識別凝血酶陰性的葡萄球菌和金黃色葡萄球菌的表面蛋白的交叉反應性單克隆抗體和多...的製作方法

2023-09-20 23:47:20 3

專利名稱：識別凝血酶陰性的葡萄球菌和金黃色葡萄球菌的表面蛋白的交叉反應性單克隆抗體和多 ...的製作方法
技術領域：
總的來說，本發明涉及來自金黃色葡萄球菌的表面蛋白質及其活性的區域例如它們的A功能域，它具有凝固酶陰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質，以及識別上述的蛋白質的抗體，具體地說，涉及分離的單克隆和多克隆抗體，它們識別來自金黃色葡萄球菌和凝固酶陰性的葡萄球菌的特定的蛋白質，以及抗金黃色葡萄球菌和凝固酶陰性的葡萄球菌的交叉反應，因此可用於疫苗，以及用於預防或治療各種各樣的由葡萄球菌的細菌引起的感染的方法。
背景技術：
對於大多數引起動物和人類的傳染病的微生物，在宿主中成功地形成集落是一個必需的過程。微生物的粘附是最後導致疾病的一系列事件的第一個重要步驟。藉助細菌表面的特定的adhesins，通過粘附到宿主組織或血清限定的移植入的生物材料，例如導管，人工的關節，血管的移植片，病原微生物在宿主中形成集落。MSCRAMMs(識別粘附的基質分子的微生物表面成分)是細胞表面adhesins的一個家族，它們識別和特定地結合到宿主的細胞外的基質的不同的組分。一旦細菌成功地粘附並且在宿主組織中形成集落，他們的生理機能被急劇改變並且分泌受損害的成分例如毒素和水解蛋白質的酶。此外，粘連的細菌常常產生生物膜並且很快地變得對大多數的抗生素的殺戮效果更具抗性。
金黃色葡萄球菌產生從皮膚上的損害例如傷口感染，膿皰病，癤到包括肺炎，膿毒性的關節炎，膿毒病，心內膜炎的威脅生命的狀況以及生物材料相關的傳染病的範圍的感染譜。已知金黃色葡萄球菌表達不同的MSCRAMMs的全部技能，它們獨立地或者協調地促進微生物的粘附到特定的宿主組織組分。另外，葡萄球菌細菌的另一個類型被識別為凝固酶陰性的細菌，包括例如已知同樣地表示MSCRAMMs的表皮葡萄球菌和溶血葡萄球菌，也對各種各樣的細菌的感染和有關的疾病負有責任。關於這一點，通常MSCRAMMs由抗體，多克隆和單克隆抗體，對免疫學的攻擊提供出色的靶。
然而，由於抗體本性非常特異的，就不同類型的葡萄球菌而言，例如一方面金黃色葡萄球菌(凝固酶陽性)和另一方面表皮葡萄球菌和溶血葡萄球菌(凝固酶-陰性的)，研製顯示不同的類型的細菌之間年的交叉反應性的抗體，仍然是值得注意的問題。這樣的交叉的反應的抗體是特別令人想望的，因為他們具有使人類和動物患者致免疫的潛在性和提供抵抗由兩種類型的葡萄球菌的細菌，即凝固酶陽性的細菌例如金黃色葡萄球菌和凝固酶陰性的細菌，例如表皮葡萄球菌和溶血葡萄球菌引起的感染潛在性。因此這樣的抗體在預防或治療各種各樣的由葡萄球菌的細菌引起的感染中非常地有效。
發明概述因此，本發明的目的之一提供識別來自凝固酶陽性的細菌例如金黃色葡萄球菌的MSCRAMMs以及來自凝固酶陰性的細菌，例如表皮葡萄球菌和溶血葡萄球菌的MSCRAMMs的單克隆抗體。
本發明的還有一個目的是識別和分離來自葡萄球菌的細菌的MSCRAMMs，以及他們的活性的區域例如A區域，該區域可用於產生與凝固酶陽性的和凝固酶陰性的葡萄球菌發生交叉反應的單克隆和多克隆抗體。
本發明的另一個目的提供可以識別來自凝固酶陰性的葡萄球菌的表面蛋白質例如DgsK蛋白質的A區域，同時識別來自金黃色葡萄球菌的表面蛋白質例如SasA蛋白質的分離的抗體。
本發明的又一方面是利用本發明的分離的蛋白質，A區域和抗體生產用於治療或預防葡萄球菌的感染的疫苗，以及提供了其中將本發明的疫苗和抗體可用於預防或治療葡萄球菌的感染的方法。
這些和其他的目的通過本發明實現，所述的發明包括識別和分離一種類型的葡萄球菌的細菌例如凝固酶陰性的或凝固酶陽性的葡萄球菌的表面蛋白質，它導致產生識別兩種類型的葡萄球菌的表面蛋白質的交叉的反應性的抗體，並且因此將抗體用於治療或預防各種各樣葡萄球菌的感染的疫苗和方法。本發明還涉及從這些表面蛋白質產生多克隆和單克隆抗體，以及它們用於預防或治療葡萄球菌的感染。
對本領域內技術人員而言，根據閱讀本說明書和/或在此處記載的參考文獻，這些具體的實施方案和在公開的發明的精神和範圍之內的其他的供替代的選擇和改變是顯而易見的，所有引用的文獻都被併入本文。
附圖簡述附

圖1是本發明的in silico預測的蛋白質的一級的結構圖。
附圖2顯示本發明的開放讀框表達的N-末端His標記的純化的重組的蛋白質的考馬斯亮藍染色凝膠。
附圖3A-3C顯示分別用抗-KesK抗體(附圖3A)，抗-KnkA抗體(附圖3B)和抗DsqA抗體(附圖3C)探測時金黃色葡萄球菌的細胞壁提取物的Western印跡。
附圖4A-4B顯示表達金黃色葡萄球菌的MSCRAMM稱為KnkA(附圖4A)和KesK(附圖4B)的乳酸乳球菌的點印跡和Western免疫印跡。
附圖5A-5D代表用康復的人血清體內檢測表達本發明重組LPXTG蛋白質，包括RrKn和RrKN2(附圖5A)，Kesk1和Kesk2A(附圖5B)，KnkA(附圖5C)和DsqA2(附圖5D)的探測情況。
附圖6顯示證實抗金黃色葡萄球菌SasA的兔多克隆抗體與從表皮葡萄球菌HB細胞表面釋放的蛋白質以及來自從表皮葡萄球菌克隆的DsgK的重組A-區域發生交叉反應的Western印跡分析結果。
優選的實施方案的詳細描述根據本發明，提供了來自凝固酶陽性的葡萄球菌的細菌例如金黃色葡萄球菌以及凝固酶陰性的葡萄球菌的細菌例如表皮葡萄球菌和溶血葡萄球菌，的特定的表面蛋白質，包括其活性片段例如這些蛋白質或可以產生識別完整的蛋白質的抗體的其他的表位區域的A區域。根據本發明，基於MSCRAMMs(識別粘附基質分子的微生物表面成分)的一些共有的特徵實施候選肽序列和蛋白質的鑑定和分離，在大多數情況下，這些特徵共價地固定到細胞壁肽葡聚糖。這些表面蛋白質具有下列的共有的特徵可用於確定和分離本發明的序列，即(i)Sec-依賴性分泌必需的N-末端信號肽(長度大約40個殘基)，(ii)一個富含脯氨酸和甘氨酸殘基或由絲氨酸和門冬氨酸鹽二肽重複組成點的跨壁功能域；(iii)蛋白質共價固定到肽葡聚糖中的五個甘氨酸交聯橋必需的LPXTG基序，(iv)一個疏水的跨膜功能區，跟隨其後的(v)幾個帶正電荷的殘基。
根據本發明，按照以上所述的特性，通過開發金黃色葡萄球菌的完整的基因組，識別了至少八個新的開放讀框編碼的蛋白質，具有分泌和固定象徵MSCRAMMs的基序(即的攜帶N-末端信號肽和跟隨疏水的功能區之後的C-末端LPXTG基序和一個帶正電的尾部)。表1闡明所識別的蛋白質的一覽表，包括在金黃色葡萄球菌基因組中的分布，蛋白質的大小和C-末端細胞壁排序序列。
表1
縮寫eMRSA-16；N，8325；C，COL；S，MSSA；J，N315，M，Mu50。
8個蛋白質中的6個存在於已經被測序的所有六個葡萄球菌的基因組中，剩餘的2個存在於5/6的這些基因組中。
根據本發明，獲得上述的蛋白質的胺基酸和編碼它的核酸序列，如下所述Ekes MRSA-SEQ ID NO1(DNA序列)；EkeS～MRSA-SEQ ID NO2(蛋白質序列)；DsqA(8325)-SEQ IDNO3(DNA序列)；DsqA(8325)-SEQ ID NO4(蛋白質序列)；KesK1(8325)-SEQ ID NO5(DNA序列)；KesK1(8325)-SEQ IDNO6(蛋白質序列)；KrkN2(8325)--SEQ ID NO7(DNA序列)；KrkN2(8325)-SEQ ID NO8(蛋白質序列)；KrkN(8325)-SEQ IDNO9(DNA序列)；KrkN(8325)-SEQ ID NO10(蛋白質序列)；RkaS(COL)-SEQ ID NO11(DNA序列)；RkaS(COL)-SEQ IDNO12(蛋白質序列)；RrkN(8325)-SEQ ID NO13(DNA序列)；RrkN(8325)-SEQ ID NO14(蛋白質序列)；KnkA(8325)-SEQ IDNO15(DNA序列)；KnkA(8325)-SEQII7 NO16(蛋白質序列)。
