抗原多肽的製作方法

2023-06-27 23:48:21

專利名稱：抗原多肽的製作方法
技術領域：
本發明涉及一種鑑定由病原微生物所表達的抗原多肽的方法；包含所述多肽的疫苗；製備所述多肽的重組方法；和針對所述多肽的治療用抗體。
微生物引起的大量致死或致弱的疾病，影響著全世界數百萬人。控制微生物的現有方法包括應用抗微生物製劑(抗生素)和消毒劑。它們被證明是有問題的，由於暴露在這些製劑的環境中所形成的顯著的選擇性壓力導致了耐藥性微生物的產生，這些微生物能夠躲避抗微生物製劑的作用。例如，近來已發現微生物對三氯生產生了耐藥性，而在家庭及工業環境中使用的許多消毒劑中都添加有三氯生。
一個越來越引起爭議的問題是許多重要的病原微生物進化出了對抗生素的耐藥株。
例如，但並不限於所舉的例子，估計全世界超過5千萬人感染了耐藥的結核病(TB)(數字來自世界衛生組織，1998)。過去使用抗生素治療結核病依賴於單一藥物(如乙硫異煙胺)，因此產生了較高頻率的耐藥性。由於這個原因，現在治療結核病採用聯合用藥。但即使聯合用藥進行治療，由至少對一種用於治療結核病的藥物產生耐藥性的菌株所引起的治療失敗的比率仍然接近50％。導致TB的分枝桿菌是一種慢性生長的細菌，需要花費很長時間消滅它。所以，聯合用藥若要產生療效，TB患者必須每天服用組合藥物至少6個月。病人通常必須每天服用兩種或更多種藥物，並需要相當長的治療期限。許多病人僅是斷續服藥而沒有完成徹底根除分枝桿菌感染的全部療程。再者，TB與愛滋病感染有密切聯繫，因而TB的產生是與免疫抑制緊密相關的。
抗TB的疫苗已經應用了許多年。作為一種對潛在的可能感染TB的大量人群的安全和廉價的預防手段，卡介苗(BCG)已在長時間內廣泛應用於全世界。BCG是從牛分枝桿菌獲得的減毒活疫苗。但是預防接種對TB感染的效果不大，所以BCG對全面控制這一疾病的作用是有限的。
金黃色葡萄球菌是病原微生物對抗生素產生耐藥性的另一個例子。金黃色葡萄球菌是一種通常寄居在約20-40％普通健康人鼻腔上皮內表面的細菌，也常在人皮膚上發現，通常不會引起疾病。但在某些場合，特別是當皮膚受損時，這種細菌能引起感染。在醫院裡這是一個特殊的問題，因為在那裡病人會進行外科手術和/或服用免疫抑制藥物。這些病人由於他們所接受的治療更易受金黃色葡萄球菌的感染。近年來已產出現了金黃色葡萄球菌的耐藥株。對甲氧苯青黴素的耐藥株已很普遍，這些耐藥株也對另外幾種抗生素有耐藥性。目前對金黃色葡萄球菌還沒有有效的預防接種措施。在美國，每年兩百萬住院感染中的13％是由金黃色葡萄球菌感染引起的。這表示260,000人感染金黃色葡萄球菌，其中60-80,000人死亡。
因此，金黃色葡萄球菌是一種重要的人類病原菌，能引起許多威脅生命的疾病，包括敗血症、心內膜炎、關節炎和中毒性休克。其致病力是由其微生物的多樣性及其致病成分的毒力決定的。致病原因是多因素的，任何單一致病成分均不能解釋一個特定的感染，見Projan，S.J. Novick，R.P.(1997)著《人類疾病中的葡萄球菌》(Crossley，K.B. Archer，G.L.，eds.)pp.55-81。
在感染初期，及發展階段，微生物的需要及其環境發生改變，相應地，金黃色葡萄球菌所產生的致病毒素隨之發生改變。在感染開始時，病原體粘附於宿主組織是關鍵性的，於是便產生了大量與在細胞表面粘附相關的蛋白質，包括膠原結合蛋白、血纖蛋白原結合蛋白和纖粘連蛋白結合蛋白。病原體還具有產生能減少吞噬作用或是幹擾細胞被循環抗體識別的因子的能力，從而逃避宿主的防禦系統。
通常，感染病灶會發展為一個膿腫，其中細菌大量滋生。金黃色葡萄球菌通過產生一定數量的感受肽能監測其自身的細胞密度。肽的積累以及所伴隨的生理變化，引起細胞的營養不足，從而使細胞所產生的致病毒素從粘附素轉變為引起浸潤和組織穿透的組分。這包括多種溶血素、蛋白酶和其他降解酶等。
在任何感染過程中，金黃色葡萄球菌所產生的致病毒素是隨環境和生理刺激而改變的。這些刺激取決於體內局部的環境並隨著感染的進程而改變。由於對體內的情況所知甚少，並且某些毒素組分似乎僅能在體內的環境下產生，所以一些可能成為疫苗的組分不能為早期的技術所發現。
近代藥物歷史中最重要的發展之一是疫苗的發展，其對各種病原微生物的感染提供了預防性保護。許多疫苗是由滅活的或減毒的病原製成的，然後注射給個體。經免疫的個體通過產生體液(抗體)和細胞(溶細胞的T細胞，CTL’s)應答而做出反應。例如，肝炎疫苗的製備是通過加熱滅活病毒和用像甲醛這樣的交聯劑處理。減毒病原的一個代表例子是由活病原弱化產生的脊髓灰質炎疫苗。
但是對特定疾病應用減毒疫苗是存在顧慮的，因為缺乏對於減毒過程的本質和條件的病理學知識的了解。這個問題對於特定病毒製劑而言尤其突出，這是因為病毒，特別是逆轉錄病毒，具有錯誤傾向的複製周期，導致含有病毒的基因產生能夠傳代的突變，其結果是被用做疫苗的抗原決定簇的發生改變。替代使用滅活的或減毒的病原的一種方法是鑑別出免疫系統對其特別敏感的病原表位。由此，許多由病原微生物在感染過程中產生的致病毒素，對於開發具有保護個體免於受到特定致病微生物感染作用的疫苗是特別有用的。
發展亞單位疫苗(疫苗其免疫原是由特定的病原微生物表達的一個蛋白質或複合體的一個片段或亞基)已經成為醫學研究的焦點。對鑑別出用於發展亞單位疫苗的候選分子的需求，顯然不僅僅是因為近來抗生素耐藥性的出現已經成為用傳統化學療法控制致病微生物的嚴重障礙。已經發展了許多方法來確定有可能用作疫苗的抗原多肽。在此舉一個方法以作說明。許多年來已知腫瘤細胞能產生一些腫瘤細胞特異性抗原，其中一些出現在腫瘤細胞表面。免疫系統識別這些抗原並將其作為外來物，然後產生針對自身抗原的抗體，這叫自身抗體或自身抗血清。
這種技術之一是重組表達克隆抗原的血清學識別，簡寫成SEREX。
典型地說，這一技術包括從腫瘤組織中提取RNA，然後從分離的總RNA中的選擇性富集mRNA。mRNA用病毒逆轉錄酶反轉錄為cDNA。這樣合成的cDNA再被克隆進入一個表達載體，並且轉化進入一個適當的細菌株。轉化的細菌被接種到一個營養適宜的瓊脂平板上，在適宜的生長條件下，亞克隆的cDNA在細菌細胞中由表達載體來表達。應用基於噬菌體的表達載體可導致細胞自然溶解，例如λ噬菌體或基於噬菌粒的載體，通過它們的溶解循環引起細胞溶解。釋放的多肽被轉移至一種合適的膜支持物(如硝酸纖維素，尼龍)，並暴露於從最初自其分離腫瘤組織的病人中獲得的自身抗血清。免疫篩選法可以用於從病人腫瘤組織中鑑定出過量表達或不恰當表達的基因。
我們使用這種技術來鑑定感染過程中由病原微生物所表達的抗原多肽。感染中產生的自身抗血清，被用於篩選一個由基因組DNA產生的表達文庫，以鑑定和克隆抗原。
本發明最廣義的方面是涉及在感染中由病原微生物表達的抗原多肽的鑑定。
按照本發明的第一方面，提供一種用於鑑定抗原多肽的方法，包括
(i)提供一個編碼病原微生物的基因或部分基因序列的核酸文庫；(ii)轉化/轉染所述核酸文庫至一種宿主細胞；(iii)提供有助於表達所述的轉化/轉染的基因或部分基因序列的條件；(iv)將所述基因/部分基因序列表達的多肽與從感染或曾經感染過所述致病微生物的動物獲得的自身抗血清相作用；以及(v)純化編碼與所述自身抗血清相結合的多肽或部分多肽的核酸。
在本發明的優選方法中，所述的文庫包含一個病原微生物的基因組DNA。
理想的所述病原微生物是細菌。
更優選地，所述的細菌微生物是從以下選擇出來的金黃色葡萄球菌；表皮葡萄球菌；糞腸球菌；結核分枝桿菌；B族鏈球菌；肺炎鏈球菌；幽門螺桿菌；淋病奈瑟氏球菌；A族鏈球菌；布氏疏螺旋體；粗球孢菌；莢膜組織胞漿菌；B型腦膜炎奈瑟氏球菌；弗氏志賀氏菌；大腸桿菌；流感嗜血菌。
更優選地，所述病原微生物是葡萄球菌屬細菌。理想的病原生物體是金黃色葡萄球菌或表皮葡萄球菌。
在本發明的一個更優選的實施方案中，所述核酸文庫是一個λ文庫，理想地，是一個λ表達文庫。
按照本發明的第二個方面，提供了含有一個DNA序列的核酸分子，所述DNA序列選自(i)如SEQ ID NO1-13中代表的DNA序列；(ii)與上述(i)中確定的SEQ ID NO1-13所示序列雜交並編碼一種致病微生物所表達的多肽的DNA序列，以及(iii)由(i)和(ii)中所確定的DNA序列的遺傳密碼所產生的簡併DNA序列。
在本發明的一個更優選的實施方案中，所述核酸分子是基因組DNA。
在本發明的一個優選的實施方案中，提供了一種分離的核酸分子，所述核酸分子在嚴格雜交條件下與SEQ ID NO1-13中所示序列退火。
嚴格的雜交/漂洗條件是本領域的公知常識。例如，核酸雜交體在0.1％×SSC，0.1％SDS中60℃下漂洗後是穩定的。本領域公知，如果已知核酸的序列，可以計算出最佳的雜交條件。例如，根據要進行雜交的核酸的GC含量，能將雜交條件確定下來。請閱Sambrook等(1989)著《分子克隆實驗手冊》。一個用於計算給定的同源核酸分子間雜交所需嚴格條件的普遍公式為Tm＝81.5℃+16.6Log[Na+]+0.41[％G+C]-0.63(％甲醯胺)按照本發明的第三個方面，提供至少一種依照本發明的方法所鑑定的多肽。
在本發明的一個優選的實施方案中，根據本發明前面所述的任一方面或實施方案，所述多肽與一種微生物的感染致病力相關。
更優選地，所述多肽至少是SEQ ID NO14-19之一或一部分。
按照本發明的第四個方面，提供了一種核酸分子，其特徵在於所述的核酸分子是一個載體的一部分，所述載體適於促進由所述核酸分子編碼的多肽的重組表達。
在本發明的一個優選的實施方案中，所說的載體是一種適應於進行原核基因表達的表達載體。或者，所述表達載體適應於進行真核基因表達。
典型地，所述的適應性包括，例如但不限於，提供介導細胞特異性表達的轉錄控制序列(啟動子序列)。這些啟動子序列可以是細胞特異性的，可誘導的或組成性的。
啟動子是本領域所知的術語，為了清楚的目的，僅舉一個例子，但不限於此例，以說明其如下的特徵。增強子元件是順式作用的核酸序列，通常位於一個基因的轉錄起始位點的5』端(增強子也可出現在基因序列的3』端或者甚至位於內含子序列內部，因此其不受位置的約束)。增強子的功能是增加與增強子連接的基因的轉錄頻率。增強子的活性是響應與增強子元件特異性結合的反式作用轉錄因子(多肽)。轉錄因子的結合/活性(請看《真核轉錄因子》，DavidS Latchman，Academic Press Ltd，San Diego著)依賴於多種環境因素包括，例如但不限於，中間代謝產物(如葡萄糖，脂質)，環境作用因子(如光，熱)。
啟動子元件還包括TATA框和RNA聚合酶起始選擇(RIS)序列，其功能是選擇轉錄起始位點。這些序列也結合於多肽上，所述多肽，和其他事物一起，促進RNA聚合酶對轉錄起始點的選擇。
適應性也包括提供選擇標誌和自動複製序列，兩者均有助於所述載體在真核細胞或原核宿主中的維持。自動維持的載體稱為附加載體。
促進載體編碼基因表達的適應性包括提供轉錄終止/多聚腺苷酸序列。還包括提供內部核糖體進入位點(IRES)，其功能是最大表達排列在二順反子或多順反子表達盒中的載體編碼基因。
這些適應性在本領域是熟知的。這裡有一系列關於表達載體的構建和重組DNA的通用技術的出版物。請看，Sambrook等(1989)著《分子克隆實驗手冊》，冷泉港實驗室，冷泉港，NY和其中的參考文獻；Marston，F(1987)著《DNA克隆技術》；一本IRL出版的實用手冊，Vol III IRL，英國牛津，《DNA克隆》F M Ausubel等著，《分子生物學中的通用方案》，John Wiley Sons，Inc.(1994)。
按照本發明的另外方面，提供了一種生產前述任一方面或實施方案所述多肽的方法，包括
(i)提供一種用本發明的載體轉化/轉染的細胞；(ii)在利於製備所述多肽的條件下培養所述細胞；以及(iii)從所述細胞或其生長環境中純化所述多肽。
在本發明的優選方法中，所述載體編碼並因此為所述重組多肽提供一種促進所述多肽純化的分泌信號。
按照本發明的第五個方面，本發明提供了一種用載體轉化或轉染的細胞或細胞系。
在本發明的一個優選的實施方案中，所述細胞是原核細胞。或者，所述細胞是選自真菌、昆蟲、兩棲動物、哺乳動物、植物的真核細胞。
按照本發明的另一方面，提供了一種疫苗，所述疫苗包含至少一種本發明的多肽。
理想地，所述疫苗還包含載體和/或佐劑。
載體和佐劑是以下述方式表述。某些多肽或肽抗原含有B-細胞表位但沒有T細胞表位。通過在多肽/肽中加入T細胞表位，或通過將多肽/肽結合到一種含有多個T細胞表位的免疫原性載體蛋白上，如血蘭蛋白或破傷風毒素，能大大增強免疫應答。結合物由抗原呈遞細胞攝取，加工，並通過人白細胞抗原(HLA’s)II類分子呈遞。這使得在T細胞的幫助下將針對源自載體表位的T細胞的特異性傳遞給對原始的抗原多肽/肽具有特異性的B細胞。這能增加抗體產生、分泌和同型轉化。
佐劑是能夠通過調節免疫細胞活性而增加對抗原的特異性免疫應答的一種物質或程序。佐劑的例子包括，例如但不限於，共刺激分子的激動抗體、弗氏佐劑、胞壁醯二肽、脂質體。因此佐劑是一種免疫調節劑。載體是一種免疫原性分子，當與第二個分子結合後會增強針對後者的免疫應答。
本發明另一方面提供了一種方法，所述方法包括通過對一種動物施用至少一種本發明的多肽或其一部分，或本發明的疫苗，使所述動物產生抵抗一種病原微生物的免疫力。
在本發明的一個優選的方法中，所述動物是人。
優選地，疫苗或抗原多肽能通過靜脈內注射，肌肉注射，或皮下注射等直接注射接種。進一步地，疫苗或抗原多肽，或許還可以口服。
優選地，疫苗能抵抗金黃色葡萄球菌。
疫苗也可能抵抗表皮葡萄球菌。
顯然，疫苗或抗原多肽對於控制或減輕除了人以外的其它動物的病情也是有效的，例如，但不限於例子，家庭寵物，家畜，馬匹。
按照本發明的其它方面，還提供了一種抗體或至少其一個有效結合部分，所述抗體結合至少一種本發明的多肽。
在本發明的一個優選實施方案中，所述抗體是一種多克隆或單克隆抗體，其中所述抗體對所述多肽具有特異性。
或者，所述抗體是一種通過重組方法產生的嵌合抗體，其包含所述抗體的可變區與人類抗體的不變或恆定區。
在本發明的另一個替代實施方案中，所述抗體是通過重組方法產生的人源化抗體，是所述抗體的互補決定區域與一種人抗體的恆定區(C)和可變(V)區的讀框的結合。
優選地，所述抗體被給予了一種標誌，包括一種傳統的標記或標籤，例如放射性和/或螢光和/或表位的標記或標籤。
優選地，針對所述的多肽的所述人源化的單克隆抗體，是由一種適合於轉化或轉染原核細胞或真核細胞的表達載體所產生的融合多肽。
抗體，也叫做免疫球蛋白，是一種對外來分子(抗原)具有特異性的蛋白分子。免疫球蛋白(Ig)是一類結構相關蛋白質，包括兩對多肽鏈，一對是輕(L)(低分子量)鏈(κ或λ)，一對是重(H)鏈(γ，α，μ，δ和ε)，所有四條鏈通過二硫鍵結合在一起。重鏈和輕鏈都有結合抗原的區域，並且在Ig分子之間是高度可變的。另外，重鏈和輕鏈包括非可變或恆定區域。
輕鏈包括兩個結構域。羧基-末端結構域在一種類型的輕鏈中是基本相同的，因此被稱為「恆定」(C)區。氨基-末端結構域在輕鏈和輕鏈間是變化的並構成抗體的結合位點。因為它的可變性，它被稱為「可變」(V)區。
Ig分子的重鏈分幾個類型，α，μ，δ，ε，和γ(其中還分幾個亞型)。裝配好的Ig分子包括一個或多個由相同的兩條重鏈和輕鏈組成的單位，並從它擁有的重鏈獲得它的名稱。這樣，Ig便分成5個型IgA，IgM，IgD，IgE和IgG(並因重鏈的不同而分成4個亞型，例如，IgG1，IgG2，IgG3和IgG4)。關於抗體結構和它們的不同功能的更詳細的解釋，可以參考《抗體的應用一本實驗室手冊》，冷泉港實驗室出版。
嵌合抗體是重組抗體，其中小鼠或大鼠抗體的所有可變區與人抗體的恆定區結合。人源化的抗體是重組雜交抗體，它融合了來自於嚙齒類抗體的可變區的互補決定區與人抗體可變區的框區。也用了人抗體的恆定區。互補決定區(CDRs)位於抗體的重鏈和輕鏈的N-末端結構域，可變區的主要變化多集中在CDRs。這些區域構成位於抗體分子表面的環狀結構。這些環狀結構在抗體和抗原之間提供結合表面。
非人類動物的抗體可引起對外來抗體產生免疫應答，並由此被從血液循環中清除。注射到人機體中的嵌合抗體和人源化抗體具有弱的抗原性，因為在重組雜交抗體中嚙齒類(即異類)抗體的量減少，而人的抗體區域不會引起免疫應答。它使得針對該抗體的免疫應答減弱，並減少了對抗體的清除。這顯然對於應用治療性抗體治療人類疾病來說是很需要的。人源化抗體被設計為具有更少「異類」抗體區域，因此與嵌合抗體相比，人源化抗體是更弱的免疫原。
本發明的另一方面是提供一種載體，所述載體適用於按照本發明表達人源化的或嵌合體的抗體。
本發明進一步提供了一種細胞或細胞系，所述細胞或細胞系用編碼本發明的人源化的或嵌合體的抗體的載體轉化或轉染。
本發明進一步提供了一種製備本發明的人源化或嵌合抗體的方法，其包括(i)提供用一個載體轉化或轉染的一種細胞，所述載體包括一種編碼本發明的人源化或嵌合體抗體的核酸分子；(ii)在有利於製造所述抗體的條件下使所述細胞增殖；以及(iii)從所述細胞中或它的生長環境中純化所述抗體。
本發明進一步提供了一種雜交瘤細胞系，其可產生如前所描述的單克隆抗體。
本發明進一步提供了一種用雜交瘤細胞系生產本發明的單克隆抗體的方法。
本發明進一步提供了製備一種能產生本發明的單克隆抗體的雜交瘤細胞系的一種方法，它包括以下步驟i)用一種免疫原免疫一種具有免疫活性的哺乳動物，所述免疫原包含至少一種具有如SEQ.ID No 14-19中所示的胺基酸序列的多肽或其片段；ii)將接受免疫的免疫活性動物的淋巴細胞與骨髓瘤細胞融合形成雜交瘤細胞；iii)根據與步驟(i)中的胺基酸序列的結合活性篩選出由步驟(ii)中的雜交瘤細胞產生的單克隆抗體；iv)培養雜交瘤細胞以使其增殖和/或分泌所述單克隆抗體；以及v)從培養上清液中回收單克隆抗體。
優選地，上述具免疫活性的哺乳動物是小鼠。或者，上述具免疫活性的哺乳動物是大鼠。
使用雜交瘤細胞產生單克隆抗體是本領域所熟知的。用於產生單克隆抗體的方法是由Kohler和Milstein，(自然256，495-497(1975))，和由Donillard和Hoffman(《免疫學概要》V.II中的「關於雜交的基本原理」，Schwartz編輯1981，該文獻列入相關參考)公布的。
本發明進一步提供了應用抗體製造一種藥物，以治療金黃色葡萄球菌引起的敗血症、食物中毒或皮膚損壞。
本發明進一步提供了應用本發明的抗體製造一種藥物，以治療表皮葡萄球菌引起的敗血症、腹膜炎或心內膜炎。
顯然，按照本發明的方法鑑定的多肽將促進治療性抗體的生產，所述抗體能夠用於一些由病原體感染引起的疾病，例如，敗血症、結核病、細菌相關性食物中毒、血液感染、腹膜炎、心內膜炎、膿毒症、腦膜炎、肺炎、胃潰瘍、淋病、鏈球菌喉炎、鏈球菌相關中毒性休克、壞死性筋膜炎、膿庖病，組織胞漿菌病、萊姆病、胃腸炎、痢疾、志賀菌病。
如前所述，病原微生物引起廣泛的不同類型的疾病。例如但不限於下表所列的一些病原微生物及其引起的疾病。
現僅舉例說明本發明的一個實施方案並以下述材料、方法、SEQ IDNO1-19和表1作為參考。
材料和方法應用一種S.aureus 8325/4基因組DNA的λZAP表達文庫。它包括從經過Sau3A部分消化的全部基因組DNA得到的2-10kb的片段。它被克隆進載體的BamH1位點。這個文庫包括＞10倍基因組的範圍。通過對在一個直徑9釐米的Petri碟上的大約20,000個噬菌斑形成單位的初篩挑選噬菌斑對文庫進行探測。鋪皿所用的細胞，對細胞的處理，鋪皿過程和緩衝液配製均嚴格參照製造商(Stratagene)提供的操作手冊進行。鋪皿細胞，大腸桿菌XL1-Blue MRF』，用噬菌體感染並放進3ml含有10mM硫酸鎂的上層LB瓊脂液中，鋪在含10mM硫酸鎂的LB平皿上。然後將平皿在42℃孵育4小時。將一個直徑8.5釐米的纖維素濾膜片(之前浸泡在10mM的IPTG中並在空氣中乾燥)放在每個平皿上，並做出定位標誌。然後將平皿進一步在37℃孵育3.5小時。將濾膜移去並用TBST緩衝液漂洗，然後在含有6％w/v脫脂奶粉和3％v/v豬血清(Sigma)的TBST中4℃下封閉過夜。血清是用於封閉濾膜上的所有蛋白A的克隆。濾膜然後用含有病人血清(1/5000稀釋)的封閉液在室溫下放置處理90分鐘。抗血清從處於嚴重金黃色葡萄球菌感染的恢復期的病人中獲得。濾膜然後在TBST中漂洗3次，每次10分鐘。第二抗體用的是結合鹼性磷酸酶的山羊抗人全IgG(Sigma)，1/30,000稀釋於封閉液中室溫孵育30分鐘。濾膜然後按上述方法漂洗，並用標準比色法程序進一步處理。
有交叉反應的噬菌斑被定位在瓊脂平皿上，被挑入0.2ml的含有0.02ml三氯甲烷的噬菌體緩衝液中。確定每個被挑入的噬菌斑的效價，然後每個平皿上大約接種200個噬菌斑。如上所述進行噬菌斑的挑選和篩選以得到單一的，純的交叉反應克隆。
然後純的克隆被接種(1μl)到平皿上，產生一個直徑0.5釐米的匯合的噬菌斑。每塊平皿上可接種30個單獨的克隆。進行一次噬菌斑挑選，並用一種適當的血清對濾膜進行探測。這樣能夠檢測克隆與其他病人的血清，未被感染的供體血清和抗蛋白A血清的交叉反應性。採用製造商的方案(Stratagene)將克隆切下以在XLOLR大腸桿菌中插入一個噬菌粒。進行一個質粒的小量製備，基因組插入的大小由限制性圖譜決定。使用針對載體序列的引物，通過DNA測序對克隆插入進行鑑定，所述引物允許測序通過插入位點。通過將所獲得得序列與公共結構域資料庫進行比較，可確定所克隆的基因的性質。
雜交溶液/條件典型地，雜交條件是用4-6倍的SSPE(20倍的SSPE含有175.3克NaCl，88.2克NaH2PO4H2O和7.4克的EDTA，溶解至1升，並且調節pH至7.4)；5-10倍的封閉溶液(50倍的封閉溶液含有5克400型的水溶性聚蔗糖，Pharmacia)，5克聚乙烯吡咯烷酮和5克牛血清白蛋白；100μg-1.0mg/ml超聲處理的鮭魚/鯡魚精DNA；0.1-1.0％的十二烷基硫酸鈉；任選地，40-60％去離子甲醯胺。雜交溫度依核酸靶序列的GC含量的不同而變化，但典型地是介於42-65℃之間。
表1在人血清中識別金黃色葡萄球菌的克隆
序列表序列表110謝菲爾德大學120抗原多肽130toxin14014116032170PatentIn Ver.2.121012112260212DNA213Staphylococcus aureus4001gatcttaatg aaagagtgac tgatgcctta gcaattgcta gttgtatcaa tgcgcatccg 60tatgtcaaag gagaactttg cgtgtccgat gacttaacgt atacgacagg ttattttgcc 120gctgctaaaa ttggttacca tcgattattt gatattaaac cagttaatac gagatatgga 180ggcagaataa tatttgtgga cgattgtatt gatttaaatc attacatatc atttttagaa 240agcacaccga agcaagttgt ttatgaaacg gtataggggt tttagtatga catcaaaaga 300tattactcaa attagtgtca ttgctgcgat tttaaccatt ttggcagttt tgaaaatacc 360gtccattata ccaggattag attttcaatt atctgcaccg gcagcattat tgatattagc 420tttctttgga attaaaaagt actttttagg tggattatta tctagcctat tattactagt 480atttggcgta tttaatccaa ttaatgtgat tatctctatt atatttagag ttatagctat 540tgcagttgtt tatttattga aaataaatgt actatcatta gttttagcaa gtgtattagg 600cagtttggta tataggctac tattatctat tattttaaat ttacctgtgt gggtagtgtt 660gttaaacgcg attccaggcg taatattcac tttaattgta gctattcctt tatatctcac 720attgagaaaa agaatggcag tattactaag ataataaatc aaaacacggt cgtcacaatt 780actgttggcg accgtgtttt actagctatt tattgttttc agtttctttt gtatctaaca 840atttcacttt gtgattttcc caatcaattt catatgttga tttaaatgtt ctagttttaa 900agtttttata atttgcgcct gcccagtaga agccattcca acgaatttgg tataaatcca 960tttcacgttg ataagttact gtaattttag attttttagc gccatcttgt ctgtgtgata 1020gtacgcttaa aaattctgga ttgaagttac ttctagataa taatggcatt tggtgttgcg 1080ctatgaagtt ttggccagcg tatgcactgc tttgtctgcc agctaagaag agttcattac 1140catatgttgg gtggaagcta tctcttccat aaggtcccca accattattc ataattttat 1200gtgcttcaac tccccagcca acatttttat aatttgtgtt gcgacttaat gttgttctgt 1260aactttcttg tttataatta attgtttcag aaaaagctgt atttccatta agtccaccag 1320ataaaccatt agagatacta atgtcaccac caaatgtata gcctaaagta ttttgaactt 1380gaaactcttc attttgattt tttggtgcat aatcaacgac gtttactgaa tcattagatt 1440gtgagcttat agatacattg tatttagctc cccaatataa ttttgaaaag tcatagtcat 1500taggattagg tttcacaaag cctgagttaa tattcccagt agctttaagt actaaagtat 1560ctttatcata acttttatct ttgatgaaat taaatgttaa aatctgtgaa attttaaatt 1620tatcagaatc tgctgtggct gttgttttgt ataaagtaac tttgtcatcg acttttttta 1680cgctgactgg tgttatttta ccttcagcat tagcagtacc agaaagtaat aataatgcca 1740tagatgtagc aacggatgat ttgactaatt tattcatttt catatcaatt ctgtcctttc 1800accttgattt catgagtctt ccaattgacc tcgtatttca cagtatagtt