Ser Thr Trp Lys Lys Phe Glu Val Tyr165 170 175Glu Asn Asn Gln Lys Leu Pro Val Arg Leu Val Ser Tyr Ser Pro Val180 185 190Pro Glu Asp His Ala Tyr Ile Arg Phe Pro Val Ser Asp Gly Thr Gln195 200 205Glu Leu Lys Ile Val Ser Ser Thr Gln Ile Asp Asp Gly Glu Glu Thr210 215 220Asn Tyr Asp Tyr Thr Lys Leu Val Phe Ala Lys Pro Ile Tyr Asn Asp225230 235 240Pro Ser Leu Val Lys Ser Asp Thr Asn Asp Ala Val Val Thr Asn Asp245250255Gln Ser Ser Ser Val Ala Ser Asn Gln Thr Asn Thr Asn Thr Ser Asn260 265270Gln Asn Ile Ser Thr Ile Asn Asn Ala Asn Asn Gln Pro Gln Ala Thr275 280285Thr Asn Met Ser Gln Pro Ala Gln Pro Lys Ser Ser Thr Asn Ala Asp290295 300Gln Ala Ser Ser Gln Pro Ala His Glu Thr Asn Ser Asn Gly Asn Thr305 310 315 320Asn Asp Lys Thr Asn Glu Ser Ser Asn Gln Ser Asp Val Asn Gln Gln325 330335
Tyr Pro Pro Ala Asp Glu Ser Leu Gln Asp Ala Ile Lys Asn Pro Ala340 345 350Ile Ile Asp Lys Glu His Thr Ala Asp Asn Trp Arg Pro Ile Asp Phe355 360 365Gln Met Lys Asn Asp Lys Gly Glu Arg Gln Phe Tyr His Tyr Ala Ser370 375 380Thr Val Glu Pro Ala Thr Val Ile Phe Thr Lys Thr Gly Pro Ile Ile385 390 395 400Glu Leu Gly Leu Lys Thr Ala Ser Thr Trp Lys Lys Phe Glu Val Tyr405 410 415Glu Gly Asp Lys Lys Leu Pro Val Glu Leu Val Ser Tyr Asp Ser Asp420 425430Lys Asp Tyr Ala Tyr Ile Arg Phe Pro Val Ser Asn Gly Thr Arg Glu435 440 445Val Lys Ile Val Ser Ser Ile Glu Tyr Gly Glu Asn Ile His Glu Asp450 455 460Tyr Asp Tyr Thr Leu Met Val Phe Ala Gln Pro Ile Thr Asn Asn Pro465470 475 480Asp Asp Tyr Val Asp Glu Glu Thr Tyr Asn Leu Gln Lys Leu Leu Ala485 490 495Pro Tyr His Lys Ala Lys Thr Leu Glu Arg Gln Val Tyr Glu Leu Glu500 505 510Lys Leu Gln Glu Lys Leu Pro Glu Lys Tyr Lys Ala Glu Tyr Lys Lys515 520 525Lys Leu Asp Gln Thr Arg Val Glu Leu Ala Asp Gln Val Lys Ser Ala530 535 540Val Thr Glu Phe Glu Asn Val Thr Pro Thr Asn Asp Gln Leu Thr Asp545550 555560
Leu Gln Glu Ala His Phe Val Val Phe Glu Ser Glu Glu Asn Ser Glu565 570 575Ser Val Met Asp Gly Phe Val Glu His Pro Phe Tyr Thr Ala Thr Leu580 585590Asn Gly Gln Lys Tyr Val Val Met Lys Thr Lys Asp Asp Ser Tyr Trp595 600 605Lys Asp Leu Ile Val Glu Gly Lys Arg Val Thr Thr Val Ser Lys Asp610 615 620Pro Lys Asn Asn Ser Arg Thr Leu Ile Phe Pro Tyr Ile Pro Asp Lys625630 635 640Ala Val Tyr Asn Ala Ile Val Lys Val Val Val Ala Asn Ile Gly Tyr645 650 655Glu Gly Gln Tyr His Val Arg Ile Ile Asn Gln Asp Ile Asn Thr Lys660 665 670Asp Asp Asp Thr Ser Gln Asn Asn Thr Ser Glu Pro Leu Asn Val Gln675 680 685Thr Gly Gln Glu Gly Lys Val Ala Asp Thr Asp Val Ala Glu Asn Ser690695 700Ser Thr Ala Thr Asn Pro Lys Asp Ala Ser Asp Lys Ala Asp Val Ile705 710 715 720Glu Pro Glu Ser Asp Val Val Lys Asp Ala Asp Asn Asn Ile Asp Lys725 730 735Asp Val Gln His Asp Val Asp His Leu Ser Asp Met Ser Asp Asn Asn740 745 750His Phe Asp Lys Tyr Asp Leu Lys Glu Met Asp Thr Gln Ile Ala Lys755 760765Asp Thr Asp Arg Asn Val Asp Lys Asp Ala Asp Asn Ser Val Gly Met770 775780
Ser Ser Asn Val Asp Thr Asp Lys Asp Ser Asn Lys Asn Lys Asp Lys785 790795800Val Ile Gln Leu Asn His Ile Ala Asp Lys Asn Asn His Thr Gly Lys805 810 815Ala Ala Lys Leu Asp Val Val Lys Gln Asn Tyr Asn Asn Thr Asp Lys820 825830Val Thr Asp Lys Lys Thr Thr Glu His Leu Pro Ser Asp Ile His Lys835840 845Thr Val Asp Lys Thr Val Lys Thr Lys Glu Lys Ala Gly Thr Pro Ser850 855 860Lys Glu Asn Lys Leu Ser Gln Ser Lys Met Leu Pro Lys Thr Gly Glu865870875 880Thr Thr Ser Ser Gln Ser Trp Trp Gly Leu Tyr Ala Leu Leu Gly Met885 890 895Leu Ala Leu Phe Ile Pro Lys Phe Arg Lys Glu Ser Lys900 905<210>7<211>1065<212>DNA<213>Staphylococcus epidermidis<400>7gaggaaaaca acatgacaaa acattattta aacagtaagt atcaatcaga acaacgttca 60tcagctatga aaaagattac aatgggtaca gcatctatca ttttaggttc ccttgtatac120ataggcgcag acagccaaca agtcaatgcg gcaacagaag ctacgaacgc aactaataat180caaagcacac aagtttctca agcaacatca caaccaatta atttccaagt gcaaaaagat240ggctcttcag agaagtcaca catggatgac tatatgcaac accctggtaa agtaattaaa300caaaataata aatattattt ccaaaccgtg ttaaacaatg catcattctg gaaagaatac360aaattttaca atgcaaacaa tcaagaatta gcaacaactg ttgttaacga taataaaaaa420gcggatacta gaacaatcaa tgttgcagtt gaacctggat ataagagctt aactactaaa480gtacatattg tcgtgccaca aattaattac aatcatagat atactacgca tttggaattt540
gaaaaagcaa ttcctacatt agctgacgca gcaaaaccaa acaatgttaa accggttcaa600ccaaaaccag ctcaacctaa aacacctact gagcaaacta aaccagttca acctaaagtt660gaaaaagtta aacctactgt aactacaaca agcaaagttg aagacaatca ctctactaaa720gttgtaagta ctgacacaac aaaagatcaa actaaaacac aaactgctca tacagttaaa780acagcacaaa ctgctcaaga acaaaataaa gttcaaacac ctgttaaaga tgttgcaaca840gcgaaatctg aaagcaacaa tcaagctgta agtgataata aatcacaaca aactaacaaa900gttacaaaac ataacgaaac gcctaaacaa gcatctaaag ctaaagaatt accaaaaact960ggtttaactt cagttgataa ctttattagc acagttgcct tcgcaacact tgccctttta1020ggttcattat ctttattact tttcaaaaga aaagaatcta aataa1065<210>8<211>354<212>PRT<213>Staphylococcus epidermidis<400>8Glu Glu Asn Asn Met Thr Lys His Tyr Leu Asn Ser Lys Tyr Gln Ser15 10 15Glu Gln Arg Ser Ser Ala Met Lys Lys Ile Thr Met Gly Thr Ala Ser20 25 30Ile Ile Leu Gly Ser Leu Val Tyr Ile Gly Ala Asp Ser Gln Gln Val3540 45Asn Ala Ala Thr Glu Ala Thr Asn Ala Thr Asn Asn Gln Ser Thr Gln50 55 60Val Ser Gln Ala Thr Ser Gln Pro Ile Asn Phe Gln Val Gln Lys Asp65 70 75 80Gly Ser Ser Glu Lys Ser His Met Asp Asp Tyr Met Gln His Pro Gly85 90 95Lys Val Ile Lys Gln Asn Asn Lys Tyr Tyr Phe Gln Thr Val Leu Asn100105110Asn Ala Ser Phe Trp Lys Glu Tyr Lys Phe Tyr Asn Ala Asn Asn Gln115 120 125
Glu Leu Ala Thr Thr Val Val Asn Asp Asn Lys Lys Ala Asp Thr Arg130 135140Thr Ile Asn Val Ala Val Glu Pro Gly Tyr Lys Ser Leu Thr Thr Lys145 150 155 160Val His Ile Val Val Pro Gln Ile Asn Tyr Asn His Arg Tyr Thr Thr165 170 175His Leu Glu Phe Glu Lys Ala Ile Pro Thr Leu Ala Asp Ala Ala Lys180 185 190Pro Asn Asn Val Lys Pro Val Gln Pro Lys Pro Ala Gln Pro Lys Thr195 200 205Pro Thr Glu Gln Thr Lys Pro Val Gln Pro Lys Val Glu Lys Val Lys210 215 220Pro Thr Val Thr Thr Thr Ser Lys Val Glu Asp Asn His Ser Thr Lys225 230 235 240Val Val Ser Thr Asp Thr Thr Lys Asp Gln Thr Lys Thr Gln Thr Ala245250255His Thr Val Lys Thr Ala Gln Thr Ala Gln Glu Gln Asn Lys Val Gln260265 270Thr Pro Val Lys Asp Val Ala Thr Ala Lys Ser Glu Ser Asn Asn Gln275 280 285Ala Val Ser Asp Asn Lys Ser Gln Gln Thr Asn Lys Val Thr Lys His290295 300Asn Glu Thr Pro Lys Gln Ala Ser Lys Ala Lys Glu Leu Pro Lys Thr305310 315320Gly Leu Thr Ser Val Asp Asn Phe Ile Ser Thr Val Ala Phe Ala Thr325 330 335Leu Ala Leu Leu Gly Ser Leu Ser Leu Leu Leu Phe Lys Arg Lys Glu340 345350Ser Lys
<210>9<211>1965<212>DNA<213>Staphylococcus epidermidis<400>9tatacaatta ggagttgttt ctacaacatg aacaaacagc aaaaagaatt taaatcattt 60tattcaatta gaaagtcatc actaggcgtt gcatctgtag caattagtac acttttatta120ttaatgtcaa atggcgaagc acaagcagca gctgaagaaa caggtggtac aaatacagaa180gcacaaccaa aaactgaagc agttgcaagt ccaacaacaa catctgaaaa agctccagaa240actaaaccag tagctaatgc tgtctcagta tctaataaag aagttgaggc ccctacttct300gaaacaaaag aagctaaaga agttaaagaa gttaaagccc ctaaggaaac aaaagaagtt360aaaccagcag caaaagccac taacaataca tatcctattt tgaatcagga acttagagaa420gcgattaaaa accctgcaat aaaagacaaa gatcatagcg caccaaactc tcgtccaatt480gattttgaaa tgaaaaagaa agatggaact caacagtttt atcattatgc aagttctgtt540aaacctgcta gagttatttt cactgattca aaaccagaaa ttgaattagg attacaatca600ggtcaatttt ggagaaaatt tgaagtttat gaaggtgaca aaaagttgcc aattaaatta660gtatcatacg atactgttaa agattatgct tacattcgct tctctgtatc aaacggaaca720aaagctgtta aaattgttag ttcaacacac ttcaataaca aagaagaaaa atacgattac780acattaatgg aattcgcaca accaatttat aacagtgcag ataaattcaa aactgaagaa840gattataaag ctgaaaaatt attagcgcca tataaaaaag cgaaaacact agaaagacaa900gtttatgaat taaataaaat tcaagataaa cttcctgaaa aattaaaggc tgagtacaag960aagaaattag aggatacaaa gaaagcttta gatgagcaag tgaaatcagc tattactgaa 1020ttcc aaatg tacaaccaac aaatgaaaaa atgactgatt tacaagatac aaaatatgtt 1080gtttatgaaa gtgttgagaa taacgaatct atgatggata cttttgttaa acaccctatt 1140aaaacaggta tgcttaacgg caaaaaatat atggtcatgg aaactactaa tgacgattac 1200tggaaagatt tcatggttga aggtcaacgt gttagaacta taagcaaaga tgctaaaaat 1260aatactagaa caattatttt cccatatgtt gaaggtaaaa ctctatatga tgctatcgtt 1320aaagttcacg taaaaacgat tgattatgat ggacaatacc atgtcagaat cgttgataaa 1380gaagcattta caaaagccaa taccgataaa tctaacaaaa aagaacaaca agataactca 1440gctaagaagg aagctactcc agctacgcct agcaaaccaa caccatcacc tgttgaaaaa 1500