根據本發明，可以從上述的蛋白質或他們的活性區域例如A區域產生分離的抗體以便能夠識別上述的蛋白質和/或上述的功能區。這些抗體可以是單克隆的或多克隆的。如果需要多克隆抗體，可以本領域內熟知的許多常規方法的任何一種製備。一種典型的方法是，可以將需要的表面蛋白質或其活性的區域注射到合適的宿主動物，例如，小鼠或兔子，在合適的時間期間之後，可以從該宿主動物分離和回收想要的抗體。對於單克隆抗體，根據本發明，可以許多合適的方法包括例如熟知的Kohler和Milstein，Nature 256495-497(1975)方法，本領域內已知的任何其他方法，例如美國專利Nos.6,331,415；5,981,216；5,807,715；和4,816,567；歐洲專利申請519,596；和PCT公開WO 00/71585公開的方法製備，所有這些專利文獻都通過在此引述而合併於本文。這些方法包括其製備如以本領域內熟知的方式進行嵌合，人性化，或人的單克隆抗體。仍然進一步地，可以從一個單鏈，例如輕或重鏈製備，以及還可以從保持完整的抗體的結合特性(例如，特異性和/或親合力)的抗體的活性的片段製備單克隆抗體。活性的片段是指具有與完全抗體相同的結合特異性的抗體片段，所述完全抗體結合到來自不同的類型的葡萄球菌細菌(即凝固酶陰性的，或凝固酶陽性的)的特定的表面蛋白質或其同系物。本文術語″抗體″包括所述的片段。另外，也可以利用本發明的單克隆或多克隆抗體製備抗血清，可以本領域內熟知的許多合適方式製備。
如上所述，可以本領域內熟知的許多合適的方式製備本發明的抗分離的表面蛋白質和/或其活性的區域的抗體，例如上述所述的完全建立的Kohler和Milstein方法，可用於產生單克隆抗體。例如，在這樣的一種方法的預備的步驟中使用，可以在長時間的時期用本發明的純化的重組MSCRAMM或其活性的部分長期給小鼠每一周腹膜內注射一次，隨後測試從免疫接種的老鼠獲得的血液以測定與純化的蛋白質的反應性。在識別老鼠與該蛋白質的反應性之後，將從小鼠脾臟分離的淋巴細胞與小鼠脊髓細胞融合以產生抗本發明的表面蛋白質的抗體陽性的雜交瘤，然後分離和培養，隨後純化和使之形成同位型。
為了產生本發明的單克隆抗體，優選的是利用本發明的重組製備的MSCRAMM′s來製備，利用本領域內熟知的許多標準方法可以製備這些重組子並且分離。例如，一種這樣的方法使用了大腸桿菌表達載體pQE-30作為克隆和表達重組蛋白質和肽的表達載體。在一個優選的方法中，利用PCR，從上面描述的序列擴增識別為DgsK或SasA的表面蛋白質的A區域，再克隆到大腸桿菌表達載體PQE-30(Qiagen)，以允許表達一種含有六個組氨酸殘基的重組融合蛋白質。
隨後將該載體轉化到大腸桿菌ATCC55151，在15-升發酵罐中生長到光密度(OD600)為0.7，用0.2mM異丙基-1β-D半乳糖苷(IPTG)誘導4小時。利用AGTechnologies中空纖維組件(孔大小0.45um)收集細胞，將細胞漿在-80C冷凍。利用2次穿過French Press，1100psi，將細胞在1×PBS(10mL緩衝液/1克的細胞漿)裂解。將裂解的細胞17,000rpm旋轉30分鐘以除去細胞碎片。將上清液經過裝備了0.1M NiCI2的5-毫升Hi俘獲Cheating(Pharmacia)柱。在上樣之後，用5個柱體積的10mM Tris，pH 8.0，100mM NaCI(緩衝液A)洗滌柱子。利用超過30個柱體積的0-100％梯度的10mM Tris，pH 8.0，100mM NaCI，200mM咪唑(Buffer B)洗脫蛋白質。SdrGN1N2N3或SdrGN2N3在-13％Buffer B(～26mM咪唑)被洗脫。監測280nm處的吸光率。在1×PBS中透析含有SdrGN1 N2N3或SdrGN2N3的餾分。
接著，然後將各個蛋白質經過一個內毒素除去方案。在該方案中所使用的緩衝液通過一個5-毫升Mono-Q瓊脂糖(Pharmacia)柱而沒有內毒素。將蛋白質平衡地分到4×15毫升試管中。各個試管的體積用緩衝液A加到9毫升。將1毫升的10％Triton X-114加入到各個試管中，在4℃旋轉保溫1小時。將試管置於37℃水浴中以使相位分離。將試管以2,000rpm旋轉10分鐘，收集各個試管的上面的含水的相，用去汙劑反覆提取。將2次提取的含水相合併，傳遞過帶有0.1M NiCl2的5-mL IDAcheating(Sigma)柱，以除去剩餘的洗滌劑。用9個柱體積的Buffer A洗滌該柱，然後用3個柱體積的Buffer B洗脫該蛋白質。將洗脫液通過5-毫升的Detoxigel(Sigma)柱，收集流出物，再上樣到柱。收集第二次過柱的流出物，在1×PBS中透析。分析純化的產物的濃度，純度和內毒素水平，然後給小鼠給藥。
在該優選的方法，以下面的方式從上面識別的重組蛋白質可以製備本發明的單克隆抗體。在該方法中，將大腸桿菌表達的和純化的重組SasA和DsgK蛋白質用於製備一組鼠的單克隆抗體，而將小鼠血清用作為多克隆抗體的來源。簡單地說，一組Balb/C或SJL小鼠接收了一系列皮下的免疫接種1-10mg溶液蛋白質或與下面表2描述的佐劑混合的蛋白質。
表2免疫接種方案RIMMS注射天數數量(微克)途徑佐劑#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常規注射天數數量(微克)途徑佐劑原始0 5 皮下FCA加強#1 14 1 腹膜內 RIBI加強#2 28 1 腹膜內 RIBI加強#3 42 1 腹膜內 RIBI在殺死時(RIMMS)或加強後幾天收集(常規的)血清，在ELISA測定中測定抗MSCRAMM蛋白質或對完整的細胞(表皮葡萄球菌和金黃色葡萄球菌)的效價。在最後的加強之後3天，去除脾臟或淋巴結，挑開成單細胞懸浮液，收集淋巴細胞。然後將淋巴細胞與P3X63Ag8.653骨髓瘤細胞系(ATCC CRL-1580)融合。根據Current Protocols in Immunology(第2章，第2單元)中的生產單克隆抗體的方案進行細胞融合，隨後塗覆平板和餵養，所述文獻都通過在此引述而合併於本文。
然後利用標準的ELISA分析方法，從融合產生的任何克隆中篩選特異的抗-SasA抗體的產生。擴增陽性的克隆，進一步通過流式血細胞計數測試完整的細菌的細胞結合測定中的活性以及Biacore分析中SasA的結合。在整個Biacore分析中，流速保持在10毫升/分鐘不變。在注射SasA或DgsK之前，藉助於RAM-Fc結合，將測試抗體吸附到晶片。在時間0，將濃度為30mg/ml的SasA或DgsK注射過晶片3分鐘，隨後解離2分鐘。對該階段的分析測定Mab/SasA和DgsK相互作用的相對關係和解離常數。