tctatttaca 1860aatgcattat ggactctatg tccgtctaaa taactgttgc cataatgcgt tgatctttta 1920atggcatgag tgacatccat gtttcttccg taagtaattt caaattcgct tgtatcgctt 1980gaaccttttt catgagatac tgtggcgata aatgaagggt taaatccact ttgtacaaga 2040ggtggtaact cactgtctgg aacgaaataa tctctaggat ctttactatg aggtttgtag 2100cctacaaata aatcgctatc aaaggctgat ttttgacctg attcagtggc gaatgaattc 2160gctttgacgc cccataaaac actttttgag ttttgttgtt ctacttcact tacataattt 2220tgttgtgtat agctaatcga tttagaatag ttaaatgatc 226021022112902212DNA213Staphylococcus aureus4002gatcgtataa tcgaaacagc accaacggat tacttatctt ggggtgtcgg tgcagtcggt 60aaccctagat tcatcaatgt tgaaatcgta cacacacacg actatgcttc atttgcacgt 120tcaatgaata actatgctga ctatgcagct acacaattac aatattatgg tttaaaacca 180gacagtgctg agtatgatgg aaatggtaca gtatggactc actacgctgt aagtaaatat 240ttaggtggta ctgaccatgc cgatccacat ggatatttaa gaagtcataa ttatagttat 300gatcaattat atgacttaat taatgaaaaa tatttaataa aaatgggtaa agtggcgcca 360tggggtacgc aatctacaac tacccctact acaccatcaa aaccaacaac accgtcgaaa 420ccatcaactg gtaaattaac agttgctgca aacaatggtg tcgcacaaat caaaccaaca 480aatagtggtt tatatactac tgtatacgac aaaactggta aagcaactaa tgaagttcaa 540aaaacatttg ctgtatctaa aacagctaca ttaggtaatc aaaaattcta tcttgttcaa 600gattacaatt ctggtaataa atttggttgg gttaaagaag gcgatgtggt ttacaacaca 660gctaaatcac ctgtaaatgt aaatcaatca tattcaatca aacctggtac gaaactttat 720acagtacctt ggggtacatc taaacaagtt gctggtagtg tgtctggctc tggaaaccaa 780acatttaagg cttcaaagca acaacaaatt gataaatcaa tttatttata tggctctgtg 840aatggtaaat ctggttgggt aagtaaagca tatttagttg atactgctaa acctacgcct 900acaccaacac ctaagccatc aacacctaca acaaataata aattaacagt ttcatcatta 960aacggtgttg ctcaaattaa tgctaaaaac aatggcttat tcactacagt ttatgacaaa 1020actggtaagc caacgaaaga agttcaaaaa acatttgctg taacaaaaga agcaagttta 1080ggtggaaaca aattctactt agttaaagat tacaatagtc caactttaat tggttgggtt 1140aaacaaggtg acgttattta taacaatgca aaatcacctg taaatgtaat gcaaacatat 1200acagtaaaac caggcactaa attatattca gtaccttggg gcacttataa acaagaagct 1260ggtgcagttt ctggtacagg taaccaaact tttaaagcga ctaagcaaca acaaattgat 1320aaatctatct atttatttgg aactgtaaat ggtaaatctg gttgggtaag taaagcatat 1380ttagctgtac ctgctgcacc taaaaaagca gtagcacaac caaaaacagc tgtaaaagct 1440tatactgtta ctaaaccaca aacgactcaa acagttagca agattgctca agttaaacca 1500aacaacactg gtattcgtgc ttctgtttat gaaaaaacag cgaaaaacgg tgcgaaatat 1560gcagaccgta cgttctatgt aacaaaagag cgtgctcatg gtaatgaaac gtatgtatta 1620ttaaacaata caagccataa catcccatta ggttggttca atgtaaaaga cttaaatgtt 1680caaaacttag gcaaagaagt taaaacgact caaaaatata ctgttaataa atcaaataac 1740ggcttatcaa tggttccttg gggtactaaa aaccaagtca ttttaacagg caataacatt 1800gctcaaggta catttaatgc aacgaaacaa gtatctgtag gcaaagatgt ttatttatac 1860ggtactatta ataaccgcac tggttgggta aatgcaaaag atttaactgc accaactgct 1920gtgaaaccaa ctacatcagc tgccaaagat tataactaca cttatgtaat taaaaatggt 1980aatggttatt actatgtaac accaaattct gatacagcta aatactcatt aaaagcattt 2040aatgaacaac cattcgcagt tgttaaagaa caagtcatta atggacaaac ttggtactat 2100ggtaaattat ctaacggtaa attagcatgg attaaatcaa ctgatttagc taaagaatta 2160attaagtata atcaaacagg tatggcatta aaccaagttg ctcaaataca agctggttta 2220caatataaac cacaagtaca acgtgtacca ggtaagtgga caggtgctaa ctttaatgat 2280gttaagcatg caatggatac gaagcgttta gctcaagatc cagcattaaa atatcaattc 2340ttacgcttag accaaccaca aaatatttct attgataaaa ttaatcaatt cttaaaaggt 2400aaaggtgtat tagaaaacca aggtgctgca tttaacaaag ctgctcaaat gtatggcatt 2460aatgaagttt atcttatctc acatgcccta ttagaaacag gtaacggtac ttctcaatta 2520gcgaaaggtg cagatgtagt gaacaacaaa gttgtaacta actcaaacac gaaataccat 2580aacgtatttg gtattgctgc atatgataac gatcctttac gtgaaggtat taaatatgct 2640aaacaagctg gttgggacac agtatcaaaa gcaatcgttg gtggtgctaa attcatcggc 2700aactcatatg taaaagctgg tcaaaataca ctttacaaaa tgagatggaa tcctgcacat 2760ccaggaacac accaatatgc tacagatgta gattgggcta acatcaatgc taaaatcatc 2820aaaggctact atgataaaat tggcgaagtc ggcaaatact tcgacatccc acaatataaa 2880taagcaacat gaacatagga tc290221032112792212DNA213Staphylococcus aureus4003gatcaactta atataatgaa ttcggcaaca gaagagcatc atcataaaga ttatattaaa 60ctatataatt taggtggcgg tgctgctaaa aaaattgcaa tagaggtttt attggggaag 120gataaagtca ttcagaaaaa atacgtgcat attttaccta gtaaagaagg gtacatgtta 180ccaattaata aaaatgtgta cgaagaatta gaaagaacga ttgagaacaa tggtcatgaa 240gctgatttga atgtacgtat gacttattat cataatgtaa gtcgcaaaca acaggaagtt 300atattaaaag gtcaaatcga ccgttttaat acttataata ataaagaaat ttatgatttg 360cagtttatct aaaaattgat ttaagagggt agttgtttat tgcgaaaaat atcattcaat 420tttaatgaaa taatggcgtc attactataa aatattactt tatgttgtaa tgcatttttc 480tataagatag aactaaaagg aggggcaaag atgcaaatta gacaaataca tcaacatgac 540tttgctcaag tggaccagtt aattagaacg gcatttgaaa atagtgaaca tggttatggt 600aatgaatcag agctagtaga ccaaattcgt ctaagtgata cgtatgacaa taccttagaa 660ttagtagctg ttcttcaaaa tgaagttgta gggcacggtt tactaagtga agtttatctt 720gataacgagg cacaacggga aattggatta gtgttagcac ctgtatctgt tgatattcat 780catcaaaata aaggtattgg gaagcgattg attcaagcat tagaacgaga agcaatatta 840aaaggatata attttatcag tgtattagga tggccgacgt attatgccaa tctaggatat 900caacgcgcaa gtatgtacga catttatcca ccatatgatg gtataccaga cgaagcgttt 960ttaattaaag aattaaaagt gaacagttta gcgggaaaaa caggtaccat aaattacaca 1020tctgcttttg aaaaaatatg atttcaagct aggattacat taggtagagt tcatattaat 1080aataaaaaat gtttgcaatc aaatcgtacg ttgtcgtttg taattcttaa aatagcaata 1140aataaaatgt ttgttagtaa agtattattg tggataataa aatatcgata caaattaatt 1200gctataatgc aattttagtg tataattcca ttaacagaga ttaaatatat ctttaaaggg 1260tatatagtta atataaaatg actttttaaa aagagggaat aaaatgaata tgaagaaaaa 1320agaaaaacac gcaattcgga aaaaatcgat tggcgtggct tcagtgcttg taggtacgtt 1380aatcggtttt ggactactca gcagtaaaga agcagatgca agtgaaaata gtgttacgca 1440atctgatagc gcaagtaacg aaagcaaaag taatgattca agtagcgtta gtgctgcacc 1500taaaacagac gacacaaacg tgagtgatac taaaacatcg tcaaacacta ataatggcga 1560aacgagtgtg gcgcaaaatc cagcacaaca ggaaacgaca caatcatcat caacaaatgc 1620aactacggaa gaaacgccgg taactggtga agctactact acgacaacga atcaagctaa 1680tacaccggca acaactcaat caagcaatac aaatgcggag gaattagtga atcaaacaag 1740taatgaaacg acttctaatg atactaatac agtatcatct gtaaattcac ctcaaaattc 1800tacaaatgcg gaaaatgttt caacaacgca agatacttca actgaagcaa caccttcaaa 1860caatgaatca gctccacaga gtacagatgc aagtaataaa gatgtagtta atcaagcggt 1920taatacaagt gcgcctagaa tgagagcatt tagtttagcg gcagtagctg cagatgcacc 1980ggcagctggc acagatatta cgaatcagtt gacgaatgtg acagttggta ttgactctgg 2040tacgactgtg tatccgcacc aagcaggtta tgtcaaactg aattatggtt tttcagtgcc 2100taattctgct gttaaaggtg acacattcaa aataactgta cctaaagaat taaacttaaa 2160tggtgtaact tcaactgcta aagtgccacc aattatggct ggagatcaag tattggcaaa 2220tggtgtaatc gatagtgatg gtaatgttat ttatacattt acagactatg taaatactaa 2280agatgatgta aaagcaactt tgaccatgcc cgcttatatt gaccctgaaa atgttaaaaa 2340gacaggtaat gtgacattgg ctactggcat aggtagtaca acagcaaaca aaacagtatt 2400agtagattat gaaaaatatg gtaagtttta taacttatct attaaaggta caattgacca 2460aatcgataaa acaaataata cgtatcgtca gacaatttat gtcaatccaa gtggagataa 2520cgttattgcg ccggttttaa caggtaattt aaaaccaaat acggatagta atgcattaat 2580agatcagcaa aatacaagta ttaaagtata taaagtagat aatgcagctg atttatctga 2640aagttacttt gtgaatccag aaaactttga ggatgtcact aatagtgtga atattacatt 2700cccaaatcca aatcaatata aagtagagtt taatacgcct gatgatcaaa ttacaacacc 2760gtatatagta gttgttaatg gtcatattga tc 279221042112478212DNA213Staphylococcus aureus4004gatcgaattg aacgaagcat ttgcttctca aacgattgca tctattaaag aagtaggtct 60agatatatca cgtacgaatg tgaatggtgg cgctattgct ttaggtcatc cattaggtgc 120tacaggcgca atgttaaccg cgcgtttact taatgaaatg ggtagacgtc ccgatagccg 180ttacggcatg gttacgatgt gtattggtgt cggcatgggt gcagctgcta tatttgaata 240tgtgcgttag aatggttgat tttggatgaa gcggattcgt tttgttattg aatgaagtag 300gctgaagttg aagccagttg aagttgaagc gggttgaagc aatttcgttt tattaatgaa 360gctgtgtgaa atatagtgat tgaacaaaaa agtggtttaa tgggatggtg gttatttccg 420ttttagaatt taacatttac acgtctaatt ttaatcattg ttttaaattt tatgaatcga 480agccctttga tttaataata tttgctaatg ctagtaactt atctgattgt tcatgtttaa 540aataaagaaa accactcaca tcagtgtgtg ttcgaactag acttgtaagt tccagttcgg 600cacgactttc taaagcaatt attattgctg tgattgtcgt atatcactta gatgtgcgtg 660gtttatttta ataggttagt aatatattag gtcatgttat gtttaagact ataatgaata 720aataatttag aaatatgctt ccgattgttc gatgctttaa ttcagttaga agcatcatag 780aatgcatgat tactgttgta aagatacgta atgttttgta ttgactgtat gtctttggat 840agagttacaa acttattttg ttactctagg cccatatgtc gcagtaccat ctgcatgtgt 900tgttacattg tatgcatttg ttttacttgg cttcttgtat gtcgggcgag ctccgtatga 960cacttgaccg tttgcatgtg ttgttacgtt gtatgcattt gttttgcttg gcttgttttg 1020tgttgggcga gcgccatatg atacttggcc gtttccatgt gttgttacgt tatatgcgtt 1080tgttttgctt ggcttgtttt gtgtcggacg agctccgtat gatacttggc cgtttgcatg 1140tgttgttaca ttgtatgcat tcgtttcgct tggcttcttg tatgtcggac gagctccgta 1200tgatacttga ccatttgcat gtgttgttac gttatatgca tttgtttctg atggcttatt 1260gaatcttggt ctcgcttcat atccaaatgt tccatcgttg tattcacgga tacctgtacc 1320agcatctcta tatttaacat atttaggtgt tttgttaaat tgcggtctcg gaccatattg 1380agaagcttct gttgtttcag ttgcttgagg tttaacttca atatcacttg attctccttg 1440agtacctttt aacgttgatt cagtaccttg tggttttatt tcaagtttag atgagctacc 1500ttcaagacct tctaaaatag ggttcgttaa cggtgggttt gtataattat tgcttaatga 1560tgggccgctt tgttccattg ttagaaaatc gggaccttga acgatttcac cttgtaccgt 1620tttattttcc atcgttggat attccggacc ttttacaatt tcacctgtaa ttgtgccctg 1680tggaatttta actaatggtt gtgcaactgg ttgtgttgtt tcttcagctt taccagccgt 1740agttttaacc tcttgttggt tatcaacttt aggtgcttga ggttcttcaa ctttcttctc 1800ttcttttact actggcgatt ttgtttcagt ttctccgtat tttttgacag ttttcttttt 1860ccaagaatca tctgcttctt taactgcttt tttcgtttct tcaactaatt tatcaaaatt 1920aggtttatta tcactatttg ttttatagtt atgtgttgta ggattatatt tcgttataga 1980tttcggtcta ttttgtttag tttccataaa gaaatcatca ataattgaat ttaagtcatc 2040aatcatttct tttttaatac gttcatttgt aattttatgt ggattgtctg tatctccaag 2100gattaagtcc agttttgctc gtaactcttt cgcgtgctcc ccataatcct tatcaccata 2160atatgataca actaatgtat caatttcaga tacgagatcg tatacttcct tagttgcttt 2220atcttcttct gctgcattaa aagttttcaa gtctgaattc ttatccttaa tatctttaac 2280ttctctgtga aaatcatcca gtgctctctt taatgcatcc tgtagttcat tgtattcttt 2340catcgaaagt tcttctaaat tatatttatg aaaattagcc atttttaaat ctgtacgagg 2400attttctttt ttataatttg cataccattg tttataatct tcatattgag atttctttct 2460ctccaaaaga tattgatc 247821052112070212DNA213Staphylococcus aureus4005tgacgctgct tttgtaaata catataattt ttccacttca tgatttaatt cgttcgcatg 60atctttgtaa tttctaccaa aagcaatcac attattcgga ggtgttactg gtggtaaaaa 120ttcaatgtca ttaaatgaaa ttttatagtc ttcagctttg ccgctatctt ctgctgctac 180aactgcttta cgtacttgtt cttgaaaatc taaagtatga ttttgttgta aaccagctaa 240caatgtttta ggatggaaat ctccttctgc aaagtcagca aatacttgtg ttaaatccca 300tacagcatct tcgcgtttta ctttaacgcc atatgaagtt ttgtcattat acttgaatga 360taagaatttc attcattctc aactcctcgt ctttatctta attcacatta taactttttt 420cgttatcaaa taacaaataa ataagtaaga caattttgaa aatgagttgt gttcattctg 480ctacaaggac tttgcactta atcgaaatta ttttttattc ttttgaaaat caaaatacta 540tagttgcaat gtaccaaatt tgaagaagta taaataacct ttaacttctt tattaagaat 600cgtttgaagc gtattttgat aatatttcat ctgtatctta tatttatttt ttaattgtgt 660accaatttct tcatctgtca tcccacggcg acgattaaat gcatcggttt tatagtctac 720aaaataatgc acaccatctt taacaaagat taagtcaatc ataccttgaa taattgagac 780gtcttcgtct ccttgtggca attggtcaac taatgcttgg ttaactacaa acggtaattc 840acgataaact tgctctgctt cagcaataat cgaatataac tcactattga taaatgtcat 900tatttcatcc atacggatat cttttttcgc atctgcttcg ataatatgtt tatcgattaa 960tccatcgata tactgatgta actcaacttc agatatgcgt tcttttttga atggtaaatg 1020ttgcatcact gtatgcatta acgtaccaat ttcattcgct tttcgtttac cttgttcact 1080tagaaattta ggtcgttcat acgttgaaaa accgatacga tattgcctta ctcgttcgta 1140acttgtgcca ctttcttctg tttcatattg tcttttcaat tcagaaacag attgttttga 1200gggcttttta gtatcattta catatggata tcgataatca agttggtgtt taatttgtgc 1260tttaacatct tcattaccat