gaatcacaaa aacaagacag ccaaaaagat gacaataaac aattaccaag tgttgaaaaa 1560gaaaatgacg catctagtga gtcaggtaaa gacaaaacgc ctgctacaaa accaactaaa 1620ggtgaagtag aatcaagtag tacaactcca actaaggtag tatctacgac tcaaaatgtt 1680gcaaaaccaa caactgcttc atcaaaaaca acaaaagatg ttgttcaaac ttcagcaggt 1740tctagcgaag caaaagatag tgctccatta caaaaagcaa acattaaaaa cacaaatgat 1800ggacacactc aaagccaaaa caataaaaat acacaagaaa ataaagcaaa atcattacca 1860caaactggtg aagaatcaaa taaagatatg acattaccat taatggcatt attagcttta 1920agtagcatcg ttgcattcgt attacctaga aaacgtaaaa actaa 1965<210>10<211>654<212>PRT<213>Staphylococcus epidermidis<400>10Tyr Thr Ile Arg Ser Cys Phe Tyr Asn Met Asn Lys Gln Gln Lys Glu1 5 10 15Phe Lys Ser Phe Tyr Ser Ile Arg Lys Ser Ser Leu Gly Val Ala Ser20 25 30Val Ala Ile Ser Thr Leu Leu Leu Leu Met Ser Asn Gly Glu Ala Gln35 40 45Ala Ala Ala Glu Glu Thr Gly Gly Thr Asn Thr Glu Ala Gln Pro Lys50 55 60Thr Glu Ala Val Ala Ser Pro Thr Thr Thr Ser Glu Lys Ala Pro Glu65 70 75 80Thr Lys Pro Val Ala Asn Ala Val Ser Val Ser Asn Lys Glu Val Glu85 9095Ala Pro Thr Ser Glu Thr Lys Glu Ala Lys Glu Val Lys Glu Val Lys100 105 110Ala Pro Lys Glu Thr Lys Glu Val Lys Pro Ala Ala Lys Ala Thr Asn115 120 125
Asn Thr Tyr Pro Ile Leu Asn Gln Glu Leu Arg Glu Ala Ile Lys Asn130 135 140Pro Ala Ile Lys Asp Lys Asp His Ser Ala Pro Asn Ser Arg Pro Ile145 150 155 160Asp Phe Glu Met Lys Lys Lys Asp Gly Thr Gln Gln Phe Tyr His Tyr165 170 175Ala Ser Ser Val Lys Pro Ala Arg Val Ile Phe Thr Asp Ser Lys Pro180 185190Glu Ile Glu Leu Gly Leu Gln Ser Gly Gln Phe Trp Arg Lys Phe Glu195200 205Val Tyr Glu Gly Asp Lys Lys Leu Pro Ile Lys Leu Val Ser Tyr Asp210215 220Thr Val Lys Asp Tyr Ala Tyr Ile Arg Phe Ser Val Ser Asn Gly Thr225 230 235 240Lys Ala Val Lys Ile Val Ser Ser Thr His Phe Asn Asn Lys Glu Glu245 250 255Lys Tyr Asp Tyr Thr Leu Met Glu Phe Ala Gln Pro Ile Tyr Asn Ser260265 270Ala Asp Lys Phe Lys Thr Glu Glu Asp Tyr Lys Ala Glu Lys Leu Leu275 280285Ala Pro Tyr Lys Lys Ala Lys Thr Leu Glu Arg Gln Val Tyr Glu Leu290 295 300Asn Lys Ile Gln Asp Lys Leu Pro Glu Lys Leu Lys Ala Glu Tyr Lys305 310 315 320Lys Lys Leu Glu Asp Thr Lys Lys Ala Leu Asp Glu Gln Val Lys Ser325 330 335Ala Ile Thr Glu Phe Gln Asn Val Gln Pro Thr Asn Glu Lys Met Thr340345350
Asp Leu Gln Asp Thr Lys Tyr Val Val Tyr Glu Ser Val Glu Asn Asn355 360 365Glu Ser Met Met Asp Thr Phe Val Lys His Pro Ile Lys Thr Gly Met370 375 380Leu Asn Gly Lys Lys Tyr Met Val Met Glu Thr Thr Asn Asp Asp Tyr385390395 400Trp Lys Asp Phe Met Val Glu Gly Gln Arg Val Arg Thr Ile Ser Lys405 410 415Asp Ala Lys Asn Asn Thr Arg Thr Ile Ile Phe Pro Tyr Val Glu Gly420 425 430Lys Thr Leu Tyr Asp Ala Ile Val Lys Val His Val Lys Thr Ile Asp435 440 445Tyr Asp Gly Gln Tyr His Val Arg Ile Val Asp Lys Glu Ala Phe Thr450 455 460Lys Ala Asn Thr Asp Lys Ser Asn Lys Lys Glu Gln Gln Asp Asn Ser465 470 475 480Ala Lys Lys Glu Ala Thr Pro Ala Thr Pro Ser Lys Pro Thr Pro Ser485 490 495Pro Val Glu Lys Glu Ser Gln Lys Gln Asp Ser Gln Lys Asp Asp Asn500 505510Lys Gln Leu Pro Ser Val Glu Lys Glu Asn Asp Ala Ser Ser Glu Ser515 520 525Gly Lys Asp Lys Thr Pro Ala Thr Lys Pro Thr Lys Gly Glu Val Glu530535 540Ser Ser Ser Thr Thr Pro Thr Lys Val Val Ser Thr Thr Gln Asn Val545 550 555 560Ala Lys Pro Thr Thr Ala Ser Ser Lys Thr Thr Lys Asp Val Val Gln565 570 575
Thr Ser Ala Gly Ser Ser Glu Ala Lys Asp Ser Ala Pro Leu Gln Lys580585 590Ala Asn Ile Lys Asn Thr Asn Asp Gly His Thr Gln Ser Gln Asn Asn595600 605Lys Asn Thr Gln Glu Asn Lys Ala Lys Ser Leu Pro Gln Thr Gly Glu610615 620Glu Ser Asn Lys Asp Met Thr Leu Pro Leu Met Ala Leu Leu Ala Leu625 630 635 640Ser Ser Ile Val Ala Phe Val Leu Pro Arg Lys Arg Lys Asn645 650<210>11<211>2406<212>DNA<213>Staphylococcus epidermidis<400>11tttataaata atttacataa aatcaatcat tttaatataa ggattatgat aatatattgg 60tgtatgacag ttaatggagg gaacgaaatg aaagctttat tacttaaaac aagtgtatgg120ctcgttttgc tttttagtgt aatgggatta tggcaagtct cgaacgcggc tgagcagcat180acaccaatga aagcacatgc agtaacaacg atagacaaag caacaacaga taagcaacaa240gtaccgccaa caaaggaagc ggctcatcat tctggcaaag aagcggcaac caacgtatca300gcatcagcgc agggaacagc tgatgataca aacagcaaag taacatccaa cgcaccatct360aacaaaccat ctacagtagt ttcaacaaaa gtaaacgaaa cacgcgacgt agatacacaa420caagcctcaa cacaaaaacc aactcacaca gcaacgttca aattatcaaa tgctaaaaca480gcatcacttt caccacgaat gtttgctgct aatgcaccac aaacaacaac acataaaata540ttacatacaa atgatatcca tggccgacta gccgaagaaa aagggcgtgt catcggtatg600gctaaattaa aaacagtaaa agaacaagaa aagcctgatt taatgttaga cgcaggagac660gccttccaag gtttaccact ttcaaaccag tctaaaggtg aagaaatggc taaagcaatg720aatgcagtag gttatgatgc tatggcagtc ggtaaccatg aatttgactt tggatacgat780cagttgaaaa agttagaggg tatgttagac ttcccgatgc taagtactaa cgtttataaa840gatggaaaac gcgcgtttaa gccttcaacg attgtaacaa aaaatggtat tcgttatgga900
attattggtg taacgacacc agaaacaaag acgaaaacaa gacctgaagg cattaaaggc960gttgaattta gagatccatt acaaagtgtg acagcggaaa tgatgcgtat ttataaagac 1020gtagatacat ttgttgttat atcacattta ggaattgatc cttcaacaca agaaacatgg 1080cgtggtgatt acttagtgaa acaattaagt caaaatccac aattgaagaa acgtattaca 1140gttattgatg gtcattcaca tacagtactt caaaatggtc aaatttataa caatgatgca 1200ttggcacaaa caggtacagc acttgcgaat atcggtaaga ttacatttaa ttatcgcaat 1260ggagaggtat cgaatattaa accgtcattg attaatgtta aagacgttga aaatgtaaca 1320ccgaacaaag cattagctga acaaattaat caagctgatc aaacatttag agcacaaact 1380gcagaggtaa ttattccaaa caataccatt gatttcaaag gagaaagaga tgacgttaga 1440acgcgtgaaa caaatttagg aaacgcgatt gcagatgcta tggaagcgta tggcgttaag 1500aatttctcta aaaagactga ctttgccgtg acaaatggtg gaggtattcg tgcctctatc 1560gcaaaaggta aggtgacacg ctatgattta atctcagtat taccatttgg aaatacgatt 1620gcgcaaattg atgtaaaagg ttcagacgtc tggacggctt tcgaacatag tttaggcgca 1680ccaacaacac aaaaggacgg taagacagtg ttaacagcga atggcggttt actacatatc 1740tctgattcaa tccgtgttta ctatgatata aataaaccgt ctggcaaacg aattaatgct 1800attcaaattt taaataaaga gacaggtaag tttgaaaata ttgatttaaa acgtgtatat 1860cacgtaacga tgaatgactt cacagcatca ggtggcgacg gatatagtat gttcggtggt 1920cctagagaag aaggtatttc attagatcaa gtactagcaa gttatttaaa aacagctaac 1980ttagctaagt atgatacgac agaaccacaa cgtatgttat taggtaaacc agcagtaagt 2040gaacaaccag ctaaaggaca acaaggtagc aaaggtagta agtctggtaa agatacacaa 2100ccaattggtg acgacaaagt gatggatcca gcgaaaaaac cagctccagg taaagttgta 2160ttgttgctag cgcatagagg aactgttagt agcggtacag aaggttctgg tcgcacaata 2220gaaggagcta ctgtatcaag caagagtggg aaacaattgg ctagaatgtc agtgcctaaa 2280ggtagcgcgc atgagaaaca gttaccaaaa actggaacta atcaaagttc aagcccagaa 2340gcgatgtttg tattattagc aggtataggt ttaatcgcga ctgtacgacg tagaaaagct 2400agctaa 2406<210>12<211>801<212>PRT