接著，對上面提到的抗體與完整細菌的結合情況進行測試。在該測試中，用兔子IgG(50mg/ml)封閉之後，收集，洗滌細菌的樣品金黃色葡萄球菌Newman，金黃色葡萄球菌60，金黃色葡萄球菌397(Sal6)，金黃色葡萄球菌Wood，金黃色葡萄球菌8325-4，甲氧苯青黴素抗性的金黃色葡萄球菌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，用濃度為2pg/ml的Mab或僅用PBS(對照)保溫。在用抗體保溫之後，將用作為檢測抗體的山羊-F(ab′)2-抗-小鼠-F(ab′)2-FITC用於孵育細菌的細胞。在抗體標記之後，用FACS管徑流式細胞計數器抽吸細菌的細胞以分析螢光射出(刺激488，發射570)。對於各個細菌的菌株，收集和檢測10,000個樣品。這些資料顯示抗金黃色SasA的抗體能夠識別凝固酶陰性的葡萄球菌的表面的同源的蛋白質。該資料支持Western印跡分析，表明抗金黃色葡萄球菌SasA的兔子多克隆抗體與從表皮葡萄球菌HB細胞表面釋放的蛋白質以及來自於從表皮葡萄球菌克隆的DsgK的重組A-區域發生交叉反應(參見附圖6和下表3)。
表3多克隆血清反應性
雖然利用重組形式的本發明的表面蛋白質生產抗體是優選的，但是該抗體可以從這些蛋白質天然分離的和純化形式或其活性區域例如A區域製備抗體，以類似於上面所述的方式利用這些蛋白質或活性的區域可以產生單克隆抗體或多克隆抗體以獲得這樣的抗體。仍然其他的常規的方法，利用重組或天然的純化蛋白質或其活性區域，可用於產生本發明的抗體，正如本領域內技術人員可以識別的。
正如本領域內技術人員可以認識到的，本發明的抗體也可以配製成可用於人或動物患者給藥的合適的藥學的組合物，以便治療或預防葡萄球菌的細菌引起的感染。含有本發明的抗體的藥物組合物，或其有效的片段，可以與本領域內常用的合適的藥物介質，賦形劑或載體結合配製，包括例如鹽水，葡萄糖，水，甘油，乙醇，其他的治療化合物和其結合使用。本領域內技術人員應該認識到，所使用的特定的介質，賦形劑或載體將依據患者和患者的狀況而改變，如本領域內技術人員將會認識到的各種各樣的給藥方式適用於本發明的組合物。在本申請中公開的以藥物組合物給藥的合適的方式包括，但不限於，局部的，口部的，肛門的，陰道的，靜脈內的，腹膜內的，肌內的，皮下的，鼻內的和真皮內的給藥。
對於局部的給藥，將組合物配製成軟膏，乳膏，凝膠，洗液，滴劑(例如眼藥水和耳朵滴劑)，或溶液(例如漱口劑)。可以將組合物埋植於傷口或外科的敷料，縫合線和氣溶膠。該組合物可以含有常規的添加劑。例如防腐劑，溶劑以促進滲透和潤滑藥。局部的製劑也可以含有常規的載體例如乳膏或油膏基底，乙醇或油醯醇。另外，通常也可以將抗體組合物，和涉及其他MSCRAMM的組合物，疫苗，方法和應用的其他的信息用於本發明，涉及前面提到的MSCRAMM和它們的活性的區域和其抗體，這些其它的MSCRAMM公開於例如美國專利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；所有這些專利都通過在此引述而合併於本文。
本發明的抗體組合物也可以與其量可有效地加強致免疫的應答的合適的佐劑一起給藥。例如，合適的佐劑可以包括廣泛用於人的明礬(磷酸鋁或氫氧化鋁)，和其他的佐劑例如皂甙和其純化的組分Quil A，弗氏完全佐劑，RIBBI佐劑，和其它用於研究和獸醫應用的佐劑，還有其他以化學方法定義的製劑例如muramyl二肽，一磷醯脂類A，磷脂軛合物例如Goodman-Snitkoff等人在J.Immunol.147410-415(1991)描述的，該文獻通過在此引述而合併於本文，蛋白脂質體內的偶合物的形成膠囊如Miller等人，J.Exp.Med.1761739-1744(1992)，該文獻通過在此引述而合併於本文，脂類載體中蛋白質形成膠囊如NovasomeTM脂類載體(Micro Vescular System，Inc.，Nashua，NH)也可以使用。
在任何情況下，本發明的識別蛋白質或如上文提出的它們的活性的區域的抗體組合物可用於預防或治療葡萄球菌的感染的方法，可用於抑制葡萄球菌的細菌與宿主組織和/或細胞的結合。根據本發明，提供了預防或治療葡萄球菌的感染的方法，包括以有效量的抗體給本文提出的表面蛋白或其活性的亞區域以便治療或預防葡萄球菌的感染。另外，這些單克隆抗體可用於損害葡萄球菌的細菌與宿主細胞的結合。
因此，根據本發明，將本發明的抗體以如上所述的任何常規的方法(例如，局部的，非腸道的，肌內的，等)給藥，因此當將有效量的抗體組合物給人或動物患者給藥時，提供了一種治療或預防人類或動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細菌粘合，抑制葡萄球菌的細菌結合到宿主細胞從而可用於治療或阻止葡萄球菌的感染的使用水平，例如抗體效價的水平。如本領域內普通技術人員認識到的，有效地治療或預防葡萄球菌的感染所必需的抗體效價的水平將依據患者的特性和健康狀況，和/或預存在的葡萄球菌的感染的嚴重程度。
除了用於治療或預防葡萄球菌的感染的方法中，本發明的抗體也可用於葡萄球菌的蛋白質的特定的檢測或作為研究工具。本文所述的術語″抗體″包括單克隆，多克隆，嵌合，單鏈，雙特異性，simianize，和人性化的或靈長目化的抗體以及Fab片段，例如保持抗體與上面詳細說明的表面蛋白質的抗體的結合特異性的那些片段，包括Fab免疫球蛋白表達文庫的產物。因此，本發明涉及單鏈的使用例如抗體的可變的重和輕鏈。這些類型的抗體或抗體片段的製備是本領域內技術人員熟知的。在本發明中，抗表面蛋白質或上面所述的它們的活性的區域的抗體可以製備，分離和/或純化，然後用於治療或免受葡萄球菌的感染。
上面描述的任何抗體可直接用可檢測的標籤進行標記以識別葡萄球菌細菌和對其定性。免疫測定中使用的標記物通常是本領域內技術人員已知的，並且包括酶，放射性同位素，螢光，發光的和產色的物質，包括有色的顆粒例如膠態的金或膠乳珠子。合適的免疫測定包括酶聯免疫吸附(ELISA)。
或者，通過與具有免疫球蛋白親合力的標記的物質可以間接標記該抗體。該抗體可以與第二物質偶合，用已標記的第三物質進行檢測，該第三物質與偶合到抗體的第二物質具有親和力。例如，可以將抗體結合到生物素，利用標記的抗生物素蛋白或鏈生抗生物素蛋白可以檢測抗體-生物素偶合物。類似地，可以將該抗體偶合到半肝素，利用標記的抗半肝素抗體檢測抗體-半肝素偶合物。這些和其他標記抗體和分析偶合物的其他方法是本領域內技術人員熟知的。
根據本發明，還提供了被設計為治療或保護其抵抗葡萄球菌感染的主動或被動免疫接種疫苗，可以利用許多本領域內熟知的常規疫苗製備方法從以上提出的表面蛋白質或活性區域製備這些疫苗。對於典型的疫苗，將致免疫量的合適的表面蛋白或其活性片段與合適的藥物學可接受載體，媒介物或賦形劑結合，合適的是可以以有效地免疫接種人或動物患者的疫苗的量給藥。本領域技術人員認識到致免疫量是指能夠在人或動物患者中產生免疫應答的蛋白質或活性片段或其亞片段的量。
除了藉助於將免疫量的本發明的蛋白質或活性片段導入或給藥產生的抗體的活性疫苗以外，也可以將本發明的分離的抗體或活性片段用於研製抗鉤環感染的被動免疫接種的疫苗。在這樣的情況下，可以將如上所述的抗體組合物，即有效量的抗體和藥物學可接受的載體，媒介物或賦形劑以適當方式給藥到人或動物患者。
因此，本發明的蛋白質或其活性片段可用作為活性疫苗，本發明的抗體可用作為被動疫苗，可用於提供合適的抗體以治療或予防葡萄球菌感染。本領域內技術人員將會認識到，可以將一秒包裝以適用於許多方式給藥，例如非腸道的(即肌內的，透皮的或皮下的)給藥或鼻咽的(即鼻內的)給藥。一種這樣的方式是將疫苗通過肌內注射到例如三角肌，但是，給藥的特定方式將依賴於待治療的細菌感染的特性和患者的狀況。優選的是將該疫苗與藥物學可接受的載體，媒介物或賦形劑結合以有助於給藥，通常該載體是有或沒有防腐劑的水，或緩衝鹽水。該疫苗可以是在給藥時重懸浮的凍幹形式或溶液形式。