tttgcatagt ttctaattga ttaaccgaac gatattcatc 1320attatctaaa atggtttctg tagacacatc ttcaaagtac acaattgaaa tatttacatt 1380cggacgacta ctatcttcaa tttgtgctat atctttttca aattttaaat catctggaat 1440tgacgcagat tgatgtttag ataaaatact ataaataaga tggaacggat ttggtgaagt 1500taatcgttca ttgacagcaa tgtgctcacc agaaatagac aattgctcta gttctagtaa 1560tgatttatca tttttcactc taccaattaa ataaagttgt tctttcgctc ttgttaatgc 1620tacatagact aatcgcattt cttctgacac aagttctttt tcggcaacag ctctatatgc 1680aaccgaagct aaagatggaa atgccatttc tttatccaca tcaaaataat ccattccgag 1740accaaattgc tgatttaaaa taactggttg tttcaaatca cgtttattaa aatcttttga 1800caatccagaa taaatgacaa atggaaactc tagaccttta ctactatgaa ttgtcatcat 1860tctaacgaca ttatcgtttg gaccaactac attttcctca ccaaaatctt tgcctctttc 1920aatcaattca tcgataaaac gaataaattg atataaacct ctaaaacttg aattctcaaa 1980ctcgatagct ttattaaata aaccataaag atttgcacgt cgtccacgtc caccaataag 2040tccactaaag tattgaataa cataatgatc207021062112394212DNA213Staphylococcus aureus4006gatcagattt attagacagt attccagata tacccacacc aaagccagaa aagacgttaa 60cacttggtaa aggtaatgga ttgttaagtg gattattaaa tgctgatggt aatgtatctt 120tgcctaaagc gggggaaacg ataaaagaac attggttgcc gatatctgta attgttggtg 180caatgggtgt actaatgatt tggttatcac gacgcaataa gttgaaaaat aaagcataat 240tatattgggg gaagagcatc tatatatttt tttaagtata taagacgtct tatttcccct 300taatttattg tgaagtatat gcaaaatgca atgaatagat tgtccatcat tttaacgtta 360taatgaattt aacgacttag aactacacaa gtaaaggaga atgaagatgt ctcgaaaaac 420ggcgctatta gttttggata tgcaagaagg tatagcgagt agtgtaccta gaataaaaaa 480tattattaaa gcgaatcaga gagcaattga agcagcaaga caacatcgaa taccagtcat 540tttcatacgt ttagtgttag ataagcattt taatgatgtc tcctcgagta ataaagtgtt 600ttcaacaatt aaagctcaag gatatgcgat tactgaagca gatgcatcta cacgaatact 660tgaagattta gcaccactag aagatgagcc gattatttct aagcgacgct ttagcgcatt 720tacaggtagt tacttggaag tttatttacg tgcaaatgat attaatcatt tagtattaac 780gggtgtctct acaagtggag ctgtattgag cacggcatta gaaagtgtag ataaagacta 840ttatattact gttttagaag atgctgttgg tgatagatca gatgataaac atgactttat 900tattgaacaa attttatcac gctcatgtga cattgaatcc gtagagtcat ggaaaagtag 960tttatagtta atataacgtc aattaaagct cggcagtaat gtttgagaat aagtacattt 1020gctcatattt ataaaatgtg tgagatggca attgaaacgg atatgatgag gaacatttga 1080acataaaata atatatttat ataaaacgac ccgaggcgtt cgaactgaat gcctcgggtt 1140taattgaata agaaatcgga cttatgaaca gaaatatgtt taagtccgaa ctccttgttt 1200atacttataa attttacggg tttaatataa tacttattta cctgtaatat atgataattc 1260ttcagcggca gctgcgttga tagttctatg agaaatgata cctaatcctt taacattgga 1320ttctgaaata acgatagaac catcactgtt aactttttca acaaatgcta catgaccgta 1380atgttgatct gcaccaaatt gtccagcctc aaatacaaca gcagcatgac gttttggtgt 1440atgacttact tgataatcac ggtattgagc tcgattattc caattatgtg catcacctaa 1500atcacctgag atagatgtac caaattgttt catacggtta tatacgtacc aagtacattg 1560gccatgtgga tatggcatac tatcagatac ctcacggaaa ggtttgaatt catctgatga 1620atcatcataa tccttgatag aacgttcata tttatctaaa tctggcatgc gttcatcgtc 1680aaactgagtt aattgatagt gtttaataat actgtttaat ttcttagcat agtttggatc 1740tgtagcatat gttttagata agtgtgatgt tgcatcttta taagaatcgg cttccgattt 1800ccatgttggt ttataaattg ttcgattgcc atcaatacca tttttaataa ggtcagagta 1860atcttttagt gattctttcg tgcttggata ttttcggaat ccagcattaa tactatacaa 1920ttgattacca tcagcttcta atgtgttaaa aggaacagaa ttcccttcaa aagcaccttt 1980gataccgaat aaattatggt ttggtgactt agctaaagca ctacgacctg agtcagattc 2040taagattgct tgggcaatca tgacagacgc ataaatatcg ttatcttgac caatgcgatg 2100tgcatcttta gcaattgatt tgacaaattg acgtgtatct tttgagtcaa caacgttaaa 2160ttgtccgcta tcatcattgt tagatatact aggatctgtt tcgaataatg atgttgcacg 2220tgtatccttt tgattaacat cgttattgaa tgattgagca ggtttagatt tatgtttcaa 2280ttcatcttgt gttggtaact gtggattctt tgtattagat ttttcatttt tgtctttttt 2340agattgagat gcataatctt tttgtgtttt ctttgcatct tcactgtatt gatc 239421072112033212DNA213Staphylococcus aureus4007gatctggaac aggtttcatt gtcggtaaaa atacaattgt taccaacaag catgtcgttg 60caggtatgga aattggtgca catattatag cgcatcccaa tggtgaatat aataatggcg 120gattttataa agttaaaaaa attgtccgtt attcaggtca agaagatatt gccattctac 180atgtggaaga taaagctgtt catccaaaaa acaggaattt taaagattac acaggcattt 240taaaaatagc atcagaagct aaagaaaatg aacgcatttc aattgttggc tatccagaac 300catatataaa taaatttcaa atgtatgagt caacaggaaa agtgctgtca gttaaaggca 360acatgattat tactgatgct ttcgtagaac caggcaactc aggttcagct gtatttaaca 420gtaaatacga agttgtaggt gttcactttg gtggaaacgg ccctggaaat aaaagtacaa 480aaggatatgg tgtttatttc tctcctgaaa ttaagaaatt cattgcagat aacacagata 540aataaatcct tacatagata aatgatttta aaaattaaca acaaactcaa caattcaaat 600catctctgtg attccattta ttcgaaatga ttaaaaaaaa taaaacttca aaaagctaac 660attataatta tacaaatact tagaggagca gaaaaatgaa taaaaatata atcatcaaaa 720gtattgcagc attgacgatt ttaacatcaa taactggtgt cggcacaaca atggttgaag 780gtattcaaca aacagccaaa gccgaaaata ctgttaaaca aattacaaat acaaatgttg 840caccatacag tggtgttaca tggatgggcg ctggaacagg atttgtagtt ggaaatcata 900caatcattac caataaacat gttacctatc acatgaaagt cggtgatgaa atcaaagcac 960atcctaatgg tttttataat aacggtggtg gactttataa agttactaag attgtagatt 1020atcctggtaa agaagatatt gcggttgtac aagttgaaga aaaatcaaca caaccaaaag 1080gtagaaaatt caaagatttc actagtaaat ttaatatagc atcagaagct aaagaaaatg 1140aacctatatc agtcattggt tatccaaatc ctaatggaaa taaactacaa atgtatgaat 1200caactggtaa agtattatca gtgaatggga atatagtgtc ttcggatgca attattcagc 1260ctggtagctc tggttcacct atattaaata gtaaacacga agctattggt gtaatctatg 1320ccggtaataa gccatcaggt gaaagcacaa gaggatttgc tgtttatttc tctcctgaaa 1380ttaagaaatt cattgcagat aatttagata aataattaaa acttagacat tcacccaatc 1440ctgacaaaat atactataac taacatttat taatatatat tgcattattt aatatgcatc 1500aaagccaatc aacgattgat tttcaccaac tcaattgttg attggtttta tttatgtatg 1560aatgaacaac tttttgacat cattaagaat ataaatgatt ttgaaagcat ttgaaagcta 1620caacatttct ataaaatttt tcaataacaa ttgcgccact aaaactcaaa atttccacca 1680ccaacatcca aattatcaac atcgcaacat aaccaaatgt tataataaat ctattacaca 1740aagagataaa ttacttatgc aaaggcggag gaatcacatg tctattactg aaaaacaacg 1800tcagcaacaa gctgaattac ataaaaaatt atggtcgatt gcgaatgatt taagagggaa 1860catggatgcg agtgaattcc gtaattacat tttaggcttg attttctatc gcttcttatc 1920tgaaaaagcc gaacaagaat atgcagatgc cttgtcaggt gaagacatca cgtatcaaga 1980agcatgggca gatgaagaat atcgtgaaga cttaaaagca gaattaattg atc 203321082112794212DNA213Staphylococcus aureus4008gatcaaacgt tgcttaactt ctttttaatg cttaaaaatt atttcaaagg cacatagaaa 60cgctatatta atctcatact cactcattat tttttgctta aattacttaa taatacttca 120ataattgtta aaaggggttt aatgtgatta tcttagaacg ccatctataa tgatgttgta 180tgattcaaat tacgtaaaaa gacaatcgaa tataatatag attggagcat acaattatga 240aaatgagaac aattgctaaa accagtttag cactagggct tttaacaaca ggcgcaatta 300cagtaacgac gcaatcggtc aaagcagaaa aaatacaatc aactaaagtt gacaaagtac 360caacgcttaa agcagagcga ttagcaatga taaacataac agcaggtgca aattcagcga 420caacacaagc agctaacaca agacaagaac gcacgcctaa actcgaaaag gcaccaaata 480ctaatgagga aaaaacctca gcttccaaaa tagaaaaaat atcacaacct aaacaagaag 540agcagaaaac gcttaatata tcagcaacgc cagcgcctaa acaagaacaa tcacaaacga 600caaccgaatc cacaacgccg aaaactaaag tgacaacacc tccatcaaca aacacgccac 660aaccaatgca atctactaaa tcagacacac cacaatctcc aaccataaaa caagcacaaa 720cagatatgac tcctaaatat gaagatttaa gagcgtatta tacaaaaccg agttttgaat 780ttgaaaagca gtttggattt atgctcaaac catggacgac ggttaggttt atgaatgtta 840ttccaaatag gttcatctat aaaatagctt tagttggaaa agatgagaaa aaatataaag 900atggacctta cgataatatc gatgtattta tcgttttaga agacaataaa tatcaattga 960aaaaatattc tgtcggtggc atcacgaaga ctaatagtaa aaaagttaat cacaaagtag 1020aattaagcat tactaaaaaa gataatcaag gtatgatttc acgcgatgtt tcagaataca 1080tgattactaa ggaagagatt tccttgaaag agcttgattt taaattgaga aaacaactta 1140ttgaaaaaca taatctttac ggtaacatgg gttcaggaac aatcgttatt aaaatgaaaa 1200acggtgggaa atatacgttt gaattacaca aaaaactgca agagcatcgt atggcagacg 1260tcatagatgg cactaatatt gataacattg aagtgaatat aaaataatca tgacattctc 1320taaatagaag ctgtcatcgg aaaaacaaga agttaagtga caacggttta catgttgctt 1380agcttctttt attatgcgta atgatgtaaa aagacgaata ttcatttgtt tgtaaaagtg 1440gcatttctat gtcttaaaag tgacgaaact tcaaatgtgc caagtgttga atcacatcaa 1500aatcattttt atttaacgaa cattatggat ttcttaattt acttaacgat gattcaaata 1560tagttaaaca aggtttaatg tgaatggagc aatacgccat ctataataaa gctgtatgat 1620tcaatgaatg taatcgaaca aatctaataa ttacgaatgg agcatacaac tatgaaaata 1680acaacgattg ctaaaacaag tttagcacta ggccttttaa caacaggtgt aatcacaacg 1740acaacgcaag cagcaaacgc gacaacacta tcttccacta aagtggaagc accacaatca 1800acaccgccct caactaaaat agaagcaccg caatcaaaac caaacgcgac aacaccgccc 1860tcaactaaag tagaagcacc gcaacaaaca gcaaatgcga caacaccgcc ttcaactaaa 1920gtgacaacac ctccatcaac aaacacgcca caaccaatgc aatctactaa atcagacaca 1980ccacaatcgc caaccacaaa acaagtacca acagaaataa atcctaaatt taaagattta 2040agagcgtatt atacgaaacc aagtttagaa tttaaaaatg agattggtat tattttaaaa 2100aaatggacga caataagatt tatgaatgtt gtcccagatt atttcatata taaaattgct 2160ttagttggta aagatgataa aaaatatggt gaaggagtac ataggaatgt cgatgtattt 2220gtcgttttag aagaaaataa ttacaatctg gaaaaatatt ctgtcggtgg tatcacaaag 2280agtaatagta aaaaagttga tcacaaagca ggagtaagaa ttactaagga agataataaa 2340ggtacaatct ctcatgatgt ttcagaattc aagattacta aagaacagat ttccttgaaa 2400gaacttgatt ttaaattgag aaaacaactt attgaaaaaa ataatctgta cggtaacgtt 2460ggttcaggta aaattgttat taaaatgaaa aacggtggaa agtacacgtt tgaattgcac 2520aaaaaattac aagaaaatcg catggcagat gtcatagatg gcactaatat tgataacatt 2580gaagtgaata taaaataatc atgacattct ctaaatagaa gctgtcatcg gaaaaacaag 2640aagttaagtg acaacggcct acatgttgct tagcttcttt tgttatgttc gatgatttga 2700gaacccgaat tttcgatggg tccaaatatg acgtggaaga gacctgaatt tatctgtaaa 2760tccctatcta tcgggtgtga agcacaacgg gatc 27942109211505212DNA213Staphylococcus aureus4009gatcatagcg caccaaactc tcgtccaatt gattttgaaa tgaaaaagaa agatggaact 60caacagtttt atcattatgc aagttctgtt aaacctgcta gagttatttt cactgattca 120aaaccagaaa ttgaattagg attacaatca ggtcaatttt ggagaaaatt tgaagtttat 180gaaggtgaca aaaagttgcc aattaaatta gtatcatacg atactgttaa agattatgct 240tacattcgct tctctgtatc aaacggaaca aaagctgtta aaattgttag ttcaacacac 300ttcaataaca aagaagaaaa atacgattac acattaatgg aattcgcaca accaatttat 360aacagtgcag ataaattcaa aactgaagaa gattataaag ctgaaaaatt attagcgcca 420tataaaaaag cgaaaacact agaaagacaa gtttatgaat taaataaaat tcaagataaa 480cttcctgaaa aattaaaggc tgagt 50521010211673212DNA213Staphylococcus aureus40010gatcaaacta aaacacaaac tgctcataca gttaaaacag cacaaactgc tcaagaacaa 60aataaagttc aaacacctgt taaagatgtt gcaacagcga aatctgaaag caacaatcaa 120gctgtaagtg ataataaatc acaacaaact aacaaagtta caaaacataa cgaaacgcct 180aaacaagcat ctaaagctaa agaattacca aaaactggtt taacttcagt tgataacttt 240attagcacag ttgccttcgc aacacttgcc cttttaggtt cattatcttt attacttttc 300aaaagaaaag aatctaaata aatcatcgtc acactcataa cttaatatat tttttatttt 360aaattttatt taacctatgt catagatatt tcataatcta taacataggt tatttttttt 420ataaaataac gttgcaatta actaacattt caatgtcaat acaagtaatc aattgataat 480gattatcagt tgataatata caattaggag ttgtttctac aacatgaaca aacagcaaaa 540agaatttaaa tcattttatt caattagaaa gtcatcacta ggcgtgcatc tgtagcaatt 600agtacacttt tattattaat gtcaaatggc gaagcacaag ccagcagctt gaagaaaaca 660ggtggtccaa ttc 673210112112238212DNA213Staphylococcus aureus40011gatcttcagc ttgatgtttt cgtttgatta aattggtaaa atagaaacgc aatccacaaa 60aatggcaagc actaaaataa tgtttggggg tgcttgtgct tttgtggatt gcggtcgatt 120atttatattg catgatttga ttaatttgat tgattatatt ggacatgatg gtgttggcgg 180gatgcgttgt tgctagtcgc gggctttgtc cactccacat atgtattaac tctttgtcgc 240cgatgtttgc tgcggctttt cttatgctac ttgttagctc attttgtatt ggataatctg 300ggatatcgcc ttcgtattgg gacatttctt cgataaacct attgttgata ccgcgtgcaa 360gctttccact aaacgctttt gtaatgactg tatctgtttc tttactattt ataattgcat 420ctcgcagtag ttctgatgca ttactgtctt gtgatgttaa aaatgcggtg cccatttgta 480ccccttctgc acctaagaca atacttgcca aaactcctct accatccata attccaccag 540cggcaatgac cggaattgaa acgacatcta caatttgtgg cactaaagat attgttccaa 600ccataggtaa ttgattttta