<213>Staphylococcus epidermidis<400>12Phe Ile Asn Asn Leu His Lys Ile Asn His Phe Asn Ile Arg Ile Met1 5 10 15Ile Ile Tyr Trp Cys Met Thr Val Asn Gly Gly Asn Glu Met Lys Ala20 2530Leu Leu Leu Lys Thr Ser Val Trp Leu Val Leu Leu Phe Ser Val Met35 40 45Gly Leu Trp Gln Val Ser Asn Ala Ala Glu Gln His Thr Pro Met Lys50 55 60Ala His Ala Val Thr Thr Ile Asp Lys Ala Thr Thr Asp Lys Gln Gln65 70 7580Val Pro Pro Thr Lys Glu Ala Ala His His Ser Gly Lys Glu Ala Ala85 9095Thr Asn Val Ser Ala Ser Ala Gln Gly Thr Ala Asp Asp Thr Asn Ser100 105110Lys Val Thr Ser Asn Ala Pro Ser Asn Lys Pro Ser Thr Val Val Ser115 120 125Thr Lys Val Asn Glu Thr Arg Asp Val Asp Thr Gln Gln Ala Ser Thr130135 140Gln Lys Pro Thr His Thr Ala Thr Phe Lys Leu Ser Asn Ala Lys Thr145 150 155 160Ala Ser Leu Ser Pro Arg Met Phe Ala Ala Asn Ala Pro Gln Thr Thr165 170 175Thr His Lys Ile Leu His Thr Asn Asp Ile His Gly Arg Leu Ala Glu180 185 190Glu Lys Gly Arg Val Ile Gly Met Ala Lys Leu Lys Thr Val Lys Glu195 200 205
Gln Glu Lys Pro Asp Leu Met Leu Asp Ala Gly Asp Ala Phe Gln Gly210 215220Leu Pro Leu Ser Asn Gln Ser Lys Gly Glu Glu Met Ala Lys Ala Met225230235 240Asn Ala Val Gly Tyr Asp Ala Met Ala Val Gly Asn His Glu Phe Asp245250 255Phe Gly Tyr Asp Gln Leu Lys Lys Leu Glu Gly Met Leu Asp Phe Pro260265270Met Leu Ser Thr Asn Val Tyr Lys Asp Gly Lys Arg Ala Phe Lys Pro275 280 285Ser Thr Ile Val Thr Lys Asn Gly Ile Arg Tyr Gly Ile Ile Gly Val290 295300Thr Thr Pro Glu Thr Lys Thr Lys Thr Arg Pro Glu Gly Ile Lys Gly305 310 315 320Val Glu Phe Arg Asp Pro Leu Gln Ser Val Thr Ala Glu Met Met Arg325 330 335Ile Tyr Lys Asp Val Asp Thr Phe Val Val Ile Ser His Leu Gly Ile340 345350Asp Pro Ser Thr Gln Glu Thr Trp Arg Gly Asp Tyr Leu Val Lys Gln355 360 365Leu Ser Gln Asn Pro Gln Leu Lys Lys Arg Ile Thr Val Ile Asp Gly370375 380His Ser His Thr Val Leu Gln Asn Gly Gln Ile Tyr Asn Asn Asp Ala385 390 395 400Leu Ala Gln Thr Gly Thr Ala Leu Ala Asn Ile Gly Lys Ile Thr Phe405 410415Asn Tyr Arg Asn Gly Glu Val Ser Asn Ile Lys Pro Ser Leu Ile Asn420 425 430
Val Lys Asp Val Glu Asn Val Thr Pro Asn Lys Ala Leu Ala Glu Gln435 440 445Ile Asn Gln Ala Asp Gln Thr Phe Arg Ala Gln Thr Ala Glu Val Ile450455 460Ile Pro Asn Asn Thr Ile Asp Phe Lys Gly Glu Arg Asp Asp Val Arg465 470 475 480Thr Arg Glu Thr Asn Leu Gly Asn Ala Ile Ala Asp Ala Met Glu Ala485 490495Tyr Gly Val Lys Asn Phe Ser Lys Lys Thr Asp Phe Ala Val Thr Asn500 505 510Gly Gly Gly Ile Arg Ala Ser Ile Ala Lys Gly Lys Val Thr Arg Tyr515 520 525Asp Leu Ile Ser Val Leu Pro Phe Gly Asn Thr Ile Ala Gln Ile Asp530 535 540Val Lys Gly Ser Asp Val Trp Thr Ala Phe Glu His Ser Leu Gly Ala545 550 555 560Pro Thr Thr Gln Lys Asp Gly Lys Thr Val Leu Thr Ala Asn Gly Gly565570 575Leu Leu His Ile Ser Asp Ser Ile Arg Val Tyr Tyr Asp Ile Asn Lys580 585590Pro Ser Gly Lys Arg Ile Asn Ala Ile Gln Ile Leu Asn Lys Glu Thr595 600 605Gly Lys Phe Glu Asn Ile Asp Leu Lys Arg Val Tyr His Val Thr Met610 615 620Asn Asp Phe Thr Ala Ser Gly Gly Asp Gly Tyr Ser Met Phe Gly Gly625630 635640Pro Arg Glu Glu Gly Ile Ser Leu Asp Gln Val Leu Ala Ser Tyr Leu645 650 655
Lys Thr Ala Asn Leu Ala Lys Tyr Asp Thr Thr Glu Pro Gln Arg Met660 665670Leu Leu Gly Lys Pro Ala Val Ser Glu Gln Pro Ala Lys Gly Gln Gln675 680 685Gly Ser Lys Gly Ser Lys Ser Gly Lys Asp Thr Gln Pro Ile Gly Asp690 695 700Asp Lys Val Met Asp Pro Ala Lys Lys Pro Ala Pro Gly Lys Val Val705 710715720Leu Leu Leu Ala His Arg Gly Thr Val Ser Ser Gly Thr Glu Gly Ser725 730 735Gly Arg Thr Ile Glu Gly Ala Thr Val Ser Ser Lys Ser Gly Lys Gln740 745 750Leu Ala Arg Met Ser Val Pro Lys Gly Ser Ala His Glu Lys Gln Leu755 760 765Pro Lys Thr Gly Thr Asn Gln Ser Ser Ser Pro Glu Ala Met Phe Val770775 780Leu Leu Ala Gly Ile Gly Leu Ile Ala Thr Val Arg Arg Arg Lys Ala785 790 795 800Ser<210>13<211>4914<212>DNA<213>Staphylococcus epidermidis<400>13agtggaaaat atggaaaaag gagtatgcaa atgagagata agaaaggacc ggtaaataaa 60agagtagatt ttctatcaaa taaattgaat aaatattcaa taagaaaatt tacagttgga120acagcatcta ttttaattgg ctcactaatg tatttgggaa ctcaacaaga ggcagaagca180gctgaaaaca atattgagaa tccaactaca ttaaaagata atgtccaatc aaaagaagtg240aagattgaag aagtaacaaa caaagacact gcaccacagg gtgtagaagc taaatctgaa300
gtaacttcaa acaaagacac aatcgaacat gaaccatcag taaaagctga agatatatca360aaaaaggagg atacaccaaa agaagtagct gatgttgctg aagttcagcc gaaatcgtca420gtcactcata acgcagagac acctaaggtt agaaaagctc gttctgttga tgaaggctct480tttgatatta caagagattc taaaaatgta gttgaatcta ccccaattac aattcaaggt540aaagaacatt ttgaaggtta cggaagtgtt gatatacaaa aaaaaccaac agatttaggg600gtatcagagg taaccaggtt taatgttggt aatgaaagta atggtttgat aggagcttta660caattaaaaa ataaaataga ttttagtaag gatttcaatt ttaaagttag agtggcaaat720aaccatcaat caaataccac aggtgctgat ggttgggggt tcttatttag taaaggaaat780gcagaagaat atttaactaa tggtggaatc cttggggata aaggtctggt aaattcaggc840ggatttaaaa ttgatactgg atacatttat acaagttcca tggacaaaac tgaaaagcaa900gctggacaag gttatagagg atacggagct tttgtgtaaa atgacagttc tggtaattca960caaatggttg gagaaaatat tgataaatca aaaactaatt ttttaaacta tgcggacaat 1020tcaactaata catcagatgg aaagtttcat gggcaacgtt taaatgatgt catcttaact 1080tatgttgctt caactggtaa aatgagagca gaatatgctg gtaaaacttg ggagacttca 1140ataacagatt taggtttatc taaaaatcag gcatataatt tcttaattac atctagtcaa 1200agatggggcc ttaatcaagg gataaatgca aatggctgga tgagaactga cttgaaaggt 1260tcagagttta cttttacacc agaagcgcca aaaacaataa cagaattaga aaaaaaagtt 1320gaagagattc cattcaagaa agaacgtaaa tttaatccgg atttagcacc agggacagaa 1380aaagtaacaa gagaaggaca aaaaggtgag aagacaataa cgacaccaac actaaaaaat 1440ccattaactg gagtaattat tagtaaaggt gaaccaaaag aagagattac aaaagatccg 1500attaatgaat taacagaata cggacctgaa acaatagcgc caggtcatcg agacgaattt 1560gatccgaagt taccaacagg agagaaagag gaagttccag gtaaaccagg aattaagaat 1620ccagaaacag gagacgtagt tagaccgccg gtcgatagcg taacaaaata tggacctgta 1680aaaggagact cgattgtaga aaaagaagag attccattcg agaaagaacg taaatttaat 1740cctgatttag caccagggac agaaaaagta acaagagaag gacaaaaagg tgagaagaca 1800ataacgacgc caacactaaa aaatccatta actggagaaa ttattagtaa aggtgaatcg 1860aaagaagaaa tcacaaaaga tccgattaat gaattaacag aatacggacc agaaacgata 1920acaccaggtc atcgagacga atttgatccg aagttaccaa caggagagaa agaggaagtt 1980
ccaggtaaac caggaattaa gaatccagaa acaggagatg tagttagacc accggtcgat 2040agcgtaacaa aatatggacc tgtaaaagga gactcgattg tagaaaaaga agagattcca 2100ttcgagaaag aacgtaaatt taatcctgat ttagcaccag ggacagaaaa agtaacaaga 2160gaaggacaaa aaggtgagaa gacaataacg acaccaacac taaaaaatcc attaactgga 2220gtaattatta gtaaaggtga accaaaagaa gaaatcacaa aagatccgat taatgaatta 2280acagaatacg gaccagaaac gataacacca ggtcatcgag acgaatttga tccgaagtta 2340ccaacaggag agaaagaaga agttccaggt aaaccaggaa ttaagaatcc agaaacagga 2400gacgtagtta gaccaccggt cgatagcgta acaaaatatg gacctgtaaa aggagactcg 2460attgtagaaa aagaagagat tccattcaag aaagaacgta aatttaatcc ggatttagca 2520ccagggacag aaaaagtaac aagagaagga caaaaaggtg agaagacaat aacgacgcca 2580acactaaaaa atccattaac tggagaaatt attagtaaag gtgaatcgaa agaagaaatc 2640acaaaagatc cgattaatga attaacagaa tacggaccag aaacgataac accaggtcat 2700cgagacgaat ttgatccgaa gttaccaaca ggagagaaag aggaagttcc aggtaaacca 2760ggaattaaga atccagaaac aggagatgta gttagaccac cggtcgatag cgtaacaaaa 2820tatggacctg taaaaggaga ctcgattgta gaaaaagaag agattccatt cgagaaagaa 2880cgtaaattta atcctgattt agcaccaggg acagaaaaag taacaagaga aggacaaaaa 2940ggtgagaaga caataacgac gccaacacta aaaaatccat taactggaga aattattagt 3000aaaggtgaat cgaaagaaga aatcacaaaa gatccgatta atgaattaac agaatacgga 3060ccagaaacga taacaccagg tcatcgagac gaatttgatc cgaagttacc aacaggagag 3120aaagaggaag ttccaggtaa accaggaatt aagaatccag aaacaggaga cgtagttaga 3180ccaccggtcg atagcgtaac aaaatatgga cctgtaaaag gagactcgat tgtagaaaaa 3240gaagaaattc cattcaagaa agaacgtaaa tttaatcctg atttagcacc agggacagaa 3300aaagtaacaa gagaaggaca aaaaggtgag aagacaataa cgacgccaac actaaaaaat 3360ccattaactg gagaaattat tagtaaaggt gaatcgaaag aagaaatcac aaaagatccg 3420attaatgaat taacagaata cggaccagaa acgataacac caggtcatcg agacgaattt 3480gatccgaagt taccaacagg agagaaagag gaagttccag gtaaaccagg aattaagaat 3540ccagaaacag gagatgtagt tagaccaccg gtcgatagcg taacaaaata tggacctgta 3600aaaggagact cgattgtaga aaaagaagaa attccattcg agaaagaacg taaatttaat 3660
