專利名稱：Hiv包膜多肽和疫苗的制作方法
技術(shù)領(lǐng)域：
本發(fā)明涉及HIV包膜多肽和包含這些多肽的疫苗。相關(guān)技術(shù)的描述獲得性免疫缺陷綜合癥(AIDS)是由命名為人免疫缺陷病毒(HIV)的逆轉(zhuǎn)錄病毒引起的。過去為研制一種基于抗體或細(xì)胞反應(yīng)誘導(dǎo)保護(hù)性免疫應(yīng)答的疫苗付出了很大的努力。最近的嘗試是利用亞單位疫苗，其中由于安全性原因，在疫苗中利用HIV蛋白質(zhì)(而非稀釋的或滅活的病毒)作為免疫原。一般地，亞單位疫苗包括gp120，即位于病毒表面的HIV包膜蛋白部分。
有關(guān)HIV包膜蛋白已有許多描述，并且人們熟知來自許多HIV菌株的氨基酸序列與編碼HIV包膜的核酸序列(Myers,G.等，1992.人逆轉(zhuǎn)錄病毒和愛滋病。核酸和氨基酸序列的編緝與分析。Los Alamos國家實(shí)驗(yàn)室，Los Alamos，新墨西哥)。HIV包膜蛋白是約160kd的糖蛋白(gp160)，其在羧基末端區(qū)錨定在膜的雙層分子上。其N端區(qū)段(gp120)伸出至圍繞毒粒的含水環(huán)境中，且C端區(qū)段(gp41)跨越膜。通過宿主細(xì)胞媒介方法，將gp160切割成gp120和整合膜蛋白gp41。由于在gp120和gp41之間沒有任何共價(jià)連接，有時從毒粒的表面釋放出游離的gp120，并感染細(xì)胞。
gp120分子包含60,000道爾頓的多肽核心，該多肽核心廣泛地被N-連接的糖基化作用所修飾，而使分子的表觀分子量增加至120,000道爾頓。gp120的氨基酸序列含有散布五個高變結(jié)構(gòu)域的五個相對保守結(jié)構(gòu)域。在gp120初級序列中的18個半胱氨酸殘基的位置，和在gp120序列中的大約24個N-連接的糖基化位點(diǎn)的13個位置，對所有g(shù)p120序列來說都是相同的。高變結(jié)構(gòu)域包括大量的氨基酸取代，插入和缺失。這些結(jié)構(gòu)域中的序列變化使得來自各種病毒分離物的gp120分子間產(chǎn)生了多達(dá)全序列30％的可變性。盡管有這一變異，所有g(shù)p120序列均保留了病毒結(jié)合病毒受體CD4的能力，且保留了與gp41相互作用而引起病毒和宿主細(xì)胞膜的融合的能力。
gp120曾是廣泛研究的目標(biāo)，因其是亞單位疫苗的疫苗候選者，并且是很有可能接近免疫進(jìn)攻的病毒蛋白質(zhì)。現(xiàn)在，利用gp120MN菌株的臨床試驗(yàn)正在進(jìn)行。然而，至今沒有任何人類疫苗試驗(yàn)具有足夠規(guī)模以確認(rèn)或否認(rèn)疫苗的有效性。
HIV-1疫苗候選者的研制由于缺乏HIV-1感染的體內(nèi)或體外模型(其精確地近似于人類自然感染的條件)而遇到障礙。已描述了幾種候選HIV-1疫苗[Berman等；病毒學(xué)雜志。7:4464-9(1992)；Haigwood等；病毒學(xué)雜志。66:172-82(1992)；Salmon-Ceron等；愛滋病研究和人逆轉(zhuǎn)錄病毒11:1479-86(1995)]廣泛激發(fā)能夠中和體外各種多樣化的HIV-1分離物的交叉反應(yīng)抗體。然而，體外試驗(yàn)與體內(nèi)保護(hù)性免疫的相關(guān)性是不確定的。雖然已對黑猩猩提供了幾種具有抗同源和異源HIV-1菌株的保護(hù)作用的疫苗，但是保護(hù)性反應(yīng)并非總是與體外的中和試驗(yàn)有聯(lián)系，其中這種體外中和試驗(yàn)在T細(xì)胞系上或在凝集素和細(xì)胞因子活化的外周血液單核細(xì)胞(PBMCs)上進(jìn)行[Bermarn等；自然,345:622-5(1990)；Bruck等；疫苗，12(12):1141-8(1994)；El-Amad等；愛滋病，9:1313-22(1995)；Girard等；病毒學(xué)雜志,69:6239-48(1995)；和Fulz等；科學(xué),256:1687-1690(1992)]。雖然在黑猩猩中成功獲得的保護(hù)作用是鼓勵人心的，并歷史地證明是可靠的疫苗有效性指示劑，但所有HIV-1感染的實(shí)驗(yàn)?zāi)Ｐ偷母腥緱l件顯著不同于人類自然感染條件。
體內(nèi)和體外的HIV-1感染實(shí)驗(yàn)所受局限性是HIV-1(為體外或體內(nèi)感染實(shí)驗(yàn)，其需要制備病毒儲備液)的體外擴(kuò)增強(qiáng)加了一種遺傳選擇，它能導(dǎo)致病毒準(zhǔn)種的光譜不同于用于建立培養(yǎng)物的臨床樣品中存在的變體的光譜[Kusumi等；病毒學(xué)雜志66:875(1992)；Meyerhans等；細(xì)胞58:901-10(1989)]。由于這些不確定性，以及所傳輸?shù)牟《镜牧浚跏紡?fù)制所涉及的位點(diǎn)和細(xì)胞類型，和病毒散布的動力學(xué)更大的不確定性，使得當(dāng)前以體外或體內(nèi)試驗(yàn)有效可靠預(yù)測疫苗功效的能力尚無把握。
曾進(jìn)入人類臨床試驗(yàn)的HIV-1疫苗候選者之一是重組gp120，其由HIV-1的MN菌株在中國倉鼠卵巢(CHO)細(xì)胞中制備(MN-rgp120)(Berman等；病毒學(xué)雜志7:4464-9(1992))。至今，大約499個成年人曾參與該疫苗的1期和2期免疫原性和安全性試驗(yàn)。所收集的數(shù)據(jù)足以證明MN-rgp120的安全性，免疫原性，并且按照0,1和6個月免疫進(jìn)度，在多于95％的免疫個體中激發(fā)了高效價(jià)的抗體[Belshe等；JAMA272(6):475-80(1994)；McElrath；癌生物學(xué)討論會6:1-11(1995)]。然而，在這些試驗(yàn)的過程中，九個接收了MN-rgp120的被接種者由高風(fēng)險(xiǎn)行為被HIV-1感染。小型試驗(yàn)(如這些)，其群體具有低的感染速率和較小規(guī)模的無效對照組，沒有足夠的統(tǒng)計(jì)效應(yīng)確認(rèn)或者否認(rèn)疫苗功效。
然而，仍然在尋求基于gp120或另外HIV蛋白質(zhì)的有效的疫苗，以產(chǎn)生抗HIV另一些菌株的保護(hù)作用，從而阻止該疾病的傳播。
背景技術(shù)：
描述重組亞單位疫苗見Berman等，PCT/US91/02250中所描述的(出版號W091/15238,19911十月7日)。也參見，例如Hu等，自然328:721-724(1987)(疫苗病毒-HIV包膜重組疫苗)；Arthur等，病毒學(xué)雜志63(12):5046-5053(1989)(純化gp120)；和Berman等，Proc.Natl.Acad.Sci.美國85:5200-5204(1988)(重組包膜糖蛋白gp120)。
已知gp120的許多序列。本文所參考的gp120序列(其來自ⅢB亞菌株HIV-1LAI)是由Muesing等測定的，見核酸結(jié)構(gòu)和人AIDS/淋巴結(jié)病逆轉(zhuǎn)錄病毒的表達(dá)，自然313:450-458(1985)。Myers等列出了來自HIV-1菌株的NY-5,Jrcsf,Z6,Z321，以HXB2的gp120的序列＂，人逆轉(zhuǎn)錄病毒及愛滋??；核酸和氨基酸序列的編緝和分析＂，Los Alamos國家實(shí)驗(yàn)室，Los Alamos，新墨西哥(1992)。泰國人分離物A244的序列由McCutchan等提供，＂泰國的HIV-1基因變體＂愛滋病研究和人類逆轉(zhuǎn)錄病毒8:1887-1895(1992)。MN1984克隆見Gurgo等所描述的＂，兩種新的美國HIV-1分離物的包膜序列＂，病毒學(xué)164:531-536(1988)。如本文所使用的，MN,MN-rgp120,MN克隆或分離物指MNGEN。MNGEN的氨基酸序列見SEQ ID NO.29。
以上每一參考文獻(xiàn)本文以其全文作為參考。
發(fā)明概要本發(fā)明提供了編碼gp120多肽(其來自用MN-rgp120的疫苗試驗(yàn)的逃逸(breakthrough)分離物)的寡核苷酸序列和所編碼的gp120多肽。利用亞單位疫苗中的一種或多種分離物的gp120多肽(通常與MN-rgp120一起)，可提供抗一些HIV菌株的保護(hù)作用，這些HIV菌株與疫苗菌株(例如；MN-rgp120)有足夠的差異，使得所說的疫苗不提供抗這些菌株的保護(hù)作用。本發(fā)明也提供了所述多肽誘導(dǎo)的抗體。
附圖簡述

圖1說明在被HIV-1感染的被接種者中抗體對MN-rgp120反應(yīng)的動力學(xué)。在所示的時間點(diǎn)收集血清，并測定與MN-rgp120(空心圈)或衍生自MN-rgp120V3結(jié)構(gòu)域的合成肽(實(shí)心圈)的抗體反應(yīng)性。箭頭表明注射的日期。加號表明第一次檢測到HIV-1感染的時間。陰影部分表明是在HIV-1感染之后所收集的數(shù)據(jù)。來自被接種者C6的數(shù)據(jù)在A組中顯示；C8在B組；C7-C組；C11-D組；C10-E組；C17-F組；和C15-G組。
圖2說明在HIV-1感染的被接種者中CD4阻斷抗體反應(yīng)的動力學(xué)。在所示的時間點(diǎn)收集血清，并測定能夠阻止[125I]標(biāo)記的MN-rgp120結(jié)合到CD4細(xì)胞表面的抗體。箭頭表明注射的日期。加號表明第一次檢測到HIV-1感染的時間。陰影部分表明是在HIV-1感染之后所收集的數(shù)據(jù)。被接種者C6的數(shù)據(jù)在A組中顯示；C8對應(yīng)于B組；C7-C組；C11-D組；C10-E組；C17-F組；和C15-G組。
圖3說明來源于逃逸病毒的包膜糖蛋白(gp120)的預(yù)測的氨基酸序列。通過PCR將原病毒的DNA序列從PBMCs擴(kuò)增，然后克隆進(jìn)PRK5表達(dá)質(zhì)粒中。對兩個來自各感染的被接種者的克隆從雙鏈質(zhì)粒DNA測序。序列編號參考gp120的起始甲硫氨酸殘基。出于比較目的，所顯示的序列從成熟(完全加工的)包膜糖蛋白的第12位氨基酸開始(對應(yīng)于gp120開放閱讀框41位)。陰影部分表明在中和表位上的序列，黑色方塊表明多態(tài)性，其被認(rèn)為對與MN-rgp120反應(yīng)性的病毒中和Mabs的結(jié)合是重要的。在序列上顯示了保守(C)區(qū)和可變(V)區(qū)。方塊表明序列同源性和多態(tài)性。
圖4說明從逃逸病毒所制備的重組gp120的免疫沉淀法。來源于七種逃逸病毒的重組gp120通過293s細(xì)胞的瞬時轉(zhuǎn)染制備。細(xì)胞用甲硫氨酸35S進(jìn)行代謝標(biāo)記，并且將生長調(diào)節(jié)細(xì)胞培養(yǎng)物上清液用MN-rgp120的多克隆抗血清進(jìn)行免疫沉淀。用SDS-PAGE展開免疫沉淀，并用放射自顯影法顯影。C8道的a和b對應(yīng)于克隆C8.3和C8.6；C6道的a和b對應(yīng)于克隆C6.1和C6.5；C7道的a和b對應(yīng)于克隆C7.2和C7.10；C17道的a和b對應(yīng)于C17.1和C17.3；C11道的a和b對應(yīng)于克隆C11.5和C11.7；C10道的a和b對應(yīng)于克隆C10.5和C10.7；C15道的a和b對應(yīng)于克隆C15.2和C15.3。
圖5說明單克隆抗體對來源于逃逸病毒的重組gp120的結(jié)合。從被質(zhì)粒瞬時轉(zhuǎn)染的293s細(xì)胞中收集生長調(diào)節(jié)細(xì)胞培養(yǎng)物上清液，其中所說的質(zhì)粒指導(dǎo)逃逸病毒包膜糖蛋白的表達(dá)。相對rgp120濃度由ELISA測定，其中利用對HSV-1糖蛋白D標(biāo)記表位(該表位位于本文所描述的所有rgp120變體的氨基末端)特異性的MAb5B6。將所產(chǎn)生的rgp120制備物置于涂布有特異于gp120的C-端保守序列的多克隆抗體的微量滴定板的孔內(nèi)。通過ELISA測定能中和與gp120反應(yīng)性的單克隆抗體的病毒的結(jié)合。A，特異于HSV-1糖蛋白D標(biāo)記表位的MAb(5B6)的結(jié)合；B，抗MN-rgp120的V3結(jié)構(gòu)域的MAb(1034)的結(jié)合；C，抗合成肽(其對應(yīng)于MN-rgp120的V3結(jié)構(gòu)域)產(chǎn)生的MAb(50.1)結(jié)合；D，已知阻斷gp120與CD4結(jié)合的人類MAb(15e)的結(jié)合。
圖6描述來源于HIV-1MN菌株的MN克隆的成熟包膜糖蛋白(gp120)(SEQ.ID.NO.29)。高變結(jié)構(gòu)域用粗體表示，并顯示了V和C區(qū)(根據(jù)Modrow等，病毒學(xué)雜志61(2):570(1987))。潛在的糖基化位點(diǎn)以(*)標(biāo)記。
發(fā)明詳述本發(fā)明提供了來源于HIV疫苗試驗(yàn)的逃逸分離物的gp120多肽。也提供了編碼逃逸分離物(其可以用于表達(dá)gp120)的gp120的新寡核苷酸序列。利用來自亞單位疫苗中的一種或多種分離物的gp120多肽(通常與MN-rgp120一起)，可提供抗一些HIV菌株的保護(hù)作用，這些HIV菌株與疫苗菌株(例如；MN-rgp120)有足夠的差異，使得所說的疫苗不提供抗這些菌株的保護(hù)作用。
在一個實(shí)施方案中，所說的疫苗基于使用來源于類MN病毒的MN-rgp120多肽(SEQ ID.NO.29)和gp120多肽，其中所說的病毒包含不存在于初始疫苗菌株中的中和表位，并且與疫苗菌株的那些病毒有足夠的差異，能在MN-rgp120接種的個體中引起HIV-1感染(即；導(dǎo)致逃逸感染)的。使用初始疫苗菌株的經(jīng)驗(yàn)性結(jié)論是存在于包含另一些中和表位的群體中的病毒(其含有充分不同于疫苗菌株的那些的附加中和表位)脫逸由疫苗菌株誘導(dǎo)的保護(hù)作用。在疫苗中利用初始的典型gp120多肽充當(dāng)篩，那么不受該疫苗菌株產(chǎn)生有效保護(hù)作用影響的病毒逃逸于該疫苗，并根據(jù)經(jīng)驗(yàn)在指定的地區(qū)檢測出另外一些菌株，其不受初始疫苗菌株的保護(hù)作用的影響。利用來源于那些逃逸分離物的gp120，通過提供另外的中和表位(其不存在于起始疫苗菌株內(nèi))以補(bǔ)充疫苗，因此創(chuàng)造更多的完全疫苗，該疫苗在所說的區(qū)域內(nèi)具有抗若干不同病毒菌株的保護(hù)作用。
原來的HIV-1疫苗策略是基于合適的候選疫苗多肽(其根據(jù)同源性序列對比研究)的選擇。然而，由于一些中和表位是依賴于構(gòu)象的，且所有這些表位的位置尚未知，這一方法不可測定所有的在特定區(qū)域內(nèi)在疫苗中應(yīng)被包括的中和表位。相反，目前該方法利用所選擇的代表菌株，和根據(jù)經(jīng)驗(yàn)決定的完全不同的菌株，因此逃逸于由初始接種計(jì)劃提供的保護(hù)屏障。在疫苗中也可包括那些菌株，以提供更多的在該區(qū)內(nèi)抗HIV菌株的完全保護(hù)。此外，那些菌株也可單獨(dú)使用，以產(chǎn)生抗逃逸病毒的保護(hù)作用。
在另一個實(shí)施方案中，本發(fā)明包括一種含有第一HIV gp120多肽序列和逃逸分離物HIV gp120多肽序列的疫苗，其中所說的多肽序列來源于用疫苗(其包含在合適載體中的第一HIV gp120多肽序列)所接種的被接種者。一種或兩種HIV gp120多肽序列的片段可被一種或兩種對應(yīng)的HIV gp120多肽序列取代。
優(yōu)選地，第一gp120多肽序列含有在一種或多種gp120多肽序列中發(fā)現(xiàn)的中和表位，其中所說的多肽序列存在于來源于初始疫苗(即，引起逃逸分離物的疫苗)施用地區(qū)的分離物中。更優(yōu)選地，第一gp120多肽序列含有至少一個或多個該區(qū)域內(nèi)共同的中和表位，以及最優(yōu)選地是第一gp120多肽序列至少含有三個最普通的中和表位中的一個。
適于用作為第一gp120多肽序列的gp120多肽序列包括gp120MN，泰國人分離物A244序列(下文為＂gp120 A244＂),gp120 MN-GNEG(SEQ ID NO.31；在本領(lǐng)域內(nèi)也稱為＂gp120 GNE6＂),以及gp120MN-GNE8(SEQ ID NO.33；在本領(lǐng)域內(nèi)也稱為＂gp120 GNE8＂)等。對北美來說尤其優(yōu)選利用gp120 MN,gp120 MN-GNEG，和gp120 MN-GNE8作為在初始疫苗中的第一gp120多肽序列。對泰國來說尤其優(yōu)選利用gp120 A244作為在初始疫苗中的第一gp120多肽序列。
在本實(shí)施方案的改進(jìn)方案中，疫苗包括與逃逸分離物HIV gp120多肽序列相組合的兩個(即第一和第二)不同的gp120多肽序列或其片段。后者來源于以第一和第二HIV gp120多肽序列兩者之一或兩者進(jìn)行接種的被接種者。
例證性疫苗包括含有g(shù)p120 MN,gp120 A244,gp120 MN-GNEG(SEQ ID.No.31)及gp120 MN-GNE8(SEQ ID NO.33)的組合體。尤其優(yōu)選gp120 MN和gp120 A244或gp120 MN-GNE8(SEQ IDNO.33)與逃逸分離物HIV gp120多肽序列的組合體。
在包含gp120 MN的疫苗中，逃逸分離物HIV gp120多肽序列可以是選自由SEQ ID No.2,4,6,8,10,12,14,16,18,20,22,24,26和28，以及它們的片段組成的組的HIV gp120多肽序列。
如本領(lǐng)域所使用的，術(shù)語＂亞單位疫苗＂在本文中是指，不含有病毒但卻含有一種或多種病毒蛋白質(zhì)或病毒蛋白質(zhì)的片段的病毒疫苗。如本文所使用的，術(shù)語＂多價(jià)＂，是指疫苗含有具有不同氨基酸序列的來源于至少兩種HIV分離物的gp120。
如本領(lǐng)域所使用的，術(shù)語＂逃逸分離物＂或＂逃逸病毒＂在本文中是指從被接種者中分離的病毒。
與本領(lǐng)域中同樣地，術(shù)語＂氨基酸序列＂，＂多肽序列＂，和＂多肽＂在本文中可交換使用，如同術(shù)語＂核酸序列＂，＂核苷酸序列＂，及＂寡核苷酸＂。來源于逃逸分離物的多肽本發(fā)明的gp120多肽對應(yīng)于在下表1中所說明的七種逃逸分離物的氨基酸序列。本發(fā)明的多肽包括表1中所說明的HIV gp120氨基酸序列(SEQ ID.NO.1,3,5,7,9,11,13,15,17,19,21,23,25和27)和它們的片段。本發(fā)明的多肽可包括由兩個或更多的HIV gp120或者gp160氨基酸序列所融合的序列。
該多肽也可以連接到另一個病毒蛋白質(zhì)上，如標(biāo)志(flag)表位氨基酸序列。本領(lǐng)域內(nèi)術(shù)語＂標(biāo)記表位＂在本文中是指包括被單克隆抗體識別的表位的氨基酸序列。標(biāo)記表位有利于利用單一單克隆抗體對許多不同的重組體蛋白質(zhì)進(jìn)行親和性純化分析，其中每一蛋白質(zhì)具有被單克隆抗體識別的標(biāo)記表位。許多氨基酸序列可作為標(biāo)記表位。皰疹單純病毒(HSV-1)的糖蛋白D(gD-1)的N端序列便于用作為標(biāo)記表位，其作用在實(shí)施例中得以詳述。標(biāo)記表位便于與HIV gp120多肽序列的N末端相融合。然而，rgp120序列中的識別中和表位的單克隆抗體可用來親和純化氨基酸序列，且可忽略標(biāo)記表位。
此外，可將各種信號序列與本發(fā)明的多肽相連接。雖然在HIV培養(yǎng)物中rgp120分泌至一定程度，宿主細(xì)胞釋放(分泌)的包膜糖蛋白的量在不同菌株間變化很大。通過將編碼信號序列的核苷酸序列連接到編碼rgp120的核苷酸序列上，把各種信號序列引入多肽序列中，以利于細(xì)胞分泌rgp120。例如，Chiron HIV gp120多肽包括來源于組織纖溶酶原激活物(TPA)的信號序列，這利于rgp120的分泌。附加信號序列為大家所熟知，其包括鼠類白血病病毒的表面蛋白gp70(Kayman等描述，病毒學(xué)雜志68:400-410(1984))的N端結(jié)構(gòu)域。
表1說明本發(fā)明七種逃逸分離物各自的兩個克隆的核苷酸和所推定的氨基酸序列。這些克隆是C6.1；C6.5；C8.3；C8.6；C15.2；C15.3；C7.2；C7.10；C11.5；C11.7；C10.5；C10.7；C17.1和C17.3。這些序列是SEQ.ID.No.1-28，每個克隆的第一個序列號是核苷酸序列，第二個是氨基酸序列。MN核苷酸序列與所推定的MN-GNE6和MN-GNE8的氨基酸序列在下文列出的序列中說明。在MN-GNE6的列出序列中，終止密碼子位于第51個氨基酸殘基位置。這一終止密碼子可被編碼對應(yīng)氨基酸(其來源于MN或MN-GNE8或其它分離物)的密碼子所取代。
表1克隆C6.1GGG GTA CCT GTG TGG AAG GAA GCA ACC ACC ACT CTA 365 Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GAG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val15 20 2510CAT AAT GTT TGG GCC ACA CAT GCT TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp30 35CCA AAC CCA CAA GAA ATG GTA TTG GAA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Met Val Leu Glu Asn Val Thr Glu15 40 45 50GAT TTT AAC ATG TGG AAA AAT GAC ATG GTA GAA CAG ATG 192Asp Phe Asn Met Trp Lys Asn Asp Met Val Glu Gln Met55 60CAT GAG GAT ATA ATC AGT TTA TGG GAT CAA AGC CTA AAA 23120His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys65 70 75CCA TGT GTA AAA TTA ACC CCA CTC TGT ATT ACT TTA AAT 270Pro Cys Val Lys Leu Thr Pro Leu Cys Ile Thr Leu Asn80 85 9025TGC ACC AAT TGG AAG AAG AAT GAT ACT AAA ACT AAT AGT 309Cys Thr Asn Trp Lys Lys Asn Asp Thr Lys Thr Asn Ser95 100AGT AGT ACT ACA ACT AAT AAT AGT AGT GCT ACA GCT AAT 348Ser Ser Thr Thr Thr Asn Asn Ser Ser Ala Thr Ala Asn30105 110 115AGT AGT AGT ACT ACA ACT AAT ACT AGT TGG GGA GAG ATA 387Ser Ser Ser Thr Thr Thr Asn Ser Ser Trp Gly Glu Ile120 125AAG GAG GGA GAA ATA AAG AAC TGC TCT TTC AAT ATC ACC 42635Lys Glu Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr130 135 140ACA ACC ATA AGA GAC AAG GTG AAG AAA GAA TAT GCA CTT 465Thr Ser Ile Arg Asp Lys Val Lys Lys Glu Tyr Ala Leu145 150 15540TTT TAT AGC CTT GAT GTA GTA CCA ATA GAA AAT GAT AAT 504Phe Tyr Ser Leu Asp Val Val Pro Ile Glu Asn Asp Asn160 165ACT AGC TAT AGG TTG AGA AGT TGT AAC ACC TCA GTC ATT 543Thr Ser Tyr Arg Leu Arg Ser Cys Asn Thr Ser Val Ile45170 175 180ACA CAA GCC TGT CCA AAG GTA ACT TTT GAG CCA ATT CCC 582Thr Gln Ala Cys Pro Lys Val Thr Phe Glu Pro Ile Pro185 190ATA CAT TAT TGT ACC CCG GCT GGT TTT GCG ATT CTG AAG 62150Ile His Tyr Cys Thr Pro Ala Gly Pne Ala Ile Leu Lys195 200 205TGT AGA GAT AAA AAG TTC AAT GGA ACA GGA CCA TGC AAA 660Cys Arg Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Lys210 215 22055AAT GTT AGC ACA GTA CAA TGT GCA CAT GGA ATT AAG CCA 699Asn Val Ser Thr Val Gln Cys Ala His Gly Ile Lys Pro225 230GTA GTG TCA ACT CAA CTG CTG TTA AAT GGC AGC CTA GCA 738Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala60225 240 245GAA GAA GAG GTA ATA ATT AGA TCT GCC AAT TTC TCA AAC 777Glu Giu Glu Val Ile Ile Arg Ser Ala Asn Phe Ser Asn250 255