ggttttaaaa atgaaccacg atgtccacct gcttcactac 660cttgagcaac gatagcatcc atacccgctt tttcattcgc aatagcttca tcaacacttg 720ttgctgtacc tataagtttg acattcgctg ctttcaacct gcttataatc tgttcgcttg 780gaattccaaa agtaaaacaa catacaggca cttgcttttt aattatcgta tcaatatgac 840acttaaattg ttgttcttcg gtaattttta caaccggctc ttctaaatgt aatgcgcgtc 900gataaggttt taaccatgca ttcatatttt caatttgact actggtatat gattgttgac 960ttggtacaaa gacatttacg ccaaaagaat ttgacgttaa ttggcgtaca taatctattt 1020catcttccaa ttgctgcgta ttaaagtaac ctgcgcctat tgtgcctaac ccaccactgt 1080tacttactga tgcaactaat ttcggtgtcg tacttcctgc catacctgct tgtataattg 1140gatattcaat acttaacatt tgagtaagtc gattcttatt ccacatagct gttcgctcct 1200tatatagata cgttgcgatt tttccgttgt tgaaattgaa tttgctgttg agaaagtttt 1260tctttttcct ttttatccat ctcatcttca atttccatac ctaataattc ttcaattaag 1320tcttcatgtg acactatcgc ttcagtacca ccaaattcgt ccaacacaat tgctaaatgt 1380tttctagaaa tagtcatctt acgtaatacc cattcagctt tattgtgttc attcacaaat 1440aatggcttag ctgaatagtt tgtaatttga ttttcttttt tattactcca agccaacaga 1500tatttagaat gaaacacccc aataatgtta tcaatatctc cctcgtacac tggatatcta 1560gtgtatggct tattcataac cgtttcataa acttcttcgt atgtcgcatt tgaagcaaat 1620gccgtcacat taattctagg tgttgtatct acatctttta cttttaaatt ttcaaaatta 1680atgacacctt ccaacctact cgtctcaatt tcatttaaag caccttcatg tccagcaatt 1740gctaacattg ttttaaattc ttcttttgaa aattgatgtt cttgaggttg gcccttagat 1800aaacttcgat taatactgtc cgtcaactta tttaaaagta atgtgatagg acggaacaca 1860atgacacaaa tattaataat tggatataca agccttgtta ttttatctgg aaatgttgca 1920gcgacagact tgggaatcac ttcggagatc aaaatgataa caactgttaa aacagctgat 1980gcaataccaa cgctaatccc ccaacgtaaa gccataattg taacaagtgt tggtaataaa 2040atattcgcga cattattccc aattagaatc gttgtaataa actcacttgg tttttcaagt 2100aactttacaa tgccttttgc ttttttatca cctttgtcag cttcagtttt aaattttgct 2160ttattggcag ccgttaatgc cgtctcgctt cctgaaaaga aaaacgaaat aaatatcaat 2220ataattatgg caatgatc 2238210122117975212DNA213Staphylococcus aureus40012gatcaaacga caattattaa ttcgttaacg tttactgaaa cagtaccaaa tagaagttat 60gcaagagcaa gtgcgaatga aatcactagt aaaacagtta gtaatgtcag tcgtactgga 120aataatgcca atgtcacagt aactgttact tatcaagatg gaacaacatc aacagtgact 180gtacctgtaa agcatgtcat tccagaaatc gttgcacatt cgcattacac tgtacaaggc 240caagacttcc cagcaggtaa tggttctagt gcatcagatt actttaagtt atctaatggt 300agtgacattg cagatgcaac tattacatgg gtaagtggac aagcgccaaa taaagataat 360acacgtattg gtgaagatat aactgtaact gcacatatct taattgatgg cgaaacaacg 420ccgattacga aaacagcaac atataaagta gtaagaactg taccgaaaca tgtctttgaa 480acagccagag gtgttttata cccaggtgtt tcagatatgt atgatgcgaa acaatatgtt 540aagccagtaa ataattcttg gtcgacaaat gcgcaacata tgaatttcca atttgttgga 600acatatggtc ctaacaaaga tgttgtaggc atatctactc gtcttattag agtgacatat 660gataatagac aaacagaaga tttaactatt ttatctaaag ttaaacctga cccacctaga 720attgacgcaa actctgtgac atataaagca ggtcttacaa accaagaaat taaagttaat 780aacgtattaa ataactcgtc agtaaaatta tttaaagcag ataatacacc attaaatgtc 840acaaatatta ctcatggtag cggttttagt tcggttgtga cagtaagtga cgcgttacca 900aatggcggaa ttaaagcaaa atcttcaatt tcaatgaaca atgtgacgta tacgacgcaa 960gacgaacatg gtcaagttgt tacagtaaca agaaatgaat ctgttgattc aaatgacagt 1020gcaacagtaa cagtgacacc acaattacaa gcaactactg aaggcgctgt atttattaaa 1080ggtggcgacg gttttgattt cggacacgta gaaagattta ttcaaaaccc gccacatggg 1140gcaacggttg catggcatga tagtccagat acatggaaga atacagtcgg taacactcat 1200aaaactgcgg ttgtaacatt acctaatggt caaggtacgc gtaatgttga agttccagtc 1260aaagtttatc cagttgctaa tgcaaaggcg ccatcacgtg atgtgaaagg tcaaaatttg 1320actaatggaa cggatgcgat gaactacatt acatttgatc caaatacaaa cacaaatggt 1380atcactgcag catgggcaaa tagacaacaa ccaaataacc aacaagcagg cgtgcaacat 1440ttaaatgtcg atgtcacata tccaggtatt tcagctgcta aacgagttcc tgttactgtt 1500aatgtatatc aatttgaatt ccctcaaact acttatacga caacggttgg aggcacttta 1560gcaagtggta cgcaagcatc aggatatgca catatgcaaa atgctactgg tttaccaaca 1620gatggattta cgtataaatg gaatcgtgat actacaggta caaatgacgc aaactggtca 1680gctatgaata aaccgaatgt ggctaaagtc gttaacgcaa aatatgacgt catctataac 1740ggacatactt ttgcaacatc tttaccagcg aaatttgtag taaaagatgt gcaaccagcg 1800aaaccaactg tgactgaaac agcggcagga gcgattacaa ttgcacctgg agcaaaccaa 1860acagtgaata cacatgccgg taacgtaacg acatacgctg ataaattagt tattaaacgt 1920aatggtaacg ttgtgacgac atttacacgt cgcaataata cgagtccatg ggtgaaagaa 1980gcatctgcag caactgtagc aggtattgct ggaactaata atggtattac tgttgcagca 2040ggtactttca accctgctga tacaattcaa gttgttgcaa cgcaaggaag cggagagaca 2100gtgagtgatg agcaacgtag tgatgatttc acagttgtcg caccacaacc gaaccaagcg 2160actactaaga tttggcaaaa tggtcatatt gatatcacgc ctaataatcc atcaggacat 2220ttaattaatc caactcaagc aatggatatt gcttacactg aaaaagtggg taatggtgca 2280gaacatagta agacaattaa tgttgttcgt ggtcaaaata atcaatggac aattgcgaat 2340aagcctgact atgtaacgtt agatgcacaa actggtaaag tgacgttcaa tgccaatact 2400ataaaaccaa attcatcaat cacaattact ccgaaagcag gtacaggtca ctcagtaagt 2460agtaatccaa gtacattaac tgcaccggca gctcatactg tcaacacaac tgaaattgtg 2520aaagattatg gttcaaatgt aacagcagct gaaattaaca atgcagttca agttgctaat 2580aaacgtactg caacgattaa aaatggcaca gcaatgccta ctaatttagc tggtggtagc 2640acaacgacga ttcctgtgac agtaacttac aatgatggta gtactgaaga agtacaagag 2700tccattttca caaaagcgga taaacgtgag ttaatcacag ctaaaaatca tttagatgat 2760ccagtaagca ctgaaggtaa aaagccaggt acaattacgc agtacaataa tgcaatgcat 2820aatgcgcaac aacaaatcaa tactgcgaaa acagaagcac aacaagtgat taataatgag 2880cgtgcaacac cacaacaagt ttctgacgca ctaactaaag ttcgtgcagc acaaactaag 2940attgatcaag ctaaagcatt acttcaaaat aaagaagata atagccaatt agtaacgtct 3000aaaaataact tacaaagttc tgtgaaccaa gtaccatcaa ctgctggtat gacgcaacaa 3060agtattgata actataatgc gaagaagcgt gaagcagaaa ctgaaataac tgcagctcaa 3120cgtgttattg acaatggcga tgcaactgca caacaaattt cagatgaaaa acatcgtgtc 3180gataacgcat taacagcatt aaaccaagcg aaacatgatt taactgcaga tacacatgcc 3240ttagagcaag cagtgcaaca attgaatcgc acaggtacaa cgactggtaa gaagccggca 3300agtattactg cttacaataa ttcgattcgt gcacttcaaa gtgacttaac aagtgctaaa 3360aatagcgcta atgctattat tcaaaagcca ataagaacag tacaagaagt gcaatctgcg 3420ttaacaaatg taaatcgtgt caatgagcga ttaacgcaag caattaatca attagtacct 3480ttagctgata atagtgcttt aaaaactgct aagacgaaac ttgatgaaga aatcaataaa 3540tcagtaacta ctgatggtat gacacaatca tcaatccaag catatgaaaa tgctaaacgt 3600gcgggtcaaa cagaatcaac aaatgcacaa aatgttatta acaatggtga tgcgactgac 3660caacaaattg ccgcagaaaa aacaaaagta gaagaaaaat ataatagctt aaaacaagca 3720attgctggat taactccaga cttggcacca ttacaaactg caaaaactca gttgcaaaat 3780gatattgatc agccaacgag tacgactggt atgacaagcg catctattgc agcatttaat 3840gaaaaacttt cagcagctag aactaaaatt caagaaattg atcgtgtatt agcctcacat 3900ccagatgttg cgacaatacg tcaaaacgtg acagcagcga atgccgctaa atcagcactt 3960gatcaagcac gtaatggctt aacagtcgat aaagcgcctt tagaaaatgc gaaaaatcaa 4020ctacaatata gtattgacac gcaaacaagt acaactggta tgacacaaga ctctataaat 4080gcatacaatg cgaagttaac agctgcacgt aataagattc aacaaatcaa tcaagtatta 4140gcaggttcac cgactgtaga acaaattaat acaaatacgt ctacagcaaa tcaagctaaa 4200tctgatttag atcatgcacg tcaagcttta acaccagata aagcgccgct tcaaactgcg 4260aaaacgcaat tagaacaaag cattaatcaa ccaacggata caacaggtat gacgaccgct 4320tcgttaaatg cgtacaacca aaaattacaa gcagcgcgtc aaaagttaac tgaaattaat 4380caagtgttga atggcaaccc aactgtccaa aatatcaatg ataaagtgac agaggcaaac 4440caagctaagg atcaattaaa tacagcacgt caaggtttaa cattagatag acagccagcg 4500ttaacaacat tacatggtgc atctaactta aaccaagcac aacaaaataa tttcacgcaa 4560caaattaatg ctgctcaaaa tcatgctgcg cttgaaacaa ttaagtctaa cattacggct 4620ttaaatactg cgatgacgaa attaaaagac agtgttgcgg ataataatac aattaaatca 4680gatcaaaatt acactgacgc aacaccagct aataaacaag cgtatgataa tgcagttaat 4740gcggctaaag gtgtcattgg agaaacgact aatccaacga tggatgttaa cacagtgaac 4800caaaaagcag catctgttaa atcgacgaaa gatgctttag atggtcaaca aaacttacaa 4860cgtgcgaaaa cagaagcaac aaatgcgatt acgcatgcaa gtgatttaaa ccaagcacaa 4920aagaatgcat taacacaaca agtgaatagt gcacaaaacg tgcaagcagt aaatgatatt 4980aaacaaacga ctcaaagctt aaatactgct atgacaggtt taaaacgtgg cgttgctaat 5040cataaccaag tcgtacaaag tgataattat gtcaacgcag atactaataa gaaaaatgat 5100tacaacaatg catacaacca tgcgaatgac attattaatg gtaatgcaca acatccagtt 5160ataacaccaa gtgatgttaa caatgcttta tcaaatgtca caagtaaaga acatgcattg 5220aatggtgaag ctaagttaaa tgctgcgaaa caagaagcga atactgcatt aggtcattta 5280aacaatttaa ataatgcaca acgtcaaaac ttacaatcgc aaattaatgg tgcgcatcaa 5340attgatgcag ttaatacaat taagcaaaat gcaacaaact tgaatagtgc aatgggtaac 5400ttaagacaag ctgttgcaga taaagatcaa gtgaaacgta cagaagatta tgcggatgca 5460gatacagcta aacaaaatgc atataacagt gcagtttcaa gtgccgaaac aatcattaat 5520caaacaacaa atccaacgat gtctgttgat gatgttaatc gtgcaacttc agctgttact 5580tctaataaaa atgcattaaa tggttatgaa aaattagcac aatctaaaac agatgctgca 5640agagcaattg atgcattacc acatttaaat aatgcacaaa aagcagatgt taaatctaaa 5700attaatgctg catcaaatat tgctggcgta aatactgtta aacaacaagg tacagattta 5760aatacagcga tgggtaactt gcaaggtgca atcaatgatg aacaaacgac gcttaatagt 5820caaaactatc aagatgcgac acctagtaag aaaacagcat acacaaatgc ggtacaagct 5880gcgaaagata ttttaaataa atcaaatggt caaaataaaa cgaaagatca agttactgaa 5940gcgatgaatc aagtgaattc tgctaaaaat aacttagatg gtacgcgttt attagatcaa 6000gcgaagcaaa cagcaaaaca gcagttaaat aatatgacgc atttaacaac tgcacaaaaa 6060acgaatttaa caaaccaaat taatagtggt actactgtcg ctggtgttca aacggttcaa 6120tcaaatgcca atacattaga tcaagccatg aatacgttaa gacaaagtat tgccaacaaa 6180gatgcgacta aagcaagtga agattacgta gatgctaata atgataagca aacagcatat 6240aacaacgcag tagctgctgc tgaaacgatt attaatgcta atagtaatcc agaaatgaat 6300ccaagtacga ttacacaaaa agcagagcaa gtgaatagtt ctaaaacggc acttaacggt 6360gatgaaaact tagctgctgc aaaacaaaat gcgaaaacgt acttaaacac attgacaagt 6420attacagatg ctcaaaagaa caatttgatt agtcaaatta ctagtgcgac aagagtgagt 6480ggtgttgata ctgtaaaaca aaatgcgcaa catctagacc aagctatggc tagcttacag 6540aatggtatta acaacgaatc tcaagtgaaa tcatctgaga aatatcgtga tgctgataca 6600aataaacaac aagagtatga taatgctatt actgcagcga aagcgatttt aaataaatcg 6660acaggtccaa acactgcgca aaatgcagtt gaagcagcat tacaacgtgt taataatgcg 6720aaagatgcat tgaatggtga tgcaaaatta attgcagctc aaaacgcagc gaaacaacat 6780ttaggtactt taacgcatat cactacagct caacgtaatg atttaacaaa tcaaatttca 6840caagctacaa acttagctgg tgttgaatct gttaaacaaa atgcgaatag tttagatggt 6900gctatgggta acttacaaac ggctatcaac gataagtcag gaacattagc gagccaaaac 6960ttcttggatg ctgatgagca aaaacgtaat gcatacaatc aagctgtatc agcagccgaa 7020accattttaa ataaacaaac tggaccgaat acagcgaaaa cagcagtcga acaagcactt 7080aataatgtta ataatgcgaa acatgcatta aatggtacgc aaaacttaaa caatgcgaaa 7140caagcagcga ttacagcaat caatggcgca tctgatttaa atcaaaaaca aaaagatgca 7200ttaaaagcac aagctaatgg tgctcaacgc gtatctaatg cacaagatgt acagcacaat 7260gcgactgaac tgaacacggc aatgggcaca ttaaaacatg ccatcgcaga taagacgaat 7320acgttagcaa gcagtaaata tgttaatgcc gatagcacta aacaaaatgc ttacacaact 7380aaagttacca atgctgaaca tattattagc ggtacgccaa cggttgttac gacaccttca 7440gaagtaacag ctgcagctaa tcaagtaaac agcgcgaaac aagaattaaa tggtgacgaa 7500agattacgtg aagcaaaaca aaacgccaat actgctattg atgcattaac acaattaaat 7560acacctcaaa aagctaaatt aaaagaacaa gtgggacaag ccaatagatt agaagacgta 7620caaactgttc aaacaaatgg acaagcattg aacaatgcaa tgaaaggctt aagagatagt 7680attgctaacg aaacaacagt caaaacaagt caaaactata cagacgcaag tccgaataac 7740caatcaacat ataatagcgc tgtgtcaaat gcgaaaggta tcattaatca aactaacaat 7800ccgactatgg atactagtgc gattacccaa gctacaacac aagtgaataa tgctaaaaat 7860ggtttaaacg gtgctgaaaa cttaagaaat gcacaaaaca ctgctaagca aaacttaaat 7920acattatcac acttaacaaa taaccaaaaa tctgccatct catcacaaat tgatc7975210132112001212DNA213Staphylococcus aureus40013gatcatggca ttgtatttaa tgcaagtcta cctttgtaca aagatgccat ccatcaaaaa 60ggatcaatgc gcagtaatga caatggtgat gatatgagta tgatggtggg tacagtgctg 120agtggctttg aatatcgagc gcaaaaagaa aagtatgata acttatataa attcttcaaa 180gaaaatgaaa agaaatatca atatacaggc tttacaaaag aggcaattaa caagacacaa 240aatgtcggat ataaaaatga atatttttat attacatact cttctagaag tttaaaagaa 300tatcgaaagt attatgaacc