cctgatttag caccagggac agaaaaagta acaagagaag gacaaaaagg tgagaagaca 3720ataacgacgc caacactaaa aaatccatta actggagaaa ttattagtaa aggtgaatcg 3780aaagaagaaa tcacaaaaga tccgattaat gaattaacag aatacggacc agaaacgata 3840acaccaggtc atcgagacga atttgatccg aagttaccaa caggagagaa agaggaagtt 3900ccaggtaaac caggaattaa gaatccagaa acaggagatg tagttagacc accggtcgat 3960agcgtaacaa aatatggacc tgtaaaagga gactcgattg tagaaaaaga agaaattcca 4020ttcgagaaag aacgtaaatt taatcctgat ttagcaccag ggacagaaaa agtaacaaga 4080gaaggacaaa aaggtgagaa gacaataacg acgccaacac taaaaaatcc attaactgga 4140gaaattatta gtaaaggtga atcgaaagaa gaaatcacaa aagatccagt taatgaatta 4200acagaattcg gtggcgagaa aataccgcaa ggtcataaag atatctttga tccaaactta 4260ccaacagatc aaacggaaaa agtaccaggt aaaccaggaa tcaagaatcc agacacagga 4320aaagtgatcg aagagccagt ggatgatgtg attaaacacg gaccaaaaac gggtacacca 4380gaaacaaaaa cagtagagat accgtttgaa acaaaacgtg agtttaatcc aaaattacaa 4440cctggtgaag agcgagtgaa acaagaagga caaccaggaa gtaagacaat cacaacacca 4500atcacagtga acccattaac aggtgaaaaa gttggcgagg gtcaaccaac agaagagatc 4560acaaaacaac cagtagataa gattgtagag ttcggtggag agaaaccaaa agatccaaaa 4620ggacctgaaa acccagagaa gccgagcaga ccaactcatc caagtggccc agtaaatcct 4680aacaatccag gattatcgaa agacagagca aaaccaaatg gcccagttca ttcaatggat 4740aaaaatgata aagttaaaaa atctaaaatt gctaaagaat cagtagctaa tcaagagaaa 4800aaacgagcag aattaccaaa aacaggttta gaaagcacgc aaaaaggttt gatctttagt 4860agtataattg gaattgctgg attaatgtta ttggctcgta gaagaaagaa ttaa 4914<210>14<211>1637<212>PRT<213>Staphylococcus epidermidis<400>14Ser Gly Lys Tyr Gly Lys Arg Ser Met Gln Met Arg Asp Lys Lys Gly1 5 10 15Pro Val Asn Lys Arg Val Asp Phe Leu Ser Asn Lys Leu Asn Lys Tyr20 25 30
Ser Ile Arg Lys Phe Thr Val Gly Thr Ala Ser Ile Leu Ile Gly Ser35 4045Leu Met Tyr Leu Gly Thr Gln Gln Glu Ala Glu Ala Ala Glu Asn Asn50 55 60Ile Glu Asn Pro Thr Thr Leu Lys Asp Asn Val Gln Ser Lys Glu Val65 70 75 80Lys Ile Glu Glu Val Thr Asn Lys Asp Thr Ala Pro Gln Gly Val Glu85 90 95Ala Lys Ser Glu Val Thr Ser Asn Lys Asp Thr Ile Glu His Glu Pro100 105 110Ser Val Lys Ala Glu Asp Ile Ser Lys Lys Glu Asp Thr Pro Lys Glu115 120 125Val Ala Asp Val Ala Glu Val Gln Pro Lys Ser Ser Val Thr His Asn130 135 140Ala Glu Thr Pro Lys Val Arg Lys Ala Arg Ser Val Asp Glu Gly Ser145 150 155 160Phe Asp Ile Thr Arg Asp Ser Lys Asn Val Val Glu Ser Thr Pro Ile165 170175Thr Ile Gln Gly Lys Glu His Phe Glu Gly Tyr Gly Ser Val Asp Ile180185 190Gln Lys Lys Pro Thr Asp Leu Gly Val Ser Glu Val Thr Arg Phe Asn195 200 205Val Gly Asn Glu Ser Asn Gly Leu Ile Gly Ala Leu Gln Leu Lys Asn210215 220Lys Ile Asp Phe Ser Lys Asp Phe Asn Phe Lys Val Arg Val Ala Asn225 230235240Asn His Gln Ser Asn Thr Thr Gly Ala Asp Gly Trp Gly Phe Leu Phe245 250255
Ser Lys Gly Asn Ala Glu Glu Tyr Leu Thr Asn Gly Gly Ile Leu Gly260265 270Asp Lys Gly Leu Val Asn Ser Gly Gly Phe Lys Ile Asp Thr Gly Tyr275 280 285Ile Tyr Thr Ser Ser Met Asp Lys Thr Glu Lys Gln Ala Gly Gln Gly290 295 300Tyr Arg Gly Tyr Gly Ala Phe Val Lys Asn Asp Ser Ser Gly Asn Ser305 310 315 320Gln Met Val Gly Glu Asn Ile Asp Lys Ser Lys Thr Asn Phe Leu Asn325 330335Tyr Ala Asp Asn Ser Thr Asn Thr Ser Asp Gly Lys Phe His Gly Gln340 345350Arg Leu Asn Asp Val Ile Leu Thr Tyr Val Ala Ser Thr Gly Lys Met355 360 365Arg Ala Glu Tyr Ala Gly Lys Thr Trp Glu Thr Ser Ile Thr Asp Leu370 375 380Gly Leu Ser Lys Asn Gln Ala Tyr Asn Phe Leu Ile Thr Ser Ser Gln385390 395 400Arg Trp Gly Leu Asn Gln Gly Ile Asn Ala Asn Gly Trp Met Arg Thr405 410415Asp Leu Lys Gly Ser Glu Phe Thr Phe Thr Pro Glu Ala Pro Lys Thr420 425 430Ile Thr Glu Leu Glu Lys Lys Val Glu Glu Ile Pro Phe Lys Lys Glu435 440 445Arg Lys Phe Asn Pro Asp Leu Ala Pro Gly Thr Glu Lys Val Thr Arg450 455 460Glu Gly Gln Lys Gly Glu Lys Thr Ile Thr Thr Pro Thr Leu Lys Asn465470 475 480
Pro Leu Thr Gly Val Ile Ile Ser Lys Gly Glu Pro Lys Glu Glu Ile485 490495Thr Lys Asp Pro Ile Asn Glu Leu Thr Glu Tyr Gly Pro Glu Thr Ile500 505 510Ala Pro Gly His Arg Asp Glu Phe Asp Pro Lys Leu Pro Thr Gly Glu515 520525Lys Glu Glu Val Pro Gly Lys Pro Gly Ile Lys Asn Pro Glu Thr Gly530 535 540Asp Val Val Arg Pro Pro Val Asp Ser Val Thr Lys Tyr Gly Pro Val545 550 555 560Lys Gly Asp Ser Ile Val Glu Lys Glu Glu Ile Pro Phe Glu Lys Glu565 570 575Arg Lys Phe Asn Pro Asp Leu Ala Pro Gly Thr Glu Lys Val Thr Arg580 585 590Glu Gly Gln Lys Gly Glu Lys Thr Ile Thr Thr Pro Thr Leu Lys Asn595 600 605Pro Leu Thr Gly Glu Ile Ile Ser Lys Gly Glu Ser Lys Glu Glu Ile610 615 620Thr Lys Asp Pro Ile Asn Glu Leu Thr Glu Tyr Gly Pro Glu Thr Ile625 630 635 640Thr Pro Gly His Arg Asp Glu Phe Asp Pro Lys Leu Pro Thr Gly Glu645650 655Lys Glu Glu Val Pro Gly Lys Pro Gly Ile Lys Asn Pro Glu Thr Gly660 665670Asp Val Val Arg Pro Pro Val Asp Ser Val Thr Lys Tyr Gly Pro Val675 680 685Lys Gly Asp Ser Ile Val Glu Lys Glu Glu Ile Pro Phe Glu Lys Glu690 695 700
Arg Lys Phe Asn Pro Asp Leu Ala Pro Gly Thr Glu Lys Val Thr Arg705710 715720Glu Gly Gln Lys Gly Glu Lys Thr Ile Thr Thr Pro Thr Leu Lys Asn725 730735Pro Leu Thr Gly Val Ile Ile Ser Lys Gly Glu Pro Lys Glu Glu Ile740 745 750Thr Lys Asp Pro Ile Asn Glu Leu Thr Glu Tyr Gly Pro Glu Thr Ile755 760 765Thr Pro Gly His Arg Asp Glu Phe Asp Pro Lys Leu Pro Thr Gly Glu770775 780Lys Glu Glu Val Pro Gly Lys Pro Gly Ile Lys Asn Pro Glu Thr Gly785 790 795 800Asp Val Val Arg Pro Pro Val Asp Ser Val Thr Lys Tyr Gly Pro Val805 810 815Lys Gly Asp Ser Ile Val Glu Lys Glu Glu Ile Pro Phe Lys Lys Glu820 825 830Arg Lys Phe Asn Pro Asp Leu Ala Pro Gly Thr Glu Lys Val Thr Arg835840 845Glu Gly Gln Lys Gly Glu Lys Thr Ile Thr Thr Pro Thr Leu Lys Asn850 855 860Pro Leu Thr Gly Glu Ile Ile Ser Lys Gly Glu Ser Lys Glu Glu Ile865 870 875 880Thr Lys Asp Pro Ile Asn Glu Leu Thr Glu Tyr Gly Pro Glu Thr Ile885890895Thr Pro Gly His Arg Asp Glu Phe Asp Pro Lys Leu Pro Thr Gly Glu900905 910Lys Glu Glu Val Pro Gly Lys Pro Gly Ile Lys Asn Pro Glu Thr Gly915 920 925
Asp Val Val Arg Pro Pro Val Asp Ser Val Thr Lys Tyr Gly Pro Val930 935 940Lys Gly Asp Ser Ile Val Glu Lys Glu Glu Ile Pro Phe Glu Lys Glu945 950 955 960Arg Lys Phe Asn Pro Asp Leu Ala Pro Gly Thr Glu Lys Val Thr Arg965970 975Glu Gly Gln Lys Gly Glu Lys Thr Ile Thr Thr Pro Thr Leu Lys Asn980 985990Pro Leu Thr Gly Glu Ile Ile Ser Lys Gly Glu Ser Lys Glu Glu Ile995 1000 1005Thr Lys Asp Pro Ile Asn Glu Leu Thr Glu Tyr Gly Pro Glu Thr1010 1015 1020Ile Thr Pro Gly His Arg Asp Glu Phe Asp Pro Lys Leu Pro Thr1025 1030 1035Gly Glu Lys Glu Glu Val Pro Gly Lys Pro Gly Ile Lys Asn Pro1040 1045 1050Glu Thr Gly Asp Val Val Arg Pro Pro Val Asp Ser Val Thr Lys1055 1060 1065Tyr Gly Pro Val Lys Gly Asp Ser Ile Val Glu Lys Glu Glu Ile1070 1075 1080Pro Phe Lys Lys Glu Arg Lys Phe Asn Pro Asp Leu Ala Pro Gly1085 1090 1095Thr Glu Lys Val Thr Arg Glu Gly Gln Lys Gly Glu Lys Thr Ile1100 1105 1110Thr Thr Pro Thr Leu Lys Asn Pro Leu Thr Gly Glu Ile Ile Ser1115 1120 1125Lys Gly Glu Ser Lys Glu Glu Ile Thr Lys Asp Pro Ile Asn Glu1130 1135 1140