另外，在某些情況下，如必要可以將本發明的抗體進行修飾，以對給藥患者的致免疫性降低，例如，如果該患者是人。通過將雜交瘤衍生的抗體的complimentarity表位移植到如Jones等人，Nature 321522-525(1986)或Tempest等人/Biotechnology 9266-273(1991)描述的人單克隆抗體而將抗體「人性化」或通過將免疫球蛋白可變區中的表面接觸的鼠骨架殘基改變為模擬同源的人骨架配體如由Padlan，Molecular Imm.28489-498(1991)描述而進行″膠合″，這些文獻都通過在此引述而合併於本文。甚至於進一步地，如果需要，本發明的單克隆抗體可以與合適的抗生素結合給藥以進一步加強所述組合物抗擊細菌感染的能力。
除了治療人或動物患者以外，本發明的組合物也可以用於停止或予防醫學裝置或其他生物材料例如移植片的感染。待用抗體，蛋白質和活性片段包被的醫學裝置或聚合物生物材料包括，但不限於鉤環，縫合線，復位心臟瓣膜，心臟的協助裝置，硬的和軟的接觸透鏡，眼內的透鏡植入片(前的室或後的室)，其他的植入片例如角膜的鑲嵌物，kerato-修補物，血管的斯滕特氏印模，epikeratophalia裝置，綠內障分流術，視網膜的鉤環，鞏膜的扣子，牙齒的修補物，thyroplastic裝置，laryngoplastic裝置，血管的移植片，軟的和硬的組織修補物，包括但不限於，泵，電的裝置包括刺激器和記錄器，聽覺的修復術，起搏器，人工的喉，牙齒的植入片，乳房的植入片，陰莖的植入片，cranio/臉的腱，人工的關節，腱，紐帶，menisci，和磁碟，人工的骨骼，人工器官包括人工胰腺，人工心臟，人工的肢，和心臟瓣膜，斯滕特氏印模，金屬絲，導向裝置絲，靜脈內的和中央的靜脈的導管，雷射和氣球血管成形術裝置，血管的和心臟裝置(管子，導管，氣球)，室的協助，血透析組合物，血氧合器，尿道的/輸尿管的/泌尿的裝置(Foley導管，斯滕特氏印模，管和氣球)，導氣管導管(氣管內的和氣管造口術管和膠管管頭)，腸內的餵養管(包括nasogastric，胃內的和空腸的管)，傷口排水管，用於引流體腔的管例如胸膜的，腹膜的，顱側的和心包的cavities，血袋，測試管，血收集管，vacutainers，注射器，針，吸移管，吸量管尖端，和血造管。
本領域內技術人員將會明白，本文所用的術語″被包被″或″包被″是指將抗體或活性片段或其衍生的藥物組合物應用到裝置的表面，優選的是與葡萄球菌細菌感染接觸的外表面。該裝置的表面不必完全被蛋白質，抗體或活性片段所覆蓋。
本發明抗體給藥的優選劑量是有效地抑制或治療葡萄球菌感染的量，易於認識到該量將大大依賴於感染的特性和患者的狀況。如上所述，用於本發明的抗體或藥物製劑的「有效量」是指無毒但足以產生需要的予防或治療效果的劑量。正如上文指出的，所需要的抗體或特定試劑的精確量將隨患者而變化，取決於患者的種類，年齡，和總的狀態，待治療的狀況的嚴重程度，所使用的特定的載體或賦形劑和給藥方式等等。因此，特定抗體組合物的「有效量″將基於特定的環境而變化，普通技術人員利用唯一的常規試驗可以測定各種應用情況下合適的有效量。可以將該劑量調節到適應於以該組合物給藥的個體，並且隨患者的年齡，重量，代謝而變化。該組合物還可以含有穩定劑或藥物學可接受的防腐劑，例如硫汞撒(乙基(2-mercaptobenzoate-S)汞鈉鹽)(Sigma Chemical Company，St.Louis，MO)。
當與合適的標記物或其他合適的可檢測的生物分子或化學物質一起使用時，本文描述單克隆抗體可用於體內或體外診斷葡萄球菌感染或檢測葡萄球菌細菌的目的。通過這樣的抗體的使用也有助於試驗室研究。各種類型的標記物和將標記物偶合到本發明的抗體的方法是本領域內技術人員熟知的，例如下面列出的一種方法。
例如，可以將抗體偶合到(直接或通過螯合劑)放射性標記物例如，但不限於32P，3H，14C，35S，125I，或131I。通過例如閃爍計數，γ射線分光光度計放射自顯影的方法可以檢測標記物。也可以使用生物發光標記物，例如螢火蟲螢光素衍生物。採用常規方法生物發光物質可以共價結合到該蛋白質，當酶例如螢光酶催化引起生物發光的與ATP的反應以散發光子時，可以檢測標記物蛋白質。也可以將螢光團用於標記蛋白質。螢光團的例子包括螢光和衍生物，藻紅蛋白，allo-藻藍蛋白，藻藍蛋白，礆性蕊香紅，和德克薩斯紅。通常螢光團可以通過螢光檢測器進行檢測。
通過對如上所述的抗體進行標記和根據本領域內技術人員熟知的方法例如免疫螢光顯微鏡，利用如Warren等人(Mol.Cell.Biol.，71326-1337，1987)描述的程序，對標記物進行檢測可以測定細胞中配體的定位。
如上所述，本發明的單克隆抗體，或活性部分或其片段特別適用於幹擾負責感染的葡萄球菌病原體和哺乳動物宿主之間的起始物理相互作用，這種對物理相互作用的幹擾可用於治療患者和in-dwelling醫學裝置予防或降低細菌感染以使它們在使用時更安全。
在本發明的另一個實施例中，提供了一種可用於分離和識別葡萄球菌細菌感染的試劑盒，包括在單個容器中的合適形式，例如凍幹的本發明的抗體，然後通過加入懷疑含有葡萄球菌細菌的含水的樣品而變成活性。這樣的試劑盒通常包括用於儲存合適形式抗體和合適的免疫檢測試劑的合適的容器，所述的免疫檢測試劑允許識別結合到本發明的表面蛋白質或抗體的複合物。通常，這些試劑盒含有本發明的抗體和識別當來自患者的樣品導入到該抗體時識別結合的抗體的工具。例如，合適的免疫檢測試劑可以包括合適的可檢測的信號或標記物，例如生物素或產生可檢測的顏色的酶等，可以連接到抗體或用於其他合適的方式中標記物以便當抗體結合到抗原時提供可檢測的結果。
簡單地說，識別和結合到本發明的表面蛋白質的本發明的抗體或其活性片段可用於治療各種各樣的人和動物患者的葡萄球菌感染，以及用於醫療或其他in-dwelling裝置。根據本發明，由於這些蛋白質的特性，以及它們含有與其他類型的葡萄球菌細菌的蛋白質共有的表位，即來自凝血酶陰性葡萄球菌的蛋白質將產生識別來自金黃色葡萄球菌的同源蛋白的抗體或者相反，本發明的抗體將顯示交叉反應性和應該有效地抗擊廣譜的葡萄球菌感染。因此，本發明提供了用於治療或抵抗廣譜葡萄球菌感染的方法和改善的方法中的組合物。
實施例提供下面的實施例，舉例說明本發明的優選的實施方式的各個方面。本領域內技術人員應該認識到實施例中公開的技術允許代表發明人發現的技術以實施本發明，因此可以認為構成了實施本發明的優選的方式。但是，本領域內技術人員應該認識到，在本發明公開的基礎上，可以在公開的特定的實施方式中進行許多改變，仍然可獲得類似的結構而不脫離本發明的精神和範圍。
實施例1.金黃色葡萄球菌的MSCRAMM′s的分離和測序已知金黃色葡萄球菌表達一類表面結合的蛋白質，所述的蛋白質通過允許細菌避免宿主防禦和通過作為adhesins對致病性起重要作用。已知這些蛋白質是MSCRAMMs(識別粘附的基質的分子的微生物表面成分)，在大多數情況下，可以共價結合到細胞壁肽葡聚糖。它們有幾個共同的特徵(i)Sec-依賴性分秘所需要的N-末端信號肽(長度約40個殘基)，(ii)富含脯氨酸和甘氨酸殘基或由絲氨酸和天冬醯胺二肽重複區組成的跨膜區，(iii)該蛋白質與肽葡聚糖中的五甘氨酸橋鍵共價結合需要的LPXTG基序，(iv)疏水的跨膜區，以及其後的(v)幾個帶正電荷的殘基。
通過開發金黃色葡萄球菌的整個基因組序列，鑑別8個編碼具有分秘和錨基基序指示MSCRAMMs的蛋白質的新的開放讀框(即攜帶N-末端的信號肽和C-末端LPXTG基序，隨後跟隨一個疏水的功能域和一個帶正電荷的尾部)。下面的表說明了所識別的蛋白質名錄，包括他們在基因組中的分布，他們的蛋白質大小，和挑選C-末端細胞壁的序列。