actgattcga aaaaatgata aagaatttaa agaaggaatg 360gaacgagcaa gaaaagaagt gaattacgct gcaaatacag atgctgttgc tacacttttt 420tctactaaga aaaactttac taaagacaat acagtagatg atgtaatcga actaagtgat 480aaattatata atttaaaaaa taaaccagat aaatctacaa tcacaataca aatagggaaa 540cccactatta atactaagaa agccttttat gatgataatc gtccaataga atatggggtg 600cacagtaaag atgaataaaa ttaatgatag ggatttaaca gaattaagta gttactgggt 660ttatcaaaat attgatataa aaaaagaatt taaagttaat ggaaaaaggt ttaaacaagt 720agacagttat aatgatgata agaatagtaa tttgaatggt gctgctgata ttaaaatata 780tgagttatta gatgataaaa gtaaaccaac tggtcaacag acaataattt atcaaggaac 840atctaatgag gcaattaatc caaataatcc attaaaatca tcggggtttg gagatgattg 900gctccaaaat gctaaattaa tgaataatga taatgaaagc acagattatt taaagcaaac 960agatcaatta tcaaatcaat ataaaataaa gttagaagat gcagatagat tatcaaatag 1020tgatttttta aaaaaatata gaatggaatc aagtaacttc aaaaacaaaa ccattgtggc 1080ggatggcggt aattcggaag gcggtgcagg agcaaaatat caaggagcga aacatccgaa 1140tgaaaaagtt gttgctactg actcagcaat gattccttat gctgcttggc agaaatttgc 1200tagaccacgc tttgataata tgattagttt taatagtacc aacgatttat taacatggtt 1260acaagatcca ttcatcaaag atatgccagg aaaacgcgtt aacattaatg atggtgtgcc 1320caggttagat actttaatag acagccatgt aggttataaa aggaagttaa atagaaaaga 1380taacacatac gatactgtac cactaatcaa aataaagtcg gtaaaagata cagaaattaa 1440aaatggaaaa aaagtaaaaa agactattaa cataacatta gatatggatg ggcgaattcc 1500aataaatgtt tggacaggag attcgattgc acgttctgga agaggaactt taattaaact 1560taatttagaa aatcttgatg cgttgagtaa actgattact ggtgaaacaa gtggtatgtt 1620agcagaatgc gtaatctttt taaatgaaag ttttaacatc tcagaaaatg aaaataaaaa 1680ttttgcagat agaaagaaac aattatcaga aggatttaag gataagatta acttatttca 1740gttagaagaa atggaaagaa ctttaattag taaaataaac tcacttgaag aagttgcaga 1800tgaaacaata gaaagtatta gtgctgttaa acacttatta cctgattttg cattggatgc 1860attaaaagaa agaattaatg agttgtttaa aggtataaaa tcttttatag aaaaagtgta 1920tgatagtata gataatgaaa ttttagaaat tttcaaaaat atagatcacg acttcagaga 1980tggagtatct gaagaaatga t 200121014211106212PRT213Staphylococcus aureus40014Asp Gln Thr Lys Thr Gln Thr Ala His Thr Val Lys Thr Ala Gln Thr1 5 10 15Ala Gln Glu Gln Asn Lys Val Gln Thr Pro Val Lys Asp Val Ala Thr20 25 30Ala Lys Ser Glu Ser Asn Asn Gln Ala Val Ser Asp Asn Lys Ser Gln35 40 45Gln Thr Asn Lys Val Thr Lys His Asn Glu Thr Pro Lys Gln Ala Ser50 55 60Lys Ala Lys Glu Leu Pro Lys Thr Gly Leu Thr Ser Val Asp Asn Phe65 70 75 80Ile Ser Thr Val Ala Phe Ala Thr Leu Ala Leu Leu Gly Ser Leu Ser85 90 95Leu Leu Leu Phe Lys Arg Lys Glu Ser Lys100 10521015211960212PRT213Staphylococcus aureus40015Asp Arg Ile Ile Glu Thr Ala Pro Thr Asp Tyr Leu Ser Trp Gly Val1 5 10 15Gly Ala Val Gly Asn Pro Arg Phe Ile Asn Val Glu Ile Val His Thr20 25 30His Asp Tyr Ala Ser Phe Ala Arg Ser Met Asn Asn Tyr Ala Asp Tyr35 40 45Ala Ala Thr Gln Leu Gln Tyr Tyr Gly Leu Lys Pro Asp Ser Ala Glu50 55 60Tyr Asp Gly Asn Gly Thr Val Trp Thr His Tyr Ala Val Ser Lys Tyr65 70 75 80Leu Gly Gly Thr Asp His Ala Asp Pro His Gly Tyr Leu Arg Ser His85 90 95Asn Tyr Ser Tyr Asp Gln Leu Tyr Asp Leu Ile Asn Glu Lys Tyr Leu100 105 110Ile Lys Met Gly Lys Val Ala Pro Trp Gly Thr Gln Ser Thr Thr Thr115 120 125Pro Thr Thr Pro Ser Lys Pro Thr Thr Pro Ser Lys Pro Ser Thr Gly130 135 140Lys Leu Thr Val Ala Ala Asn Asn Gly Val Ala Gln Ile Lys Pro Thr145 150 155 160Asn Ser Gly Leu Tyr Thr Thr Val Tyr Asp Lys Thr Gly Lys Ala Thr165 170 175Asn Glu Val Gln Lys Thr Phe Ala Val Ser Lys Thr Ala Thr Leu Gly180 185 190Asn Gln Lys Phe Tyr Leu Val Gln Asp Tyr Asn Ser Gly Asn Lys Phe195 200 205Gly Trp Val Lys Glu Gly Asp Val Val Tyr Asn Thr Ala Lys Ser Pro210 215 220Val Asn Val Asn Gln Ser Tyr Ser Ile Lys Pro Gly Thr Lys Leu Tyr225 230 235 240Thr Val Pro Trp Gly Thr Ser Lys Gln Val Ala Gly Ser Val Ser Gly245 250 255Ser Gly Asn Gln Thr Phe Lys Ala Ser Lys Gln Gln Gln Ile Asp Lys260 265 270Ser Ile Tyr Leu Tyr Gly Ser Val Asn Gly Lys Ser Gly Trp Val Ser275 280 285Lys Ala Tyr Leu Val Asp Thr Ala Lys Pro Thr Pro Thr Pro Thr Pro290 295 300Lys Pro Ser Thr Pro Thr Thr Asn Asn Lys Leu Thr Val Ser Ser Leu305 310 315 320Asn Gly Val Ala Gln Ile Asn Ala Lys Asn Asn Gly Leu Phe Thr Thr325 330 335Val Tyr Asp Lys Thr Gly Lys Pro Thr Lys Glu Val Gln Lys Thr Phe340 345 350Ala Val Thr Lys Glu Ala Ser Leu Gly Gly Asn Lys Phe Tyr Leu Val355 360 365Lys Asp Tyr Asn Ser Pro Thr Leu Ile Gly Trp Val Lys Gln Gly Asp370 375 380Val Ile Tyr Asn Asn Ala Lys Ser Pro Val Asn Val Met Gln Thr Tyr385 390 395 400Thr Val Lys Pro Gly Thr Lys Leu Tyr Ser Val Pro Trp Gly Thr Tyr405 410 415Lys Gln Glu Ala Gly Ala Val Ser Gly Thr Gly Asn Gln Thr Phe Lys420 425 430Ala Thr Lys Gln Gln Gln Ile Asp Lys Ser Ile Tyr Leu Phe Gly Thr
435 440 445Val Asn Gly Lys Ser Gly Trp Val Ser Lys Ala Tyr Leu Ala Val Pro450 455 460Ala Ala Pro Lys Lys Ala Val Ala Gln Pro Lys Thr Ala Val Lys Ala465 470 475 480Tyr Thr Val Thr Lys Pro Gln Thr Thr Gln Thr Val Ser Lys Ile Ala485 490 495Gln Val Lys Pro Asn Asn Thr Gly Ile Arg Ala Ser Val Tyr Glu Lys500 505 510Thr Ala Lys Asn Gly Ala Lys Tyr Ala Asp Arg Thr Phe Tyr Val Thr515 520 525Lys Glu Arg Ala His Gly Asn Glu Thr Tyr Val Leu Leu Asn Asn Thr530 535 540Ser His Asn Ile Pro Leu Gly Trp Phe Asn Val Lys Asp Leu Asn Val545 550 555 560Gln Asn Leu Gly Lys Glu Val Lys Thr Thr Gln Lys Tyr Thr Val Asn565 570 575Lys Ser Asn Asn Gly Leu Ser Met Val Pro Trp Gly Thr Lys Asn Gln580 585 590Val Ile Leu Thr Gly Asn Asn Ile Ala Gln Gly Thr Phe Asn Ala Thr595 600 605Lys Gln Val Ser Val Gly Lys Asp Val Tyr Leu Tyr Gly Thr Ile Asn610 615 620Asn Arg Thr Gly Trp Val Asn Ala Lys Asp Leu Thr Ala Pro Thr Ala625 630 635 640Val Lys Pro Thr Thr Ser Ala Ala Lys Asp Tyr Asn Tyr Thr Tyr Val645 650 655Ile Lys Asn Gly Asn Gly Tyr Tyr Tyr Val Thr Pro Asn Ser Asp Thr660 665 670Ala Lys Tyr Ser Leu Lys Ala Phe Asn Glu Gln Pro Phe Ala Val Val675 680 685Lys Glu Gln Val Ile Asn Gly Gln Thr Trp Tyr Tyr Gly Lys Leu Ser690 695 700Asn Gly Lys Leu Ala Trp Ile Lys Ser Thr Asp Leu Ala Lys Glu Leu705 710 715 720Ile Lys Tyr Asn Gln Thr Gly Met Ala Leu Asn Gln Val Ala Gln Ile725 730 735Gln Ala Gly Leu Gln Tyr Lys Pro Gln Val Gln Arg Val Pro Gly Lys740 745 750Trp Thr Gly Ala Asn Phe Asn Asp Val Lys His Ala Met Asp Thr Lys755 760 765Arg Leu Ala Gln Asp Pro Ala Leu Lys Tyr Gln Phe Leu Arg Leu Asp770 775 780Gln Pro Gln Asn Ile Ser Ile Asp Lys Ile Asn Gln Phe Leu Lys Gly785 790 795 800Lys Gly Val Leu Glu Asn Gln Gly Ala Ala Phe Asn Lys Ala Ala Gln805 810 815Met Tyr Gly Ile Asn Glu Val Tyr Leu Ile Ser His Ala Leu Leu Glu820 825 830Thr Gly Asn Gly Thr Ser Gln Leu Ala Lys Gly Ala Asp Val Val Asn835 840 845Asn Lys Val Val Thr Asn Ser Asn Thr Lys Tyr His Asn Val Phe Gly850 855 860Ile Ala Ala Tyr Asp Asn Asp Pro Leu Arg Glu Gly Ile Lys Tyr Ala865 870 875 880Lys Gln Ala Gly Trp Asp Thr Val Ser Lys Ala Ile Val Gly Gly Ala885 890 895Lys Phe Ile Gly Asn Ser Tyr Val Lys Ala Gly Gln Asn Thr Leu Tyr900 905 910Lys Met Arg Trp Asn Pro Ala His Pro Gly Thr His Gln Tyr Ala Thr915 920 925Asp Val Asp Trp Ala Asn Ile Asn Ala Lys Ile Ile Lys Gly Tyr Tyr930 935 940Asp Lys Ile Gly Glu Val Gly Lys Tyr Phe Asp Ile Pro Gln Tyr Lys945 950 955 96021016211386212PRT213Staphylococcus aureus40016Asp Gln Tyr Ser Glu Asp Ala Lys Lys Thr Gln Lys Asp Tyr Ala Ser1 5 10 15Gln Ser Lys Lys Asp Lys Asn Glu Lys Ser Asn Thr Lys Asn Pro Gln20 25 30Leu Pro Thr Gln Asp Glu Leu Lys His Lys Ser Lys Pro Ala Gln Ser35 40 45Phe Asn Asn Asp Val Asn Gln Lys Asp Thr Arg Ala Thr Ser Leu Phe50 55 60Glu Thr Asp Pro Ser Ile Ser Asn Asn Asp Asp Ser Gly Gln Phe Asn65 70 75 80Val Val Asp Ser Lys Asp Thr Arg Gln Phe Val Lys Ser Ile Ala Lys85 90 95Asp Ala His Arg Ile Gly Gln Asp Asn Asp Ile Tyr Ala Ser Val Met100 105 110Ile Ala Gln Ala Ile Leu Glu Ser Asp Ser Gly Arg Ser Ala Leu Ala115 120 125Lys Ser Pro Asn His Asn Leu Phe Gly Ile Lys Gly Ala Phe Glu Gly130 135 140Asn Ser Val Pro Phe Asn Thr Leu Glu Ala Asp Gly Asn Gln Leu Tyr145 150 155 160Ser Ile Asn Ala Gly Phe Arg Lys Tyr Pro Ser Thr Lys Glu Ser Leu165 170 175Lys Asp Tyr Ser Asp Leu Ile Lys Asn Gly Ile Asp Gly Asn Arg Thr180 185 190Ile Tyr Lys Pro Thr Trp Lys Ser Glu Ala Asp Ser Tyr Lys Asp Ala195 200 205Thr Ser His Leu Ser Lys Thr Tyr Ala Thr Asp Pro Asn Tyr Ala Lys210 215 220Lys Leu Asn Ser Ile Ile Lys His Tyr Gln Leu Thr Gln Phe Asp Asp225 230 235 240Glu Arg Met Pro Asp Leu Asp Lys Tyr Glu Arg Ser Ile Lys Asp Tyr245 250 255Asp Asp Ser Ser Asp Glu Phe Lys Pro Phe Arg Glu Val Ser Asp Ser260 265 270Met Pro Tyr Pro His Gly Gln Cys Thr Trp Tyr Val Tyr Asn Arg Met275 280 285Lys Gln Phe Gly Thr Ser Ile Ser Gly Asp Leu Gly Asp Ala His Asn290 295 300Trp Asn Asn Arg Ala Gln Tyr Arg Asp Tyr Gln Val Ser His Thr Pro305 310 315 320Lys Arg His Ala Ala Val Val Phe Glu Ala Gly Gln Phe Gly Ala Asp325 330 335Gln His Tyr Gly His Val Ala Phe Val Glu Lys Val Asn Ser Asp Gly340 345 350Ser Ile Val Ile Ser Glu Ser Asn Val Lys Gly Leu Gly Ile Ile Ser355 360 365His Arg Thr Ile Asn Ala Ala Ala Ala Glu Glu Leu Ser Tyr Ile Thr370 375 380Gly Lys38521017211325212PRT213Staphylococcus aureus40017Met Lys Met Asn Lys Leu Val Lys Ser Ser Val Ala Thr Ser Met Ala1 5 10 15Leu Leu Leu Leu Ser Gly Thr Ala Asn Ala Glu Gly Lys Ile Thr Pro20 25 30Val Ser Val Lys Lys Val Asp Asp Lys Val Thr Leu Tyr Lys Thr Thr35 40 45Ala Thr Ala Asp Ser Asp Lys Phe Lys Ile Ser Gln Ile Leu Thr Phe50 55 60Asn Phe Ile Lys Asp Lys Ser Tyr Asp Lys Asp Thr Leu Val Leu Lys65 70 75 80Ala Thr Gly Asn Ile Asn Ser Gly Phe Val Lys Pro Asn Pro Asn Asp85 90 95Tyr Asp Phe Ser Lys Leu Tyr Trp Gly Ala Lys Tyr Asn Val Ser Ile100 105 110Ser Ser Gln Ser Asn Asp Ser Val Asn Val Val Asp Tyr Ala Pro Lys115 120 125Asn Gln Asn Glu Glu Phe Gln Val Gln Asn Thr Leu Gly Tyr Thr Phe130 135 140Gly Gly Asp Ile Ser Ile Ser Asn Gly Leu Ser Gly Gly Leu Asn Gly145 150 155 160Asn Thr Ala Phe Ser Glu Thr Ile Asn Tyr Lys Gln Glu Ser Tyr Arg165 170 175Thr Thr Leu Ser Arg Asn Thr Asn Tyr Lys Asn Val Gly Trp Gly Val180 185 190Glu Ala His Lys Ile Met Asn Asn Gly Trp Gly Pro Tyr Gly Arg Asp195 200 205Ser Phe His Pro Thr Tyr Gly Asn Glu Leu Phe Leu Ala Gly Arg Gln210 215 220Ser Ser Ala Tyr Ala Gly Gln Asn Phe Ile Ala Gln His Gln Met Pro225 230 235 240Leu Leu Ser Arg Ser Asn Phe Asn Pro Glu Phe Leu Ser Val Leu Ser245 250 255His Arg Gln Asp Gly Ala Lys Lys Ser Lys Ile Thr Val Thr Tyr Gln260 265 270Arg Glu Met Asp Leu Tyr Gln Ile Arg Trp Asn Gly Phe Tyr Trp Ala275 280 285Gly Ala Asn Tyr Lys Asn Phe Lys Thr Arg Thr Phe Lys Ser Thr Tyr290 295 300Glu Ile Asp Trp Glu Asn His Lys Val Lys Leu Leu Asp Thr Lys Glu305 310 315 320Thr Glu Asn Asn Lys32521018211157212PRT213Staphylococcus aureus40018Ser Phe Asn Tyr Ser Lys Ser Ile Ser Tyr Thr Gln Gln Asn Tyr Val1 5 10 15Ser Glu Val Glu Gln Gln Asn Ser Lys Ser Val Leu Trp Gly Val Lys20 25 30Ala Asn Ser Phe Ala Thr Glu Ser Gly Gln Lys Ser Ala Phe Asp Ser35 40 45Asp Leu Phe Val Gly Tyr Lys Pro His Ser Lys Asp Pro Arg Asp Tyr50 55 60Phe Val Pro Asp Ser Glu Leu Pro Pro Leu Val Gln Ser Gly Phe Asn65 70 75 80Pro Ser Phe Ile Ala Thr Val Ser His Glu Lys Gly Ser Ser Asp Thr85 90 95Ser Glu Phe Glu Ile Thr Tyr Gly Arg Asn Met Asp Val Thr His Ala100 105 110Ile Lys Arg Ser Thr His Tyr Gly Asn Ser Tyr Leu Asp Gly His Arg115 120 125Val His Asn Ala Phe Val Asn Arg Asn Tyr Thr Val Lys Tyr Glu Val130 135 140Asn Trp Lys Thr His Glu Ile Lys Val Lys Gly Gln Asn145 150 15521019211345212PRT213Staphylococcus aureus40019Ile Ile Ala Ile Ile Ile Leu Ile Phe Ile Ser Phe Phe Phe Ser Gly1 5 10 15Ser Glu Thr Ala Leu Thr Ala Ala Asn Lys Ala Lys Phe Lys Thr Glu20 25 30Ala Asp Lys Gly Asp Lys Lys Ala Lys Gly Ile Val Lys Leu Leu Glu35 40 45Lys Pro Ser Glu Phe Ile Thr Thr Ile Leu Ile Gly Asn Asn Val Ala50 55 60Asn Ile Leu Leu Pro Thr Leu Val Thr Ile Met Ala Leu Arg Trp Gly65 70 75 80Ile Ser Val Gly Ile Ala Ser Ala Val Leu Thr Val Val Ile Ile Leu85 90 95Ile Ser Glu Val Ile Pro