Leu Thr Glu Tyr Gly Pro Glu Thr Ile Thr Pro Gly His Arg Asp1145 1150 1155Glu Phe Asp Pro Lys Leu Pro Thr Gly Glu Lys Glu Glu Val Pro1160 1165 1170Gly Lys Pro Gly Ile Lys Asn Pro Glu Thr Gly Asp Val Val Arg1175 1180 1185Pro Pro Val Asp Ser Val Thr Lys Tyr Gly Pro Val Lys Gly Asp1190 1195 1200Ser Ile Val Glu Lys Glu Glu Ile Pro Phe Glu Lys Glu Arg Lys1205 1210 1215Phe Asn Pro Asp Leu Ala Pro Gly Thr Glu Lys Val Thr Arg Glu1220 1225 1230Gly Gln Lys Gly Glu Lys Thr Ile Thr Thr Pro Thr Leu Lys Asn1235 1240 1245Pro Leu Thr Gly Glu Ile Ile Ser Lys Gly Glu Ser Lys Glu Glu1250 1255 1260Ile Thr Lys Asp Pro Ile Asn Glu Leu Thr Glu Tyr Gly Pro Glu1265 1270 1275Thr Ile Thr Pro Gly His Arg Asp Glu Phe Asp Pro Lys Leu Pro1280 1285 1290Thr Gly Glu Lys Glu Glu Val Pro Gly Lys Pro Gly Ile Lys Asn1295 1300 1305Pro Glu Thr Gly Asp Val Val Arg Pro Pro Val Asp Ser Val Thr1310 1315 1320Lys Tyr Gly Pro Val Lys Gly Asp Ser Ile Val Glu Lys Glu Glu1325 1330 1335Ile Pro Phe Glu Lys Glu Arg Lys Phe Asn Pro Asp Leu Ala Pro1340 1345 1350
Gly Thr Glu Lys Val Thr Arg Glu Gly Gln Lys Gly Glu Lys Thr1355 1360 1365Ile Thr Thr Pro Thr Leu Lys Asn Pro Leu Thr Gly Glu Ile Ile1370 1375 1380Ser Lys Gly Glu Ser Lys Glu Glu Ile Thr Lys Asp Pro Val Asn1385 1390 1395Glu Leu Thr Glu Phe Gly Gly Glu Lys Ile Pro Gln Gly His Lys1400 1405 1410Asp Ile Phe Asp Pro Asn Leu Pro Thr Asp Gln Thr Glu Lys Val1415 1420 1425Pro Gly Lys Pro Gly Ile Lys Asn Pro Asp Thr Gly Lys Val Ile1430 1435 1440Glu Glu Pro Val Asp Asp Val Ile Lys His Gly Pro Lys Thr Gly1445 1450 1455Thr Pro Glu Thr Lys Thr Val Glu Ile Pro Phe Glu Thr Lys Arg1460 1465 1470Glu Phe Asn Pro Lys Leu Gln Pro Gly Glu Glu Arg Val Lys Gln1475 1480 1485Glu Gly Gln Pro Gly Ser Lys Thr Ile Thr Thr Pro Ile Thr Val1490 1495 1500Asn Pro Leu Thr Gly Glu Lys Val Gly Glu Gly Gln Pro Thr Glu1505 1510 1515Glu Ile Thr Lys Gln Pro Val Asp Lys Ile Val Glu Phe Gly Gly1520 1525 1530Glu Lys Pro Lys Asp Pro Lys Gly Pro Glu Asn Pro Glu Lys Pro1535 1540 1545Ser Arg Pro Thr His Pro Ser Gly Pro Val Asn Pro Asn Asn Pro1550 1555 1560
Gly Leu Ser Lys Asp Arg Ala Lys Pro Asn Gly Pro Val His Ser1565 1570 1575Met Asp Lys Asn Asp Lys Val Lys Lys Ser Lys Ile Ala Lys Glu1580 1585 1590Ser Val Ala Asn Gln Glu Lys Lys Arg Ala Glu Leu Pro Lys Thr1595 1600 1605Gly Leu Glu Ser Thr Gln Lys Gly Leu Ile Phe Ser Ser Ile Ile1610 1615 1620Gly Ile Ala Gly Leu Met Leu Leu Ala Arg Arg Arg Lys Asn1625 1630 1635<210>15<211>1923<212>DNA<213>Staphylococcus epidermidis<400>15ggaaggagta tgttgatggc taaatatcga gggaaaccgt ttcaattata tgtaaagtta 60tcgtgttcga caatgatggc gacaagtatc attttaacga atatcttgcc gtacgatgcc120caagctgcat ctgaaaagga tactgaaatt acaaaagaga tattatctaa gcaagattta180ttagacaaag ttgacaaggc aattcgtcaa attgagcaat taaaacagtt atcggcttca240tctaaagaac attataaagc acaactaaat gaagcgaaaa cagcatcgca aatagatgaa300atcataaaac gagctaatga gttggatagc aaagacaata aaagttctca cactgaaatg360aacggtcaaa gtgatataga cagtaaatta gatcaattgc ttaaagattt aaatgaggtt420tcttcaaatg ttgatagggg tcaacaaagt ggcgaggacg atcttaatgc aatgaaaaat480gatatgtcac aaacggctac aacaaaacat ggagaaaaag atgataaaaa tgatgaagca540atggtaaata aggcgttaga agacctagac catttgaatc agcaaataca caaatcgaaa600gatgcatcga aagatacatc ggaagatcca gcagtgtcta caacagataa taatcatgaa660gtagctaaaa cgccaaataa tgatggttct ggacatgttg tgttaaataa attcctttca720aatgaagaga atcaaagcca tagtaatcga ctcactgata aattacaagg aagcgataaa780attaatcatg ctatgattga aaaattagct aaaagtaatg cctcaacgca acattacaca840tatcataaac tgaatacgtt acaatcttta gatcaacgta ttgcaaatac gcaacttcct900
aaaaatcaaa aatcagactt aatgagcgaa gtaaataaga cgaaagagcg tataaaaagt960caacgaaata ttattttgga agaacttgca cgtactgatg ataaaaagta tgctacacaa 1020agcattttag aaagtatatt taataaagac gaggcagtta aaattctaaa agatatacgt 1080gttgatggta aaacagatca acaaattgca gatcaaatta ctcgtcatat tgatcaatta 1140tctctgacaa cgagtgatga tttattaacg tcattgattg atcaatcaca agataagtcg 1200ctattgattt ctcaaatttt acaaacgaaa ttaggaaaag ctgaagcaga taaattggct 1260aaagattgga cgaataaagg attatcaaat cgccaaatcg ttgaccaatt gaagaaacat 1320tttgcatcaa ctggcgacac gtcttcagat gatatattaa aagcaatttt gaataatgcc 1380aaagataaaa aacaagcaat tgaaacgatt ttagcaacac gtatagaaag acaaaaggca 1440aaattactgg cagatttaat tactaaaata gaaacagatc aaaataaaat ttttaattta 1500gttaaatcgg cattgaatgg taaagcggat gatttattga atttacaaaa gagactcaat 1560caaacgaaaa aagatataga ttatatttta tcaccaatag taaatcgtcc aagtttacta 1620gatcgattga ataaaaatgg gaaaacgaca gatttaaata agttagcaaa tttaatgaat 1680caaggatcag atttattaga cagtattcca gatataccca caccaaagcc agaaaagacg 1740ttaacacttg gtaaaggtaa tggattgtta agtggattat taaatgctga tggtaatgta 1800tctttgccta aagcggggga aacgataaaa gaacattggt tgccgatatc tgtaattgtt 1860ggtgcaatgg gtgtactaat gatttggtta tcacgacgca ataagttgaa aaataaagca 1920taa 1923<210>16<211>640<212>PRT<213>Staphylococcus epidermidis<400>16Gly Arg Ser Met Leu Met Ala Lys Tyr Arg Gly Lys Pro Phe Gln Leu1 5 10 15Tyr Val Lys Leu Ser Cys Ser Thr Met Met Ala Thr Ser Ile Ile Leu202530Thr Asn Ile Leu Pro Tyr Asp Ala Gln Ala Ala Ser Glu Lys Asp Thr35 40 45
Glu Ile Thr Lys Glu Ile Leu Ser Lys Gln Asp Leu Leu Asp Lys Val50 55 60Asp Lys Ala Ile Arg Gln Ile Glu Gln Leu Lys Gln Leu Ser Ala Ser65 70 75 80Ser Lys Glu His Tyr Lys Ala Gln Leu Asn Glu Ala Lys Thr Ala Ser8590 95Gln Ile Asp Glu Ile Ile Lys Arg Ala Asn Glu Leu Asp Ser Lys Asp100 105110Asn Lys Ser Ser His Thr Glu Met Asn Gly Gln Ser Asp Ile Asp Ser115 120 125Lys Leu Asp Gln Leu Leu Lys Asp Leu Asn Glu Val Ser Ser Asn Val130135140Asp Arg Gly Gln Gln Ser Gly Glu Asp Asp Leu Asn Ala Met Lys Asn145150 155 160Asp Met Ser Gln Thr Ala Thr Thr Lys His Gly Glu Lys Asp Asp Lys165 170175Asn Asp Glu Ala Met Val Asn Lys Ala Leu Glu Asp Leu Asp His Leu180 185190Asn Gln Gln Ile His Lys Ser Lys Asp Ala Ser Lys Asp Thr Ser Glu195 200 205Asp Pro Ala Val Ser Thr Thr Asp Asn Asn His Glu Val Ala Lys Thr210 215220Pro Asn Asn Asp Gly Ser Gly His Val Val Leu Asn Lys Phe Leu Ser225230 235240Asn Glu Glu Asn Gln Ser His Ser Asn Arg Leu Thr Asp Lys Leu Gln245 250 255Gly Ser Asp Lys Ile Asn His Ala Met Ile Glu Lys Leu Ala Lys Ser260 265270
Asn Ala Ser Thr Gln His Tyr Thr Tyr His Lys Leu Asn Thr Leu Gln275 280 285Ser Leu Asp Gln Arg Ile Ala Asn Thr Gln Leu Pro Lys Asn Gln Lys290295 300Ser Asp Leu Met Ser Glu Val Asn Lys Thr Lys Glu Arg Ile Lys Ser305310315 320Gln Arg Asn Ile Ile Leu Glu Glu Leu Ala Arg Thr Asp Asp Lys Lys325 330 335Tyr Ala Thr Gln Ser Ile Leu Glu Ser Ile Phe Asn Lys Asp Glu Ala340 345350Val Lys Ile Leu Lys Asp Ile Arg Val Asp Gly Lys Thr Asp Gln Gln355 360365Ile Ala Asp Gln Ile Thr Arg His Ile Asp Gln Leu Ser Leu Thr Thr370 375 380Ser Asp Asp Leu Leu Thr Ser Leu Ile Asp Gln Ser Gln Asp Lys Ser385390 395 400Leu Leu Ile Ser Gln Ile Leu Gln Thr Lys Leu Gly Lys Ala Glu Ala405410 415Asp Lys Leu Ala Lys Asp Trp Thr Asn Lys Gly Leu Ser Asn Arg Gln420 425430Ile Val Asp Gln Leu Lys Lys His Phe Ala Ser Thr Gly Asp Thr Ser435 440 445Ser Asp Asp Ile Leu Lys Ala Ile Leu Asn Asn Ala Lys Asp Lys Lys450 455 460Gln Ala Ile Glu Thr Ile Leu Ala Thr Arg Ile Glu Arg Gln Lys Ala465 470 475 480Lys Leu Leu Ala Asp Leu Ile Thr Lys Ile Glu Thr Asp Gln Asn Lys485 490 495