縮寫eMRSA-16；N，8325；C，COL；S，MSSA；J，N315，M，Mu50。
8個蛋白質中的6個存在於已經被測序的所有六個葡萄球菌的基因組中，剩餘的2個存在於5/6的這些基因組中。
下面是DNA和蛋白質序列的名錄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一級結構分析將生物信息方法(bioinformatic approach)用於一級結構和功能預測(附圖1)。蛋白質RrkN和DsqA具有與先前描述的MSCRAMMs相似的結構組織。RrkN的結構類似於分別為金黃色葡萄球菌和表皮葡萄球菌的Pls/Aap蛋白質。它在N-末端含有200個殘基的區域顯示與Pis和Aap有40％相同。該蛋白質的C-末端突出地由128個殘基重複區域組成，菌株與菌株之間重複區的數量是變化的。這些重複也存在於Pis和Aap。推定的sar同源區和fnbpA和fnbpB正好位於基因組上RrkN的上遊。
DsqA的結構的組織相似於蛋白質的Sdr家族。它含有典型的A區域，其後跟隨TYYFTDVK基序，該基序類似於在所有Sdr蛋白質中存在的保守的TYTFTVYVD基序。該基序的功能至今沒有被測定。2個88個殘基的重複區域居留在該蛋白質的中心，其後跟隨類似於在所Sdr蛋白質中存在的SD-重複基序的C-末端SX-重複基序。該重複區的大小隨菌株變化。在基因組上DsqA鄰接secY和secA。DsqA同系物(＞90％同一性)也存在於表皮葡萄球菌。
KnkA的序列中不含有重複區域。次級的結構預測分析顯示該蛋白質主要地由α-螺旋。
RkaS的序列中不含有重複區域。BLAST分析顯示它類似於5′核苷酸酶UDP-糖水解酶。編碼RkaS的基因正好地位於fromoryx的上遊，mec元件的插入位點。
KesK在該蛋白質的N-末端含有二個140個殘基的重複區域，有38％相同。水療法繪製圖分析(Kyte和Doolittle，1982)顯示在該蛋白質的中心點有一個大的親水的區域(殘基500-560)。
EkeS在該蛋白質的N-末端含有二個300個殘基的重複區域，有38％相同。Blast分析顯示該蛋白質的N-末端(殘基1-1268，攜帶兩種重複)與FmtB有49％等同，所述的FmtBan是具有17個串聯的重複的LPXTG蛋白質。FmtB被推測間接參與甲氧苯青黴素抗性，因為fmt8的滅活去除了甲氧苯青黴素抗性。這似乎是由於通過增加細胞壁前體葡萄糖胺-1-磷酸鹽的產生可以減輕對細胞壁組成的影響如甲氧苯青黴素敏感性(Komatsuzawa等人，2000)。
KrkN和KrkN2在基因組上互相鄰接。
表達分析由於與蛋白質資料庫之序列沒有同源性，不能預知這些蛋白質中每一個的推定的功能，因此以分子的近似值為依據。利用Qiagen pQE-30表達系統，四個開放讀框的獨特的區域可以在大腸桿菌中表達為重組his-標記的融合蛋白質。附圖2表示純化的N-末端his-標記的融合蛋白的考馬斯亮藍染色的SDS-PAGE凝膠。將重組蛋白質RrkN1，DsqA2，KesK1和KnkA用於在兔子中產生抗體。金黃色葡萄球菌細胞壁提取物的Western印跡分析顯示KesK，KnkA和DsqA被表達並且細胞壁相關(附圖3)。菌株eMRSA-16代表aknkA-陰性的菌株，因為它缺乏knkA基因。65kDa的免疫反應譜帶與菌株8325-4的指數和靜止的時期細胞的細胞壁提取物起反應(附圖3，B)。菌株eMRSA-16中沒有該譜帶暗示它代表了knkA的基因產物。
在指數和靜止期的培養物中，利用抗-KesK抗體進行的菌株8325-4的細胞壁成分的Western免疫印跡識別了150kDa免疫反應譜帶。表達質粒(pKS80)上全長KesK的表達獲得的從乳酸乳球菌的細胞壁成分釋放的類似大小的免疫反應蛋白質暗示該150kDa譜帶代表kesK基因產物(數據未顯示)。需要金黃色葡萄球菌的kesK敲出突變體來證實細胞壁釋放的KesK蛋白質的大小。
利用抗-DsqA抗體，金黃色葡萄球菌菌株MSSA和eMRSA-16的細胞壁成分的Western免疫印跡結果識別了一個130kDa免疫反應譜帶。在靜止期細胞中表達水平更高。
在乳酸乳球菌中異源表達以前已經將金黃色葡萄球菌的的表面蛋白質在乳酸乳球菌(L.lactis)中的異源表達用作為研究蛋白質功能的工具(Sinha等人，2000)。在該研究中，也將該替代系統用於在乳酸乳球菌表面表達各個in silico-預知的MSCRAMMs以查出功能。已經將KesK和KnkA克隆到乳酸乳球菌，並且通過點印跡顯示是在表面表達的(附圖4)。陰性的對照中沒有觀察到交叉反應(pKS80質粒沒有插入物)，顯示這是一個特異反應。通過用溶菌酶和mutanolysin消化產生了攜帶pKS-KnkA和pKS-KesK的乳酸乳球菌的細胞壁和原生質體成分，並且用於分別利用抗-KnkA和抗-KesK的抗體進行的Western印跡研究。與在金黃色葡萄球菌中觀察到的不同，在乳酸乳球菌細胞壁成分中沒有檢測到KnkA，但是發現與原生質體成分相關。KnkA的固定用基序不同於一致LPXTG序列，其中含有一個丙氨酸殘基而不是蘇氨酸(即LPKAG)(表1)。最近已經公開了金黃色葡萄球菌含有二個sortase基因，srtA和srtB(Pallen，2001)。由第二個sortase基因加工LPXTG基序的變異形式是可能的，乳酸乳球菌中沒有該基因。這也可以解釋在原生質體成分中觀察到的KnkA蛋白質大小的輕微的增加，而細胞壁分類揀選信號沒有被裂解。
在乳酸乳球菌的細胞壁成分中探測到KesK，但是遷移的分子量小於從金黃色葡萄球菌細胞壁釋放的KesK蛋白質。在乳酸乳球菌表面表達的大多數MSCRAMMs被證明在細胞壁提取程序中發生蛋白水解(Louise O′Brien，個人通訊)。因此，從乳酸乳球菌表面釋放KesK蛋白質表示截短形式的KesK是可能的。已經證明用溶菌酶和mutanolysin縮短消化時間以限制蛋白水解的程度。
in silico-預測的MSCRAMMs體內表達在ELISA分析中對來自金黃色葡萄球菌感染痊癒的33個患者的康復的人-時期血清進行測試，以檢測其識別純化的N-末端his-標記的融合蛋白質的能力。將從兒童和健康的供血者採掘的血清用作為陰性的對照。將陽性的反應其值作為等同於或高於陰性的對照的二倍。附圖5A-5D闡明在27-42％患者中識別了所有的蛋白質，這暗示這些蛋白質在體內表達並且在宿主感染期間是致免疫的。
引證文獻Komatsuzawa，H.，Ohta，K.，Sugai，M.，Fujiwara，T.，Glanzmann，P.，Berger-Bachi，B.，Suginaka，H.(2000)編碼細胞壁相關的蛋白質的fmt B基因的Tn551-介導的插入失活消除了金黃色葡萄球菌的甲氧苯青黴素抗性。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)異源表達的金黃色葡萄球菌粘連蛋白結合蛋白質足以侵入宿主細胞Infect.Immun.686871-6878。
Pallen，M.J.，Lam，A.C.，Antonio，M.，Dunbar，K.(2000)sortases的阻礙-一種豐富的反應物？Trends.Microbiol.997-101。