Lys Ser Val Ala Ala Thr Phe Pro Asp Lys100 105 110Ile Thr Arg Leu Val Tyr Pro Ile Ile Asn Ile Cys Val Ile Val Phe115 120 125Arg Pro Ile Thr Leu Leu Leu Asn Lys Leu Thr Asp Ser Ile Asn Arg130 135 140Ser Leu Ser Lys Gly Gln Pro Gln Glu His Gln Phe Ser Lys Glu Glu145 150 155 160Phe Lys Thr Met Leu Ala Ile Ala Gly His Glu Gly Ala Leu Asn Glu165 170 175Ile Glu Thr Ser Arg Leu Glu Gly Val Ile Asn Phe Glu Asn Leu Lys180 185 190Val Lys Asp Val Asp Thr Thr Pro Arg Ile Asn Val Thr Ala Phe Ala195 200 205Ser Asn Ala Thr Tyr Glu Glu Val Tyr Glu Thr Val Met Asn Lys Pro210 215 220Tyr Thr Arg Tyr Pro Val Tyr Glu Gly Asp Ile Asp Asn Ile Ile Gly225 230 235 240Val Phe His Ser Lys Tyr Leu Leu Ala Trp Ser Asn Lys Lys Glu Asn245 250 255Gln Ile Thr Asn Tyr Ser Ala Lys Pro Leu Phe Val Asn Glu His Asn260 265 270Lys Ala Glu Trp Val Leu Arg Lys Met Thr Ile Ser Arg Lys His Leu275 280 285Ala Ile Val Leu Asp Glu Phe Gly Gly Thr Glu Ala Ile Val Ser His290 295 300Glu Asp Leu Ile Glu Glu Leu Leu Gly Met Glu Ile Glu Asp Glu Met305 310 315 320Asp Lys Lys Glu Lys Glu Lys Leu Ser Gln Gln Gln Ile Gln Phe Gln325 330 335Gln Arg Lys Asn Arg Asn Val Ser Ile340 34521020211133212PRT213Staphylococcus aureus40020Met Asn Lys Gln Gln Lys Glu Phe Lys Ser Phe Tyr Ser Ile Arg Lys1 5 10 15Ser Ser Leu Gly Val Ala Ser Val Ala Ile Ser Thr Leu Leu Leu Leu20 25 30Met Ser Asn Gly Glu Ala Gln Ala Ala Ala Glu Glu Thr Gly Gly Thr35 40 45Asn Thr Glu Ala Gln Pro Lys Thr Glu Ala Val Ala Ser Pro Thr Thr50 55 60Thr Ser Glu Lys Ala Pro Glu Thr Lys Pro Val Ala Asn Ala Val Ser65 70 75 80Val Ser Asn Lys Glu Val Glu Ala Pro Thr Ser Glu Thr Lys Glu Ala85 90 95Lys Glu Val Lys Glu Val Lys Ala Pro Lys Glu Thr Lys Glu Val Lys100 105 110Pro Ala Ala Lys Ala Thr Asn Asn Thr Tyr Pro Ile Leu Asn Gln Glu115 120 125Leu Ile Arg Ser Asp13021021211205212PRT213Staphylococcus aureus40021Asp His Gly Ile Val Phe Asn Ala Ser Leu Pro Leu Tyr Lys Asp Ala1 5 10 15Ile His Gln Lys Gly Ser Met Arg Ser Asn Asp Asn Gly Asp Asp Met20 25 30Ser Met Met Val Gly Thr Val Leu Ser Gly Phe Glu Tyr Arg Ala Gln35 40 45Lys Glu Lys Tyr Asp Asn Leu Tyr Lys Phe Phe Lys Glu Asn Glu Lys50 55 60Lys Tyr Gln Tyr Thr Gly Phe Thr Lys Glu Ala Ile Asn Lys Thr Gln65 70 75 80Asn Val Gly Tyr Lys Asn Glu Tyr Phe Tyr Ile Thr Tyr Ser Ser Arg85 90 95Ser Leu Lys Glu Tyr Arg Lys Tyr Tyr Glu Pro Leu Ile Arg Lys Asn100 105 110Asp Lys Glu Phe Lys Glu Gly Met Glu Arg Ala Arg Lys Glu Val Asn115 120 125Tyr Ala Ala Asn Thr Asp Ala Val Ala Thr Leu Phe Ser Thr Lys Lys130 135 140Asn Phe Thr Lys Asp Asn Thr Val Asp Asp Val Ile Glu Leu Ser Asp145 150 155 160Lys Leu Tyr Asn Leu Lys Asn Lys Pro Asp Lys Ser Thr Ile Thr Ile165 170 175Gln Ile Gly Lys Pro Thr Ile Asn Thr Lys Lys Ala Phe Tyr Asp Asp180 185 190Asn Arg Pro Ile Glu Tyr Gly Val His Ser Lys Asp Glu195 200 20521022211510212PRT213Staphylococcus aureus40022Asp His Tyr Val Ile Gln Tyr Phe Ser Gly Leu Ile Gly Gly Arg Gly1 5 10 15Arg Arg Ala Asn Leu Tyr Gly Leu Phe Asn Lys Ala Ile Glu Phe Glu20 25 30Asn Ser Ser Phe Arg Gly Leu Tyr Gln Phe Ile Arg Phe Ile Asp Glu35 40 45Leu Ile Glu Arg Gly Lys Asp Phe Gly Glu Glu Asn Val Val Gly Pro50 55 60Asn Asp Asn Val Val Arg Met Met Thr Ile His Ser Ser Lys Gly Leu65 70 75 80Glu Phe Pro Phe Val Ile Tyr Ser Gly Leu Ser Lys Asp Phe Asn Lys85 90 95Arg Asp Leu Lys Gln Pro Val Ile Leu Asn Gln Gln Phe Gly Leu Gly100 105 110Met Asp Tyr Phe Asp Val Asp Lys Glu Met Ala Phe Pro Ser Leu Ala115 120 125Ser Val Ala Tyr Arg Ala Val Ala Glu Lys Glu Leu Val Ser Glu Glu130 135 140Met Arg Leu Val Tyr Val Ala Leu Thr Arg Ala Lys Glu Gln Leu Tyr145 150 155 160Leu Ile Gly Arg Val Lys Asn Asp Lys Ser Leu Leu Glu Leu Glu Gln165 170 175Leu Ser Ile Ser Gly Glu His Ile Ala Val Asn Glu Arg Leu Thr Ser180 185 190Pro Asn Pro Phe His Leu Ile Tyr Ser Ile Leu Ser Lys His Gln Ser195 200 205Ala Ser Ile Pro Asp Asp Leu Lys Phe Glu Lys Asp Ile Ala Gln Ile210 215 220Glu Asp Ser Ser Arg Pro Asn Val Asn Ile Ser Ile Val Tyr Phe Glu225 230 235 240Asp Val Ser Thr Glu Thr Ile Leu Asp Asn Asp Glu Tyr Arg Ser Val245 250 255Asn Gln Leu Glu Thr Met Gln Asn Gly Asn Glu Asp Val Lys Ala Gln260 265 270Ile Lys His Gln Leu Asp Tyr Arg Tyr Pro Tyr Val Asn Asp Thr Lys275 280 285Lys Pro Ser Lys Gln Ser Val Ser Glu Leu Lys Arg Gln Tyr Glu Thr290 295 300Glu Glu Ser Gly Thr Ser Tyr Glu Arg Val Arg Gln Tyr Arg Ile Gly305 310 315 320Phe Ser Thr Tyr Glu Arg Pro Lys Phe Leu Ser Glu Gln Gly Lys Arg325 330 335Lys Ala Asn Glu Ile Gly Thr Leu Met His Thr Val Met Gln His Leu340 345 350Pro Phe Lys Lys Glu Arg Ile Ser Glu Val Glu Leu His Gln Tyr Ile355 360 365Asp Gly Leu Ile Asp Lys His Ile Ile Glu Ala Asp Ala Lys Lys Asp370 375 380Ile Arg Met Asp Glu Ile Met Thr Phe Ile Asn Ser Glu Leu Tyr Ser385 390 395 400Ile Ile Ala Glu Ala Glu Gln Val Tyr Arg Glu Leu Pro Phe Val Val405 410 415Asn Gln Ala Leu Val Asp Gln Leu Pro Gln Gly Asp Glu Asp Val Ser420 425 430Ile Ile Gln Gly Met Ile Asp Leu Ile Phe Val Lys Asp Gly Val His435 440 445Tyr Phe Val Asp Tyr Lys Thr Asp Ala Phe Asn Arg Arg Arg Gly Met450 455 460Thr Asp Glu Glu Ile Gly Thr Gln Leu Lys Asn Lys Tyr Lys Ile Gln465 470 475 480Met Lys Tyr Tyr Gln Asn Thr Leu Gln Thr Ile Leu Asn Lys Glu Val485 490 495Lys Gly Tyr Leu Tyr Phe Phe Lys Phe Gly Thr Leu Gln Leu500 505 51021023211124212PRT213Staphylococcus aureus40023Met Lys Phe Leu Ser Phe Lys Tyr Asn Asp Lys Thr Ser Tyr Gly Val1 5 10 15Lys Val Lys Arg Glu Asp Ala Val Trp Asp Leu Thr Gln Val Phe Ala20 25 30Asp Phe Ala Glu Gly Asp Phe His Pro Lys Thr Leu Leu Ala Gly Leu35 40 45Gln Gln Asn His Thr Leu Asp Phe Gln Glu Gln Val Arg Lys Ala Val50 55 60Val Ala Ala Glu Asp Ser Gly Lys Ala Glu Asp Tyr Lys Ile Ser Phe65 70 75 80Asn Asp Ile Glu Phe Leu Pro Pro Val Thr Pro Pro Asn Asn Val Ile85 90 95Ala Phe Gly Arg Asn Tyr Lys Asp His Ala Asn Glu Leu Asn His Glu100 105 110Val Glu Lys Leu Tyr Val Phe Thr Lys Ala Ala Ser115 12021024211180212PRT213Staphylococcus aureus40024Ser Gly Thr Gly Phe Ile Val Gly Lys Asn Thr Ile Val Thr Asn Lys1 5 10 15His Val Val Ala Gly Met Glu Ile Gly Ala His Ile Ile Ala His Pro20 25 30Asn Gly Glu Tyr Asn Asn Gly Gly Phe Tyr Lys Val Lys Lys Ile Val35 40 45Arg Tyr Ser Gly Gln Glu Asp Ile Ala Ile Leu His Val Glu Asp Lys50 55 60Ala Val His Pro Lys Asn Arg Asn Phe Lys Asp Tyr Thr Gly Ile Leu65 70 75 80Lys Ile Ala Ser Glu Ala Lys Glu Asn Glu Arg Ile Ser Ile Val Gly85 90 95Tyr Pro Glu Pro Tyr Ile Asn Lys Phe Gln Met Tyr Glu Ser Thr Gly100 105 110Lys Val Leu Ser Val Lys Gly Asn Met Ile Ile Thr Asp Ala Phe Val115 120 125Glu Pro Gly Asn Ser Gly Ser Ala Val Phe Asn Ser Lys Tyr Glu Val130 135 140Val Gly Val His Phe Gly Gly Asn Gly Pro Gly Asn Lys Ser Thr Lys145 150 155 160Gly Tyr Gly Val Tyr Phe Ser Pro Glu Ile Lys Lys Phe Ile Ala Asp165 170 175Asn Thr Asp Lys18021025211239212PRT213Staphylococcus aureus40025Met Asn Lys Asn Ile Ile Ile Lys Ser Ile Ala Ala Leu Thr Ile Leu1 5 10 15Thr Ser Ile Thr Gly Val Gly Thr Thr Met Val Glu Gly Ile Gln Gln20 25 30Thr Ala Lys Ala Glu Asn Thr Val Lys Gln Ile Thr Asn Thr Asn Val35 40 45Ala Pro Tyr Ser Gly Val Thr Trp Met Gly Ala Gly Thr Gly Phe Val50 55 60Val Gly Asn His Thr Ile Ile Thr Asn Lys His Val Thr Tyr His Met65 70 75 80Lys Val Gly Asp Glu Ile Lys Ala His Pro Asn Gly Phe Tyr Asn Asn85 90 95Gly Gly Gly Leu Tyr Lys Val Thr Lys Ile Val Asp Tyr Pro Gly Lys100 105 110Glu Asp Ile Ala Val Val Gln Val Glu Glu Lys Ser Thr Gln Pro Lys115 120 125Gly Arg Lys Phe Lys Asp Phe Thr Ser Lys Phe Asn Ile Ala Ser Glu130 135 140Ala Lys Glu Asn Glu Pro Ile Ser Val Ile Gly Tyr Pro Asn Pro Asn145 150 155 160Gly Asn Lys Leu Gln Met Tyr Glu Ser Thr Gly Lys Val Leu Ser Val165 170 175Asn Gly Asn Ile Val Ser Ser Asp Ala Ile Ile Gln Pro Gly Ser Ser180 185 190Gly Ser Pro Ile Leu Asn Ser Lys His Glu Ala Ile Gly Val Ile Tyr195 200 205Ala Gly Asn Lys Pro Ser Gly Glu Ser Thr Arg Gly Phe Ala Val Tyr210 215 220Phe Ser Pro Glu Ile Lys Lys Phe Ile Ala Asp Asn Leu Asp Lys225 230 23521026211470212PRT213Staphylococcus aureus40026Met Gly Cys Thr Val Lys Met Asn Lys Ile Asn Asp Arg Asp Leu Thr1 5 10 15Glu Leu Ser Ser Tyr Trp Val Tyr Gln Asn Ile Asp Ile Lys Lys Glu20 25 30Phe Lys Val Asn Gly Lys Arg Phe Lys Gln Val Asp Ser Tyr Asn Asp35 40 45Asp Lys Asn Ser Asn Leu Asn Gly Ala Ala Asp Ile Lys Ile Tyr Glu50 55 60Leu Leu Asp Asp Lys Ser Lys Pro Thr Gly Gln Gln Thr Ile Ile Tyr65 70 75 80Gln Gly Thr Ser Asn Glu Ala Ile Asn Pro Asn Asn Pro Leu Lys Ser85 90 95Ser Gly Phe Gly Asp Asp Trp Leu Gln Asn Ala Lys Leu Met Asn Asn100 105 110Asp Asn Glu Ser Thr Asp Tyr Leu Lys Gln Thr Asp Gln Leu Ser Asn115 120 125Gln Tyr Lys Ile Lys Leu Glu Asp Ala Asp Arg Leu Ser Asn Ser Asp130 135 140Phe Leu Lys Lys Tyr Arg Met Glu Ser Ser Asn Phe Lys Asn Lys Thr145 150 155 160Ile Val Ala Asp Gly Gly Asn Ser Glu Gly Gly Ala Gly Ala Lys Tyr165 170 175Gln Gly Ala Lys His Pro Asn Glu Lys Val Val Ala Thr Asp Ser Ala180 185 190Met Ile Pro Tyr Ala Ala Trp Gln Lys Phe Ala Arg Pro Arg Phe Asp195 200 205Asn Met Ile Ser Phe Asn Ser Thr Asn Asp Leu Leu Thr Trp Leu Gln210 215 220Asp Pro Phe Ile Lys Asp Met Pro Gly Lys Arg Val Asn Ile Asn Asp225 230 235 240Gly Val Pro Arg Leu Asp Thr Leu Ile Asp Ser His Val Gly Tyr Lys245 250 255Arg Lys Leu Asn Arg Lys Asp Asn Thr Tyr Asp Thr Val Pro Leu Ile260 265 270Lys Ile Lys Ser Val Lys Asp Thr Glu Ile Lys Asn Gly Lys Lys Val275 280 285Lys Lys Thr Ile Asn Ile Thr Leu Asp Met Asp Gly Arg Ile Pro Ile290 295 300Asn Val Trp Thr Gly Asp Ser Ile Ala Arg Ser Gly Arg Gly Thr Leu305 310 315 320Ile Lys Leu Asn Leu Glu Asn Leu Asp Ala Leu Ser Lys Leu Ile Thr325 330 335Gly Glu Thr Ser Gly Met Leu Ala Glu Cys Val Ile Phe Leu Asn Glu340 345 350Ser Phe Asn Ile Ser Glu Asn Glu Asn Lys Asn Phe Ala Asp Arg Lys355 360 365Lys Gln Leu Ser Glu Gly Phe Lys Asp Lys Ile Asn Leu Phe Gln Leu370 375 380Glu Glu Met Glu Arg Thr Leu Ile Ser Lys Ile Asn Ser Leu Glu Glu385 390 395 400Val Ala Asp Glu Thr Ile Glu Ser Ile Ser Ala Val Lys His Leu Leu405 410 415Pro Asp Phe Ala Leu Asp Ala Leu Lys Glu Arg Ile Asn Glu Leu Phe420 425 430Lys Gly Ile Lys Ser Phe Ile Glu Lys Val Tyr Asp Ser Ile Asp Asn435 440 445Glu Ile Leu Glu Ile Phe Lys Asn Ile Asp His Asp Phe Arg Asp Gly450 455 460Val Ser Glu Glu Met Met465 47021027211306212PRT213Staphylococcus aureus40027Met Lys Lys Lys Asp Gly Thr Gln Gln Phe Tyr His Tyr Ala Ser Ser1 5 10 15Val Lys Pro Ala Arg Val Ile Phe Thr Asp Ser Lys Pro Glu Ile Glu20 25 30Leu Gly Leu Gln Ser Gly Gln Phe Trp Arg Lys Phe Glu Val Tyr Glu35 40 45Gly Asp Lys Lys Leu Pro Ile Lys Leu Val Ser Tyr Asp Thr Val Lys50 55 60Asp Tyr Ala Tyr Ile Arg Phe Ser Val Ser Asn Gly Thr Lys Ala Val65 70 75 80Lys Ile Val Ser Ser Thr His Phe Asn Asn Lys Glu Glu Lys Tyr Asp85 90 95Tyr Thr Leu Met Glu Phe Ala Gln Pro Ile Tyr Asn Ser Ala Asp Lys100 105 110Phe Lys Thr Glu Glu Asp Tyr Lys Ala Glu Lys Leu Leu Ala Pro Tyr115 120 125Lys Lys Ala Lys Thr Leu Glu Arg Gln Val Tyr Glu Leu Asn Lys Ile130 135 140Gln Asp Lys Leu Pro Glu Lys Leu Lys Ala Glu Tyr Lys Lys Lys Leu145 150 155 160Glu Asp Thr Lys Lys Ala Leu Asp Glu Gln Val Lys Ser Ala Ile Thr165 170 175Glu Phe Gln Asn Val Gln Pro Thr Asn Glu Lys Met Thr Asp Leu Gln180 185 190Asp Thr Lys Tyr Val Val Tyr Glu Ser Val Glu Asn Asn Glu Ser Met195 200 205Met Asp Thr Phe Val Lys His Pro Ile Lys Thr Gly Met Leu Asn Gly210 215 220Lys Lys Tyr Met Val Met Glu Thr Thr Asn Asp Asp Tyr Trp Lys Asp225 230 235 240Phe Met Val Glu Gly Gln Arg Val Arg Thr Ile Ser Lys Asp Ala Lys245 250 255Asn Asn Thr Arg Thr Ile Ile Phe Pro Tyr Val Glu Gly Lys Thr Leu260 265 270Tyr Asp Ala Ile Val Lys Val His Val Lys Thr Ile Asp Tyr Asp Gly275 280 285Gln Tyr His Val Arg Ile Val Asp Lys Glu Ala Phe Thr Lys Ala His290 295 300Thr Asp305210282112659212PRT213Staphylococcus aureus40028Asp Gln Thr Thr Ile Ile Asn Ser Leu Thr Phe Thr Glu Thr Val Pro1 5 10 15Asn Arg Ser Tyr Ala Arg Ala Ser Ala Asn