AAT GCT AAA ATC ATA ATA GTA CAG TTG AGG GAA CCT GTA 816Asn Ala Lys Ile Ile Ile Val Gln Leu Arg Glu Pro Val260 265 270GAA ATT AAT TGT ACA AGA CCC AGC AAC AAT ACA ATA AAA 855Glu Ile Asn Cys Thr Arg Pro Ser Asn Asn Thr Ile Lys275 280 285GGT ATA CAC ATA GGA CCA GGG AGA GCA TTT TAT GCA ACA 894Gly Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr290 29510GGA GAC ATA CGA GGA GAT ATA AGA CAA GCA CAT TGT AAC 933Gly Asp Ile Arg Gly Asp Ile Arg Gln Ala His Cys Asn300 305 310ATT AGT GGA GCA AAA TGG AAT AAC ACT TTA AAG AAG GTA 972Ile Ser Gly Ala Lys Trp Ash Asn Thr Leu Lys Lys Val15315 320GTT AAA AAA TTA AAA GAA CAA TTT CCA AAT AAA ACA ATA 1011Val Lys Lys Leu Lys Glu Gln Phe Pro Asn Lys Thr Ile325 330 335GTC TTT AAC CAT TCC TCA GGA GGG GAC CCA GAA ATT GTA 105020Val Phe Asn His Ser Ser Gly Gly Asp Pro Glu Ile Val340 345 350ATG CAC AGT TTT AAT TGT CAA GGG GAA TTT TTC TAC TGT 1089Met His Ser Phe Asn Cys Gln Gly Glu Phe Phe Tyr Cys355 36025AAT ACA ACA AAG CTG TTT AAT AGT ACT TGG AAT GAT ACT 1128Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Asp Thr365 370 375ACA GAG TCA AAT AAC AAT GAT AGT ACT ATT ACA CTC CCA 1167Thr Glu Ser Asn Asn Asn Asp Ser Thr Ile Thr Leu Pro30380 385TGC AGA ATA AAA CAA ATT ATA AAC ATG TGG CAG GAA ATA 1206Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Ile390 395 400GGA AAA GCA ATG TAT GCC CCT CCC ACC AGA GGA GAA ATT 124535Gly Lys Ala Met Tyr Ala Pro Pro Thr Arg Gly Glu Ile405 410 415AAA TGT TCA TCA AAT ATT ACA GGA CTA CTG TTA ATA AGA 1284Lys Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Ile Arg420 42540GAT GGT GGT ATT AAC ACT AGC GAT GCC ACC GAG ACC TTC 1323Asp Gly Gly Ile Asn Thr Ser Asp Ala Thr Glu Thr Phe430 435 440AGA CCG GGA GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT 1362Arg Pro Gly Gly Gly Asp Mer Arg Asp Asn Trp Arg Ser45445 450GAA TTA TAT AAA TAT AAA GTA GTG AAA ATT GAG CCA TTA 1401Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu455 460 465GGA GTA GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG 144050Gly Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln470 475 480AGA GAA AAA AGA GCA GTA ACA CTA GGA GCT ATG TTC CTT 1479Arg Glu Lys Arg Ala Val Thr Leu Gly Ala Met Phe Leu485 49055GGG TTC TTA GGA GCA TAA AGC TTC 1503Gly Phe Leu Gly Ala Xaa Ser Phe495 500 501克隆C6.560GGG GTA CCT GTA TGG AAA GAA GCA ACC ACC ACT CTA 36Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GAG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val65 15 20 25
CAT AAT GTT TGG GCC ACA CAT GCT TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp30 35CCA AAC CCA CAA GAA ATG GTA TTG GAA AAT GTG ACA GAA 1535 Pro Asn Pro Gln Glu Met Val Leu Glu Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT GAC ATG GTA GAA CAG ATG 192Asp Phe Asn Met Trp Lys Asn Asp Met Val Glu Gln Met55 6010CAT GAG ANT ATA ATC AGT TTA TGG GAT CAA AGC CTA AAA 231His Glu Xaa Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys65 70 75CCA TGT GTA AAA TTA ACC CCA CTC TGT ATT ACT TTA AAT 270Pro Cys Val Lys Leu Thr Pro Leu Cys Ile Thr Leu Asn1580 85 90TGC ACC AAT TGG AAG GAG AAT GAT ACT AAA ACT AAT AGT 309Cys Thr Asn Trp Lys Glu Asn Asp Thr Lys Thr Asn Ser95 100AGT AGT ACT ACA ACT AAT AAT AGT AGT GCT ACA GCT AAT 34820Ser Ser Thr Thr Thr Asn Asn Ser Ser Ala Thr Ala Asn105 110 115AGT AGT AGT ACT ACA ACT AAT AGT AGT TGG GGA GAG ATA 387Ser Ser Ser Thr Thr Thr Asn Ser Ser Trp Gly Glu Ile120 12525AAG GAG GGA GAA ATA AAG AAC TGC TCT TTC AAT ATC ACC 426Lys Glu Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr130 135 140ACA GGC ATA AGA GAC AAG GTG AAG AAA GAA TAT GCA CTT 465Thr Gly Ile Arg Asp Lys Val Lys Lys Glu Tyr Ala Leu30145 150 155TTT TAT AGC CTT GAT GTA GTA CCA ATA GAA AAT GAT AAT 504Phe Tyr Ser Leu Asp Val Val Pro Ile Glu Asn Asp Asn160 165ACT AGC TAT AGG TTG AGA AGT TGT AAC ACC TCA GTC ATT 54335Thr Ser Tyr Arg Leu Arg Ser Cys Asn Thr Ser Val Ile170 175 180ACA CAA GCC TGT CCA AAG GTA ACT TTT GAG CCA ATT CCC 582Thr Gln Ala Cys Pro Lys Val Thr Phe Glu Pro Ile Pro185 19040ATA CAT TAT TGT ACC CCG GCT GGT TTT GCG ATT CTG AAG 621Ile His Tyr Cys Thr Pro Ala Gly Phe Ala Ile Leu Lys195 200 205TGT AAA GAT AAA AAG TTC AAT GGA ACA GGA CCA TGC AAA 660Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Lys45210 215 220AAT GTT AGC ACA GTA CAA TGT ACA CAT GGA ATT AAG CCA 699Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys Pro225 230GTA GTG TCA ACT CAA CTG CTG TTA AAT GGC AGC CTA GCA 73850Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala235 240 245GAA GAA GAG GTA ATA ATT AGA TCT GCC AAT TTC TCA AAC 777Glu Glu Glu Val Ile Ile Arg Ser Ala Asn Phe Ser Asn250 25555AAT GCT AAA ATC ATA ATA GTA CAG TTG AAG GAA CCT GTA 816Asn Ala Lys Ile Ile Ile Val Gln Leu Lys Glu Pro Val260 265 27GAA ATT AAT TGT ACA AGA CCC AGC AAC AAT ACA ATA AAA 855Glu Ile Asn Cys Thr Arg Pro Ser Asn Asn Thr Ile Lys60275 280 285GGT ATA CAC ATA GGA CCA GGG AGA GCA TTT TAT GCA ACA 894Gly Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr290 295
GGA GAC ATA CGA GGA GAT ATA AGA CAA GCA CAT TGT AAC 933Gly Asp Ile Arg Gly Asp Ile Arg Gln Ala His Cys Asn300 305 310ATT AGT GGA GCA AAA TGG AAT AAC ACT TTA AAG AAG GTA 9725 Ile Ser Gly Ala Lys Trp Asn Asn Thr Leu Lys Lys Val315 320GTT ATA AAA TTA AAA GAA CAA TTT CCA AAT AAA ACA ATA 1011Val Ile Lys Leu Lys Glu Gln Phe Pro Asn Lys Thr Ile325 330 33510GTC TTT AAC CAT TCC TCA GGA GGG GAC CCA GAA ATT GTA 1050Val Phe Asn His Ser Ser Gly Gly Asp Pro Glu Ile Val340 345 350ATG CAC AGT TTT AAT TGT CAA GGG GAA TTT TTC TAC TGT 1089Met His Ser Phe Asn Cys Gln Gly Glu Phe Phe Tyr Cys15355 360AAT ACA ACG AAG CTG TTT AAT AGT ACT TGG AAT GAT ACT 1128Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Asp Thr365 370 375ACA GAG TCA AAT AAC AAT GAT AGT ACT ATT ACA CTC CCA 116720Thr Glu Ser Asn Asn Asn Asp Ser Thr Ile Thr Leu Pro380 385TGC AGA ATA AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA 1206Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val390 395 40025GGA AAA GCA ATG TAT GCC CCT CCC ATC AGA GGA GAA ATT 1245Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile405 410 415AAA TGT TCA TCA AAT ATT ACA GGA CTA CTG TTA ACA AGA 1284Lys Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg30420 425GAT GGT GGT ATT AAC ACT AGC GAT GCC ACC GAG ACC TTC 1323Asp Gly Gly Ile Asn Thr Ser Asp Ala Thr Glu Thr Phe430 435 440AGA CCG GGA GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT 136235Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg Ser445 450GAA TTA TAT AAA TAT AAA GTA GTG AAA ATT GAG CCA TTA 1401Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu455 460 46540GGA GTA GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG 1440Gly Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln470 475 480AGA GAA AAA AGA GCA GTA ACA CTA GGA GCT ATG TTC CTT 1479Arg Glu Lys Arg Ala Val Thr Leu Gly Ala Met Phe Leu45485 490GGG TTC TTG GGA GCA TAA AGC TTC 1503Gly Phe Leu Gly Ala Xaa Ser Phe495 500 50150 克隆C8.3G GTA CCT GTA TGG AAA GAA GCA ACC ACC ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT CAT ACA GAG GTA CAT 7655Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His15 20 25AAT GTT TGG GCT ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 3560AAC CCA CAA GAA GTA GTA TTG GAA AAT GTA ACA GAA AAT 154Asn Pro Gln Glu Val Val Leu Glu Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His65 55 60
GAG GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA AAG CCA 232Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA AAT TGC 2715 Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90ACT AAT TTG GAG AAT GCT AAT AAT ACC GAG AAT GCT AAT 310Thr Asn Leu Glu Asn Ala Asn Asn Thr Glu Asn Ala Asn95 10010AAT ACC AAT AAT TAT ACC TTG GGG ATG GAG AGA GGT GAA 349Asn Thr Asn Asn Tyr Thr Leu Gly Met Glu Arg Gly Glu105 110 115ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA AGC TTA AGA 388Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Leu Arg15120 125GAT AAG GTG AAA AAA GAA TAT GCA TTG TTT TAT AAA CTT 427Asp Lys Val Lys Lys Glu Tyr Ala Leu Phe Tyr Lys Leu130 135 140GAT GTA GTA CAA ATA GAT AAT AGT ACC AAC TAT AGG CTG 46620Asp Val Val Gln Ils Asp Asn Ser Thr Asn Tyr Arg Leu145 150 155ATA AGT TGT AAT ACC TCA GTC ATT ACA CAG GCC TGT CCA 505Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro160 16525AAG GTA TCC TTT GAG CTA ATT CCC ATA CAT TAT TGT GCC 544Lys Val Ser Phe Glu Leu Ile Pro Ile His Tyr Cys Ala170 175 180CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG AAG 583Pro Ala Gly Pha Ala Ile Leu Lys Cys Lys Asp Lys Lys30185 190TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGC ACA GTA 622Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val195 200 205CAA TGT ACA CAT GGA ATT AGA CCA GTA GTA TCA ACT CAA 66135Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln210 215 220CTA CTG TTA AAT GGC AGT CTA GCA GAA GAA GAG ATA GTA 700Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Ile Val225 23040ATT AGA TCT GAA AAT ATC ACA GAC AAT GCT AAA ACC ATA 739Ile Arg Ser Glu Asn Ile Thr Asp Asn Ala Lys Thr Ile235 240245ATA GTG CAG CTA AAT GAA TCT ATA GTG ATT AAT TGT ACA 778Ile Val Gln Leu Asn Glu Ser Ile Val Ile Asn Cys Thr45250 255AGA CCC AAT AAC AAC ACA AGA AAA AGT ATA AAT ATA GGA 817Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Asn Ile Gly260 265 270CCA GGG AGA GCA TTC TAT ACA ACA GGA GAC ATA ATA GGA 85650Pro Gly Arg Ala Phe Tyr Thr Thr Gly Asp Ile Ile Gly275 280 285GAT ATA AGA CAA GCA CAT TGT AAC CTT AGT AAA ACA CAA 895Asp Ile Arg Gln Ala His Cys Asn Leu Ser Lys Thr Gln290 29555TGG GAA AAA ACG TTA AGA CAG ATA GCT ATA AAA TTA GAA 934Trp Glu Lys Thr Leu Arg Gln Ile Ala Ile Lys Leu Glu300 305 310GAA AAA TTT AAG AAT AAA ACA ATA GCC TTT AAT AAA TCC 973Glu Lys Phe Lys Asn Lys Thr Ile Ala Phe Asn Lys Ser60315 320TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT AAT 1012Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe Asn325 330 335
TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA AAA CTG 1051Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Lys Leu340 345 350TTT AAT AGT ACC TGG AAT TTA ACA CAA CCG TTT AGT AAT 10905 Phe Asn Ser Thr Trp Asn Leu Thr Gln Pro Phe Ser Asn355 360ACC GGG AAT CGT ACT GAA GAG TTA AAT ATT ACA CTC CCA 1129Thr Gly Asn Arg Thr Glu Glu Leu Asn Ile Thr Leu Pro365 370 37510TGC AGA ATA AAA CAA ATC ATA AAC TTG TGG CAG GAA GTA 1168Cys Arg Ile Lys Gln Ile Ile Asn Leu Trp Gln Glu Val380 385GGC AAA GCA ATG TAT GCC CCT CCC ATC AGA GGA CAA ATT 1207Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile15390 395 400AGA TGT TCA TCA AAT ATT ACA GGG CTA CTA TTA ACA AGA 1246Arg Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg405 410 415GAT GGT GGA AGT AAC ACC GGT GAC AAC AGG ACT GAG ACC 128520Asp Gly Gly Ser Asn Thr Gly Asp Asn Arg Thr Glu Thr420 425TTT AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG AGA 1324Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg430 435 44025AGT GAA TTA TAT AAA TAT AAA GTA GTA AGA ATT GAA CCA 1363Ser Glu Leu Tyr Lys Tyr Lys Val Val Arg Ile Glu Pro445 450TTA GGA GTA GCA CCC ACC CAG GCA AAG AGA AGA GTG GTG 1402Leu Gly Val Ala Pro Thr Gln Ala Lys Arg Arg Val Val30455 460 465CAA AGA GAA AAA AGA GCA GTG GGG ATA GGA GCT ATG TTC 1441Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Met Phe470 475 480CTT GGG TTC TTG GGA GAT AA 146135Leu Gly Phe Leu Gly Asp485 486克隆C8.6G GTA CCT GTG TGG AAA GAA GCA ACC ACC ACT CTA TTT 3740Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT GAT ACA GAG GTA CAT 76Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His15 20 2545AAT GTT TGG GCT ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AAC CCA CAA GAA GTA GTA TTG GAA AAT GTA ACA GAA AAT 154Asn Pro Gln Glu Val Val Leu Glu Asn Val Thr Glu Asn50 40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His55 60GAG GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA AAG CCA 23255Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA AAT TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 9060ACT AAT TTG GAG AAT GCT AAT AAT ACC GAG AAT GCT AAT 310Thr Asn Leu Glu Asn Ala Asn Asn Thr Glu Asn Ala Asn95 100AAT ACC AAT AAT TAT ACC TTG GGG ATG GAG AGA GGT GAA 349Asn Thr Asn Asn Tyr Thr Leu Gly Met Glu Arg Gly Glu65105 110 115
AGA AAA AAC TGC TCT TTC AAT ATC ACC ACA AGC TTA AGA 388Arg Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Leu Arg120 125GAT AAG GGG AAA AAA GAA TAT GCA TTG TTT TAT AAA CTT 4275 Asp Lys Gly Lys Lys Glu Tyr Ala Leu Phe Tyr Lys Leu130 135 140GAT GTA GTA CAA ATA GAT AAT AGT ACC AAC TAT AGG CTG 466Asp Val Val Gln Ile Asp Asn Ser Thr Asn Tyr Arg Leu145 150 15510ATA AGT TGT AAT ACC TCA GTC ATT ACA CAG GCC TGT CCA 505Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro160 165AAG GTA TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT GCC 544Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala15170 175 180CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG AAG 583Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys185 190TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGG ACA GTA 62220Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Arg Thr Val195 200 205CAA TGT ACA CAT GGA ATT AGA CCA GTA GTA TCA ACT CAA 661Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln210 215 22025CTA CTG TTA AAT GGC AGT CTA GCA GAA GAA GAG ATA GTA 700Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Ile Val225 230ATT AGA TCT GAA AAT ATC ACA GAC AAT GCT AAA ACC ATA 739Ile Arg Ser Glu Asn Ile Thr Asp Asn Ala Lys Thr Ile30235 240 245ATA GTG CAG CTA AAT GAA TCT ATA GTG ATT AAT TGT ACA 778Ile Val Gln Leu Asn Glu Ser Ile Val Ile Asn Cys Thr250 255AGA CCC AAT AAC AAC ACA AGA AAA AGT ATA AAT ATA GGA 81735Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Asn Ile GLy260 265 270CCA GGG AGA GCA TTC TAT ACA ACA GGA GAC ATA ATA GGA 856Pro Gly Arg Ala Phe Tyr Thr Thr Gly Asp Ile Ile Gly275 280 28540GAT ATA AGA CAA GCA CAT TGT AAC CTT AGT AAA ACA CAA 895Asp Ile Arg Gln Ala His Cys Asn Leu Ser Lys Thr Gln290 295TGG GAA AAA ACG TTA AGA CAG ATA GCT ATA AAA TTA GAA 934Trp Glu Lys Thr Leu Arg Gln Ile Ala Ile Lys Leu Glu45300 305 310GAA AAA TTT AAG AAT AAA ACA ATA GCC TTT AAT AAA TCC 973Glu Lys Phe Lys Asn Lys Thr Ile Ala Phe Asn Lys Ser315 320TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT AAT 101250Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe Asn325 330 335TGT GGA GGG GGA TTT TTC TAC TGT AGT ACG AGA AAA CTG 1051Cys Gly Gly Gly Phe Phe Tyr Cys Ser Thr Arg Lys Leu340 345 35055TTT AAT AGT ACC TGG AAT TTA ACA CAA CCG TTT AGT AAT 1090Phe Asn Ser Thr Trp Asn Leu Thr Gln Pro Phe Ser Asn355 360ACC GGG GAT CGT ACT GAA GAG TTA AAT ATT ACA CTC CCA 1129Thr Gly Asp Arg Thr Glu Glu Leu Asn Ile Thr Leu Pro60365 370 375TGC AGA ATA AAA CAA ATC ATA AAC TTG TGG CAG GAA GTA 1168Cys Arg Ile Lys Gln Ile Ile Asn Leu Trp Gln Glu Val380 385
GGC AAA GCA ATG TAT GCC CCT CCC ATC AGA GGA CAA ATT 1207Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile390 395 400AGA TGT TCA TCA AAT ATT ACA GGG CTA CTA TTA AGG AGA 12465 Arg Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Arg Arg405 410 415
GAT GGT GGA AGT AAC ACC AGT GAC AAC CAG ACT GAG ACC 1285Asp Gly Gly Ser Asn Thr Ser Asp Asn Gln Thr Glu Thr420 42510TTT AGA CCT GGG GGA GGA GAT ATG AGG GAC AAG TGG AGA 1324Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Lys Trp Arg430 435 440AGT GAA TTA TAT AAA TAT AAA GTA GTA AGA ATT GAA CCA 1363Ser Glu Leu Tyr Lys Tyr Lys Val Val Arg Ile Glu Pro15445 450TTA GGA GTA GCA CCC ACC CAG GCA AAG AGA AGA GTG GTG 1402Leu Gly Val Ala Pro Thr Gln Ala Lys Arg Arg Val Val455 460 465CAA AGA GAA AAA AGA GCA GTG GGG ATA GGA GCT ATG TTC 144120Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Met Phe470 475 480CTT AGG TTC TTA GGA GAT AAA GCT TCT AGA GTC 1474Leu Arg Phe Leu Gly Asp Lys Ala Ser Arg Val485 490 49125克隆 C15.2CTC GAG GTA CCT GTA TGG AAA GAA GCA ACT ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 1030CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT AAT ACA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asn Thr Glu15 20 25AAA CAT AAT GTT TGG GCC ACA CAC GCC TGT GTA CCC ACA 114Lys His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr35 30 35GAT CCC AAC CCA CAA GAA GTA GTA TTG GGA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Val Leu Gly Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA 19240Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln55 60ATG CAT GAA GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA 231Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 7545AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAT GAT TTA GGG AAT GCT ACT AAT ACC AAT 309Asn Cys Thr Asp Asp Leu Gly Asn Ala Thr Asn Thr Asn50 95 100AGT AGT GCC ACT ACC AAT AGT AGT AGT TGG GAA GAA ATG 348Ser Ser Ala Thr Thr Asn Ser Ser Ser Trp Glu Glu Met105 110 115AAG GGG GAA ATG AAA AGA TGC TCT TTC AAT ATC ACC ACA 38755Lys Gly Glu Met Lys Arg Cys Ser Phe Asn Ile Thr Thr120 125AGC ATA AGA GAT AAG ATT AAG AAA GAA CAT GCA CTT TTC 426Ser Ile Arg Asp Lys Ile Lys Lys Glu His Ala Leu Phe130 135 14060TAT AGA CTT GAT GTA GTA CCA ATA GAT AAT GAT AAT ACC 465Tyr Arg Leu Asp Val Val Pro Ile Asp Asn Asp Asn Thr145150 155
ACA TAT AGG TTG ATA AAT TGT AAT ACC TCA GTC ATT ACA 504Thr Tyr Arg Leu Ile Asn Cys Asn Thr Ser Val Ile Thr160 165CAG GCC TGT CCA AAG GTA TCA TTT GAG CCA ATT CCC ATA 5435 Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile170 175 180CAT TTT TGT GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT 582His Phe Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys185 19010AAT AAT AAG ACG TTC GAG GGA AAA GGA CCA TGT AAA AAT 621Asn Asn Lys Thr Phe Glu Gly Lys Gly Pro Cys Lys Asn195 200 205GTC AGT ACA GTA CAA TGC ACA CAT GGA ATT AGG CCA GTA 660Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro Val15210 215 220GTG TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA GAA 699Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu225 230GAA GAG GTA ATA ATT AGA TCT GAC AAT ATC ACA GAC AAT 73820Glu Glu Val Ile Ile Arg Ser Asp Asn Ile Thr Asp Asn235 240 245ACT AAA ACC ATT ATA GTA CAG CTA AAC GAA TCT GTA GTA 777Thr Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Val Val250 25525ATT AAT TGT ACA AGA CCC AAC AAC AAT ACA AGA AAA AGT 816Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser260 265 270ATA CAT ATA GGA CCA GGG AGT GCA TTT TTT GCA ACA GGA 855Ile His Ile Gly Pro Gly Ser Ala Phe Phe Ala Thr Gly30275 280 285GAA ATA ATA GGA GAT ATA AGA CAA GCA CAC TGT AAC CTT 894Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Leu290 295AGT AGA ACA CAA TGG AAT AAC ACT TTA GGA AAG ATA GTC 93335Ser Arg Thr Gln Trp Asn Asn Thr Leu Gly Lys Ile Val300 305 310ATA AAA TTA AGA GAA CAA TTT AGA AAA CAA TTT GGA GAA 972Ile Lys Leu Arg Glu Gln Phe Arg Lys Gln Phe Gly Glu315 32040AAA ACA ATA GTC TTT AAT CGA TCC TCA GGA GGG GAC CCG 1011Lys Thr IIe Val Phe Asn Arg Ser Ser Gly Gly Asp Pro325 330 335GAA ATT GCA ATG CAC AGT TTT AAT TGT GGA GGG GAA TTT 1050Glu Ile Ala Met His Ser Phe Asn Cys Gly Gly Glu Phe45340 345 350TTC TAC TGT AAC ACA ACA GCA CTG TTT AAT AGT ACC TGG 1089Phe Tyr Cys Asn Thr Thr Ala Leu Phe Asn Ser Thr Trp355 360AAT GTT ACT AAA GGG TTG AAT AAC ACT GAA GGA AAT AGC 112850Asn Val Thr Lys Gly Leu Asn Asn Thr Glu Gly Asn Ser365 370 375ACA GGA GAT GAA AAT ATC ATA CTC CCA TGT AGA ATA AAA 1167Thr Gly Asp Glu Asn Ile Ile Leu Pro Cys Arg Ile Lys380 38555CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA ATG 1206Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met390 395 400TAT GCC CCT CCC ATC AGT GGA CAA ATT AGA TGT TCA TCA 1245Tyr Ala Pro Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser60405 410 415AAC ATT ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT AGT 1284Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Ser420 425
AAG AAC GAG AGC ATC ACC ACC GAG GTC TTC AGA CCT GGA 1323Lys Asn Glu Ser Ile Thr Thr Glu Val Phe Arg Pro Gly430 435 440GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT GAA TTA TAT 1362Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr445 450AAA TAT AAA GTA GTA AAA ATT GAA CCA TTA GGA GTA GCG 1401Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala455 460 46510CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG AGA GAA AAA 1440Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys470 475 480AGA GCA GTG GGA ACA ATA GGA GCT ATG TTC CTT GGG TTC 1479Arg Ala Val Gly Thr Ile Gly Ala Met Phe Leu Gly Phe15485 490TTG GGA GCA TAA AGC TTC TAG AGT CGA CCT GCA 1512Leu Gly Ala Xaa Ser Phe Xaa Ser Arg Pro Ala495 500 50420克隆 C15.3CTC GAG GTA CCT GTG TGG AAA GAA GCA ACT ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT AAT ACA GAG 7525Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asn Thr Glu15 20 25AAA CAT AAT GTT TGG GCC ACA CAC GCC TGT GTA CCC ACA 114Lys His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr30 3530GAT CCC AAC CCA CAA GAA GTA GTA TTG GGA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Val Leu Gly Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA 192Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln35 55 60ATG CAT GAA GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA 231Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA 27040Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAT GAT TTA GGG AAT GCT ACT AAT ACC AAT 309Asn Cys Thr Asp Asp Leu Gly Asn Ala Thr Asn Thr Asn95 10045AGC AGT GCC ACT ACC AAT AGT AGT AGT TGG GAA GAA ATG 348Ser Ser Ala Thr Thr Asn Ser Ser Ser Trp Glu Glu Met105 110 115AAG GGG GAA ATG AAA AGG TGC TCT TTC AAT ATC ACC ACA 387Lys Gly Glu Met Lys Arg Cys Ser Phe Asn Ile Thr Thr50120 125AGC ATA AGA GAT AAG ATT AAG AAA GAA CAT GCA CTT TTC 426Ser Ile Arg Asp Lys Ile Lys Lys Glu His Ala Leu Phe130 135 140TAT AGA CTT GAT GTA GTA CCA ATA GAT AAT GAT AAT ACC 46555Tyr Arg Leu Asp Val Val Pro Ile Asp Asn Asp Asn Thr145 150 155ACA TAT AGG TTG ATA AAT TGT AAT ACC TCA GTC ATT ACA 504Thr Tyr Arg Leu Ile Asn Cys Ash Thr Ser Val Ile Thr160 16560CAG GCC TGT CCA AAG GTA TCA TTT GAG CCA ATT CCC ATA 543Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile170 175 180CAT TTT TGT GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT 582His Phe Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys65185 190