實施例2從金黃色葡萄球菌和從凝固酶陰性的葡萄球菌分離交叉反應的蛋白質並且進行測序最近已經證明表皮葡萄球菌含有結構上與金黃色葡萄球菌MSCRAMM蛋白質相關的表面蛋白質(US09/386,962)。從金黃色葡萄球菌中分離一個蛋白質特別有利，因為它與表皮葡萄球菌緊密同源。該蛋白質被稱為DsqA或SasA(金黃色葡萄球菌)和DgsK(表皮葡萄球菌)。它們的特徵是包括一個大約500個胺基酸殘基的典型的″A″區域，其後跟隨約有40％相同有，各為88個殘基的二個B重複區，和一個獨特的SXSX二肽重複區，該區域的長度隨菌株而變化。金黃色葡萄球菌的DsqA/SasA的A區域內含有一個180殘基的區域，該區域與金黃色葡萄球菌蛋白質的RrkN，Pls和表皮葡萄球菌蛋白質Aap的A區域內的一個相似的大小的區域有40％一致性。DsqA/SasA和DgsK蛋白質的A區域在胺基酸水平上有46％的一致性，BB重複區是50％一致性。所包括的主動和被動免疫接種策略；疫苗，識別金黃色葡萄球菌和凝固酶陰性的葡萄球菌的蛋白質的多克隆抗體和單克隆抗體是本發明的主題。
與凝固酶陰性的葡萄球菌和金黃色葡萄球菌發生交叉反應的抗體的特異性例子凝固酶陰性的葡萄球菌DgsK A-區域胺基酸序列(SEQID NO17)ASETPITSEISSNSETVANQNSTTIKNSQKETVNSTSLESNHSNSTNKQMSSEVTNTAQSSEKAGISQQSSETSNQSSKLNTYASTDHVESTTINNDNTAQQDQNKSSNVTSKSTQSNTSSSEKNISSNLTQSIETKATDSLATSEARTSTNQISNLTSTSTSNQSSPTSFANLRTFSRFTVLNTMAAPTTTSTTTTSSLTSNSVWNKDNFNEHMNLSGSATYDPKTGIATLTPDAYSQKGAISLNTRLDSNRSFRFIGKVNLGNRYEGYSPDGVAGGDGIGFAFSPGPLGQIGKEGAAVGIGGLNNAFGFKLDTYHNTSTPRSDAKAKADPRNVGGGGAFGAFVSTDRNGMATTEESTAAKLNVQPTDNSFQDFVIDYNGDTKVMTVTYAGQTFTRNLTDWIKNSGGTTFSLSMTASTGGAKNLQQVQFGTFEYTESAVAKVRYVDANTGKDIIPPPKTIAGEVDGTVNIDKQLNNFKNLGYSYVGTDALKAPNYTETSGTPTLKLTNSSQTVIYKFKDVQ
金黃色葡萄球菌SasA A-區域胺基酸序列(SEQ ID NO18)ASDAPLTSELNTQSETVGNQNSTTIEASTSTADSTSVTKNSSSVQTSNSDTVSSEKSEKVTSTTNSTSNQQEKLTSTSESTSSKNTTSSSDTKSVASTSSTEQPINTSTNQSTASNNTSQSTTPSSVNLNKTSTTSTSTAPVKLRTFSRLAMSTFASAATTTAVTANTITVNKDNLKQYMTTSGNATYDQSTGIVTLTQDAYSQKGAITLGTRIDSNKSFHFSGKVNLGNKYEGHGNGGDGIGFAFSPGVLGETGLNGAAVGIGGLSNAFGFKLDTYHNTSKPNSAAKANADPSNVAGGGAFGAFVTTDSYGVATTYTSSSTADNAAKLNVQPTNNTFQDFDINYNGDTKVMTVKYAGQTWTRNISDWIAKSGTTNFSLSMTASTGGATNLQQVQFGTFEYTESAVTQVRYVDVTTGKDIIPPKTYSGNVDQVVTIDNQQSALTAKGYNYTSVDSSYASTYNDTNKTVKMTNAGQSVTYYFTDWAap蛋白質的完整序列和它的編碼DNA(指示有A區域存在)顯示如下表皮葡萄球菌Aap蛋白質(加下劃線的為A-區域)(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從A-區域到表皮葡萄球菌Aap(胺基酸55-600)(SEQ IDNO21)55EKQVDPITQANQNDSSERSLENTNQPTVNNEAPQMSSTLQAEEGSNAEAPQSEPTKAEEGGNAEAAQSEPTKAEEGGNAEAPQSEPTKAEEGGNAEAAQSEPTKTEEGSNVKAAQSEPTKAEEGSNAEAPQSEPTKTEEGSNAKAAQSEPTKAEEGGNAEAAQSEPTKTEEGSNAEAPQSEPTKAEEGGNAEAPQSEPTKTEEGGNAEAPNVPTIKANSDNDTQTQFSEAPTRNDLARKEDIPAVSKNEELQSSQPNTDSKIEPTTSEPVNLNYSSPFMSLLSMPADSSSNNTKNTIDIPPTTVKGRDNYDFYGRVDIESNPTDLNAT
NLTRYNYGQPPGTTTAGAVQFKNQVSFDKDFDFNIRVANNRQSNTTGADGWGFMFSKKDGDDFLKNGGILREKGTPSAAGFRIDTGYYNNDPLDKIQKQAGQGYRGYGTFVKNDSQGNTSKVGSGTPSTDFLNYADNTTNDLDGKFHGQKLNNVNLKYNASNQTFTATYAGKTWTATLSELGLSPTDSYNFLVTSSQYGNGNSGTYASGVMRADLDGA600蛋白質的生產和純化利用PCR，從上面描述的序列擴增識別為DgsK或SasA的表面蛋白質的A區域，再克隆到大腸桿菌表達載體PQE-30(Qiagen)，以允許表達一種含有六個組氨酸殘基的重組融合蛋白質。隨後將該載體轉化到大腸桿菌ATCC 55151，在15-升發酵罐中生長到光密度(OD600)為0.7，用0.2mM異丙基-1β-D半乳糖苷(IPTG)誘導4小時。利用AG Technologies中空纖維組件(孔大小0.45um)收集細胞，將細胞漿在-80C冷凍。利用2次穿過French Press，1100psi，將細胞在1×PBS(10mL緩衝液/1克的細胞漿)裂解。將裂解的細胞17,000rpm旋轉30分鐘以除去細胞碎片。將上清液經過裝備了0.1M NiCI2的5-mL Hi俘獲Cheating(Pharmacia)柱。在上樣之後，用5個柱體積的10mM Tris，pH 8.0，100mM NaCI(緩衝液A)洗滌柱子。利用超過30個柱體積的0-100％梯度的10mM Tris，pH 8.0，100mM NaCI，200mM咪唑(Buffer B)洗脫蛋白質。SdrGN1N2N3或SdrGN2N3在～13％Buffer B(～26mM咪唑)被洗脫。監測280nm處的吸光率。在1×PBS中透析含有SdrGN1N2N3或SdrGN2N3的餾分。
然後將各個蛋白質經過一個內毒素除去方案。在該方案中所使用的緩衝液通過一個5-mL Mono-Q瓊脂糖(Pharmacia)柱而沒有內毒素。將蛋白質平衡地分到4×15毫升試管中。各個試管的體積用緩衝液A加到9毫升。將1毫升的10％Triton X-114加入到各個試管中，在4℃旋轉保溫1小時。將試管置於37℃水浴中以使相位分離。將試管以2,000rpm旋轉10分鐘，收集各個試管的上面的含水的相，用去汙劑反覆提取。將2次提取的含水相合併，傳遞過帶有0.1M NiCl2的5-毫升IDA cheating(Sigma)柱，以除去剩餘的洗滌劑。用9個柱體積的Buffer A洗滌該柱，然後用3個柱體積的Buffer B洗脫該蛋白質。將洗脫液通過5-毫升的Detoxigel(Sigma)柱，收集流出物，再上樣到柱。收集第二次過柱的流出物，在1×PBS中透析。分析純化的產物的濃度，純度和內毒素水平，然後給小鼠給藥。