Glu Ile Thr Ser Lys Thr20 25 30Val Ser Asn Val Ser Arg Thr Gly Asn Asn Ala Asn Val Thr Val Thr35 40 45Val Thr Tyr Gln Asp Gly Thr Thr Ser Thr Val Thr Val Pro Val Lys50 55 60His Val Ile Pro Glu Ile Val Ala His Ser His Tyr Thr Val Gln Gly65 70 75 80Gln Asp Phe Pro Ala Gly Asn Gly Ser Ser Ala Ser Asp Tyr Phe Lys85 90 95Leu Ser Asn Gly Ser Asp Ile Ala Asp Ala Thr Ile Thr Trp Val Ser100 105 110Gly Gln Ala Pro Asn Lys Asp Asn Thr Arg Ile Gly Glu Asp Ile Thr115 120 125Val Thr Ala His Ile Leu Ile Asp Gly Glu Thr Thr Pro Ile Thr Lys130 135 140Thr Ala Thr Tyr Lys Val Val Arg Thr Val Pro Lys His Val Phe Glu145 150 155 160Thr Ala Arg Gly Val Leu Tyr Pro Gly Val Ser Asp Met Tyr Asp Ala165 170 175Lys Gln Tyr Val Lys Pro Val Asn Asn Ser Trp Ser Thr Asn Ala Gln180 185 190His Met Asn Phe Gln Phe Val Gly Thr Tyr Gly Pro Asn Lys Asp Val195 200 205Val Gly Ile Ser Thr Arg Leu Ile Arg Val Thr Tyr Asp Asn Arg Gln210 215 220Thr Glu Asp Leu Thr Ile Leu Ser Lys Val Lys Pro Asp Pro Pro Arg225 230 235 240Ile Asp Ala Asn Ser Val Thr Tyr Lys Ala Gly Leu Thr Asn Gln Glu245 250 255Ile Lys Val Asn Asn Val Leu Asn Asn Ser Ser Val Lys Leu Phe Lys260 265 270Ala Asp Asn Thr Pro Leu Asn Val Thr Asn Ile Thr His Gly Ser Gly275 280 285Phe Ser Ser Val Val Thr Val Ser Asp Ala Leu Pro Asn Gly Gly Ile290 295 300Lys Ala Lys Ser Ser Ile Ser Met Asn Asn Val Thr Tyr Thr Thr Gln305 310 315 320Asp Glu His Gly Gln Val Val Thr Val Thr Arg Asn Glu Ser Val Asp325 330 335Ser Asn Asp Ser Ala Thr Val Thr Val Thr Pro Gln Leu Gln Ala Thr340 345 350Thr Glu Gly Ala Val Phe Ile Lys Gly Gly Asp Gly Phe Asp Phe Gly355 360 365His Val Glu Arg Phe Ile Gln Asn Pro Pro His Gly Ala Thr Val Ala370 375 380Trp His Asp Ser Pro Asp Thr Trp Lys Asn Thr Val Gly Asn Thr His385 390 395 400Lys Thr Ala Val Val Thr Leu Pro Asn Gly Gln Gly Thr Arg Asn Val405 410 415Glu Val Pro Val Lys Val Tyr Pro Val Ala Asn Ala Lys Ala Pro Ser420 425 430Arg Asp Val Lys Gly Gln Asn Leu Thr Asn Gly Thr Asp Ala Met Asn435 440 445Tyr Ile Thr Phe Asp Pro Asn Thr Asn Thr Asn Gly Ile Thr Ala Ala450 455 460Trp Ala Asn Arg Gln Gln Pro Asn Asn Gln Gln Ala Gly Val Gln His465 470 475 480Leu Asn Val Asp Val Thr Tyr Pro Gly Ile Ser Ala Ala Lys Arg Val485 490 495Pro Val Thr Val Asn Val Tyr Gln Phe Glu Phe Pro Gln Thr Thr Tyr500 505 510Thr Thr Thr Val Gly Gly Thr Leu Ala Ser Gly Thr Gln Ala Ser Gly515 520 525Tyr Ala His Met Gln Asn Ala Thr Gly Leu Pro Thr Asp Gly Phe Thr530 535 540Tyr Lys Trp Asn Arg Asp Thr Thr Gly Thr Asn Asp Ala Asn Trp Ser545 550 555 560Ala Met Asn Lys Pro Asn Val Ala Lys Val Val Asn Ala Lys Tyr Asp565 570 575Val Ile Tyr Asn Gly His Thr Phe Ala Thr Ser Leu Pro Ala Lys Phe580 585 590Val Val Lys Asp Val Gln Pro Ala Lys Pro Thr Val Thr Glu Thr Ala595 600 605Ala Gly Ala Ile Thr Ile Ala Pro Gly Ala Asn Gln Thr Val Asn Thr610 615 620His Ala Gly Asn Val Thr Thr Tyr Ala Asp Lys Leu Val Ile Lys Arg625 630 635 640Asn Gly Asn Val Val Thr Thr Phe Thr Arg Arg Asn Asn Thr Ser Pro645 650 655Trp Val Lys Glu Ala Ser Ala Ala Thr Val Ala Gly Ile Ala Gly Thr660 665 670Asn Asn Gly Ile Thr Val Ala Ala Gly Thr Phe Asn Pro Ala Asp Thr675 680 685Ile Gln Val Val Ala Thr Gln Gly Ser Gly Glu Thr Val Ser Asp Glu690 695 700Gln Arg Ser Asp Asp Phe Thr Val Val Ala Pro Gln Pro Asn Gln Ala705 710 715 720Thr Thr Lys Ile Trp Gln Asn Gly His Ile Asp Ile Thr Pro Asn Asn725 730 735Pro Ser Gly His Leu Ile Asn Pro Thr Gln Ala Met Asp Ile Ala Tyr740 745 750Thr Glu Lys Val Gly Asn Gly Ala Glu His Ser Lys Thr Ile Asn Val755 760 765Val Arg Gly Gln Asn Asn Gln Trp Thr Ile Ala Asn Lys Pro Asp Tyr770 775 780Val Thr Leu Asp Ala Gln Thr Gly Lys Val Thr Phe Asn Ala Asn Thr785 790 795 800Ile Lys Pro Asn Ser Ser Ile Thr Ile Thr Pro Lys Ala Gly Thr Gly805 810 815His Ser Val Ser Ser Asn Pro Ser Thr Leu Thr Ala Pro Ala Ala His820 825 830Thr Val Asn Thr Thr Glu Ile Val Lys Asp Tyr Gly Ser Asn Val Thr835 840 845Ala Ala Glu Ile Asn Asn Ala Val Gln Val Ala Asn Lys Arg Thr Ala850 855 860Thr Ile Lys Asn Gly Thr Ala Met Pro Thr Asn Leu Ala Gly Gly Ser865 870 875 880Thr Thr Thr Ile Pro Val Thr Val Thr Tyr Asn Asp Gly Ser Thr Glu885 890 895Glu Val Gln Glu Ser Ile Phe Thr Lys Ala Asp Lys Arg Glu Leu Ile900 905 910Thr Ala Lys Asn His Leu Asp Asp Pro Val Ser Thr Glu Gly Lys Lys915 920 925Pro Gly Thr Ile Thr Gln Tyr Asn Asn Ala Met His Asn Ala Gln Gln930 935 940Gln Ile Asn Thr Ala Lys Thr Glu Ala Gln Gln Val Ile Asn Asn Glu945 950 955 960Arg Ala Thr Pro Gln Gln Val Ser Asp Ala Leu Thr Lys Val Arg Ala965 970 975Ala Gln Thr Lys Ile Asp Gln Ala Lys Ala Leu Leu Gln Asn Lys Glu980 985 990Asp Asn Ser Gln Leu Val Thr Ser Lys Asn Asn Leu Gln Ser Ser Val99510001005Asn Gln Val Pro Ser Thr Ala Gly Met Thr Gln Gln Ser Ile Asp Asn101010151020Tyr Asn Ala Lys Lys Arg Glu Ala Glu Thr Glu Ile Thr Ala Ala Gln1025 103010351040Arg Val Ile Asp Asn Gly Asp Ala Thr Ala Gln Gln Ile Ser Asp Glu104510501055Lys His Arg Val Asp Asn Ala Leu Thr Ala Leu Asn Gln Ala Lys His106010651070Asp Leu Thr Ala Asp Thr His Ala Leu Glu Gln Ala Val Gln Gln Leu107510801085Asn Arg Thr Gly Thr Thr Thr Gly Lys Lys Pro Ala Ser Ile Thr Ala109010951100Tyr Asn Asn Ser Ile Arg Ala Leu Gln Ser Asp Leu Thr Ser Ala Lys1105 111011151120Asn Ser Ala Asn Ala Ile Ile Gln Lys Pro Ile Arg Thr Val Gln Glu112511301135Val Gln Ser Ala Leu Thr Asn Val Asn Arg Val Asn Glu Arg Leu Thr114011451150Gln Ala Ile Asn Gln Leu Val Pro Leu Ala Asp Asn Ser Ala Leu Lys115511601165Thr Ala Lys Thr Lys Leu Asp Glu Glu Ile Asn Lys Ser Val Thr Thr117011751180Asp Gly Met Thr Gln Ser Ser Ile Gln Ala Tyr Glu Asn Ala Lys Arg1185 119011951200Ala Gly Gln Thr Glu Ser Thr Asn Ala Gln Asn Val Ile Asn Asn Gly120512101215Asp Ala Thr Asp Gln Gln Ile Ala Ala Glu Lys Thr Lys Val Glu Glu122012251230Lys Tyr Asn Ser Leu Lys Gln Ala Ile Ala Gly Leu Thr Pro Asp Leu123512401245Ala Pro Leu Gln Thr Ala Lys Thr Gln Leu Gln Asn Asp Ile Asp Gln125012551260Pro Thr Ser Thr Thr Gly Met Thr Ser Ala Ser Ile Ala Ala Phe Asn1265 127012751280Glu Lys Leu Ser Ala Ala Arg Thr Lys Ile Gln Glu Ile Asp Arg Val128512901295Leu Ala Ser His Pro Asp Val Ala Thr Ile Arg Gln Asn Val Thr Ala130013051310Ala Asn Ala Ala Lys Ser Ala Leu Asp Gln Ala Arg Asn Gly Leu Thr131513201325Val Asp Lys Ala Pro Leu Glu Asn Ala Lys Asn Gln Leu Gln Tyr Ser133013351340Ile Asp Thr Gln Thr Ser Thr Thr Gly Met Thr Gln Asp Ser Ile Asn1345 135013551360Ala Tyr Asn Ala Lys Leu Thr Ala Ala Arg Asn Lys Ile Gln Gln Ile136513701375Asn Gln Val Leu Ala Gly Ser Pro Thr Val Glu Gln Ile Asn Thr Asn138013851390Thr Ser Thr Ala Asn Gln Ala Lys Ser Asp Leu Asp His Ala Arg Gln139514001405Ala Leu Thr Pro Asp Lys Ala Pro Leu Gln Thr Ala Lys Thr Gln Leu141014151420Glu Gln Ser Ile Asn Gln Pro Thr Asp Thr Thr Gly Met Thr Thr Ala1425 143014351440Ser Leu Asn Ala Tyr Asn Gln Lys Leu Gln Ala Ala Arg Gln Lys Leu144514501455Thr Glu Ile Asn Gln Val Leu Asn Gly Asn Pro Thr Val Gln Asn Ile146014651470Asn Asp Lys Val Thr Glu Ala Asn Gln Ala Lys Asp Gln Leu Asn Thr147514801485Ala Arg Gln Gly Leu Thr Leu Asp Arg Gln Pro Ala Leu Thr Thr Leu149014951500His Gly Ala Ser Asn Leu Asn Gln Ala Gln Gln Asn Asn Phe Thr Gln1505 151015151520Gln Ile Asn Ala Ala Gln Asn His Ala Ala Leu Glu Thr Ile Lys Ser152515301535Asn Ile Thr Ala Leu Asn Thr Ala Met Thr Lys Leu Lys Asp Ser Val154015451550Ala Asp Asn Asn Thr Ile Lys Ser Asp Gln Asn Tyr Thr Asp Ala Thr155515601565Pro Ala Asn Lys Gln Ala Tyr Asp Asn Ala Val Asn Ala Ala Lys Gly157015751580Val Ile Gly Glu Thr Thr Asn Pro Thr Met Asp Val Asn Thr Val Asn1585 159015951600Gln Lys Ala Ala Ser Val Lys Ser Thr Lys Asp Ala Leu Asp Gly Gln160516101615Gln Asn Leu Gln Arg Ala Lys Thr Glu Ala Thr Asn Ala Ile Thr His162016251630Ala Ser Asp Leu Asn Gln Ala Gln Lys Asn Ala Leu Thr Gln Gln Val163516401645Asn Ser Ala Gln Asn Val Gln Ala Val Asn Asp Ile Lys Gln Thr Thr165016551660Gln Ser Leu Asn Thr Ala Met Thr Gly Leu Lys Arg Gly Val Ala Asn1665 167016751680His Asn Gln Val Val Gln Ser Asp Asn Tyr Val Asn Ala Asp Thr Asn
168516901695Lys Lys Asn Asp Tyr Asn Asn Ala Tyr Asn His Ala Asn Asp Ile Ile170017051710Asn Gly Asn Ala Gln His Pro Val Ile Thr Pro Ser Asp Val Asn Asn171517201725Ala Leu Ser Asn Val Thr Ser Lys Glu His Ala Leu Asn Gly Glu Ala173017351740Lys Leu Asn Ala Ala Lys Gln Glu Ala Asn Thr Ala Leu Gly His Leu1745 175017551760Asn Asn Leu Asn Asn Ala Gln Arg Gln Asn Leu Gln Ser Gln Ile Asn176517701775Gly Ala His Gln Ile Asp Ala Val Asn Thr Ile Lys Gln Asn Ala Thr178017851790Asn Leu Asn Ser Ala Met Gly Asn Leu Arg Gln Ala Val Ala Asp Lys179518001805Asp Gln Val Lys Arg Thr Glu Asp Tyr Ala Asp Ala Asp Thr Ala Lys181018151820Gln Asn Ala Tyr Asn Ser Ala Val Ser Ser Ala Glu Thr Ile Ile Asn1825 183018351840Gln Thr Thr Asn Pro Thr Met Ser Val Asp Asp Val Asn Arg Ala Thr184518501855Ser Ala Val Thr Ser Asn Lys Asn Ala Leu Asn Gly Tyr Glu Lys Leu186018651870Ala Gln Ser Lys Thr Asp Ala Ala Arg Ala Ile Asp Ala Leu Pro His187518801885Leu Asn Asn Ala Gln Lys Ala Asp Val Lys Ser Lys Ile Asn Ala Ala189018951900Ser Asn Ile Ala Gly Val Asn Thr Val Lys Gln Gln Gly Thr Asp Leu1905 191019151920Asn Thr Ala Met Gly Asn Leu Gln Gly Ala Ile Asn Asp Glu Gln Thr192519301935Thr Leu Asn Ser Gln Asn Tyr Gln Asp Ala Thr Pro Ser Lys Lys Thr194019451950Ala Tyr Thr Asn Ala Val Gln Ala Ala Lys Asp Ile Leu Asn Lys Ser195519601965Asn Gly Gln Asn Lys Thr Lys Asp Gln Val Thr Glu Ala Met Asn Gln197019751980Val Asn Ser Ala Lys Asn Asn Leu Asp Gly Thr Arg Leu Leu Asp Gln1985 199019952000Ala Lys Gln Thr Ala Lys Gln Gln Leu Asn Asn Met Thr His Leu Thr200520102015Thr Ala Gln Lys Thr Asn Leu Thr Asn Gln Ile Asn Ser Gly Thr Thr202020252030Val Ala Gly Val Gln Thr Val Gln Ser Asn Ala Asn Thr Leu Asp Gln203520402045Ala Met Asn Thr Leu Arg Gln Ser Ile Ala Asn Lys Asp Ala Thr Lys205020552060Ala Ser Glu Asp Tyr Val Asp Ala Asn Asn Asp Lys Gln Thr Ala Tyr2065 207020752080Asn Asn Ala Val Ala Ala Ala Glu Thr Ile Ile Asn Ala Asn Ser Asn208520902095Pro Glu Met Asn Pro Ser Thr Ile Thr Gln Lys Ala Glu Gln Val Asn210021052110Ser Ser Lys Thr Ala Leu Asn Gly Asp Glu Asn Leu Ala Ala Ala Lys211521202125Gln Asn Ala Lys Thr Tyr Leu Asn Thr Leu Thr Ser Ile Thr Asp Ala213021352140Gln Lys Asn Asn Leu Ile Ser Gln Ile Thr Ser Ala Thr Arg Val Ser2145 215021552160Gly Val Asp Thr Val Lys Gln Asn Ala Gln His Leu Asp Gln Ala Met216521702175Ala Ser Leu Gln Asn Gly Ile Asn Asn Glu Ser Gln Val Lys Ser Ser218021852190Glu Lys Tyr Arg Asp Ala Asp Thr Asn Lys Gln Gln Glu Tyr Asp Asn219522002205Ala Ile Thr Ala Ala Lys Ala Ile Leu Asn Lys Ser Thr Gly Pro Asn221022152220Thr Ala Gln Asn Ala Val Glu Ala Ala Leu Gln Arg Val Asn Asn Ala2225 223022352240Lys Asp Ala Leu Asn Gly Asp Ala Lys Leu Ile Ala Ala Gln Asn Ala224522502255Ala Lys Gln His Leu Gly Thr Leu Thr His Ile Thr Thr Ala Gln Arg226022652270Asn Asp Leu Thr Asn Gln Ile Ser Gln Ala Thr Asn Leu Ala Gly Val227522802285Glu Ser Val Lys Gln Asn Ala Asn Ser Leu Asp Gly Ala Met Gly Asn229022952300Leu Gln Thr Ala Ile Asn Asp Lys Ser Gly Thr Leu Ala Ser Gln Asn2305 231023152320Phe Leu Asp Ala Asp Glu Gln Lys Arg Asn Ala Tyr Asn Gln Ala Val232523302335Ser Ala Ala Glu Thr Ile Leu Asn Lys Gln Thr Gly Pro Asn Thr Ala234023452350Lys Thr Ala Val Glu Gln Ala Leu Asn Asn Val Asn Asn Ala Lys His235523602365Ala Leu Asn Gly Thr Gln Asn Leu Asn Asn Ala Lys Gln Ala Ala Ile237023752380Thr Ala Ile Asn Gly Ala Ser Asp Leu Asn Gln Lys Gln Lys Asp Ala2385 239023952400Leu Lys Ala Gln Ala Asn Gly Ala Gln Arg Val Ser Asn Ala Gln Asp240524102415Val Gln His Asn Ala Thr Glu Leu Asn Thr Ala Met Gly Thr Leu Lys242024252430His Ala Ile Ala Asp Lys Thr Asn Thr Leu Ala Ser Ser Lys Tyr Val243524402445Asn Ala Asp Ser Thr Lys Gln Asn Ala Tyr Thr Thr Lys Val Thr Asn245024552460Ala Glu His Ile Ile Ser Gly Thr Pro Thr Val Val Thr Thr Pro Ser2465 247024752480Glu Val Thr Ala Ala Ala Asn Gln Val Asn Ser Ala Lys Gln Glu