AAT AAT AAG ACG TTC GAG GGA AAA GGA CCA TGT AAA AAT 621Asn Asn Lys Thr Phe Glu Gly Lys Gly Pro Cys Lys Asn195 200 205GTC AGT ACA GTA CAA TGC ACA CAT GGA ATT AGG CCA GTA 6605 Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro Val210 215 220GTG TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA GAA 699Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu225 23010GAA GAG GTA ATA ATT AGA TCT GGC AAT ATC ACA GAC AAT 738Glu Glu Val Ile Ile Arg Ser Gly Asn Ile Thr Asp Asn235 240 245ACT AAA ACC ATT ATA GTA CAG CTA AAC GAA TCT GTA GTA 777Thr Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Val Val15250 255ATT AAT TGT ACA AGA TCC AAC AAC AAT ACA AGA AAA AGT 816Ile Asn Cys Thr Arg Ser Asn Asn Asn Thr Arg Lys Ser260 265 270ATA CAT ATA GGA CCA GGG AGT GCA TTT TTT GCA ACA GGA 85520Ile His Ile Gly Pro Gly Ser Ala Phe Phe Ala Thr Gly275 280 285GAA ATA ATA GGA GAT ATA AGA CAA GCA CAC TGT AAC CTT 894Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Leu290 29525AGT AGA ACA CAA TGG AAT AAC ACT TTA GGA AAG ATA GTC 933Ser Arg Thr Gln Trp Asn Asn Thr Leu Gly Lys Ile Val300 305 310ATA AAA TTA AGA GAA CAA TTT AGA AAA CAA TTT GGA GAA 972Ile Lys Leu Arg Glu Gln Phe Arg Lys Gln Phe Gly Glu30315 320AAA ACAATA GTC TTT AAT CGA TCC TCA GGA GGG GAC CCG 1011Lys Thr Ile Val Phe Asn Arg Ser Ser Gly Gly Asp Pro325 330 335GAA ATT GCA ATG CAC AGT TTT AAT TGT GGA GGG GAA TTT 105035Glu Ile Ala Met His Ser Phe Asn Cys Gly Gly Glu Phe340 345 350TTC TAC TGT AAC ACA ACA GCA CTG TTT AAT AGT ACC TGG 1089Phe Tyr Cys Asn Thr Thr Ala Leu Phe Asn Ser Thr Trp355 36040AAT GTT ACT AAA GGG TTG AAT AAC ACT GAA GGA AAT AGC 1128Asn Val Thr Lys Gly Leu Asn Asn Thr Glu Gly Asn Ser365 370 375ACA GGG GAT GAA AAT ATC ATA CTC CCA TGT AGA ATA AAA 1167Thr Gly Asp Glu Asn Ile Ile Leu Pro Cys Arg Ile Lys45380 385CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA ATG 1206Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met390 395 400TAT GCC CCT CCC ATC AGT GGA CAA ATT AGA TGT TCA TCA 124550Tyr Ala Pro Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser405 410 415AAT ATT ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT AGT 1284Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Ser420 42555AAG AAC GAG AGC ATC ACC ACC GAG GTC TTC AGA CCT GGA 1323Lys Asn Glu Ser Ile Thr Thr Glu Val Phe Arg Pro Gly430 435 440GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT GAA TTA TAT 1362Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr60445 450AAA TAT AAA GTA GTA AAA ATT GAA CCA TTA GGA GTA GCG 1401Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala455 460 465
CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG AGA GAA AAA 1440Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys470 475 480AGA GCA GTG GGA ACA ATA GGA GCT ATG TTC CTT GGG TTC 1475Arg Ala Val Gly Thr Ile Gly Ala Met Phe Leu Gly Phe485 490TTA GGA GCA TAA AGC TTC TAG A 1501Leu Gly Ala Xaa Ser Phe Xaa495 50010克隆 C7.2GG GAA TTC GGA TCC GGG GTA CCT GTG TGG AAG GAA GCA 38Glu Phe Gly Ser Gly Val Pro Val Trp Lys Glu Ala1 51015ACC ACC ACT CTA TTC TGT GCA TCA GAT GCT AGA GCA TAT 77Thr Thr Thr Leu Phe Cys Ala Ser Asp Ala Arg Ala Tyr15 20 25GAC ACA GAG GTA CAT AAT GTT TGG GCC ACA CAT GCC TGT 116Asp Thr Glu Val His Asn Val Trp Ala Thr His Ala Cys20 30 35GTA CCC ACA GAC CCT AGT CCA CAA GAA GTA GTT TTG GAA 155Val Pro Thr Asp Pro Ser Pro Gln Glu Val Val Leu Glu40 45 50AAT GTG ACA GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG 19425Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met55 60GTA GAA CAA ATG CAT GAG GAT ATA ATT AGT TTA TGG GAT 233Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp65 70 7530CAA AGC TTA AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT 272Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys80 85 90GTT ACT TTA AAT TGC AGT GAT TAT AGG AAT GCT ACT GAT 311Val Thr Leu Asn Cys Ser Asp Tyr Arg Asn Ala Thr Asp35 95 100TAT AAG AAT GCT ACT GAT ACC ACT AGT AGT AAC GAGGGA 350Tyr Lys Asn Ala Thr Asp Thr Thr Ser Ser Asn Glu Gly105 110 115AAG ATG GAG AGA GGA GAA ATA AAA AAC TGC TCT TTC AAT 38940Lys Met Glu Arg Gly Glu Ile Lys Asn Cys Ser Phe Asn120 125ATT ACC ACA AGC ATA AAA AAT AAG ATG CAG AAA GAA TAT 428Ile Thr Thr Ser Ile Lys Asn Lys Met Gln Lys Glu Tyr130 135 14045GCA CTT TTC TAT AAA CTT GAT ATA GTA CCA ATA GAT AAT 467Ala Leu Phe Tyr Lys Leu Asp Ile Val Pro Ile Asp Asn145 150 155ACA AGC TAT ACA TTG ATA AGT TGT AAC ACC TCA GTC ATT 506Thr Ser Tyr Thr Leu Ile Ser Cys Asn Thr Ser Val Ile50160 165ACA CAG GCC TGT CCA AAG GTA TCC TTT GAA CCA ACT CCC 545Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Thr Pro170 175 180ATA CAT TAT TGT GCT CCG GCT GGT TTT GCG ATT CTA AAG 58455Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys185 190TGT AAT GAT AAG AAG TTC AGT GGA AAA GGA GAA TGT AAA 623Cys Asn Asp Lys Lys Phe Ser Gly Lys Gly Glu Cys Lys195 200 20560AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AGG CCA 662Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro210 215 220GTA GTA TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA 701Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala65225 230
GAA GAA GAG GTG GTA ATT AGA TCT GAC AAT TTC ATA GAC 740Glu Glu Glu Val Val Ile Arg Ser Asp Asn Phe Ile Asp235 240 245AAT ACT AAA ACC ATA ATA GTA CAG CTG AAA GAA TCT GTA 7795 Asn Thr Lys Thr Ile Ile Val Gln Leu Lys Glu Ser Val250 255GAA ATT AAT TGT ATA AGA CCC AAC AAT AAT ACA AGA AAA 818Glu Ile Asn Cys Ile Arg Pro Asn Asn Asn Thr Arg Lys260 265 27010GGT ATA CAT ATA GGA CCA GGG AGA GCA TGG TAT GCA ACA 857Gly Ile His Ile Gly Pro Gly Arg Ala Trp Tyr Ala Thr275 280 285GGA GAA ATA GTA GGA GAT ATA AGA AAG GCA TAT TGT AAC 896Gly Glu Ile Val Gly Asp Ile Arg Lys Ala Tyr Cys Asn15290 295ATT AGT AGA ACA AAA TGG AAT AAC ACT TTA ATA CAG ATA 935Ile Ser Arg Thr Lys Trp Asn Asn Thr Leu Ile Gln Ile300 305 310GCT AAC AAA TTA AAA GAA AAA TAT AAT ACA ACA ATA AGC 97420Ala Asn Lys Leu Lys Glu Ly5 Tyr Asn Thr Thr Ile Ser315 320TTT AAT CGA TCC TCA GGA GGG GAC CCA GAA ATT GTA ACG 1013Phe Asn Arg Ser Ser Gly Gly Asp Pro Glu Ile Val Thr325 330 33525CAT AGT TTT AAT TGT GGA GGG GAG TTT TTC TAC TGT GAT 1052His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asp340 345 350TCA ACA CAA CTG TTT AAT AGT ACT TGG AAT TTA AAT GGT 1091Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Leu Asn Gly30355 360ACT TGG AAT TTT ACT GCA GGG TCA AAT GAA ACT GAA GGC 1130Thr Trp Asn Phe Thr Ala Gly Ser Asn Glu Thr Glu Gly365 370 375AAT ATC ACA CTC CCA TGC AGA ATA AAA CAA ATT ATA AAC 116935Asn Ile Thr Leu Pro Cys Arg Ile Lys Gln Ile Ile Asn380 385AGG TGG CAG GAA GTA GGG AAA GCA ATG TAT GCC CCT CCC 1208Arg Trp Gln Glu Val Gly Lys Ala Met Tyr Ala Pro Pro390 395 40040ATC AGT GGA CAA ATA AAA TGC TCA TCA AAC ATT ACA GGG 1247Ile Ser Gly Gln Ile Lys Cys Ser Ser Asn Ile Thr Gly405 410 415ATG ATA TTA ACA AGG GAT GGT GGT AAC GAG AAC AAT AAT 1285Met Ile Leu Thr Arg Asp Gly Gly Asn Glu Asn Asn Asn45420 425GAG AGC AGT ACT ACT GAG ACC TTC AGA CCG GGA GGA GGA 1325Glu Ser Ser Thr Thr Glu Thr Phe Arg Pro Gly Gly Gly430 435 440GAT ATG AGG AAC AAT TGG AGA AGT GAA TTA TAT AAA TAT 136450Asp Met Arg Asn Asn Trp Arg Ser Glu Leu Tyr Lys Tyr445 450AAA GTA GTA AAA ATT GAA CCA TTA GGA GTA GCA CCC ACC 1403Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala Pro Thr455 460 46555AAG GCA AAG AGA AGA GTG GTG CAG AGA GAA AAA AGA GCA 1442Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys Arg Ala470 475 480GTG GGA GCG CTA GGA GCT ATG TTC CTT GGG TTC TTA GGA 1481Val Gly Ala Leu Gly Ala Met Phe Leu Gly Phe Leu Gly60485 490GCA TAA AGC TTC TAG ACC GAC TCT AGA GGA TCC 1514Ala Xaa Ser Phe Xaa Thr Asp Ser Arg Gly Ser495 500 504
克隆 C7.10G CTA CCT GTC TGG AAG GAA GCA ACC ACC ACT CTA TTC 37Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe1 5 105 TGT GCA TCA GAT GCT AGA GCA TAT GAC ACA GAG GTA CAT 76Cys Ala Ser Asp Ala Arg Ala Tyr Asp Thr Glu Val His1520 25AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC CCT 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro10 3035AGT CCA CAA GAA GTA TTT TTG GGA AAT GTG ACA GAA AAT 154Ser Pro Gln Glu Val Phe Leu Gly Asn Val Thr Glu Asn404550TTT AAT ATG TGG AAA AAT AAC ATG GTA GAA CAA ATG TAT 19315Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met Tyr55 60GAG GAT ATA ATT AGT TTA TGG CAT CAA AGC TTA AAG CCA 232Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro65 70 7520TGT GTA AAA TTA ACC CCA CTC TGI GTT ACT TTA AAT TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90AGT GAT TAT AGG AAT GCT ACT GAT TAT AAG AAT GCT ACT 310Ser Asp Tyr Arg Asn Ala Thr Asp Tyr Lys Asn Ala Thr25 95 100GAT ACC ACT AGT AGT AAC GAG GGA AAG ATG GAG AGA GGA 349Asp Thr Thr Ser Ser Asn Glu Gly Lys Mec GIu Arg Gly105 110 115GAA ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA AGC ATA 38830Glu Ile Lys Asn Cys Ser phs Asn Ile Thr Thr Ser Ile120 125AAA AAT AAG ATG CAG AAA GAA TAT GCA CTT TTC TAT AAA 427Lys Asn Lys Met Gln Lys Glu Tyr Ala Leu Phe Tyr Lys130135 l4035CTT AAT ATA GTA CCA ATA GAT AAT ACA AGC TAT ACA TTG 466Leu Asn Ile Val Pro Ile Asp Asn Thr Ser Tyr Thr Leu145 150 l55ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCC TGT CCA 505Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro40160 165AAG GTA TCC TTT GAA CCA ATT CCC ATA CAT TAT TGT GCT 544Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ale170 175 180CCG GCT GGT TTT GCG ATT CTA AAG TGT AAT GAT AAG AAG 58345Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys Lys185 190TTC AGT GGA AAA GGA GAA TGT AAA AAT GTC AGC ACA GTA 622Phs Ser Gly Lys Gly Glu Cys Lys Asn Val Ser Thr Vsl195 200 20550CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT CAA 661Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln210 215 220CTG CTG TTA AAT GGC AGT CTA GCA GAA GAA GAG GTG GTA 700Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Val55225 230ATT AGA TCT GAC AAT TTC ACA GAC AAT ACT AAA ACC ATA 739Ile Arg Ser Asp Asn Phe Thr Asp Asn Thr Lys Thr Ile235 240 245ATA GTA CAG CTG AAA GAA TCT GTA GAA ATT AAT TGT ATA 77860Ile Val Gln Leu Lye Glu Ser Val Glu Ile Asn Cys Ile250 255AGA CCC AAC AAT AAT ACA AGA AAA GGT ATA CAT ATA GGA 817Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Ile Gly260 265 270