Ile Phe Asn Leu Val Lys Ser Ala Leu Asn Gly Lys Ala Asp Asp Leu500 505510Leu Asn Leu Gln Lys Arg Leu Asn Gln Thr Lys Lys Asp Ile Asp Tyr515520 525Ile Leu Ser Pro Ile Val Asn Arg Pro Ser Leu Leu Asp Arg Leu Asn530 535 540Lys Asn Gly Lys Thr Thr Asp Leu Asn Lys Leu Ala Asn Leu Met Asn545 550 555 560Gln Gly Ser Asp Leu Leu Asp Ser Ile Pro Asp Ile Pro Thr Pro Lys565 570 575Pro Glu Lys Thr Leu Thr Leu Gly Lys Gly Asn Gly Leu Leu Ser Gly580 585590Leu Leu Asn Ala Asp Gly Asn Val Ser Leu Pro Lys Ala Gly Glu Thr595600 605Ile Lys Glu His Trp Leu Pro Ile Ser Val Ile Val Gly Ala Met Gly610 615 620Val Leu Met Ile Trp Leu Ser Arg Arg Asn Lys Leu Lys Asn Lys Ala625 630 635 640<210>17<211>522<212>PRT<213>Staphylococcus epidermidis<400>17Ala Ser Glu Thr Pro Ile Thr Ser Glu Ile Ser Ser Asn Ser Glu Thr1 5 10 15Val Ala Asn Gln Asn Ser Thr Thr Ile Lys Asn Ser Gln Lys Glu Thr20 25 30Val Asn Ser Thr Ser Leu Glu Ser Asn His Ser Asn Ser Thr Asn Lys35 40 45
Gln Met Ser Ser Glu Val Thr Asn Thr Ala Gln Ser Ser Glu Lys Ala50 55 60Gly Ile Ser Gln Gln Ser Ser Glu Thr Ser Asn Gln Ser Ser Lys Leu65 70 7580Asn Thr Tyr Ala Ser Thr Asp His Val Glu Ser Thr Thr Ile Asn Asn85 90 95Asp Asn Thr Ala Gln Gln Asp Gln Asn Lys Ser Ser Asn Val Thr Ser100105 110Lys Ser Thr Gln Ser Asn Thr Ser Ser Ser Glu Lys Asn Ile Ser Ser115 120 125Asn Leu Thr Gln Ser Ile Glu Thr Lys Ala Thr Asp Ser Leu Ala Thr130 135 140Ser Glu Ala Arg Thr Ser Thr Asn Gln Ile Ser Asn Leu Thr Ser Thr145 150 155 160Ser Thr Ser Asn Gln Ser Ser Pro Thr Ser Phe Ala Asn Leu Arg Thr165 170 175Phe Ser Arg Phe Thr Val Leu Asn Thr Met Ala Ala Pro Thr Thr Thr180185 190Ser Thr Thr Thr Thr Ser Ser Leu Thr Ser Asn Ser Val Val Val Asn195 200 205Lys Asp Asn Phe Asn Glu His Met Asn Leu Ser Gly Ser Ala Thr Tyr210215220Asp Pro Lys Thr Gly Ile Ala Thr Leu Thr Pro Asp Ala Tyr Ser Gln225 230 235 240Lys Gly Ala Ile Ser Leu Asn Thr Arg Leu Asp Ser Asn Arg Ser Phe245250 255Arg Phe Ile Gly Lys Val Asn Leu Gly Asn Arg Tyr Glu Gly Tyr Ser260 265 270
Pro Asp Gly Val Ala Gly Gly Asp Gly Ile Gly Phe Ala Phe Ser Pro275 280 285Gly Pro Leu Gly Gln Ile Gly Lys Glu Gly Ala Ala Val Gly Ile Gly290 295 300Gly Leu Asn Asn Ala Phe Gly Phe Lys Leu Asp Thr Tyr His Asn Thr305310 315 320Ser Thr Pro Arg Ser Asp Ala Lys Ala Lys Ala Asp Pro Arg Asn Val325 330 335Gly Gly Gly Gly Ala Phe Gly Ala Phe Val Ser Thr Asp Arg Asn Gly340 345350Met Ala Thr Thr Glu Glu Ser Thr Ala Ala Lys Leu Asn Val Gln Pro355 360 365Thr Asp Asn Ser Phe Gln Asp Phe Val Ile Asp Tyr Asn Gly Asp Thr370 375 380Lys Val Met Thr Val Thr Tyr Ala Gly Gln Thr Phe Thr Arg Asn Leu385 390 395 400Thr Asp Trp Ile Lys Asn Ser Gly Gly Thr Thr Phe Ser Leu Ser Met405 410 415Thr Ala Ser Thr Gly Gly Ala Lys Asn Leu Gln Gln Val Gln Phe Gly420425430Thr Phe Glu Tyr Thr Glu Ser Ala Val Ala Lys Val Arg Tyr Val Asp435 440 445Ala Asn Thr Gly Lys Asp Ile Ile Pro Pro Lys Thr Ile Ala Gly Glu450 455 460Val Asp Gly Thr Val Asn Ile Asp Lys Gln Leu Asn Asn Phe Lys Asn465 470 475 480Leu Gly Tyr Ser Tyr Val Gly Thr Asp Ala Leu Lys Ala Pro Asn Tyr485490495
Thr Glu Thr Ser Gly Thr Pro Thr Leu Lys Leu Thr Asn Ser Ser Gln500 505 510Thr Val Ile Tyr Lys Phe Lys Asp Val Gln515 520<210>18<211>485<212>PRT<213>Staphylococcus epidermidis<400>18Ala Ser Asp Ala Pro Leu Thr Ser Glu Leu Asn Thr Gln Ser Glu Thr1 5 10 15Val Gly Asn Gln Asn Ser Thr Thr Ile Glu Ala Ser Thr Ser Thr Ala20 25 30Asp Ser Thr Ser Val Thr Lys Asn Ser Ser Ser Val Gln Thr Ser Asn35 40 45Ser Asp Thr Val Ser Ser Glu Lys Ser Glu Lys Val Thr Ser Thr Thr50 55 60Asn Ser Thr Ser Asn Gln Gln Glu Lys Leu Thr Ser Thr Ser Glu Ser65 7075 80Thr Ser Ser Lys Asn Thr Thr Ser Ser Ser Asp Thr Lys Ser Val Ala85 90 95Ser Thr Ser Ser Thr Glu Gln Pro Ile Asn Thr Ser Thr Asn Gln Ser100 105110Thr Ala Ser Asn Asn Thr Ser Gln Ser Thr Thr Pro Ser Ser Val Asn115 120 125Leu Asn Lys Thr Ser Thr Thr Ser Thr Ser Thr Ala Pro Val Lys Leu130 135 140Arg Thr Phe Ser Arg Leu Ala Met Ser Thr Phe Ala Ser Ala Ala Thr145150 155160
Thr Thr Ala Val Thr Ala Asn Thr Ile Thr Val Asn Lys Asp Asn Leu165 170 175Lys Gln Tyr Met Thr Thr Ser Gly Asn Ala Thr Tyr Asp Gln Ser Thr180 185190Gly Ile Val Thr Leu Thr Gln Asp Ala Tyr Ser Gln Lys Gly Ala Ile195 200 205Thr Leu Gly Thr Arg Ile Asp Ser Asn Lys Ser Phe His Phe Ser Gly210 215 220Lys Val Asn Leu Gly Asn Lys Tyr Glu Gly His Gly Asn Gly Gly Asp225230 235240Gly Ile Gly Phe Ala Phe Ser Pro Gly Val Leu Gly Glu Thr Gly Leu245 250 255Asn Gly Ala Ala Val Gly Ile Gly Gly Leu Ser Asn Ala Phe Gly Phe260 265270Lys Leu Asp Thr Tyr His Asn Thr Ser Lys Pro Asn Ser Ala Ala Lys275 280 285Ala Asn Ala Asp Pro Ser Asn Val Ala Gly Gly Gly Ala Phe Gly Ala290295 300Phe Val Thr Thr Asp Ser Tyr Gly Val Ala Thr Thr Tyr Thr Ser Ser305 310 315 320Ser Thr Ala Asp Asn Ala Ala Lys Leu Asn Val Gln Pro Thr Asn Asn325330 335Thr Phe Gln Asp Phe Asp Ile Asn Tyr Asn Gly Asp Thr Lys Val Met340345 350Thr Val Lys Tyr Ala Gly Gln Thr Trp Thr Arg Asn Ile Ser Asp Trp355360 365Ile Ala Lys Ser Gly Thr Thr Asn Phe Ser Leu Ser Met Thr Ala Ser370375 380
Thr Gly Gly Ala Thr Asn Leu Gln Gln Val Gln Phe Gly Thr Phe Glu385390 395400Tyr Thr Glu Ser Ala Val Thr Gln Val Arg Tyr Val Asp Val Thr Thr405 410 415Gly Lys Asp Ile Ile Pro Pro Lys Thr Tyr Ser Gly Asn Val Asp Gln420 425430Val Val Thr Ile Asp Asn Gln Gln Ser Ala Leu Thr Ala Lys Gly Tyr435 440445Asn Tyr Thr Ser Val Asp Ser Ser Tyr Ala Ser Thr Tyr Asn Asp Thr450 455 460Asn Lys Thr Val Lys Met Thr Asn Ala Gly Gln Ser Val Thr Tyr Tyr465 470475480Phe Thr Asp Val Val485<210>19<211>1245<212>PRT<213>Staphylococcus epidermidis<400>19Met Gly Lys Arg Arg Gln Gly Pro Ile Asn Lys Lys Val Asp Phe Leu1 5 10 15Pro Asn Lys Leu Asn Lys Tyr Ser Ile Arg Lys Phe Thr Val Gly Thr20 25 30Ala Ser Ile Leu Leu Gly Ser Thr Leu Ile Phe Gly Ser Ser Ser His35 40 45Glu Ala Lys Ala Ala Glu Glu Lys Gln Val Asp Pro Ile Thr Gln Ala50 55 60Asn Gln Asn Asp Ser Ser Glu Arg Ser Leu Glu Asn Thr Asn Gln Pro6570 75 80
Thr Val Asn Asn Glu Ala Pro Gln Met Ser Ser Thr Leu Gln Ala Glu85 90 95Glu Gly Ser Asn Ala Glu Ala Pro Gln Ser Glu Pro Thr Lys Ala Glu100 105 110Glu Gly Gly Asn Ala Glu Ala Ala Gln Ser Glu Pro Thr Lys Ala Glu115 120 125Glu Gly Gly Asn Ala Glu Ala Pro Gln Ser Glu Pro Thr Lys Ala Glu130 135 140Glu Gly Gly Asn Ala Glu Ala Ala Gln Ser Glu Pro Thr Lys Thr Glu145150 155160Glu Gly Ser Asn Val Lys Ala Ala Gln Ser Glu Pro Thr Lys Ala Glu165 170175Glu Gly Ser Asn Ala Glu Ala Pro Gln Ser Glu Pro Thr Lys Thr Glu180 185 190Glu Gly Ser Asn Ala Lys Ala Ala Gln Ser Glu Pro Thr Lys Ala Glu195 200 205Glu Gly Gly Asn Ala Glu Ala Ala Gln Ser Glu Pro Thr Lys Thr Glu210215 220Glu Gly Ser Asn Ala Glu Ala Pro Gln Ser Glu Pro Thr Lys Ala Glu225 230 235 240Glu Gly Gly Asn Ala Glu Ala Pro Gln Ser Glu Pro Thr Lys Thr Glu245 250255Glu Gly Gly Asn Ala Glu Ala Pro Asn Val Pro Thr Ile Lys Ala Asn260 265 270Ser Asp Asn Asp Thr Gln Thr Gln Phe Ser Glu Ala Pro Thr Arg Asn275280 285Asp Leu Ala Arg Lys Glu Asp Ile Pro Ala Val Ser Lys Asn Glu Glu290 295 300