單克隆抗體的生產將大腸桿菌表達的和純化的重組SasA和DsgK蛋白質用於製備一組鼠的單克隆抗體，而將小鼠血清用作為多克隆抗體的來源。簡單地說，一組Balb/C或SJL小鼠接收了一系列皮下的免疫接種1-10mg溶液蛋白質或與下面表描述的佐劑混合的蛋白質。
免疫接種方案RIMMS注射天數數量(微克)途徑佐劑#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
常規注射 天數數量(微克)途徑佐劑原始 0 5 皮下FCA加強#114 1 腹膜內 RIBI加強#228 1 腹膜內 RIBI加強#342 1 腹膜內 RIBI在殺死時(RIMMS)或加強後幾天收集(常規的)血清，在ELISA測定中測定抗MSCRAMM蛋白質或對完整的細胞(表皮葡萄球菌和金黃色葡萄球菌)的效價。在最後的加強之後3天，去除脾臟或淋巴結，挑開成單細胞懸浮液，收集淋巴細胞。然後將淋巴細胞與P3X63Ag8.653骨髓瘤細胞系(ATCC CRL-1580)融合。根據Current Protocols in Immunology(第2章，第2單元)中的生產單克隆抗體的方案進行細胞融合，隨後塗覆平板和餵養。
然後利用標準的ELISA分析方法，從融合產生的任何克隆中篩選特異的抗-SasA抗體的產生。擴增陽性的克隆，進一步通過流式血細胞計數測試完整的細菌的細胞結合測定中的活性以及Biacore分析中SasA的結合。
Biacore分析在整個分析中，流速保持在10毫升/分鐘不變。在注射SasA或DgsK之前，藉助於RAM-Fc結合，將測試抗體吸附到晶片。在時間0，將濃度為30mg/ml的SasA或DgsK注射過晶片3分鐘，隨後解離2分鐘。該階段的分析測定了Mab/SasA或DgsK相互作用的相對關係和分離動力學。
與完整細菌的結合用兔子IgG(50mg/ml)封閉之後，收集，洗滌細菌的樣品金黃色葡萄球菌Newman，金黃色葡萄球菌60，金黃色葡萄球菌397(Sal6)，金黃色葡萄球菌Wood，金黃色葡萄球菌8325-4，甲氧苯青黴素抗性的金黃色葡萄球菌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，用濃度為2pg/ml的Mab或僅用PBS(對照)保溫。在用抗體保溫之後，將用作為檢測抗體的山羊-F(ab′)2-抗-小鼠-F(ab′)2-FITC用於孵育細菌的細胞。在抗體標記之後，用FACS caliber流式細胞計數器抽吸細菌的細胞以分析螢光射出(刺激488，發射570)。對於各個細菌的菌株，收集和檢測10,000個樣品。這些資料顯示抗金黃色SasA的抗體能夠識別凝固酶陰性的葡萄球菌的表面的同源的蛋白質。該資料支持Western印跡分析，表明抗金黃色葡萄球菌SasA的兔子多克隆抗體與從表皮葡萄球菌HB細胞表面釋放的蛋白質以及來自於從表皮葡萄球菌克隆的DsgK的重組A-區域發生交叉反應(參見附圖6和下表)。
多克隆血清反應性
序列表110蒂莫西·J·福斯特等人120識別凝血酶陰性的葡萄球菌和金黃色葡萄球菌的表面蛋白的交叉反應性單克隆抗體和多克隆抗體130P07263US01/BAS140US 10/172,5021412002-06-17150US 60/298,0981512001-06-1516029170Patent In version 3.121012116609212DNA213Staphylococcus epidermidis4001acaacacagc 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 660921022112189212PRT213Staphylococcus epidermidis4002Met 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 218521032116852212DNA213Staphylococcus epidermidis4003tctaatgaat 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 685221042112283212PRT
213Staphylococcus epidermidis4004Ser 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 2275228021052112730212DNA213Staphylococcus epidermidis4005ttattatcaa 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 atctaaataa27302106211909212PRT213Staphylococcus epidermidis4006Leu 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 90521072111065212DNA213Staphylococcus epidermidis4007gaggaaaaca 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 aataa10652108211354212PRT213Staphylococcus epidermidis4008Glu 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
21092111965212DNA213Staphylococcus epidermidis4009tatacaatta 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 196521010211654212PRT213Staphylococcus epidermidis40010Tyr 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 650210112112406212DNA213Staphylococcus epidermidis40011tttataaata 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 240621012211801212PRT
213Staphylococcus epidermidis40012Phe 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 800Ser210132114914212DNA213Staphylococcus epidermidis40013agtggaaaat 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 4914210142111637212PRT213Staphylococcus epidermidis40014Ser 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 1635210152111923212DNA213Staphylococcus epidermidis40015ggaaggagta 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 192321016211640212PRT213Staphylococcus epidermidis40016Gly 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 