CCA GGG AGA GCA TGG TAT GCA ACA GGA GAA ATA GTA GGA 856Pro Gly Arg Ala Trp Tyr Ala Thr Gly Glu Ile Val Gly275 280 285GAT ATA AGA CAG GCA TAT TGT AAC ATT AGT AGA ACA AAA 8955 Asp Ile Arg Gln Ala Tyr Cys Asn Ile Ser Arg Thr Lys290 295TGG AAT AAC ACT TTA ATA CAG ATA GCT AAC AAA TTA AAA 934Trp Asn Asn Thr Leu Ile Gln Ile Ala Asn Lys Leu Lys300 305 31010GAA AAA TAT AAT ACA ACA ATA AGC TTT AAT CGA TCC TCA 973Glu Lys Tyr Asn Thr Thr Ile Ser Phe Asn Arg Ser Ser315 320GGA GGG GAC CCA GAA ATT GTA ACC CAT AGT TTT AAT TGT 1012Gly Gly Asp Pro Glu Ile Val Thr His Ser Phe Asn Cys15325 330 335GGA GGG GAA TTT TTC TAC TGT AAT TCA ACA CAA CTG TTT 1051Gly Gly Glu Phe Phe Tyr Cys Asn Ser Thr Gln Leu Phe340 345 350AAT AGT ACT TGG AAT TTA AAT GGT ACT TGG AAT TTT ACT 109020Asn Ser Thr Trp Asn Leu Asn Gly Thr Trp Asn Phe Thr355 360GCA GGG TCA AAT GAA ACT GAA GGC AAT ATC ACA CTC CCA 1129Ala Gly Ser Asn Glu Thr Glu Gly Asn Ile Thr Leu Pro365 370 37525TGC AGA ATA AAA CAA ATT ATA AAC AGG TGG CAG GAA GTA 1168Cys Arg Ile Lys Gln Ile Ile Asn Arg Trp Gln Glu Val380 385GGA AAA GCA ATG TAT GCC CCT CCC ATC AGT GGA CAA ATA 1207Gly Lys Ala Met Tyr Ala Pro Pro Ile Ser Gly Gln Ile30390 395 400AGA TGC TCA TCA AAC ATT ACA GGG ATG ATA TTA ACA AGG 1246Arg Cys Ser Ser Asn Ile Thr Gly Met Ile Leu Thr Arg405 410 415GAT GGT GGT AAC GAG AAC AAT AAT GAG AGC AGT ACT ACT 128535Asp Gly Gly Asn Glu Asn Asn Asn Glu Ser Ser Thr Thr420 425GAG ACC TTC AGA CCG GGA GGA GGA GAT ATG AGG AAC AAT 1324Glu Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asn Asn430 435 44040TGG AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA AAA ATT 1363Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile445 450GAG CCA TTA GGA GTA GCA CCC ACC GAC TCT AGA GGA TCC 1402Glu Pro Leu Gly Val Ala Pro Thr Asp Ser Arg Gly Ser45455 460 465TCT AGA 1408Ser Arg46950 克隆 C11.5GAG GTA CCT GTG TGG AAA GAA GCA ACC ACT ACT CTA 36Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GGG GTG 7555Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Gly Val15 20 25CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp30 3560CCC AAC CCA CAA GAA ATA GAA TTG GTA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Ile Glu Leu Val Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT AAA ATG GTA GAC CAG ATG 192Asp Phe Asn Met Trp Lys Asn Lys Met Val Asp Gln Met65 55 60
CAT GAG GAT ATA ATC AGT TTA TGG GAT GAA AGC CTA AAG 231His Glu Asp Ile Ile Ser Leu Trp Asp Glu Ser Leu Lys65 70 75CCA TGT GTA AAG TTA ACC CCA CTT TGT GTT ACT CTA AAC 2705 Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn80 85 90TGC AGT GAT GTG AAC AAT TCC ACA AAT CCT AAT GAT ACT 309Cys Ser Asp Val Asn Asn Ser Thr Asn Pro Asn Asp Thr95 10010AAT ACT AAT TCC ACT AAT ACT ACT TCC TCT ACT CCT ACG 348Asn Thr Asn Ser Thr Asn Thr Thr Ser Ser Thr Pro Thr105 110 115GCC ACT ACT AGT AGC GAG GAA AAG ATG GAG AAG GGA GAA 387Ala Thr Thr Ser Ser Glu Glu Lys Met Glu Lys Gly Glu15120 125ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA CAC ATG AAA 426Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr His Met Lys130 135 140GAT AAG GCA CAG AAA GAA TAT GCA CTT TTT TAT AAA CTT 46520Asp Lys Ala Gln Lys Glu Tyr Ala Leu Phe Tyr Lys Leu145 150 155GAT ATA GTA CCA ATA GAT GAT AAT AAT GCC AGC TAT AGG 504Asp Ile Val Pro Ile Asp Asp Asn Asn Ala Ser Tyr Arg160 16525TTG ATA AGT TGT AAT ACC TCA GAC ATT ACA CAG GCC TGT 543Leu Ile Ser Cys Asn Thr Ser Asp Ile Thr Gln Ala Cys17C 175 180CCA AAG GTG ACC TTT GAG CCA ATT CCC ATA CAT TAT TGT 582Pro Lys Val Thr Phe Glu Pro Ile Pro Ile His Tyr Cys30l85 190GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG 621Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys195 200 205AAG TTC AAT GGA ACA GGA CCA TGT TCA AAG GTC AGC ACA 66035Lys Phe Asn Gly Thr Gly Pro Cys Ser Lys Val Ser Thr210 215 220GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 699Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr225 23040CAA CTG TTG TTA AAT GGC AGT CTT GCA GAA GAA GAA GTA 738Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val235 240 245GTA ATT AGA TCT GTC AAT TTC ACA GAC AAT GCT AAA ATC 777Val Ile Arg Ser Val Asn Pne Thr Asp Asn Ala Lys Ile45250 255ATA ATA GTA CAG CTG AAA GAA CCT GTA GCA ATT AAT TGT 816Ile Ile Val Gln Leu Lys Glu Pro Val Ala Ile Asn Cys260 265 270ACA AGA CCC AAC AAC AAT ACA AGA AAA GGT ATA CAT CTA 85550Thr Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Leu275 280 285GGA CCA GGG AGC ACA TTT TAT ACA ACA GGA GAA ATA ATA 894Gly Pro Gly Ser Thr Pne Tyr Thr Thr Gly Glu Ile Ile290 29555GGA GAC ATA AGA AAA GCA TAT TGC AAG ATT AGT AAA GAA 933Gly Asp Ile Arg Lys Ala Tyr Cys Lys Ile Ser Lys Glu300 305 310AAA TGG AAT AAC ACT TTA AGA CAG GTA GTT AAA AAA TTA 972Lys Trp Asn Asn Thr Leu Arg Gln Val Val Lys Lys Leu60315 320AGA GAA CAA TTT GGG AAT AAA ACA ATA ATT TTT AAT CGA 1011Arg Glu Gln Phe Gly Asn Lys Thr Ils Ile Phs Asn Arg325 330 335
TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT 1050Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe340 345 350AAC TGT GGA GGG GAG TTT TTC TAC TGT AAT ACA ACA CAA 1089Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Gln355 360CTG TTT AAT AGT ACT TGG AAT AAT ACT GAA GGG ACA AAT 1128Leu Phe Asn Ser Thr Trp Asn Asn Thr Glu Gly Thr Asn365 370 37510AGC ACT GAA GGA AAT AGC ACA ATC ACA CTC CCA TGC AGA 1167Ser Thr Glu Gly Asn Ser Thr Ile Thr Leu Pro Cys Arg380 385ATA AAA CAA ATT ATA AAT ATG TGG CAG GAA GTA GGA AAA 1206Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys15390 395 400GCA ACG TAT GCC CCT CCC ATC AGA GGA CGA ATT AGA TGC 1245Ala Thr Tyr Ala Pro Pro Ile Arg Gly Arg Ile Arg Cys405 410 415ATA TCA AAT ATT ACA GGA CTG CTA TTA ACA AGA GAT GGT 128420Ile Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly420 425GGT AGG AAT GTC ACA AAC AAT ACC GAA ACC TTC AGA CCT 1323Gly Arg Asn Val Thr Asn Asn Thr Glu Thr Phe Arg Pro430 435 44025GGA GGA GGA GAC ATG AGG GAC AAT TGG AGA AGT GAA TTA 1362Gly Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu445 450TAT AAA TAT AAA GTA GTA AAA GTT GAA CCA TTA GGA ATA 1401Tyr Lys Tyr Lys Val Val Lys Val Glu Pro Leu Gly Ile30455 460 465GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAC AGA GAC 1440Ala Pro Thr Lys Ala Lys Arg Arg Val Val His Arg Asp470 475 480AAA AGA GCA GCA CTA GGA GCC TTG TTC CTT GGG TTC TTA 147935Lys Arg Ala Ala Leu Gly Ala Leu Phe Leu Gly Phe Leu485 490GGA GCA TAA AAG CTT CTA GA 1499Gly Ala Xaa Lys Leu Leu495 49940克隆 C11.7GAG GTA CCT GTA TGG AAA GAA GGA ACC ACT ACT CTA 36Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 1045TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GAG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val15 20 25CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp50 30 35CCC AAC CCA CAA GAA ATA GAA TTG GTA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Ile Glu Leu Val Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT AAA ATG GTA GAC CAG ATG 19255Asp Phe Asn Met Trp Lys Asn Lys Met Val Asp Gln Met55 60CAT GAG GAT ATA ATC AGT TTA TGG GAT GAA AGC CTA AAG 231His Glu Asp Ile Ile Ser Leu Trp Asp Glu Ser Leu Lys65 70 7560CCA TGT GTA AAG TTA ACC CCA CTT TGT GTT ACT CTA AAC 270Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn80 85 90TGC AGT GAT GTG AAC AAT TCC ACA AAT CCT AAT GAT ACT 309Cys Ser Asp Val Asn Asn Ser Thr Asn Pro Asn Asp Thr65 95 100
AAT ACT AAT TCC ACT AAT ACT ACT TCC TCT ACT CCT ACG 348Asn Thr Asn Ser Thr Asn Thr Thr Ser Ser Thr Pro Thr105 110 115GCC ACT ACT AGT AGC GAG GAA AAG ATG GAG AAG GGA GAA 3875 Ala Thr Thr Ser Ser Glu Glu Lys Met Glu Lys Gly Glu120 125ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA CAC ATG AAA 426Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr His Met Lys130 135 14010GAT AAG GTA CAG AAA GAA TAT GCA CTT TTT TAT AAA CTT 465Asp Lys Val Gln Lys Glu Tyr Ala Leu Phe Tyr Lys Leu145 150 155GAT ATA GTA CCA ATA GAT GAT AAT AAT ACC AGC TAT AGG 504Asp Ile Val Pro Ile Asp Asp Asn Asn Thr Ser Tyr Arg15160 165TTG ATA AGT TGT AAT ACC TCA GTC ATT ACA CAG GCC TGT 543Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys170 175 180CCA ATG GTG ACC TTT GAG CCA ATT CCC ATA CAT TAT TGT 58220Pro Met Val Thr Phe Glu Pro Ile Pro Ile Mis Tyr Cys185 190GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG 621Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys195 200 20525AAG TTC AAT GGA ACA GGA CCA TGT TCA AAG GTC AGC ACA 660Lys Phe Asn Gly Thr Gly Pro Cys Ser Lys Val Ser Thr210 215 220GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 699Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr30225 230CAA CTG TTG TTA AAT GGC AGT CTT GCA GAA GAA GAA GTA 738Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val235 240 245GTA ATT AGA TCT GTC AAT TTC ACA GAC AAT GCT AAA ATC 77735Val Ile Arg Ser Val Asn Phe Thr Asp Asn Ala Lys Ile250 255ATA ATA GTA CAG CTG AAA GAA CCT GTA GCA ATT AAT TGT 816Ile Ile Val Gln Leu Lys Glu Pro Val Ala Ile Asn Cys260 265 27040ACA AGA CCC AAC AAC AAT ACA AGA AAA GGT ATA CAT CTA 855Thr Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Leu275 280 285GGA CCA GGG AGC ACA TTT TAT ACA ACA GGA GAA ATA ATA 894Gly Pro Gly Ser Thr Phe Tyr Thr Thr Gly Glu Ile Ile45290 295GGA GAC ATA AGA AAA GCA TAT TGC AAG ATT AGT AAA GAA 933Gly Asp Ile Arg Lys Ala Tyr Cys Lys Ile Ser Lys Glu300 305 310AAA TGG AAT AAC ACT TTA AGA CAG GTA GTT AAA AAA TTA 97250Lys Trp Asn Asn Thr Leu Arg Gln Val Val Lys Lys Leu315 320AGA GAA CAA TTT GGG AAT AAA ACA ATA ATT TTT AAT CGA 1011Arg Glu Gln Phe Gly Asn Lys Thr Ile Ile Phe Asn Arg325 330 33555TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT 1050Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe340 345 350AAC TGT GGA GGG GAG TTT TTC TAC TGT AAT ACA ACA CAA 1089Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Gln60355 360CTG TTT AAT AGT ACT TGG AAT AAT ACT GAA GGG ACA AAT 1128Leu Phe Asn Ser Thr Trp Asn Asn Thr Glu Gly Thr Asn365 370 375
AGC ACT GAA GGA AAT AGC ACA ATC ACA CTC CCA TGC AGA 1167Ser Thr Glu Gly Asn Ser Thr Ile Thr Leu Pro Cys Arg380 385ATA AAA CAA ATT ATA AAT ATG TGG CAG GAA GTA GGA AAA 12065 Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys390395400GCA ACG TAT GCC CCT CCC ATC AGA GGA CGA ATT AGA TGC 1245Ala Thr Tyr Ala Pro Pro Ile Arg Gly Arg Ile Arg Cys405 410 41510ATA TCA AAT ATT ACA GGA CTG CTA TTA ACA AGA GAT GGT 1284Ile Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly420 425GGT AGG AAT GTC ACA AAC AAT ACC GAN NCC TTC AGA CCT 1323Gly Arg Asn Val Thr Asn Asn Thr Xaa Xaa Phe Arg Pro15430 435 440GGA GGA GGA GAC ATG AGG GAC AAT TGG AGA AGT GAA TTA 1362Gly Gly GIy Asp Met Arg Asp Asn Trp Arg Ser Glu Leu445 450TAT AAA TAT AAA GTA GTA AAA GTT GAA CCA TTA GGA ATA 140120Tyr Lys Tyr Lys Val Val Lys Val Glu Pro Leu Gly Ile455 460 465GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAC AGA GAC 1440Ala Pro Thr Lys Ala Lys Arg Arg Val Val His Arg Asp470 475 48025AAA AGA GCA GCA CTA GGA GCT TTG TTC CTT GGG TTC TTA 1479Lys Arg Ala Ala Leu Gly Ala Leu Phe Leu Gly Phe Leu485 490GGA GCA TAA AAG CTT CTA GA 1499Gly Ala Xaa Lys Leu Leu30 495 499克隆C10.5G GTA CCT GTG TGG AAA GAA GCA AAC ACA ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe35 15 10TGT GCA TCA GAT GCT AAA GCA TAT GAT AGA GAA GTA CAT 76Cys Ala Ser Asp Ala Lys Ala Tyr Asp Arg Glu Val His15 20 25AAT GTT TGG GCA ACA CAT GCC TGT GTA CCC ACA GAC CCC 11540Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AAC CCA CAA GAA ATA GTA TTG GGA AAT GTG ACA GAA AAT 154Asn Pro Gln Glu Ile Val Leu Gly Asn Val Thr Glu Asn40 45 5045TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His55 60GAG GAT ATA ATC AAT TTA TGG GAT CAA AGC TTA AAG CCA 232Glu Asp Ile Ile Asn Leu Trp Asp Gln Ser Leu Lys Pro50 65 70 75TGT GTA AAG TTA ACT CCA CTC TGT GTT ACT TTA AAG TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Lys Cys80 85 90AAG GAT CTG GAG AGG AAT ACT ACC TAT AAT AGC ACT ATT 31055Lys Asp Leu Glu Arg Asn Thr Thr Tyr Asn Ser Thr Ile95 100ACC AAT AAT AGT AGT TTG GAG GGA CTA AGA GAA CAA ATG 349Thr Asn Asn Ser Ser Leu Glu Gly Leu Arg Glu Gln Met105 110 11560ACA AAC TGC TCT TTC AAC ATC ACC ACA AGT ATA AGA GAT 388Thr Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp120 125AAG GTG CAG AAA GAA TAT GCA CTT TTG TAT AAA CTT GAT 427Lys Val Gln Lys Glu Tyr Ala Leu Leu Tyr Lys Leu Asp65 130 135 140