Leu248524902495Asn Gly Asp Glu Arg Leu Arg Glu Ala Lys Gln Asn Ala Asn Thr Ala250025052510Ile Asp Ala Leu Thr Gln Leu Asn Thr Pro Gln Lys Ala Lys Leu Lys251525202525Glu Gln Val Gly Gln Ala Asn Arg Leu Glu Asp Val Gln Thr Val Gln253025352540Thr Asn Gly Gln Ala Leu Asn Asn Ala Met Lys Gly Leu Arg Asp Ser2545 255025552560Ile Ala Asn Glu Thr Thr Val Lys Thr Ser Gln Asn Tyr Thr Asp Ala256525702575Ser Pro Asn Asn Gln Ser Thr Tyr Asn Ser Ala Val Ser Asn Ala Lys258025852590Gly Ile Ile Asn Gln Thr Asn Asn Pro Thr Met Asp Thr Ser Ala Ile259526002605Thr Gln Ala Thr Thr Gln Val Asn Asn Ala Lys Asn Gly Leu Asn Gly261026152620Ala Glu Asn Leu Arg Asn Ala Gln Asn Thr Ala Lys Gln Asn Leu Asn2625 263026352640Thr Leu Ser His Leu Thr Asn Asn Gln Lys Ser Ala Ile Ser Ser Gln264526502655Ile Asp Arg21029211496212PRT213Staphylococcus aureus40029Met Asn Met Lys Lys Lys Glu Lys His Ala Ile Arg Lys Lys Ser Ile1 5 10 15Gly Val Ala Ser Val Leu Val Gly Thr Leu Ile Gly Phe Gly Leu Leu20 25 30Ser Ser Lys Glu Ala Asp Ala Ser Glu Asn Ser Val Thr Gln Ser Asp35 40 45Ser Ala Ser Asn Glu Ser Lys Ser Asn Asp Ser Ser Ser Val Ser Ala50 55 60Ala Pro Lys Thr Asp Asp Thr Asn Val Ser Asp Thr Lys Thr Ser Ser65 70 75 80Asn Thr Asn Asn Gly Glu Thr Ser Val Ala Gln Asn Pro Ala Gln Gln85 90 95Glu Thr Thr Gln Ser Ser Ser Thr Asn Ala Thr Thr Glu Glu Thr Pro100 105 110Val Thr Gly Glu Ala Thr Thr Thr Thr Thr Asn Gln Ala Asn Thr Pro115 120 125Ala Thr Thr Gln Ser Ser Asn Thr Asn Ala Glu Glu Leu Val Asn Gln130 135 140Thr Ser Asn Glu Thr Thr Phe Asn Asp Thr Asn Thr Val Ser Ser Val145 150 155 160Asn Ser Pro Gln Asn Ser Thr Asn Ala Glu Asn Val Ser Thr Thr Gln165 170 175Asp Thr Ser Thr Glu Ala Thr Pro Ser Asn Asn Glu Ser Ala Pro Gln180 185 190Ser Thr Asp Ala Ser Asn Lys Asp Val Val Asn Gln Ala Val Asn Thr195 200 205Ser Ala Pro Arg Met Arg Ala Phe Ser Leu Ala Ala Val Ala Ala Asp210 215 220Ala Pro Ala Ala Gly Thr Asp Ile Thr Asn Gln Leu Thr Asn Val Thr225 230 235 240Val Gly Ile Asp Ser Gly Thr Thr Val Tyr Pro His Gln Ala Gly Tyr245 250 255Val Lys Leu Asn Tyr Gly Phe Ser Val Pro Asn Ser Ala Val Lys Gly260 265 270Asp Thr Phe Lys Ile Thr Val Pro Lys Glu Leu Asn Leu Asn Gly Val
275 280 285Thr Ser Thr Ala Lys Val Pro Pro Ile Met Ala Gly Asp Gln Val Leu290 295 300Ala Asn Gly Val Ile Asp Ser Asp Gly Asn Val Ile Tyr Thr Phe Thr305 310 315 320Asp Tyr Val Asn Thr Lys Asp Asp Val Lys Ala Thr Leu Thr Met Pro325 330 335Ala Tyr Ile Asp Pro Glu Asn Val Lys Lys Thr Gly Asn Val Thr Leu340 345 350Ala Thr Gly Ile Gly Ser Thr Thr Ala Asn Lys Thr Val Leu Val Asp355 360 365Tyr Glu Lys Tyr Gly Lys Phe Tyr Asn Leu Ser Ile Lys Gly Thr Ile370 375 380Asp Gln Ile Asp Lys Thr Asn Asn Thr Tyr Arg Gln Thr Ile Tyr Val385 390 395 400Asn Pro Ser Gly Asp Asn Val Ile Ala Pro Val Leu Thr Gly Asn Leu405 410 415Lys Pro Asn Thr Asp Ser Asn Ala Leu Ile Asp Gln Gln Asn Thr Ser420 425 430Ile Lys Val Tyr Lys Val Asp Asn Ala Ala Asp Leu Ser Glu Ser Tyr435 440 445Phe Val Asn Pro Glu Asn Phe Glu Asp Val Thr Asn Ser Val Asn Ile450 455 460Thr Phe Pro Asn Pro Asn Gln Tyr Lys Val Glu Phe Asn Thr Pro Asp465 470 475 480Asp Gln Ile Thr Thr Pro Tyr Ile Val Val Val Asn Gly His Ile Asp485 490 49521030211541212PRT213Staphylococcus aureus40030Asp Gln Tyr Leu Leu Glu Arg Lys Lys Ser Gln Tyr Glu Asp Tyr Lys1 5 10 15Gln Trp Tyr Ala Asn Tyr Lys Lys Glu Asn Pro Arg Thr Asp Leu Lys20 25 30Met Ala Asn Phe His Lys Tyr Asn Leu Glu Glu Leu Ser Met Lys Glu35 40 45Tyr Asn Glu Leu Gln Asp Ala Leu Lys Arg Ala Leu Asp Asp Phe His
50 55 60Arg Glu Val Lys Asp Ile Lys Asp Lys Asn Ser Asp Leu Lys Thr Phe65 70 75 80Asn Ala Ala Glu Glu Asp Lys Ala Thr Lys Glu Val Tyr Asp Leu Val85 90 95Ser Glu Ile Asp Thr Leu Val Val Ser Tyr Tyr Gly Asp Lys Asp Tyr100 105 110Gly Glu His Ala Lys Glu Leu Arg Ala Lys Leu Asp Leu Ile Leu Gly115 120 125Asp Thr Asp Asn Pro His Lys Ile Thr Asn Glu Arg Ile Lys Lys Glu130 135 140Met Ile Asp Asp Leu Asn Ser Ile Ile Asp Asp Phe Phe Met Glu Thr145 150 155 160Lys Gln Asn Arg Pro Lys Ser Ile Thr Lys Tyr Asn Pro Thr Thr His165 170 175Asn Tyr Lys Thr Asn Ser Asp Asn Lys Pro Asn Phe Asp Lys Leu Val180 185 190Glu Glu Thr Lys Lys Ala Val Lys Glu Ala Asp Asp Ser Trp Lys Lys195 200 205Lys Thr Val Lys Lys Tyr Gly Glu Thr Glu Thr Lys Ser Pro Val Val210 215 220Lys Glu Glu Lys Lys Val Glu Glu Pro Gln Ala Pro Lys Val Asp Asn225 230 235 240Gln Gln Glu Val Lys Thr Thr Ala Gly Lys Ala Glu Glu Thr Thr Gln245 250 255Pro Val Ala Gln Pro Leu Val Lys Ile Pro Gln Gly Thr Ile Thr Gly260 265 270Glu Ile Val Lys Gly Pro Glu Tyr Pro Thr Met Glu Asn Lys Thr Val275 280 285Gln Gly Glu Ile Val Gln Gly Pro Asp Phe Leu Thr Met Glu Gln Ser290 295 300Gly Pro Ser Leu Ser Asn Asn Tyr Thr Asn Pro Pro Leu Thr Asn Pro305 310 315 320Ile Leu Glu Gly Leu Glu Gly Ser Ser Ser Lys Leu Glu Ile Lys Pro325 330 335Gln Gly Thr Glu Ser Thr Leu Lys Gly Thr Gln Gly Glu Ser Ser Asp340 345 350Ile Glu Val Lys Pro Gln Ala Thr Glu Thr Thr Glu Ala Ser Gln Tyr355 360 365Gly Pro Arg Pro Gln Phe Asn Lys Thr Pro Lys Tyr Val Lys Tyr Arg370 375 380Asp Ala Gly Thr Gly Ile Arg Glu Tyr Asn Asp Gly Thr Phe Gly Tyr385 390 395 400Glu Ala Arg Pro Arg Phe Asn Lys Pro Ser Glu Thr Asn Ala Tyr Asn405 410 415Val Thr Thr His Ala Asn Gly Gln Val Ser Tyr Gly Ala Arg Pro Thr420 425 430Tyr Lys Lys Pro Ser Glu Thr Asn Ala Tyr Asn Val Thr Thr His Ala435 440 445Asn Gly Gln Val Ser Tyr Gly Ala Arg Pro Thr Gln Asn Lys Pro Ser450 455 460Lys Thr Asn Ala Tyr Asn Val Thr Thr His Gly Asn Gly Gln Val Ser465 470 475 480Tyr Gly Ala Arg Gln Ala Gln Asn Lys Pro Ser Lys Thr Asn Ala Tyr485 490 495Asn Val Thr Thr His Ala Asn Gly Gln Val Ser Tyr Gly Ala Arg Pro500 505 510Thr Tyr Lys Lys Pro Ser Lys Thr Asn Ala Tyr Asn Val Thr Thr His515 520 525Ala Asp Gly Thr Ala Thr Tyr Gly Pro Arg Val Thr Lys530 535 54021031211356212PRT213Staphylococcus aureus40031Met Lys Met Arg Thr Ile Ala Lys Thr Ser Leu Ala Leu Gly Leu Leu1 5 10 15Thr Thr Gly Ala Ile Thr Val Thr Thr Gln Ser Val Lys Ala Glu Lys20 25 30Ile Gln Ser Thr Lys Val Asp Lys Val Pro Thr Leu Lys Ala Glu Arg35 40 45Leu Ala Met Ile Asn Ile Thr Ala Gly Ala Asn Ser Ala Thr Thr Gln50 55 60Ala Ala Asn Thr Arg Gln Glu Arg Thr Pro Lys Leu Glu Lys Ala Pro65 70 75 80Asn Thr Asn Glu Glu Lys Thr Ser Ala Ser Lys Ile Glu Lys Ile Ser85 90 95Gln Pro Lys Gln Glu Glu Gln Lys Thr Leu Asn Ile Ser Ala Thr Pro100 105 110Ala Pro Lys Gln Glu Gln Ser Gln Thr Thr Thr Glu Ser Thr Thr Pro115 120 125Lys Thr Lys Val Thr Thr Pro Pro Ser Thr Asn Thr Pro Gln Pro Met130 135 140Gln Ser Thr Lys Ser Asp Thr Pro Gln Ser Pro Thr Ile Lys Gln Ala145 150 155 160Gln Thr Asp Met Thr Pro Lys Tyr Glu Asp Leu Arg Ala Tyr Tyr Thr165 170 175Lys Pro Ser Phe Glu Phe Glu Lys Gln Phe Gly Phe Met Leu Lys Pro180 185 190Trp Thr Thr Val Arg Phe Met Asn Val Ile Pro Asn Arg Phe Ile Tyr195 200 205Lys Ile Ala Leu Val Gly Lys Asp Glu Lys Lys Tyr Lys Asp Gly Pro210 215 220Tyr Asp Asn Ile Asp Val Phe Ile Val Leu Glu Asp Asn Lys Tyr Gln225 230 235 240Leu Lys Lys Tyr Ser Val Gly Gly Ile Thr Lys Thr Asn Ser Lys Lys245 250 255Val Asn His Lys Val Glu Leu Ser Ile Thr Lys Lys Asp Asn Gln Gly260 265 270Met Ile Ser Arg Asp Val Ser Glu Tyr Met Ile Thr Lys Glu Glu Ile275 280 285Ser Leu Lys Glu Leu Asp Phe Lys Leu Arg Lys Gln Leu Ile Glu Lys290 295 300His Asn Leu Tyr Gly Asn Met Gly Ser Gly Thr Ile Val Ile Lys Met305 310 315 320Lys Asn Gly Gly Lys Tyr Thr Phe Glu Leu His Lys Lys Leu Gln Glu325 330 335His Arg Met Ala Asp Val Ile Asp Gly Thr Asn Ile Asp Asn Ile Glu340 345 350Val Asn Ile Lys35521032211313212PRT213Staphylococcus aureus40032Met Glu His Thr Thr Met Lys Ile Thr Thr Ile Ala Lys Thr Ser Leu1 5 10 15Ala Leu Gly Leu Leu Thr Thr Gly Val Ile Thr Thr Thr Thr Gln Ala20 25 30Ala Asn Ala Thr Thr Leu Ser Ser Thr Lys Val Glu Ala Pro Gln Ser35 40 45Thr Pro Pro Ser Thr Lys Ile Glu Ala Pro Gln Ser Lys Pro Asn Ala50 55 60Thr Thr Pro Pro Ser Thr Lys Val Glu Ala Pro Gln Gln Thr Ala Asn65 70 75 80Ala Thr Thr Pro Pro Ser Thr Lys Val Thr Thr Pro Pro Ser Thr Asn85 90 95Thr Pro Gln Pro Met Gln Ser Thr Lys Ser Asp Thr Pro Gln Ser Pro100 105 110Thr Thr Lys Gln Val Pro Thr Glu Ile Asn Pro Lys Phe Lys Asp Leu115 120 125Arg Ala Tyr Tyr Thr Lys Pro Ser Leu Glu Phe Lys Asn Glu Ile Gly130 135 140Ile Ile Leu Lys Lys Trp Thr Thr Ile Arg Phe Met Asn Val Val Pro145 150 155 160Asp Tyr Phe Ile Tyr Lys Ile Ala Leu Val Gly Lys Asp Asp Lys Lys165 170 175Tyr Gly Glu Gly Val His Arg Asn Val Asp Val Phe Val Val Leu Glu180 185 190Glu Asn Asn Tyr Asn Leu Glu Lys Tyr Ser Val Gly Gly Ile Thr Lys195 200 205Ser Asn Ser Lys Lys Val Asp His Lys Ala Gly Val Arg Ile Thr Lys210 215 220Glu Asp Asn Lys Gly Thr Ile Ser His Asp Val Ser Glu Phe Lys Ile225 230 235 240Thr Lys Glu Gln Ile Ser Leu Lys Glu Leu Asp Phe Lys Leu Arg Lys245 250 255Gln Leu Ile Glu Lys Asn Asn Leu Tyr Gly Asn Val Gly Ser Gly Lys260 265 270Ile Val Ile Lys Met Lys Asn Gly Gly Lys Tyr Thr Phe Glu Leu His275 280 285Lys Lys Leu Gln Glu Asn Arg Met Ala Asp Val Ile Asp Gly Thr Asn290 295 300Ile Asp Asn Ile Glu Val Asn Ile Lys305 310
權利要求
1.包含一個DNA序列的分離的核酸分子，所述DNA序列選自由以下所構成的組(i)SEQ ID NO1-13中所示的DNA序列；(ii)與上述(i)中確定的SEQ ID NO1-13所示序列雜交並編碼由一種病原微生物表達的多肽的DNA序列；和(iii)由(i)和(ii)中所限定的DNA序列的遺傳密碼所導致的簡併的DNA序列。
2.如權利要求1所述的分離的核酸分子，其是基因組DNA。
3.如權利要求1或2所述的分離的核酸分子，所述核酸分子在嚴格的雜交條件下與SEQ ID NO1-13所示的序列退火。
4.一種載體，其包含如權利要求1-3任一項所述的核酸分子。
5.如權利要求4所述的載體，其適於重組表達由所述核酸編碼的多肽。
6.如權利要求4或5所述的載體，其是一種適合原核基因表達的表達載體。
7.如權利要求4或5所述的載體，其中所述載體是一種適合真核基因表達的表達載體。
8.如權利要求4至7任一項所述的載體，其中載體的適用性包括提供啟動子序列。
9.如權利要求8所述的載體，其中啟動子序列提供細胞特異性的，可誘導的或組成性的表達。
10.一種鑑別抗原多肽的方法，包括(i)提供一個核酸文庫，其編碼一種病原微生物的基因或部分基因序列；(ii)轉化/轉染所述文庫到一種宿主細胞中；(iii)將由所述基因/部分基因序列表達的多肽與從感染或曾經感染所述病原微生物的動物得到的自體抗血清接觸；和(iv)純化編碼與所述自身抗血清結合的多肽或部分多肽的核酸。
11.如權利要求10所述的方法，其中所述文庫包含一種病原微生物的基因組DNA。
12.如權利要求10或11所述的方法，其中所述病原微生物是細菌。
13.如權利要求10至12任何一項所述的方法，其中所述細菌微生物選自以下金黃色葡萄球菌；表皮葡萄球菌；糞腸球菌；結核分枝桿菌；B族鏈球菌；肺炎鏈球菌；幽門螺桿菌；淋病奈瑟氏球菌；A族鏈球菌；布氏疏螺旋體；粗球孢菌；莢膜組織胞漿菌；B型腦膜炎奈瑟氏球菌；弗氏志賀氏菌；大腸桿菌；流感嗜血菌。
14.如權利要求13所述的方法，其中所述病原微生物是金黃色葡萄球菌。
15.如權利要求13所述的方法，其中所述病原微生物是表皮葡萄球菌。
16.如權利要求10至15任何一項所述的方法，其中所述核酸文庫是λ文庫。
17.由如權利要求10至16任何一項所述的方法鑑定的多肽。
18.如權利要求17所述的多肽，其選自由SEQ ID NO14-19組成的組中。
19.一種製備如權利要求17或18所述多肽的方法，包括(i)提供一種細胞及細胞培養條件，所述細胞被權利要求4至9任何一項所述的載體轉化/轉染；和(ii)從所述細胞中或其生長環境中純化所述多肽。
20.如權利要求19所述的方法，其中所述載體編碼並因此為所述重組多肽提供一種促進所述多肽純化的分泌信號。
21.由權利要求4至9任何一項所述的載體轉化或轉染的細胞。
22.如權利要求21所述的細胞，其是一種原核細胞。
23.如權利要求21所述的細胞，其是選自真菌細胞、昆蟲細胞、兩棲動物細胞、哺乳動物細胞、植物細胞的一種真核細胞。
24.一種含有權利要求16或17所述的至少一種多肽的疫苗。
25.權利要求24所述的疫苗，其進一步包含一種載體和/或佐劑。
26.一種免疫動物抵抗一種病原微生物的方法，所述方法包含給該動物施用前述任一權利要求所述的至少一種多肽或其一部分，或前述任一權利要求所述的疫苗。
27.如權利要求26所述的方法，其中所述動物是人。
28.如權利要求26或27所述的方法，其中所述疫苗或抗原多肽是通過直接靜脈注射，肌肉內注射或皮下注射給予的。
29.如權利要求25或26所述的方法，其中所述疫苗或抗原多肽是口服給予的。
30.如權利要求26至29任何一項所述的方法，其中所述疫苗是抵抗葡萄球菌屬細菌的疫苗。
31.如權利要求30所述的方法，其中所述疫苗是抵抗金黃色葡萄球菌的疫苗。
32.如權利要求30所述的方法，其中所述疫苗是抵抗表皮葡萄球菌的疫苗。
33.一種抗體或至少是其一個有效部分，其至少結合權利要求16或17所述的多肽的一個選擇部分。
34.如權利要求33所述的抗體，其是一種單克隆抗體。
35.如權利要求33或34所述的抗體，其中所述的有效部分包括Fab片段。
36.如權利要求33至35任何一項所述的抗體，其是一種嵌合抗體。
37.如權利要求33至35任何一項所述的抗體，其是一種人源化抗體。
38.如權利要求33至37任何一項所述的抗體，其中所述抗體被提供一種標誌，標記或標籤。
39.如權利要求38所述的抗體，其中所述抗體被提供一種選自放射性標記，螢光標記，附加表位的標誌。
40.權利要求34至39任何一項所述的抗體，所述抗體以融合多肽的形式產生。
41.一種適合於表達權利要求34至40任何一項所述的抗體的載體。
42.一種用權利要求41所述的載體轉化或轉染的細胞。
43.一種生產權利要求34或40所述的抗體的方法，包括i)提供一種用權利要求41所述的載體轉化或轉染的細胞及細胞培養條件；和ii)從所述細胞或其生長環境中純化所述抗體。
44.一種產生權利要求34所述的抗體的雜交瘤細胞系。
45.權利要求33至40任何一項所述的抗體在製備用於治療與金黃色葡萄球菌相關的敗血症、食物中毒或皮膚損害的藥物中的用途。
46.權利要求33至40任何一項所述的抗體在製備用於治療與表皮葡萄球菌相關的敗血症、腹膜炎或心內膜炎的藥物中的用途。
47.製備一種產生權利要求34所述的單克隆抗體的雜交瘤細胞系的方法，其步驟包括i)用一種免疫原免疫一種有免疫活性的哺乳動物，所述免疫原包含至少一種具有SEQ ID NO14-19的胺基酸序列的多肽或其片段；ii)將被免疫的具有免疫活性的哺乳動物的淋巴細胞與骨髓瘤細胞融合形成雜交瘤細胞；iii)篩選由步驟(ii)中的雜交瘤細胞產生的、具有結合(i)中胺基酸序列的活性的單克隆抗體；iv)培養雜交瘤細胞使其增殖和/或分泌所述單克隆抗體；和v)從培養上清液中回收單克隆抗體。
48.權利要求47所述的方法，其中所述免疫活性哺乳動物是小鼠。
49.權利要求47所述的方法，其中所述免疫活性哺乳動物是大鼠。
全文摘要
本發明涉及一種鑑定由病原微生物所表達的抗原多肽的方法；包含所述多肽的疫苗；製備所述多肽的重組方法；和針對所述多肽的治療用抗體。
文檔編號A61P9/00GK1437653SQ0181154
公開日2003年8月20日 申請日期2001年6月20日 優先權日2000年6月20日
發明者西蒙·福斯特, 菲利普·麥克道爾, 基爾斯蒂·布魯梅爾, 西蒙·克拉克 申請人:謝菲爾德大學, 比奧辛內克斯公司