64021017211522212PRT213Staphylococcus epidermidis40017Ala 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 52021018211485212PRT213Staphylococcus epidermidis40018Ala 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 Val485210192111245212PRT213Staphylococcus epidermidis40019Met 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 1245210202113765212DNA213Staphylococcus epidermidis40020atgggcaaac 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 376521021211546212PRT213Staphylococcus epidermidis40021Glu 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 Ala5452102221136212PRT213Staphylococcus aureus40022Leu 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 Ser352102321143212PRT213Staphylococcus aureus40023Leu 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 402102421135212PRT
213Staphylococcus aureus40024Leu 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 Lys352102521138212PRT213Staphylococcus aureus40025Leu 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 Lys352102621136212PRT213Staphylococcus aureus40026Leu 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 Asn352102721134
212PRT213Staphylococcus aureus40027Leu 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 Ser2102821133212PRT213Staphylococcus aureus40028Leu 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 30Asn2102921139212PRT213Staphylococcus aureus40029Leu 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權利要求
1.一種分離的抗體，其與葡萄球菌的表面蛋白質結合，所述蛋白質選自於下列組SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21。
2.根據權利要求1所述的抗體，其中抗體針對所述表面蛋白質的A區域。
3.根據權利要求1所述的抗體，其中抗體用於治療或預防人類或動物的金黃色葡萄球菌感染。
4.根據權利要求1所述的抗體，其中抗體適用於人類或動物的非腸道的，口部的，鼻內的，皮下的，aerosolized或靜脈內的給藥。
5.根據權利要求1所述的抗體，其中所述的抗體是單克隆抗體。
6.根據權利要求1所述的抗體，其中所述的抗體是多克隆抗體。
7.根據權利要求5所述的抗體，其中單克隆抗體是一種選自於下列組的類型鼠的，嵌合的，人性化的，和人的單克隆抗體。
8.根據權利要求5所述的抗體，其中抗體是一種單鏈單克隆抗體。
9.根據權利要求1所述的抗體，它包括具有類似於一種抗體的結合特異性的抗體片段，所述的抗體結合到具有選自於下列組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質。
10.根據權利要求1所述的抗體，它針對具有選自於下列組SEQID NOS.2、4、6、8、10、12、14、16、17、18、19和21的胺基酸序列的蛋白質。
11.根據權利要求1所述的抗體，其中所述的表面蛋白質具有由選自於下列組的核酸序列SEQ ID NOS.1、3、5、7、9、11、13、15、20和編碼Aap蛋白質的A區域或其簡併序列的核酸序列編碼的一個胺基酸序列的核酸序列。
12.含有權利要求1的抗體的分離的抗血清。
13.一種診斷試劑盒，包括權利要求1的抗體和用於檢測該抗體引起的結合的工具。
14.根據權利要求13所述的診斷試劑盒，其中所述的用於檢測結合的工具包括連接到所述的抗體的一種可檢測標記物。
15.一種用於診斷金黃色葡萄球菌感染的方法，包括將權利要求1的抗體加入到被懷疑感染了金黃色葡萄球菌的樣品中，測定抗體是否已經結合到該樣品。
16.一種用於治療或預防金黃色葡萄球菌感染的藥物組合物，包括有效量的權利要求1的抗體和藥學的可接受的介質，載體或賦形劑。
17.一種治療或預防金黃色葡萄球菌感染的方法，包括將有效量的權利要求1的抗體給人或動物患者給藥。
18.一種誘導免疫應答的方法，包括以致免疫量的選自於下列組SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21的胺基酸序列的分離的蛋白質給人類或動物給藥。
19.根據權利要求1所述的分離的抗體，它具有結合到由SEQ IDNOS.1、3、5、7、9、11、13、15、20的核酸序列和編碼Aap蛋白質的A區域的核酸序列或其簡併序列編碼的胺基酸序列的能力。
20.一種來自DsqA蛋白質的A區域的分離的活性片段。
21.根據權利要求1所述的分離的抗體，進一步包括生理學上的可接受的抗生素。
22.一種用於治療或預防金黃色葡萄球菌感染的疫苗，包括選自於下列組SEQ ID NOS.2、4、6、8、10、12、14、16、17、18、19和21的蛋白質序列，和一種藥學的可接受的介質，載體或賦形劑，所述的蛋白質的量可有效地激發免疫應答。
全文摘要
提供了與凝血酶陽性的葡萄球菌細菌，例如溶血葡萄球菌發生交叉反應的多克隆和單克隆抗體，所述的抗體能夠識別凝固酶陽性的和凝固酶陰性的葡萄球菌細菌的表面蛋白質。從基於金黃色葡萄球菌和凝固酶陰性的葡萄球菌之間共有的特性分離的表面蛋白質可以製備抗體，這些重組表面蛋白質可用於製備本發明的抗體。還提供了利用這些蛋白質和抗體用於治療或抵抗各種各樣的葡萄球菌的感染的疫苗和方法。
文檔編號A61P31/04GK1543569SQ02816001
公開日2004年11月3日 申請日期2002年6月17日 優先權日2001年6月15日
發明者蒂莫西·J·福斯特, 菲奧那·羅切, 約瑟夫·M·帕蒂, 傑夫·T·哈欽斯, 皮特羅·斯皮茲利, 馬克·帕倫, M 帕蒂, 斯皮茲利, 羅切, T 哈欽斯, 帕倫, 蒂莫西 J 福斯特 申請人:英希比泰克斯公司, 都柏林伊莉莎白女皇神學院, 帕維研究學院