GTA GTA CCA ATA GAA GAA GAT GAC AAT ACT AGC TAT AGA 466Val Val Pro Ile Glu Glu Asp Asp Asn Thr Ser Tyr Arg145 150 155TTG ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCT TGT 5055Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys160 165CCA AAG ACA TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT 544Pro Lys Thr Ser Phe Glu Pro Ile Pro Ile His Tyr Cys170 175 18010GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAT GAT AAG 583Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys185 190AAG TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGC ACA 622Lys Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr15195 200 205GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 661Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr210 215 220CAA CTG TTG TTA AAT GGC AGT CTA GCA GAA GAA GAG GTA 70020Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val225 230GTA ATC AGA TCT GCC AAT TTC ACA GAC AAT GCT AAA ACC 739Val Ile Arg Ser Ala Asn Phe Thr Asp Asn Ala Lys Thr235 240 24525ATA ATA GTA CAT CTA AAT GAA ACT GTA AAA ATT AAT TCT 778Ile Ile Val His Leu Asn GIu Thr Val Lys Ile Asn Cys250 255ACA AGA CTT GGC AAC AAT ACA AGA AAA AGT ATA AAT ATA 817Thr Arg Leu Gly Asn Asn Thr Arg Lys Ser Ile Asn Ile30250 265 270GGA CCA GGG AGA GTA CTC TAT GCA ACA GGA GAA ATA ATA 856Gly Pro Gly Arg Val Leu Tyr Ala Thr Gly Glu Ile Ile275 280 285GGA GAC ATA AGA CAA GCA CAT TGT AAC ATT AGT AGA GCA 89535Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Arg Ala290 295CAA TGG AAT AAG ACT TTA GAA AAG GTA GTT GAC AAA TTA 934Gln Trp Asn Lys Thr Leu Glu Lys Val Val Asp Lys Leu300 305 31040AGA AAA CAA TTT GGG GAT AAT ACA ACA ATA GCT TTT AAT 973Arg Lys Gln Phe Gly Asp Asn Thr Thr Ile Ala Phe Asn315 320CGA TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC ACT 1012Arg Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Thr45325 330 335TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA 1051Phe Asn Cys GIy Gly Glu Phe Phe Tyr Cys Asn Thr Thr340 345 350CAA CTG TTT AAT AGT ACT TGG AAT AAT ACT TGG AAG GAT 109050Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr Trp Lys Asp355 360CCT AAC AGG AGT GAC AAT ATC ACA CTC CCA TGC AGA ATA 1129Pro Asn Arg Ser Asp Asn Ile Thr Leu Pro Cys Arg Ile365 370 37555AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA 1168Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala380 385ATG TAC GCC CCT CCC ATC AGA GGG GAA ATT AGA TGT TCA 1207Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile Arg Cys Ser60390 395 400TCA AAT ATC ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT 1246Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly GIy405 410 415
AAT GAC GAT GGT AAT GAC ACG ACC ACA AAC AGG ACC GAG 1285Asn Asp Asp Gly Asn Asp Thr Thr Thr Asn Arg Thr Glu420 425ATC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1324Ile Phe Arg Pro Gly GIy Gly Asp Met Arg Asp Asn Trp430 435 440AGA AGT GAA TTA TAT AGA TAT AAA GTA GTA AAA ATT GAA 1363Arg Ser Glu Leu Tyr Arg Tyr Lys Val Val Lys Ile Glu445 45010CCA TTA GGA ATA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1402Pro Leu Gly Ile Ala Pro Thr Arg Ala Lys Arg Arg Val455 4G0 465GTG CAG AGA GAA AAA AGA GCA GTA GGA CTA GGA GCT TTG 1441Val Gln Arg Glu Lys Arg Ala Val Gly Leu Gly Ala Leu15470 475 480TTC CTT GGG T TCTTAGGAG CATAAAGCTT CTAGA 1475Phe Leu Gly48320 克隆 C10.7G GTA CTT GTG TGG AAA GAA GCA AAC ACA ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT GAT AGA GAA GTA CAT 7625Cys Ala Ser Asp Ala Lys Ala Tyr Asp Arg Glu Val His15 20 25AAT GTT TGG GCA ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 3530AAC CCA CAA GAA ATA GTA TTG GGA AAT GTG ACA GAA AAT 154Asn Pro Gln Glu Ile Val Leu Gly Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His35 55 60GAG GAT ATA ATC AAT TTA TGG GAT CAA AGC TTA AAG CCA 232Glu Asp Ile Ile Asn Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAG TTA ACT CCA CTC TGT GTT ACT TTA AAG TGC 27140Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Lys Cys80 85 90AAG GAT CTG GAG AGG AAT ACT ACC TAT AAT AGC ACT ATT 310Lys Asp Leu Glu Arg Asn Thr Thr Tyr Asn Ser Thr Ile95 10045ACC AAT AAT AGT AGT TTG GAG GGA CTA AGA GAA CAA ATG 349Thr Asn Asn Ser Ser Leu Glu Gly Leu Arg Glu Gln Met105 110 115ACA AAC TGC TCT TTC AAC ATC ACC ACA AGT ATA AGA GAT 388Thr Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp50120 125AAG GTG CAG AAA GAA TAT GCA CTT TTG TAT AAA CTT GAT 427Lys Val Gln Lys Glu Tyr Ala Leu Leu Tyr Lys Leu Asp130 135 140GTA GTA CCA ATA GAA GAA GAT GAC AAT ACT AGC TAT AGA 46655Val Val Pro Ile Glu Glu Asp Asp Asn Thr Ser Tyr Arg145 150 155TTG ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCT TGT 505Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys160 16560CCA AAG ACA TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT 544Pro Lys Thr Ser Phe Glu Pro Ile Pro Ile His Tyr Cys170 175 180GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAT GAT AAG 583Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys65185 190
AAG TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGC ACA 622Lys Phe Asn Gly Thr Gly Pro Cys Lys Ash Val Ser Thr195 200 205GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 6615Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr210 215 220CAA CTG TTG TTA AAT GGC AGT CTA GCA GAA GAA GAG GTA 700Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val225 23010GTA ATC AGA TCT GCC AAT TTC ACA GAC AAT GCT AAA ACC 739Val Ile Arg Ser Ala Asn Phe Thr Asp Ash Ala Lys Thr235 240 245ATA ATA GTA CAT CTA AAT GAA ACT GTA AAA ATT AAT TGT 778Ile Ile Val His Leu Asn Glu Thr Val Lys Ile Asn Cys15250 256ACA AGA CTT GGC AAC AAT ACA AGA AAA AGT ATA AAT ATA 817Thr Arg Leu Gly Asn Asn Thr Arg Lys Ser Ile Asn Ile260 265 270GGA CCA GGG AGA GTA CTC TAT GCA ACA GGA GAA ATA ATA 85620Gly Pro Gly Arg Val Leu Tyr Ala Thr Gly Glu Ile Ile275 280 285GGA GAC ATA AGA CAA GCA CAT TGT AAC ATT AGT AGA GCA 895Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Arg Ala290 29525CAA TGG AAT AAG ACT TTA GAA AAG GTA GTT GAC AAG TTA 934Gln Trp Asn Lys Thr Leu Glu Lys Val Val Asp Lys Leu300 305 310AGA AAA CAA TTT GGG GAT AAT ACA ACA ATA GCT TTT AAT 973Arg Lys Gln Phe Gly Asp Asn Thr Thr Ile Ala Phe Asn30315 320CGA TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC ACT 1012Arg Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Thr325 330 335TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA 105135Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr340 345 350CAA CTG TTT AAT AGT ACT TGG AAT AAT ACT TGG AAG GAT 1090Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr Trp Lys Asp355 36040CCT AAC AGG AGT GAC AAT ATC ACA CTC CCA TGC AGA ATA 1129Pro Asn Arg Ser Asp Asn Ile Thr Leu Pro Cys Arg Ile365 370 375AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA 1168Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala45380 385ATG TAC GCC CCT CCC ATC AGA GGG GAA ATT AGA TGT TCA 1207Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile Arg Cys Ser390 395 400TCA AAT ATC ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT 124650Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly405 410 415AAT GAC GAT GGT AAT GAC ACG ACC ACA AAC AGG ACC GAG 1285Asn Asp Asp Gly Asn Asp Thr Thr Thr Asn Arg Thr Glu420 42555ATC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1324Ile Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp430 435 440AGA AGT GAA TTA TAT AGA TAT AAA GTA GTA AAA ATT GAA 1363Arg Ser Glu Leu Tyr Arg Tyr Lys Val Val Lys Ile Glu60445 450CCA TTA GGA ATA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1402Pro Leu Gly Ile Ala Pro Thr Arg Ala Lys Arg Arg Val455 460 465
GTG CAG AGA GAA AAA AGA GCA GTA GGA CTA GGA GCT TTG 1441Val Gln Arg Glu Lys Arg Ala Val Gly Leu Gly Ala Leu470 475 480TTC CTT GGG TTC TTG GGA GCA TAA AGC TTC TAG A 1475Phe Leu GIy Phe Leu Gly Ala Xaa Ser Phe Xaa485 490 491克隆 C17.1CTC GAG GTA CCT GTG TGG AAA GAA GCA ACC ACC ACT 3610Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT GAT TCA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Ser Glu15 20 2515GCA CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA 114Ala His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr30 35GAC CCC AAC CCA CAA GAA GTA GAA TTG GAA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Glu Leu Glu Asn Val Thr20 40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG 192Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln55 60ATG CAT GGG GAT ATA ATT AGT TTA TGG GAT CAA AGC CTA 23125Met His Gly Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACG TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 9030AAT TGC ACT GAC CCA AAT GTT ACT AAT AGC GAG AGA ACG 309Asn Cys Thr Asp Pro Asn Val Thr Asn Ser Glu Arg Thr95 100ATA GAG GGG GGA GAA ATA AAA AAT TGC TCT TTC AAT ATC 348Ile Glu Gly Gly Glu Ile Lys Asn Cys Ser Phe Asn lle35105 110 115ACC ACA AAC ATA AGA GAT AGG TTT CAG AAA GAA TAT GCA 387Thr Thr Asn Ile Arg Asp Arg Phe Gln Lys Glu Tyr Ala120 125CTT TTT TAT AAA CTT GAT GTA ATA CCA TTA GGT AAT GAT 42640Leu Phe Tyr Lys Leu Asp Val Ile Pro Leu Gly Asn Asp130 135 140AAT ACT AGC TAT AGG TTG ATA AGT TGT AAC ACC TCA GTC 465Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val145 150 15545ATT ACA CAG GCC TGT CCA AAG GTA TCC TTT GAG CCA ATT 504Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile160 165CCC ATA CAT TAT TGT GCC CCG GCT GGT TTT GCG ATT CTA 543Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu50170 175 180AAG TGT AAA GAT AAG AAG TTC AAT GGA ACA GGA CCA TGT 582Lys Cys Lys Asp Lys Lys Phe Aso Gly Thr Gly Pro Cys185 190ACA AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AAG 62155Thr Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys195 200 205CCA GTA GTA TCA ACT CAA CTG TTG TTA AAT GGC AGT CTA 660Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu210 215 22060GCA GAA GAA GAC ATA GTA ATT AGA TCC GCC AAT CTC ACA 699Ala Glu Glu Asp I1e Val Ile Arg Ser Ala Asn Leu Thr225 230GAC AAT GCT AAA AAC ATA ATA GTA CAG CTG AAT GAA TCT 738Asp Asn Ala Lys Asn Ile Ile Val Gln Leu Asn Glu Ser65235 240 245