Leu Gln Ser Ser Gln Pro Asn Thr Asp Ser Lys Ile Glu Pro Thr Thr305 310 315 320Ser Glu Pro Val Asn Leu Asn Tyr Ser Ser Pro Phe Met Ser Leu Leu325 330 335Ser Met Pro Ala Asp Ser Ser Ser Asn Asn Thr Lys Asn Thr Ile Asp340 345 350Ile Pro Pro Thr Thr Val Lys Gly Arg Asp Asn Tyr Asp Phe Tyr Gly355 360 365Arg Val Asp Ile Glu Ser Asn Pro Thr Asp Leu Asn Ala Thr Asn Leu370 375380Thr Arg Tyr Asn Tyr Gly Gln Pro Pro Gly Thr Thr Thr Ala Gly Ala385 390 395 400Val Gln Phe Lys Asn Gln Val Ser Phe Asp Lys Asp Phe Asp Phe Asn405 410415Ile Arg Val Ala Asn Asn Arg Gln Ser Asn Thr Thr Gly Ala Asp Gly420 425430Trp Gly Phe Met Phe Ser Lys Lys Asp Gly Asp Asp Phe Leu Lys Asn435 440 445Gly Gly Ile Leu Arg Glu Lys Gly Thr Pro Ser Ala Ala Gly Phe Arg450 455 460Ile Asp Thr Gly Tyr Tyr Asn Asn Asp Pro Leu Asp Lys Ile Gln Lys465 470 475480Gln Ala Gly Gln Gly Tyr Arg Gly Tyr Gly Thr Phe Val Lys Asn Asp485 490495Ser Gln Gly Asn Thr Ser Lys Val Gly Ser Gly Thr Pro Ser Thr Asp500 505 510Phe Leu Asn Tyr Ala Asp Asn Thr Thr Asn Asp Leu Asp Gly Lys Phe515520 525
His Gly Gln Lys Leu Asn Asn Val Asn Leu Lys Tyr Asn Ala Ser Asn530 535 540Gln Thr Phe Thr Ala Thr Tyr Ala Gly Lys Thr Trp Thr Ala Thr Leu545 550 555 560Ser Glu Leu Gly Leu Ser Pro Thr Asp Ser Tyr Asn Phe Leu Val Thr565 570575Ser Ser Gln Tyr Gly Asn Gly Asn Ser Gly Thr Tyr Ala Ser Gly Val580 585 590Met Arg Ala Asp Leu Asp Gly Ala Thr Leu Thr Tyr Thr Pro Lys Ala595600 605Val Asp Gly Asp Pro Ile Ile Ser Thr Lys Glu Ile Pro Phe Asn Lys610 615 620Lys Arg Glu Phe Asp Pro Asn Leu Ala Pro Gly Thr Glu Lys Val Val625630635 640Gln Lys Gly Glu Pro Gly Ile Glu Thr Thr Thr Thr Pro Thr Tyr Val645 650655Asn Pro Asn Thr Gly Glu Lys Val Gly Glu Gly Glu Pro Thr Glu Lys660 665 670Ile Thr Lys Gln Pro Val Asp Glu Ile Val His Tyr Gly Gly Glu Glu675 680 685Ile Lys Pro Gly His Lys Asp Glu Phe Asp Pro Asn Ala Pro Lys Gly690 695 700Ser Gln Thr Thr Gln Pro Gly Lys Pro Gly Val Lys Asn Pro Asp Thr705 710 715 720Gly Glu Val Val Thr Pro Pro Val Asp Asp Val Thr Lys Tyr Gly Pro725 730735Val Asp Gly Asp Pro Ile Thr Ser Thr Glu Glu Ile Pro Phe Asp Lys740 745750
Lys Arg Glu Phe Asn Pro Asp Leu Lys Pro Gly Glu Glu Arg Val Lys755 760 765Gln Lys Gly Glu Pro Gly Thr Lys Thr Ile Thr Thr Pro Thr Thr Lys770 775 780Asn Pro Leu Thr Gly Glu Lys Val Gly Glu Gly Glu Pro Thr Glu Lys785790795 800Ile Thr Lys Gln Pro Val Asp Glu Ile Thr Glu Tyr Gly Gly Glu Glu805 810 815Ile Lys Pro Gly His Lys Asp Glu Phe Asp Pro Asn Ala Pro Lys Gly820825830Ser Gln Glu Asp Val Pro Gly Lys Pro Gly Val Lys Asn Pro Gly Thr835 840 845Gly Glu Val Val Thr Pro Pro Val Asp Asp Val Thr Lys Tyr Gly Pro850 855 860Val Asp Gly Asp Pro Ile Thr Ser Thr Glu Glu Ile Pro Phe Asp Lys865 870 875 880Lys Arg Glu Phe Asn Pro Asp Leu Lys Pro Gly Glu Glu Arg Val Lys885890 895Gln Lys Gly Glu Pro Gly Thr Lys Thr Ile Thr Thr Pro Thr Thr Lys900 905 910Asn Pro Leu Thr Gly Glu Lys Val Gly Glu Gly Glu Pro Thr Glu Lys915 920925Ile Thr Lys Gln Pro Val Asp Glu Ile Val His Tyr Gly Gly Glu Gln930 935 940Ile Pro Gln Gly His Lys Asp Glu Phe Asp Pro Asn Ala Pro Val Asp945 950 955 960Ser Lys Thr Glu Val Pro Gly Lys Pro Gly Val Lys Asn Pro Asp Thr965 970 975
Gly Glu Val Val Thr Pro Pro Val Asp Asp Val Thr Lys Tyr Gly Pro980 985 990Val Asp Gly Asp Ser Ile Thr Ser Thr Glu Glu Ile Pro Phe Asp Lys99510001005Lys Arg Glu Phe Asp Pro Asn Leu Ala Pro Gly Thr Glu Lys Val1010 1015 1020Val Gln Lys Gly Glu Pro Gly Thr Lys Thr Ile Thr Thr Pro Thr1025 1030 1035Thr Lys Asn Pro Leu Thr Gly Glu Lys Val Gly Glu Gly Lys Ser1040 1045 1050Thr Glu Lys Val Thr Lys Gln Pro Val Asp Glu Ile Val Glu Tyr1055 1060 1065Gly Pro Thr Lys Ala Glu Pro Gly Lys Pro Ala Glu Pro Gly Lys1070 1075 1080Pro Ala Glu Pro Gly Lys Pro Ala Glu Pro Gly Thr Pro Ala Glu1085 1090 1095Pro Gly Lys Pro Ala Glu Pro Gly Thr Pro Ala Glu Pro Gly Lys1100 1105 1110Pro Ala Glu Pro Gly Lys Pro Ala Glu Pro Gly Lys Pro Ala Glu1115 1120 1125Pro Gly Lys Pro Ala Glu Pro Gly Thr Pro Ala Glu Pro Gly Thr1130 1135 1140Pro Ala Glu Pro Gly Lys Pro Ala Glu Pro Gly Thr Pro Ala Glu1145 1150 1155Pro Gly Lys Pro Ala Glu Pro Gly Thr Pro Ala Glu Pro Gly Lys1160 1165 1170Pro Ala Glu Ser Gly Lys Pro Val Glu Pro Gly Thr Pro Ala Gln1175 1180 1185
Ser Gly Ala Pro Glu Gln Pro Asn Arg Ser Met His Ser Thr Asp1190 1195 1200Asn Lys Asn Gln Leu Pro Asp Thr Gly Glu Asn Arg Gln Ala Asn1205 1210 1215Glu Gly Thr Leu Val Gly Ser Leu Leu Ala Ile Val Gly Ser Leu1220 1225 1230Phe Ile Phe Gly Arg Arg Lys Lys Gly Asn Glu Lys1235 1240 1245<210>20<211>3765<212>DNA<213>Staphylococcus epidermidis<400>20atgggcaaac gtagacaagg tcctattaat aaaaaagtgg attttttacc taacaaatta 60aacaagtatt ctataagaaa attcactgtt ggtacggcct caatattact tggttcgaca120cttatttttg gaagtagtag ccatgaagcg aaagctgcag aagaaaaaca agttgatcca180attacacaag ctaatcaaaa tgatagtagt gaaagatcac ttgaaaacac aaatcaacct240actgtaaaca atgaagcacc acagatgtct tctacattgc aagcagaaga aggaagcaat300gcagaagcac ctcaatctga gccaacgaag gcagaagaag gaggcaatgc agaagcagct360caatctgagc caacgaaggc agaagaagga ggcaatgcag aagcacctca atctgagcca420acgaaggcag aagaaggagg caatgcagaa gcagctcaat ctgagccaac gaagacagaa480gaaggaagca acgtaaaagc agctcaatct gagccaacga aggcagaaga aggaagcaat540gcagaagcac ctcaatctga gccaacgaag acagaagaag gaagcaacgc aaaagcagct600caatctgagc caacgaaggc agaagaagga ggcaatgcag aagcagctca atctgagcca660acgaagacag aagaaggaag caatgcagaa gcacctcaat ctgagccaac gaaggcagaa720gaaggaggca atgcagaagc acctcaatct gagccaacga agacagaaga aggaggcaat780gcagaagcac cgaatgttcc aactatcaaa gctaattcag ataatgatac acaaacacaa840ttttcagaag cccctacaag aaatgaccta gctagaaaag aagatatccc tgctgtttct900aaaaacgagg aattacaatc atcacaacca aacactgaca gtaaaataga acctacaact960tcagaacctg tgaatttaaa ttatagttct ccgtttatgt ccttattaag catgcctgct 1020
gatagttcat ccaataacac taaaaataca atagatatac cgccaactac ggttaaaggt 1080agagataatt acgattttta cggtagagta gatatcgaaa gtaatcctac agatttaaat 1140gcgacaaatt taacgagata taattatgga cagccacctg gtacaacaac agctggtgca 1200gttcaattta aaaatcaagt tagttttgat aaagatttcg actttaacat tagagtagca 1260aacaatcgtc aaagtaatac aactggtgca gatggttggg gctttatgtt cagcaagaaa 1320gatggggatg atttcctaaa aaacggtggt atcttacgtg aaaaaggtac acctagtgca 1380gctggtttca gaattgatac aggatattat aataacgatc cattagataa aatacagaaa 1440caagctggtc aaggctatag agggtatggg acatttgtta aaaatgactc ccaaggtaat 1500acttctaaag taggatcagg tactccatca acagattttc ttaactacgc agataatact 1560actaatgatt tagatggtaa attccatggt caaaaattaa ataatgttaa tttgaaatat 1620aatgcttcaa atcaaacttt tacagctact tatgctggta aaacttggac ggctacgtta 1680tctgaattag gattgagtcc aactgatagt tacaattttt tagttacatc aagtcaatat 1740ggaaatggta atagtggtac atacgcaagt ggcgttatga gagctgattt agatggtgca 1800acattgacat acactcctaa agcagtcgat ggagatccaa ttatatcaac taaggaaata 1860ccatttaata agaaacgtga atttgatcca aacttagccc caggtacaga aaaagtagtc 1920caaaaaggtg aaccaggaat tgaaacaaca acaacaccaa cttatgtcaa tcctaataca 1980ggagaaaaag ttggcgaagg tgaaccaaca gaaaaaataa caaaacaacc agtggatgaa 2040atcgttcatt atggtggcga agaaatcaag ccaggccata aggatgaatt tgatccaaat 2100gcaccgaaag gtagtcaaac aacgcaacca ggtaagccgg gggttaaaaa tcctgataca 2160ggcgaagtag ttactccacc tgtggatgat gtgacaaaat atggtccagt tgatggagat 2220ccgatcacgt caacggaaga aattccattc gacaagaaac gtgaattcaa tcctgattta 2280aaaccaggtg aagagcgtgt taaacaaaaa ggtgaaccag gaacaaaaac aattacaaca 2340ccaacaacta agaacccatt aacaggggaa aaagttggcg aaggtgaacc aacagaaaaa 2400ataacaaaac aaccagtaga tgaaatcaca gaatatggtg gcgaagaaat caagccaggc 2460cataaggatg aatttgatcc aaatgcaccg aaaggtagcc aagaggacgt tccaggtaaa 2520ccaggagtta aaaaccctgg aacaggcgaa gtagtcacac caccagtgga tgatgtgaca 2580aaatatggtc cagttgatgg agatccgatc acgtcaacgg aagaaattcc attcgacaag 2640aaacgtgaat tcaatcctga tttaaaacca ggtgaagagc gcgttaaaca gaaaggtgaa 2700