GTA ACA ATG AAT TGT ACA AGA CCC AAC AAC AAT ACA ATG 777Val Thr Met Asn Cys Thr Arg Pro Asn Asn Asn Thr Met250 255AAA AGT ATA CAT ATA GGA CCA GGC AGA GCA TTT TAT GCA 8165 Lys Ser Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala260 265 270ACA GGA AAC ATA ATA GGA GAT ATA AGA CAA GCA CAT TGT 855Thr Gly Asn Ile Ile Gly Asp Ile Arg Gln Ala His Cys275 280 28510AAC ATT AGT GGA ACA AAA TGG AAT GAC ACT TTG AAA AAG 894Asn Ile Ser Gly Thr Lys Trp Asn Asp Thr Leu Lys Lys290 295ATA GCT ATA AAA TTA AGA GAA CAA TTT AAT AAG ACA ATA 933Ile Ala Ile Lys Leu Arg Glu Gln Phe Asn Lys Thr Ile15300 305 310GTC TTT AAT CAA TCC TCA GGA GGG GAC CCA GAA ATT GCA 972Val Phe Asn Gln Ser Ser Gly Gly Asp Pro Glu Ile Ala315 320ACG CTC AGT TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT 101120Thr Leu Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys325 330 335AAT TCA ACA CAA CTG TTT AAT AGT ACT TGG AAT AGT ACT 1050Asn Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Ser Thr340 345 35025GGG TCA AAT AAC ACT AAA GGA AAT GAC ACA ATC ACA CTC 1089Gly Ser Asn Asn Thr Lys Gly Asn Asp Thr Ile Thr Leu355 360CCA TGC AGA ATA AGA CAA ATT ATA AAC ATG TGG CAG AAA 1128Pro Cys Arg Ile Arg Gln Ils Ile Asn Met Trp Gln Lys30365 370 375ATA GGA AAA GCA ATG TAT GCC CCT CCC ATC AAA GGG CAA 1167Ile Gly Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln380 385ATT AGA TGT TCA TCA AAT ATT ACA GGG CTA ATA TTA ACA 120635Ile Arg Cys Ser Ser Asn Ile Thr Gly Leu Ile Leu Thr390 395 400AGA GAT GGT GGT AAC AAC AAC ATG AGC AAG ACC ACC GAG 1245Arg Asp Gly Gly Asn Asn Asn Met Ser Lys Thr Thr Glu405 410 41540ACC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1284Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp420 425AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA AAA ATT GAA 1323Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu45430 435 440CCA TTA GGA GTA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1362Pro Leu Gly Val Ala Pro Thr Arg Ala Lys Arg Arg Val445 450GTG CAG AGA GAA AAA AGA GCA GTG GGA ATA GGA GCT GTG 140150Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Val455 460 465TTC CTT GGG TTC TTG GGA GCA TAA AGC TTC TAG A 1435Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa470 475 47855克隆 C17.3CTC GAG GTA CCT GTG TGG AAA GAA GCA ACC ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 1060CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT GAT TCA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Ser Glu15 20 25GCA CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA 114Ala His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr65 30 35
GAC CCC AAC CCA CAA GAA GTA GAA TTG GAA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Glu Leu Glu Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG 1925 Glu Asn Phe Asn Met Trp Lys Asn Asn Mer Val Glu Gln55 60ATG CAT GGG GAT ATA ATT AGT TTA TGG GAT CAA AGC CTA 231Met His Gly Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 7510AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACG TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAC CCA AAT GTT ACT AAT AGC GAG AGA ACG 309Asn Cys Thr Asp Pro Asn Val Thr Asn Ser Glu Arg Thr15 95 100ATA GAG GGG GGA GAA ATA AAA AAT TGC TCT TTC AAT ATC 348Ile Glu Gly Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile105 110 115ACC ACA AAC ATA AGA GAT AGG TTT CAG AAA GAA TAT GCA 3S720Thr Thr Asn Ile Arg Asp Arg Phe Gln Lys Glu Tyr Ala120 125CTT TTT TAT AAA CTT GAT GTA ATA CCA TTA GGT AAT GAT 426Leu Phe Tyr Lys Leu Asp Val Ile Pro Leu Gly Asn Asp130 135 14025AAT ACT AGC TAT AGG TTG ATA AGT TGT AAC ACC TCA GTC 465Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val145 150 155ATT ACA CAG GCC TGT CCA AAG GTA TCC TTT GAG CCA ATT 504Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile30160 165CCC ATA CAT TAT TGT GCC CCG GCT GGT TTT GCG ATT CTA 543Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu170 175 180AAG TGT AAA GAT AAG AAG TTC AAT GGA ACA GGA CCA TGT 55235Lys Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys185 190ACA AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AAG 621Thr Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys195 200 20540CCA GTA GTA TCA ACT CAA CTG TTG TTA AAT GGC AGT CTA 660Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu210 215 220GCA GAA GAA GAC ATA GTA ATT AGA TCC GCC AAT CTC ACA 699Ala Glu Glu Asp Ile Val Ile Arg Ser Ala Asn Leu Thr45225 230GAC AAT GCT AAA AAC ATA ATA GTA CAG CTG AAT GAA TCT 738Asp Asn Ala Lys Asn Ile Ile Val Gln Leu Asn Glu Ser235 240 245GTA ACA ATG AAT TGT ACA AGA CCC AAC AAC AAT ACA ATG 77750Val Thr Met Asn Cys Thr Arg Pro Asn Asn Asn Thr Met250 255AAA AGT ATA CAT ATA GGA CCA GGC AGA GCA TTT TAT GCA 815Lys Ser Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala260 265 27055ACA GGA AAC ATA ATA GGA GAT ATA AGA CAA GCA CAT TGT 855Thr Gly Asn Ile Ile Gly Asp Ile Arg Gln Ala His Cys275 280 285AAC ATT AGT GGA ACA AAA TGG AAT GAC ACT TTG AAA AAG 894Asn Ile Ser Gly Thr Lys Trp Asn Asp Thr Leu Lys Lys60290 295ATA GCT ATA AAA TTA AGA GAA CAA TTT AAT AAG ACA ATA 933Ile Ala Ile Lys Leu Arg Glu Gln Phe Asn Lys Thr Ile300 305 310
GTC TTT AAT CAA TCC TCA GGA GGG GAC CCA GAA ATT GCA 972Val Phe Asn Gln Ser Ser Gly Gly Asp Pro Glu Ile Ala315 320ACG CTC AGT TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT 10115 Thr Leu Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys325 330 335AAT TCA ACA CAA CTG TTT AAT AGT ACT TGG AAT AGT ACT 1050Asn Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Ser Thr340 345 35010GGG TCA AAT AAC ACT AAA GGA AAT GAC ACA ATC ACA CTC 1089Gly Ser Asn Asn Thr Lys Gly Asn Asp Thr Ile Thr Leu355 360CCA TGC AGA ATA AGA CAA ATT ATA AAC ATG TGG CAG AAA l128Pro Cys Arg Ile Arg Gln Ile Ile Asn Met Trp Gln Lys15365 370 375ATA GGA AAA GCA ATG TAT GCC CCT CCC ATC AAA GGG CAA 1167Ile Gly Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln380 385ATT AGA TGT TCA TCA AAT ATT ACA GGG CTA ATA TTA ACA 120620Ile Arg Cys Ser Ser Asn Ile Thr Gly Leu Ile Leu Thr390 395 400AGA GAT GGT GGT AAC AAC AAC ATG AGC AAG ACC ACC GAG 1245Arg Asp Gly Gly Asn Asn Asn Met Ser Lys Thr Thr Glu405 410 41525ACC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1284Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp420 425AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA AAA ATT GAA 1323Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu30430 435 440CCA TTA GGA GTA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1362Pro Leu Gly Val Ala Pro Thr Arg Ala Lys Arg Arg Val445 450GTG CAG AGA GAA AAA AGA GCA GTG GGA ATA GGA GCT GTG 140135Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Val455 460 465TTC CTT GGG TTC TTG GGA GCA TAA AGC TTC TAG A 1435Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa470 475 478
除了表1中所列出的，圖3顯示七個分離物各自克隆的氨基酸序列的序列對比。方框內(nèi)顯示各克隆所對應(yīng)的殘基。在圖中，將該氨基酸序列與MN-rgp120(SEQ.ID.NO.29)進(jìn)行序列對比。
在一個實(shí)施方案中，本發(fā)明的gp120多肽與逃逸分離物之一的序列具有相同的氨基酸序列。在另一個實(shí)施方案中，如下文詳述，氨基酸序列是截短的。另一個實(shí)施方案中，本發(fā)明的gp120多肽序列在逃逸分離物的序列中，含有一種或多種氨基酸的取代，插入或缺失(改變)。通常，除了不位于截短的氨基酸序列的氨基酸序列和消除表位外，本發(fā)明的gp120多肽將包括在逃逸分離物中的氨基酸序列中的改變，這些改變不改變多肽具有的與該分離物的氨基酸序列相同的誘導(dǎo)中和抗體的能力。
一般來說，本發(fā)明的gp120多肽的氨基酸序列中的取代是保守取代，尤其是gp120的V2,V3，及C4結(jié)構(gòu)域中的氨基酸殘基，這些結(jié)構(gòu)域中含有中和表位。然而，非保守取代位于特別是不含中和表位的的結(jié)構(gòu)域中。
保守取代以類似大小和特性的氨基酸取代另一氨基酸。例如，疏水殘基或親水殘基分別被另一個疏水殘基或親水殘基所代替。氨基酸可分成下列組帶正電荷的的殘基(K,R和H)；帶負(fù)電荷的殘基(D和E)；酰胺類(N和Q)；芳香類(F,Y和W)；疏水殘基(P,G,V,L,I,和M)；和不帶電荷的殘基(S和T)。通常，在小組內(nèi)的殘基可被該小組另一成員所代替。
在一個實(shí)施方案中，在gp120的V2,V3及C4結(jié)構(gòu)域中的臨界氨基酸殘基與逃逸分離物序列所對應(yīng)的殘基是完全相同的。gp120的V2,V3，及C4結(jié)構(gòu)域中的臨界氨基酸殘基在實(shí)驗(yàn)部分得以描述。在另一實(shí)施方案中，gp120的V2,V3，及C4結(jié)構(gòu)域中的所有氨基酸殘基與逃逸分離物序列所對應(yīng)的殘基是完全相同的。來源于逃逸分離物的編碼gp120的寡核苷酸本發(fā)明也提供了來源于逃逸分離物的編碼gp120的新寡核苷酸(其可用于表達(dá)gp120)。本發(fā)明的寡核苷酸編碼本發(fā)明的多肽。寡核苷酸可以是DNA或RNA，通常是DNA。雖然由于基因密碼子的簡并性，許多核苷酸序列能編碼相同的氨基酸，但簡而言之，本發(fā)明的寡核苷酸包括表1中所說明的逃逸分離物的核苷酸序列(SEQ ID No.2,4,6,8,10,12,14,16,18,20,22,24,26和28)。通常，本發(fā)明的寡核苷酸小于約5千堿基(kb)，優(yōu)選為小于約3kb。
為了表達(dá)所編碼的氨基酸序列，可將寡核苷酸插入到轉(zhuǎn)錄單位中。而轉(zhuǎn)錄單位可插入到用于細(xì)胞系的制備的質(zhì)粒中，或插入到病毒中(例如；疫苗)或直接用作為疫苗DNA。合適的用于疫苗蛋白質(zhì)生產(chǎn)的轉(zhuǎn)錄單位為大家所熟知。優(yōu)選的表達(dá)載體，命名為psvI685，在SEQID NO.32中說明。載體包括連接到連接序列上的HSV-1gD1信號序列。將要表達(dá)的gp120核苷酸序列始于基因的Kpn I位點(diǎn)。由于所有g(shù)p120或gp160序列含有這一位點(diǎn)，任何gp120核苷酸序列均可類似地插入到載體中，并且被表達(dá)。載體從SV40開始以polyA尾結(jié)束。
除了對本發(fā)明的多肽序列的表達(dá)有用之外，本發(fā)明的寡核苷酸也可用于對HIV分離物檢測的診斷學(xué)上。例如，編碼中和表位的寡核苷酸或其部分可用于支鏈DNA診斷學(xué)上或在原位雜交研究中做探針。疫苗的制備利用在合適載體中的本發(fā)明的gp120多肽(其來源于所選擇的逃逸分離物)來制備亞單位疫苗。多肽可單獨(dú)使用，但通常與多價(jià)疫苗一起施用，這種亞單位疫苗包括gp120MN。除了本發(fā)明的一種或多種gp120多肽之外，一般地疫苗包括MN多肽(在下文中為MN-rgp120)。疫苗通常包括大約3至5個不同的gp120多肽，但有30個或者更多的不同的gp120多肽可加以利用。
用于疫苗的gp120多肽的制備方法為大家所熟知，并在下文中得以描述。除了所選擇的HTV分離物的利用外，該方法中所制備的gp120亞單位疫苗不同于現(xiàn)有技術(shù)的gp120亞單位疫苗。
如同現(xiàn)有技術(shù)的gp120亞單位疫苗，在可引起抗體形成所要求的純度和足夠的濃度下，將gp120與生理上可接受的載體相混合。生理上可接受載體在疫苗中所使用的劑量與濃度對接受者無毒。一般地，疫苗配制成注射劑，通常用于肌內(nèi)或皮下注射。用于注射的合適載體包括無菌水，但是優(yōu)選為生理鹽溶液，如正常的鹽水或緩沖的含鹽溶液，如磷酸緩沖鹽水或環(huán)乳酸。疫苗一般地含有佐劑。有用的佐劑包括QS21(皂樹肥皂草，由劍橋Biotech,Worcester,MA提供)，它能刺激細(xì)胞毒性T-細(xì)胞，以及明礬(鋁氫氧化物佐劑)。也利用不同的能提高細(xì)胞的或局部免疫性的佐劑。尤其是，在疫苗中可包括免疫加強(qiáng)因子如細(xì)胞因子。合適的加強(qiáng)免疫的細(xì)胞因子的例子包括白細(xì)胞介素，如白細(xì)胞介素(IL-2)和白細(xì)胞介素-12(IL-12)，以及腫瘤壞死因子-α(TNF-α)存在于疫苗中的附加賦形劑包括低分子量的多肽(少于約10個殘基)，蛋白質(zhì)，氨基酸，碳水化合物其包括葡萄糖或葡聚糖，螯合劑如EDTA，及其它賦形劑，這些物質(zhì)能穩(wěn)定蛋白質(zhì)或阻止微生物生長。
疫苗也可含有其它HIV蛋白質(zhì)。尤其是，gp41或gp41的胞外部分或HIV-1核心蛋白質(zhì)如P24,P17，和P55可存在于疫苗中。雖然gp41的氨基酸序列比gp120的更保守，但gp41含有中和表位。優(yōu)選地，疫苗中存在的任何gp41來自疫苗中存在的HIV分離物。用于疫苗中的來自分離物的gp160可代替疫苗中的gp120或與來自分離物的gp120一起使用。此外，來自不同分離物的gp160與疫苗中的那些相比，可另外存在于疫苗中。
按照本發(fā)明的疫苗也可含有與工程病毒組合的一種或多種可溶性gp120多肽序列或其片段，其中所說的病毒特異性地用于表達(dá)具有引起細(xì)胞毒性T-細(xì)胞效應(yīng)能力的蛋白質(zhì)。合適的工程病毒來源于例如，金絲雀痘病毒，疫苗病毒，稀釋的皰疹病毒(如，皰疹單純病毒)，和水痘帶狀皰疹。如上述，那些典型的工程病毒修飾得表達(dá)任何HIV蛋白質(zhì)(其能誘導(dǎo)細(xì)胞毒性T-細(xì)胞效應(yīng))。通常，在用gp120/人工病毒疫苗進(jìn)行免疫后，施用一劑量或多劑量的gp120多肽序列促進(jìn)免疫應(yīng)答。若需要，病毒可人工做成表達(dá)一種或多種本發(fā)明的gp120多肽序列，或其片段，并與可溶性gp120多肽序列一起或單獨(dú)用于疫苗中。
一般地，疫苗制劑包括在約1.0ml載體內(nèi)的總共大約為300-600μg的gp120。優(yōu)選疫苗制劑包括在兩倍600μg明礬中利用兩倍重量的gp120多肽。然而，也利用更小量的疫苗制劑(例如；每劑量50μg)，一般地與明礬或其它佐劑一起使用。疫苗中任何分離物的gp120的量根據(jù)gp120的免疫原性發(fā)生相應(yīng)變化。例如，來自一些HIV菌株的gp120比來自MN菌株的gp120(SEQ ID NO.29)免疫原性要低一些。如果混合使用具有不同免疫原性的兩種菌株，對每種病毒的量進(jìn)行根據(jù)經(jīng)驗(yàn)的滴定以測定疫苗中每一菌株的百分比。對于具有類似免疫原性的分離物，在疫苗中將采用大約相等量的每種分離物的gp120。例如，在一優(yōu)選實(shí)施方案中，疫苗包括來源于MN和本發(fā)明的一種菌株的gp120，其在約1.0ml的載體中，且濃度為每菌株約300μg。當(dāng)疫苗包括來自30個分離物的gp120時，從約10μg到約50μg的分離物均可加以利用。在多價(jià)疫苗中測定免疫原性蛋白質(zhì)的相對量的方法為大家所熟知，并用來，例如，進(jìn)行檢測在多價(jià)小兒麻痹癥疫苗中的各種分離物的相對比例。
本發(fā)明的疫苗以與現(xiàn)有技術(shù)HIV gp120亞單位施用方法相同的方式進(jìn)行施用。尤其是，根據(jù)方案，通常在0,1，及6,8或12月施用疫苗。一個優(yōu)選的方案包括在0,1,6和12月施用疫苗。在免疫處理后，可進(jìn)行年度或雙年度的增強(qiáng)處理。然而，在免疫過程期間和此后，可以分析中和抗體水平并相應(yīng)調(diào)整方案。
對未感染的個體施用疫苗。此外，可對血清反應(yīng)陽性的個體施用疫苗，如同用現(xiàn)有技術(shù)HIV疫苗，以加強(qiáng)對病毒的免疫應(yīng)答。目前也在探索將編碼用于疫苗的gp120菌株的DNA利用合適載體進(jìn)行施用，以使其在宿主中得以表達(dá)。按這一方法，除去重復(fù)免疫的需要，gp120可由受感染的宿主產(chǎn)生。下文描述gp120表達(dá)載體的制備。
如上述，雖然本文所描述gp120分離物可用作為疫苗，但該氨基酸序列也可單獨(dú)或與制劑中相同類型的氨基酸序列組合使用而作為免疫原，以引起識別存在免疫原中的種種分離物的抗體。免疫原以與病毒相同的方式進(jìn)行配制，且其包括相同的賦形劑等。由這些免疫原誘導(dǎo)的抗體可用于診斷中以檢測免疫原中的HIV菌株或親和純化菌株。gp120多肽序列和趨化因子受體若CD4是HIV-1的初級細(xì)胞內(nèi)受體，它不足以滿足HIV-1進(jìn)入細(xì)胞內(nèi)。已確定出協(xié)同CD4所需要的協(xié)同受體。這些協(xié)同受體是七個跨膜G-蛋白偶聯(lián)受體家族的趨化因子的成員。根據(jù)氨基端半胱氨酸的間距，將趨化因子超家族亞分成兩個組。CXC趨化因子主要涉及中性白細(xì)胞介導(dǎo)的炎癥，CC趨化因子易涉及慢性炎癥。已確定了至少五個CC趨化因子受體，命名為CC-CKR1-5的趨化因子受體(本領(lǐng)域也稱為CCR1-5)，以及至少四個CXC趨化因子受體，命名為CXC-CKR1-4的趨化因子受體(本領(lǐng)域也稱為CXCR-1-4)。
CXC-CKR4(CXCR-4)(其也稱為α-趨化因子受體fusin)是T-細(xì)胞-毒性的HIV-1菌株進(jìn)入的協(xié)同因子。CC-CKR-5(CC-R5)(其也稱為β-趨化因子受體)與其相關(guān)家族的成員如CC-CKR-2b和CC-CKR3一起充當(dāng)巨噬細(xì)胞-毒性的HIV-1菌株進(jìn)入的協(xié)同因子。T-細(xì)胞-毒性的菌株可感染原始T-細(xì)胞和T-細(xì)胞系，但不是巨噬細(xì)胞，而巨噬細(xì)胞-毒性的菌株可感染巨噬細(xì)胞和原始T-細(xì)胞，但不是T-細(xì)胞系。在Deng等自然381:661-666(1996)中更充分地討論了T-細(xì)胞-和巨噬細(xì)胞-毒性的菌株，本文以其整體一并參考。T-細(xì)胞-毒性的菌株的例子包括實(shí)驗(yàn)室分離物，如ⅢB和MN。巨噬細(xì)胞-毒性的菌株包括原始分離物，這些分離物包括但不限于來源于用基于gp120的疫苗進(jìn)行免疫的被接種者的A244,GNE6,GNE8，以及逃逸病毒。二元-毒性的菌株可利用協(xié)同受體的兩種類型，該菌株經(jīng)CXC-CKR-4或經(jīng)一種或多種CC-CKR家族的成員，優(yōu)選為CC-CKR-5,CC-CKR-2b，或CC-CKR-3進(jìn)入細(xì)胞。雖然本發(fā)明不被任何理論約束或限制，但認(rèn)為T-細(xì)胞-毒性和巨噬細(xì)胞-毒性的HIV-1菌株的進(jìn)入提供了一個統(tǒng)一的對各病毒菌株間的細(xì)胞趨性的不同，及對許多CD4-轉(zhuǎn)染的非靈長類細(xì)胞的抗HIV-1感染的抗性，以及對人類群體中的部分個體的抗HIV-1-感染抗性的解釋。
因此，一個實(shí)施方案是包含(1)第一gp120多肽序列或其片段(其來自巨噬細(xì)胞-毒性的HIV-1菌株)和/或第二gp120多肽序列或其片段(其來自T-細(xì)胞毒性菌株)，和(2)逃逸分離物HIV gp120多肽序列或其片段(其來自用第一/或第二HIV gp120多肽序列接種的被接種者)相組合的一種疫苗。優(yōu)選地，疫苗包括至少兩種結(jié)合在不同趨化因子受體上的gp120多肽序列。在一個實(shí)施方案中，疫苗包含結(jié)合在不同趨化因子受體上的第一和第二gp120多肽序列。此外，與被第一和第二gp120多肽序列的二者之一或兩者結(jié)合的趨化因子受體相比，逃逸分離物gp120多肽序列可結(jié)合不同的趨化因子受體。