ccaggaacaa aaacaattac aacgccaaca actaagaacc cattaacagg agaaaaagtt 2760ggcgaaggtg aaccaacaga aaaaataaca aaacaaccag tggatgagat tgttcattat 2820ggtggtgaac aaataccaca aggtcataaa gatgaatttg atccaaatgc acctgtagat 2880agtaaaactg aagttccagg taaaccagga gttaaaaatc ctgatacagg tgaagttgtt 2940accccaccag tggatgatgt gacaaaatat ggtccagttg atggagattc gattacgtca 3000acggaagaaa ttccgtttga taaaaaacgc gaatttgatc caaacttagc gccaggtaca 3060gagaaagtcg ttcaaaaagg tgaaccagga acaaaaacaa ttacaacgcc aacaactaag 3120aacccattaa caggagaaaa agttggcgaa ggtaaatcaa cagaaaaagt cactaaacaa 3180cctgttgacg aaattgttga gtatggtcca acaaaagcag aaccaggtaa accagcggaa 3240ccaggtaaac cagcggaacc aggtaaacca gcggaaccag gtacgccagc agaaccaggt 3300aaaccagcgg aaccaggtac gccagcagaa ccaggtaaac cagcggaacc aggtaaacca 3360gcggaaccag gtaaaccagc ggaaccaggt aaaccagcgg aaccaggtac gccagcagaa 3420ccaggtacgc cagcagaacc aggtaaacca gcggaaccag gtacgccagc agaaccaggt 3480aaaccagcgg aaccaggtac gccagcagaa ccaggtaaac cagcggaatc aggtaaacca 3540gtggaaccag gtacgccagc acaatcaggt gcaccagaac aaccaaatag atcaatgcat 3600tcaacagata ataaaaatca attacctgat acaggtgaaa atcgtcaagc taatgaggga 3660actttagtcg gatctctatt agcaattgtc ggatcattgt tcatatttgg tcgtcgtaaa 3720aaaggtaatg aaaaataatt tcatataaaa actttctgcc attaa 3765<210>21<211>546<212>PRT<213>Staphylococcus epidermidis<400>21Glu Lys Gln Val Asp Pro Ile Thr Gln Ala Asn Gln Asn Asp Ser Ser1 5 10 15Glu Arg Ser Leu Glu Asn Thr Asn Gln Pro Thr Val Asn Asn Glu Ala20 25 30Pro Gln Met Ser Ser Thr Leu Gln Ala Glu Glu Gly Ser Asn Ala Glu35 40 45
Ala Pro Gln Ser Glu Pro Thr Lys Ala Glu Glu Gly Gly Asn Ala Glu50 55 60Ala Ala Gln Ser Glu Pro Thr Lys Ala Glu Glu Gly Gly Asn Ala Glu65 70 75 80Ala Pro Gln Ser Glu Pro Thr Lys Ala Glu Glu Gly Gly Asn Ala Glu85 90 95Ala Ala Gln Ser Glu Pro Thr Lys Thr Glu Glu Gly Ser Asn Val Lys100 105110Ala Ala Gln Ser Glu Pro Thr Lys Ala Glu Glu Gly Ser Asn Ala Glu115 120 125Ala Pro Gln Ser Glu Pro Thr Lys Thr Glu Glu Gly Ser Asn Ala Lys130 135140Ala Ala Gln Ser Glu Pro Thr Lys Ala Glu Glu Gly Gly Asn Ala Glu145 150 155160Ala Ala Gln Ser Glu Pro Thr Lys Thr Glu Glu Gly Ser Asn Ala Glu165170 175Ala Pro Gln Ser Glu Pro Thr Lys Ala Glu Glu Gly Gly Asn Ala Glu180 185190Ala Pro Gln Ser Glu Pro Thr Lys Thr Glu Glu Gly Gly Asn Ala Glu195 200205Ala Pro Asn Val Pro Thr Ile Lys Ala Asn Ser Asp Asn Asp Thr Gln210 215 220Thr Gln Phe Ser Glu Ala Pro Thr Arg Asn Asp Leu Ala Arg Lys Glu225 230 235 240Asp Ile Pro Ala Val Ser Lys Asn Glu Glu Leu Gln Ser Ser Gln Pro245250 255Asn Thr Asp Ser Lys Ile Glu Pro Thr Thr Ser Glu Pro Val Asn Leu260 265270
Asn Tyr Ser Ser Pro Phe Met Ser Leu Leu Ser Met Pro Ala Asp Ser275 280 285Ser Ser Asn Asn Thr Lys Asn Thr Ile Asp Ile Pro Pro Thr Thr Val290295 300Lys Gly Arg Asp Asn Tyr Asp Phe Tyr Gly Arg Val Asp Ile Glu Ser305310315 320Asn Pro Thr Asp Leu Asn Ala Thr Asn Leu Thr Arg Tyr Asn Tyr Gly325330 335Gln Pro Pro Gly Thr Thr Thr Ala Gly Ala Val Gln Phe Lys Asn Gln340 345350Val Ser Phe Asp Lys Asp Phe Asp Phe Asn Ile Arg Val Ala Asn Asn355360 365Arg Gln Ser Asn Thr Thr Gly Ala Asp Gly Trp Gly Phe Met Phe Ser370 375380Lys Lys Asp Gly Asp Asp Phe Leu Lys Asn Gly Gly Ile Leu Arg Glu385390395 400Lys Gly Thr Pro Ser Ala Ala Gly Phe Arg Ile Asp Thr Gly Tyr Tyr405 410 415Asn Asn Asp Pro Leu Asp Lys Ile Gln Lys Gln Ala Gly Gln Gly Tyr420 425 430Arg Gly Tyr Gly Thr Phe Val Lys Asn Asp Ser Gln Gly Asn Thr Ser435 440445Lys Val Gly Ser Gly Thr Pro Ser Thr Asp Phe Leu Asn Tyr Ala Asp450 455 460Asn Thr Thr Asn Asp Leu Asp Gly Lys Phe His Gly Gln Lys Leu Asn465 470 475 480Asn Val Asn Leu Lys Tyr Asn Ala Ser Asn Gln Thr Phe Thr Ala Thr485 490 495
Tyr Ala Gly Lys Thr Trp Thr Ala Thr Leu Ser Glu Leu Gly Leu Ser500 505510Pro Thr Asp Ser Tyr Asn Phe Leu Val Thr Ser Ser Gln Tyr Gly Asn515 520 525Gly Asn Ser Gly Thr Tyr Ala Ser Gly Val Met Arg Ala Asp Leu Asp530 535 540Gly Ala545<210>22<211>36<212>PRT<213>Staphylococcus aureus<400>22Leu Pro Asn Thr Gly Ser Glu Glu Met Asp Leu Pro Leu Lys Glu Leu1 5 10 15Ala Leu IIe Thr Gly Ala Ala Leu Leu Ala Arg Arg Arg Ser Lys Lys20 25 30Glu Lys Glu Ser35<210>23<211>43<212>PRT<213>Staphylococcus aureus<400>23Leu Pro Asp Thr Gly Asp Ser Ile Lys Gln Asn Gly Leu Leu Gly Gly1 5 10 15Val Met Thr Leu Leu Val Gly Leu Gly Leu Met Lys Arg Lys Lys Lys20 25 30Lys Asp Glu Asn Asp Gln Asp Asp Ser Gln Ala35 40<210>24<211>35<212>PRT
<213>Staphylococcus aureus<400>24Leu Pro Lys Thr Gly Glu Thr Thr Ser Ser Gln Ser Trp Trp Gly Leu1 5 10 15Tyr Ala Leu Leu Gly Met Leu Ala Leu Phe Ile Pro Lys Phe Arg Lys20 25 30Glu Ser Lys35<210>25<211>38<212>PRT<213>Staphylococcus aureus<400>25Leu Pro Lys Thr Gly Leu Thr Ser Val Asp Asn Phe Ile Ser Thr Val1 5 10 15Ala Phe Ala Thr Leu Ala Leu Leu Gly Ser Leu Ser Leu Leu Leu Phe20 25 30Lys Arg Lys Glu Ser Lys35<210>26<211>36<212>PRT<213>Staphylococcus aureus<400>26Leu Pro Gln Thr Gly Glu Glu Ser Asn Lys Asp Met Thr Leu Pro Leu1 5 10 15Met Ala Leu Ile Ala Leu Ser Ser Ile Val Ala Phe Val Leu Pro Arg202530Lys Arg Lys Asn35<210>27<211>34
<212>PRT<213>Staphylococcus aureus<400>27Leu Pro Lys Thr Gly Thr Asn Gln Ser Ser Ser Pro Glu Ala Met Phe1 5 10 15Val Leu Leu Ala Gly Ile Gly Leu Ile Ala Thr Val Arg Arg Arg Lys20 25 30Ala Ser<210>28<211>33<212>PRT<213>Staphylococcus aureus<400>28Leu Pro Lys Thr Gly Leu Glu Ser Thr Gln Lys Gly Leu Ile Phe Ser1 5 10 15Ser Ile Ile Gly Ile Ala Gly Leu Met Leu Leu Ala Arg Arg Arg Lys20 25 30Asn<210>29<211>39<212>PRT<213>Staphylococcus aureus<400>29Leu Pro Lys Ala Gly Glu Thr Ile Lys Glu His Trp Leu Pro Ile Ser1 510 15Val Ile Val Gly Ala Met Gly Val Leu Met Ile Trp Leu Ser Arg Arg20 25 30Asn Lys Leu Lys Asn Lys Ala3权利要求
1.一种分离的抗体，其与葡萄球菌的表面蛋白质结合，所述蛋白质选自于下列组SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21。
2.根据权利要求1所述的抗体，其中抗体针对所述表面蛋白质的A区域。
3.根据权利要求1所述的抗体，其中抗体用于治疗或预防人类或动物的金黄色葡萄球菌感染。
4.根据权利要求1所述的抗体，其中抗体适用于人类或动物的非肠道的，口部的，鼻内的，皮下的，aerosolized或静脉内的给药。
5.根据权利要求1所述的抗体，其中所述的抗体是单克隆抗体。
6.根据权利要求1所述的抗体，其中所述的抗体是多克隆抗体。
7.根据权利要求5所述的抗体，其中单克隆抗体是一种选自于下列组的类型鼠的，嵌合的，人性化的，和人的单克隆抗体。
8.根据权利要求5所述的抗体，其中抗体是一种单链单克隆抗体。
9.根据权利要求1所述的抗体，它包括具有类似于一种抗体的结合特异性的抗体片段，所述的抗体结合到具有选自于下列组SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21的序列的葡萄球菌的表面蛋白质。
10.根据权利要求1所述的抗体，它针对具有选自于下列组SEQID NOS.2、4、6、8、10、12、14、16、17、18、19和21的氨基酸序列的蛋白质。
11.根据权利要求1所述的抗体，其中所述的表面蛋白质具有由选自于下列组的核酸序列SEQ ID NOS.1、3、5、7、9、11、13、15、20和编码Aap蛋白质的A区域或其简并序列的核酸序列编码的一个氨基酸序列的核酸序列。
12.含有权利要求1的抗体的分离的抗血清。
13.一种诊断试剂盒，包括权利要求1的抗体和用于检测该抗体引起的结合的工具。
14.根据权利要求13所述的诊断试剂盒，其中所述的用于检测结合的工具包括连接到所述的抗体的一种可检测标记物。
15.一种用于诊断金黄色葡萄球菌感染的方法，包括将权利要求1的抗体加入到被怀疑感染了金黄色葡萄球菌的样品中，测定抗体是否已经结合到该样品。
16.一种用于治疗或预防金黄色葡萄球菌感染的药物组合物，包括有效量的权利要求1的抗体和药学的可接受的介质，载体或赋形剂。
17.一种治疗或预防金黄色葡萄球菌感染的方法，包括将有效量的权利要求1的抗体给人或动物患者给药。
18.一种诱导免疫应答的方法，包括以致免疫量的选自于下列组SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21的氨基酸序列的分离的蛋白质给人类或动物给药。
19.根据权利要求1所述的分离的抗体，它具有结合到由SEQ IDNOS.1、3、5、7、9、11、13、15、20的核酸序列和编码Aap蛋白质的A区域的核酸序列或其简并序列编码的氨基酸序列的能力。
20.一种来自DsqA蛋白质的A区域的分离的活性片段。
21.根据权利要求1所述的分离的抗体，进一步包括生理学上的可接受的抗生素。
22.一种用于治疗或预防金黄色葡萄球菌感染的疫苗，包括选自于下列组SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21的蛋白质序列，和一种药学的可接受的介质，载体或赋形剂，所述的蛋白质的量可有效地激发免疫应答。
全文摘要
提供了与凝血酶阳性的葡萄球菌细菌，例如溶血葡萄球菌发生交叉反应的多克隆和单克隆抗体，所述的抗体能够识别凝固酶阳性的和凝固酶阴性的葡萄球菌细菌的表面蛋白质。从基于金黄色葡萄球菌和凝固酶阴性的葡萄球菌之间共有的特性分离的表面蛋白质可以制备抗体，这些重组表面蛋白质可用于制备本发明的抗体。还提供了利用这些蛋白质和抗体用于治疗或抵抗各种各样的葡萄球菌的感染的疫苗和方法。
文档编号A61P31/04GK1543569SQ02816001
公开日2004年11月3日 申请日期2002年6月17日 优先权日2001年6月15日
发明者蒂莫西·J·福斯特, 菲奥那·罗切, 约瑟夫·M·帕蒂, 杰夫·T·哈钦斯, 皮特罗·斯皮兹利, 马克·帕伦, M 帕蒂, 斯皮兹利, 罗切, T 哈钦斯, 帕伦, 蒂莫西 J 福斯特 申请人:英希比泰克斯公司, 都柏林伊丽莎白女皇神学院, 帕维研究学院