一種優(yōu)選的T-細(xì)胞毒性菌株是實(shí)驗(yàn)室分離物，最優(yōu)選為MN。用于本發(fā)明的優(yōu)選巨噬細(xì)胞-毒性病毒是GNE6和GNE8，它們是本文所揭示的逃逸病毒的代表物，且由于它們的gp120誘導(dǎo)產(chǎn)生能識別gp120序列(例如，V3結(jié)構(gòu)域)的抗體則不同于MN，其中所說的gp120序列涉及與CC趨化因子受體如CXC-CKR-5的結(jié)合。
如上述，在一個實(shí)施方案中，在疫苗中可通過施用一種或多種趨化因子受體結(jié)合的gp120多肽序列或其片段(其含有合適的趨化因子受體結(jié)合的結(jié)構(gòu)域)以阻止HIV感染。優(yōu)選地，疫苗也包含一種或多種CD4-結(jié)合的gp120多肽序列或其適當(dāng)?shù)钠巍＿@些疫苗可誘導(dǎo)抗HIV的抗體，它們能抑制病毒gp120-趨化因子受體或-CD4的結(jié)合過程。此外，這些gp120多肽可通過與HIV感染有關(guān)的一種或多種協(xié)同受體(如CD4或趨化因子受體)相結(jié)合直接阻止HIV感染，這樣為治療HIV感染提供了預(yù)防和治療作用。優(yōu)選地，這一方面有用的gp120多肽序列含有T-細(xì)胞結(jié)合(TCB)的結(jié)構(gòu)域。
趨化因子受體-結(jié)合性gp120多肽的各種運(yùn)用及關(guān)于CC趨化因子受體家族的討論如下。然而，本領(lǐng)域技術(shù)人員將發(fā)現(xiàn)這一討論同樣適用于在HIV感染中充當(dāng)協(xié)同因子的CXC趨化因子受體。
如用作疫苗或免疫原所描述的，gp120多肽可用作含有一種或多種gp120多肽的組合物。對于可能被感染或需要這種治療(如利用本文所描述的施用劑量和途徑及方法進(jìn)行治療)的患者而言，可預(yù)防性地或治療性地施用該組合物。然而，優(yōu)選慢性施用且因此調(diào)整劑量。據(jù)記載在體內(nèi)施用也可誘導(dǎo)能結(jié)合病毒gp120且進(jìn)一步抑制病毒與CC-CKR的結(jié)合的抗體。
gp120多肽也可用于篩選試驗(yàn)以確定CC-CKR的拮抗劑。例如，由于候選拮抗劑能抑制gp120與CC-CKR CC-CKR受體的結(jié)合因而可篩選出候選拮抗劑，其中所說的受體是分離的并附著在物體表面上(例如，塑料培養(yǎng)皿)或經(jīng)重組或天然地表達(dá)在細(xì)胞的表面上。拮抗劑也可結(jié)合gp120或結(jié)合受體。理想的候選拮抗劑包括能結(jié)合受體的gp120化合物，gp120小肽(長度為5-20個氨基酸，優(yōu)選為7-10個氨基酸)或gp120的肽模擬物(peptidomimetics)，和結(jié)合gp120的單克隆抗體，以及能結(jié)合gp120或受體的有機(jī)小分子。
由gp120多肽誘導(dǎo)的抗體也可用來誘導(dǎo)能結(jié)合CC趨化因子的抗自生類型抗體。這些抗自生類型的抗體可通過其與抗gp120多肽抗體的結(jié)合且其能抑制gp120與CC-CKR受體的結(jié)合而進(jìn)行篩選。這些抗自生類型抗體通過結(jié)合CC-CKR受體上而模擬gp120。這樣的抗體，優(yōu)選人類抗體，可由許多方法獲得，如來自于復(fù)合文庫(例如，Burton等，免疫學(xué)進(jìn)展(1994)57:191-280)的人類抗體。現(xiàn)在有可能制備轉(zhuǎn)基因動物(例如，小鼠)，在免疫方面，這些轉(zhuǎn)基因動物在缺乏內(nèi)源性免疫球蛋白產(chǎn)物的情況下能夠產(chǎn)生所有的人類抗體。例如，嵌合的和胚系突變體小鼠中的抗體重鏈連接區(qū)(JH)的基因發(fā)生純合缺失，則可完全抑制內(nèi)源性抗體的產(chǎn)生。如Jakobovitis等，美國國家科學(xué)院院報(bào)90:2551(1993)；Jakobovits等，自然362:255-258(1993)；Bruggermann等，年度免疫學(xué)7:33(1993)所描述的，將人胚系免疫球蛋白的基因排列轉(zhuǎn)移到這樣的胚系突變體小鼠中，則可導(dǎo)致挑戰(zhàn)抗原的人類抗體的產(chǎn)生。
此外，可利用McCafferty等，自然348:552-553(1990)所描述的噬菌體顯示技術(shù)，在體外由未接受免疫的供體提供的免疫球蛋白可變(V)結(jié)構(gòu)域全部基因來制備人類抗體和抗體片段。根據(jù)這一技術(shù)，閱讀框中的任何的抗體V結(jié)構(gòu)域基因被關(guān)閉成絲狀噬菌體的主要或次要的外被蛋白質(zhì)基因，如M13或fd，并且在噬菌體微粒的表面作為功能抗體片段。因?yàn)榻z狀微粒含有噬菌體基因組的單鏈DNA拷貝，基于抗體的功能性質(zhì)的選擇導(dǎo)致編碼具有那些性質(zhì)的抗體的基因的選擇。如Johnson等，結(jié)構(gòu)生物學(xué)的當(dāng)前觀點(diǎn)3:564-571(1993)所綜述的，可以各種形式進(jìn)行噬菌體顯示操作。
V-基因片斷的幾種來源均可用于噬菌體顯示操作。Clackson等，自然352:624-628(1991)從來源于免疫小鼠脾的V-基因小型隨機(jī)復(fù)合文庫中分離出抗oxazolone抗體的多樣化的排列基因?？蓸?gòu)建來源于未免疫的人供體(或早期細(xì)胞)的全部V基因組。目前已有闡明，對應(yīng)于多樣化排列的抗原(包括自體抗原)的抗體可通過按照Marks等，分子生物學(xué)雜志222:581-537(1991)，或Griffith等，EMBO J.12:725-734(1993)中所描述的技術(shù)進(jìn)行分離。
在天然的免疫應(yīng)答中，抗體基因以高速率積累突變(體細(xì)胞高變)。一些引起更高親和性的變化以及具有高親和性表面的免疫球蛋白的B細(xì)胞，在爾后的抗原攻擊期間，優(yōu)選復(fù)制并得以分化。這一自然過程可通過利用被稱為的＂鏈重組＂技術(shù)(Marks等，生物技術(shù)10:779-783[19921])進(jìn)行模擬。在這一方法中，通過噬菌體顯示技術(shù)所獲得的＂原始＂人類抗體的親和性可通過用V結(jié)構(gòu)域基因(其來源于未免疫供體)的自然發(fā)生的變體(全部)的全部基因依次取代重鏈和輕鏈的V結(jié)構(gòu)域的基因得以提高。這一技術(shù)使得可在nM范圍內(nèi)產(chǎn)生具有親和性的的抗體和抗體片段。用于制備大噬菌體抗體全體的策略見Waterhouse等，核酸研究21:2265-2266(1993)。
因此，如同上述的利用gp120單克隆抗體(其抑制gp120結(jié)合在受體上)，在復(fù)合文庫或其他系統(tǒng)中結(jié)合CC-CKR的抗體可通過篩選在噬菌體表面表達(dá)的抗體或其片段獲得。
除利用gp-120抗體篩選抗體之外，隨機(jī)或復(fù)合肽文庫也可利用任何本發(fā)明的gp120抗體或gp120化合物進(jìn)行篩選。有效的方法可從含有1百萬至十億種不同序列的收集大量肽的文庫中確定出肽配體，其中所說的不同序列可利用單克隆抗體或靶分子進(jìn)行篩選。這一技術(shù)的功效來自于與文庫中大序列所偶聯(lián)的氨基酸的化學(xué)多樣性。參見實(shí)施例，Scott等，Cur.Open.生物技術(shù)5(1):40-8(1994)；Kenan等，生物化學(xué)科學(xué)潮流(1994)19(2):57-64。因此，如本文討論產(chǎn)生的單克隆抗體(優(yōu)選為人類單克隆或其片段)對于抑制或治療HIV感染或疾病，及確定附加藥物的篩選試驗(yàn)有利用價(jià)值。gp120的制備如同現(xiàn)有技術(shù)的HIVgp120亞單位疫苗，用于疫苗的gp120可用任何合適的方法進(jìn)行制備。重組制備的或化學(xué)合成的gp120，出于安全考慮優(yōu)選從HIV直接分離的gp120。用于制備gp120重組體的方法描述如下。
編碼來源于逃逸分離物的gp120和能夠表達(dá)gp120的寡核苷酸可由常規(guī)方法制備。例如，可合成核苷酸序列。此外，編碼gp120的另一個HIV核苷酸序列可用作骨架，并且如同通過定點(diǎn)誘變改變可改變?nèi)魏尾煌臍埢６c(diǎn)誘變的描述見Kunkel等，美國國家科學(xué)院院報(bào)82:488-492(1985)和Zoller等，核酸研究10:6487-6500(1982)，且其為大家所熟知。
在一個優(yōu)選實(shí)施方案中，核苷酸序列存在于含有編碼gp120并在所編碼蛋白質(zhì)的表達(dá)啟動子的轉(zhuǎn)錄和翻譯控制之下的DNA的表達(dá)構(gòu)建體中。在哺乳動物的細(xì)胞中，啟動子可以是用于表達(dá)的真核啟動子。若人們希望擴(kuò)充啟動子或在原核宿主中產(chǎn)生gp120，啟動子可是原核啟動子。通常，利用強(qiáng)啟動子以提供高水平的轉(zhuǎn)錄和表達(dá)。
表達(dá)構(gòu)建體可以是載體的一部分，其在適當(dāng)?shù)募?xì)胞宿主中能在染色體外穩(wěn)定存在或整合到宿主基因組中。通常，為表達(dá)構(gòu)建體提供標(biāo)記，這便于對含有構(gòu)建體的宿主的選擇。標(biāo)記可位于相同或不同的DNA分子上，合乎需要的為相同的DNA分子。
可將表達(dá)構(gòu)建體連接到為所需宿主細(xì)胞識別的復(fù)制系統(tǒng)上。各種復(fù)制系統(tǒng)包括病毒復(fù)制系統(tǒng)諸如來源于逆轉(zhuǎn)錄病毒，猴病毒，牛乳頭瘤病毒，或類似的那些系統(tǒng)。此外，構(gòu)建體可連接到擴(kuò)增基因上，例如DHFR基因，那么可產(chǎn)生gp120 DNA若干拷貝。將構(gòu)建體導(dǎo)入到宿主內(nèi)隨構(gòu)建體而發(fā)生變化，且可采用任何方便的方法來完成。為表達(dá)蛋白質(zhì)可采用許多原核與真核宿主。
優(yōu)選地，gp120在哺乳動物細(xì)胞中表達(dá)，因?yàn)檫@些細(xì)胞提供了與天然gp120相同的糖基化和二硫鍵。gp120和gp120片段(其作為與單純皰疹病毒1(HSV-1)的糖蛋白D(gD-1)的N端序列相結(jié)合的融合蛋白)在哺乳動物中的表達(dá)分別見Lasky L.A.等1986所描述的(被重組體包膜糖蛋白的抗體中和的愛滋病逆轉(zhuǎn)錄病毒)科學(xué)233:209-212和Haffar,O.K.等，1991(HIV-1gp160的胞質(zhì)尾區(qū)含有與細(xì)胞膜相連的區(qū)域)病毒學(xué)180:439-441。用于表達(dá)gp120的優(yōu)選方法見實(shí)施例的描述。在實(shí)施例中，異源信號序列便于蛋白質(zhì)的表達(dá)。然而，也可利用天然信號序列表達(dá)蛋白質(zhì)。
通過常規(guī)的方法可產(chǎn)生具有一種表1中所說明的氨基酸序列的分離的，純化的gp120多肽。例如，這些可化學(xué)合成蛋白質(zhì)。在一個優(yōu)選的實(shí)施方案中，利用本發(fā)明的表達(dá)構(gòu)建體，在哺乳動物細(xì)胞中表達(dá)蛋白質(zhì)。用常規(guī)方法可純化被表達(dá)的蛋白質(zhì)。一個優(yōu)選的純化方法見實(shí)施例中的描述。gp120片段本發(fā)明也提供了適于誘導(dǎo)用于疫苗的抗體的的gp120片段。如本文所使用的，一種截?cái)嗟膅p120沒有完整的gp120序列的部分(其始于gp120的氨基或羧基末端的任一端)。本發(fā)明的截短gp120序列沒有C5結(jié)構(gòu)域。gp120的C5結(jié)構(gòu)域是分子的一個主要的免疫原性位點(diǎn)。然而，對該區(qū)的抗體不中和病毒。因此，從誘導(dǎo)用于血清分類的抗體的免疫原中消除gp120的這一部分是有利的。
在另一個實(shí)施方案中，截短的gp120序列還沒有g(shù)p120V5結(jié)構(gòu)域，從羧基末端區(qū)至約453個氨基酸殘基。保持在序列中的V5結(jié)構(gòu)域部分為制備表達(dá)構(gòu)建體的便利的限制酶切位點(diǎn)。然而，沒有整個gp120V5結(jié)構(gòu)域的截短gp120也適于用于誘導(dǎo)抗體。
此外，也可從截短的gp120序列中除去gp120的氨基末端部分。尤其是，截短的gp120序列可沒有g(shù)p120信號序列。截短的gp120序列可沒有g(shù)p120C1結(jié)構(gòu)域，從由羧基末端至第111個氨基酸殘基，且除去了大多數(shù)C1結(jié)構(gòu)域但是保存一個方便的限制酶切位點(diǎn)。然而，經(jīng)過V2半胱氨酸殘基(其形成一個二硫鍵)的C1結(jié)構(gòu)域部分可另外被除去，那么截短的gp120序列從gp120C1結(jié)構(gòu)域的羧基末端至第117個氨基酸殘基。在一個優(yōu)選實(shí)施方案中，截短的gp120序列從gp120氨基末端至C1結(jié)構(gòu)域的第111個殘基和第453個殘基至gp120的羧基末端。
如上述，可由重組工程制備截短的gp120序列。方便地，將編碼截短的gp120序列的DNA連接到編碼信號序列的異源DNA序列上。
人們認(rèn)為，在本文的教導(dǎo)下，對于本發(fā)明的內(nèi)容在具體問題或情況中的應(yīng)用在本領(lǐng)域普通技術(shù)人員的能力之內(nèi)。本發(fā)明的產(chǎn)品和其分離、使用及制造的代表性方法的實(shí)施例如下，但是本發(fā)明不限于這些。本文所引用的所有文獻(xiàn)在此一并作為參考。
實(shí)施例材料和方法收集自志愿者的樣品。血液樣品從MN-rgp120免疫個體收集，這些個體在參加國家衛(wèi)生協(xié)會(NIH)發(fā)起的HIV-1疫苗Ⅰ期(NIH方案AVEG016)和Ⅱ期(NIH方案AVEG201)試驗(yàn)時，受到HIV-1的感染。研究過程中的受試驗(yàn)者人口統(tǒng)計(jì)以及研究方法設(shè)計(jì)由McElrath(生物癌癥討論會，6:1-11,1995)和McElrath等(愛滋病的國家協(xié)同疫苗發(fā)展組織的第八年度會議摘要，Bethseda,MD216,1996)進(jìn)行了描述。按照參與機(jī)構(gòu)協(xié)會考察委員會批準(zhǔn)的一個討論通過方案獲得樣品。試驗(yàn)階段，HIV-1的感染時間根據(jù)NIH艾滋病疫苗評估網(wǎng)提供的數(shù)據(jù)具體確定，試驗(yàn)中PCR和/或血清分析用于檢測HIV-1的感染。
用于克隆HIV-1包膜糖蛋白的樣品的制備。通過FICOLL-HYPAQUE梯度離心法，從肝素化靜脈血液制備HIV-1感染疫苗的外周血液單核細(xì)胞(PBMCs)。確定細(xì)胞數(shù)量和生存力。分離后，在磷酸鹽緩沖鹽水中洗滌PBMCs兩次，并以6x106細(xì)胞/ml的密度懸浮于PCR溶解緩沖液(50mM KCl,10mM Tris(PH8.4),2.5mM MgCl2,0.1mg/ml明膠(Sigma),0.45％NONIDET p40清洗劑，0.45％TWEEN20清洗劑(兩種清洗劑都可從美國生化公司買到)和溶解細(xì)胞的0.06mg/ml的蛋白酶K(Gibco BRL))中。在50-60℃條件下培養(yǎng)溶解物1小時，其后在95℃條件下滅活蛋白酶10分鐘。轉(zhuǎn)移和凍結(jié)溶解產(chǎn)物，在-70℃條件下儲存以備使用。
聚合酶鏈?zhǔn)椒磻?yīng)(PCR)擴(kuò)增。利用下述的嵌合引物對樣品進(jìn)行兩輪的PCR擴(kuò)增反應(yīng)。在第一輪反應(yīng)中，將25μl的PBMC溶解產(chǎn)物等分試樣(包含大約1μg的基因組DNA)與等量的PCR反應(yīng)混合物(包含400μM的每種dNTP、200Cμg/ml BSA(Sigma化學(xué)公司，RIA級)以及在50mM KCl、20mM Tris(pH8.4)和3mM MgCl2中的大約100pmole的每種引物)混合。95℃條件下變性步驟開始10分鐘之后，在55℃的浸透步驟中加入5個單位的標(biāo)記聚合酶(AMPLITAQ,PekinElmer Cetus)，并用礦物油覆蓋樣品。
PCR流程如下2個55℃下1分鐘、72℃下2.5分鐘和94℃下1分鐘的循環(huán)，隨后是28個55℃下30秒、72℃下45秒和94℃下2.5分鐘的循環(huán)，以及一個72℃下5分鐘的延伸步驟。
利用以上或PCR優(yōu)化試劑盒(Invitrogen)中描述的試劑和流程，在最后100μl的反應(yīng)量中用合適的嵌合引物再擴(kuò)增第一輪反應(yīng)物的10μl等分試樣。用QIAQUICK-旋轉(zhuǎn)體(Riagen公司)純化PCR反應(yīng)物。第一輪反應(yīng)中使用的引物對是正向的120.os.F(在HIVPV22 6248-6270位置的同源序列5′-gggaattcggatccAGAGCAGAAGACAGTGGCAATGA)(SEQ.ID.NO.34)或者JM11A(在6048-6074位置的5′-ctcgag-CTCCTGAAGACAGTCAGACTCATCAAG)(SEQ.ID.NO.35)[Kusumi等；病毒學(xué)雜志66:875(1992)]與反向的120.os.R(在7836-7859位置的5′-ggtctagaagctttaGC CCATAGTGCTTCCTGCTGC)(SEQ.ID.NO.36)組合。內(nèi)部的嵌合引物是在6389-6410位置的120.BX.F(5′-gggcggatcctcgaGGTACCTGTRTGGAAAGAAGCA.R:A或G)(SEQ.ID.NO.37)和120.is.R(在7819-7841位置的5′-ggtctagaagctttaTGCTC CYAAGAACCCAAGGAA；Y:T或C)(SEQ.ID.NO.38)。異源引物序列以小寫字母表示。
PCR產(chǎn)物的亞克隆和作為融合蛋白的重組包膜糖蛋白的表達(dá)。HIV-1包膜糖蛋白gp120序列作為嵌合的基因和融合蛋白被克隆和表達(dá)，其中與成熟gp120序列的氨基酸13相對應(yīng)，來自單純皰疹病毒類型1(HSV-1)成熟N端的信號序列和27個氨基酸與gp120基因的N端序列融合。通過人工引入異源PCR引物末端的限制位點(diǎn)和人工引入HSV-1gD的N端序列的XhoI位點(diǎn)組合，將包含逃逸樣品gp120序列的PCR產(chǎn)物作為嵌合基因克隆進(jìn)pRKS表達(dá)質(zhì)粒。
用測序酶和dGTP試劑盒(美國生化公司)對所得的雙鏈DNA進(jìn)行測序。來自糖蛋白D的序列用來加強(qiáng)表達(dá)并為方便的蛋白質(zhì)分析提供標(biāo)記表位，如以下文章中所描述的Berman等；病毒學(xué)雜志.7:4464-9(1932)；Nakamura等；艾滋病和人類逆轉(zhuǎn)錄病毒1875-85(1992)；以及Nakamura等；病毒學(xué)雜志67:6179-91(1993)。
簡言之，通過PCR反應(yīng)所產(chǎn)生的分離DNA片段連接進(jìn)入一質(zhì)粒(pRK.gD-5,pRKgDstop)，在讀框中，該質(zhì)粒用來將gp120片段融合于1型單純皰疹病毒(gD-1)的糖蛋白D(gD)基因5′序列并朝向翻譯終止密碼子的3′端。gD-1基因片段編碼成熟形式HSV-1蛋白質(zhì)的信號序列和25位氨基酸。為了能夠?qū)崿F(xiàn)在哺乳動物細(xì)胞中的表達(dá)，嵌合的基因片段經(jīng)克隆進(jìn)入pRKS表達(dá)質(zhì)粒(Eaten等，生物化學(xué)291:8343-8347(1986))，該質(zhì)粒包含一具有克隆位點(diǎn)和翻譯終止密碼子的多接頭，克隆位點(diǎn)和翻譯終止密碼子位于巨細(xì)胞病毒促進(jìn)劑和猴病毒40病毒聚腺苷酸化位點(diǎn)之間。
利用磷酸鈣技術(shù)(Graham等，病毒學(xué)52:456-467(1973))，將所得的質(zhì)粒轉(zhuǎn)染到293s早期人類腎細(xì)胞系(Graham等，基因病毒雜志36:59-77(1977)。轉(zhuǎn)染48小時后，收集生長調(diào)節(jié)細(xì)胞培養(yǎng)物介質(zhì)，通過ELISA方法或特定的放射性免疫沉淀法檢測可溶性蛋白質(zhì)。放射性免疫沉淀法，是將細(xì)胞培養(yǎng)上清液中的代謝標(biāo)記蛋白進(jìn)行十二烷基硫酸鈉聚丙烯酰胺凝膠電泳(PAGE)分離，并通過如Berman(病毒學(xué)雜志63:3489-3498(1989))和Laemmli(自然，227:680-685(1970))描述的放射自顯影術(shù)使其顯現(xiàn)。
血清測定。利用Berman等(病毒學(xué)雜志7:4464-9(1992))、Nakamura等(愛滋病和人類逆轉(zhuǎn)錄病毒8:1875-85(1992))和Nakamura等(病毒學(xué)雜志67:6179-91(1993))描述的測定方法進(jìn)行血清測定，目的是檢測rgp120抗體，與gp120V3結(jié)構(gòu)域序列對應(yīng)的合成gp120V3結(jié)構(gòu)域肽抗體，以及能夠阻斷MN-rgp120結(jié)合到細(xì)胞表面CD4的抗體。利用連續(xù)三倍稀釋的血清液確定與gp120和V3肽結(jié)合的抗體的終點(diǎn)滴度。定義終點(diǎn)稀釋液滴度為比產(chǎn)生1∶50稀釋的、集合的正常人類血清光密度值高兩倍的光密度值?？贵w滴度由計(jì)算機(jī)程序計(jì)算，這種程序在抗體稀釋液之間進(jìn)行插值運(yùn)算。陽性對照標(biāo)準(zhǔn)血清的變異的中間測定(inter-assay)系數(shù)是35％。
單克隆抗體從逃逸病毒結(jié)合到rgp120。一種類似于Moore等(愛滋病3:155-63(1989))描述的ELISA用來測量把各種單克隆抗體(MAbs)從逃逸病毒結(jié)合到rgp120s上。簡言之，在Nunc-免疫平板(Maxisorp，鑒定過的)上涂上(100μg,4℃條件下在5μg/ml PBS中放置過夜)一層親和純化的抗在gp120(D7324，國際酶，F(xiàn)allbrook，加拿大)C末端上的肽的綿羊多克隆抗血清。在用PBS-0.05％TWEEN-20的清洗劑洗滌一次之后，在室溫下用PBS-1.0％BSA封阻平板30-60分鐘。將293s細(xì)胞的細(xì)胞培養(yǎng)上清液稀釋以便含有等量的gD-rgp120融合蛋白，加入到平板上并在室溫下培養(yǎng)2小時，隨后用PBS-0.05％TWEEN-20清洗劑洗滌三次。將各種MAbs在PBS-1.0％BSA中稀釋，將稀釋的MAbs加到每孔中并在室溫下培養(yǎng)1小時。
洗滌平板3次，在室溫下用辣根過氧化物酶軛和物(山羊抗鼠類或者抗人類IgG,Cappel)第二抗體培養(yǎng)1小時。洗滌3次之后展開平板，在平板讀數(shù)計(jì)中讀出OD492(492nm處的光密度)。通過稀釋標(biāo)準(zhǔn)化生長調(diào)節(jié)的細(xì)胞培養(yǎng)上清液，稀釋方法以MAb5B6與HSV-1糖蛋白D融合蛋白的特異性結(jié)合為基礎(chǔ)。
病毒中和測定。在細(xì)胞病變測定中測定疫苗血清通過HIV-1MN抑制MT4細(xì)胞感染的能力，測定中用一種測熱指示劑染料定量細(xì)胞生存力，如Robertson等所描述的方法(病毒學(xué)方法雜志20:195-202(1988))。簡言之，以澄清的上清液從慢性傳染的H9/HIV-1細(xì)胞培養(yǎng)液制備HIV-1MN的病毒貯存液(從Dr.Michael Norcross，美國食品與藥品管理署獲得)。離心受HIV-1MN慢性感染的H9細(xì)胞并重懸于十分之一的原體積量培養(yǎng)物中。正如Wrin等(病毒學(xué)雜志69:39-48(1995))描述的快速旋轉(zhuǎn)細(xì)胞方法，通過機(jī)械剪切作用釋放出細(xì)胞關(guān)聯(lián)病毒。
將足以確保在7天中完全溶解殺死細(xì)胞的數(shù)量的病毒與試驗(yàn)抗血清的三倍連續(xù)稀釋液一起培養(yǎng)，隨后用于在10％的FCS/RPMI-1640細(xì)胞培養(yǎng)介質(zhì)中攻擊MT4T-淋巴細(xì)胞。培養(yǎng)物在37℃、5％CO2條件下培養(yǎng)7天，然后用MTT染色檢測細(xì)胞生存力，如Robertson等所描述的方法(病毒學(xué)雜志methods20:195-202(1988))。通過測定570-650nm處的OD值定量化病毒中和終點(diǎn)，然后計(jì)算終點(diǎn)滴度作為抗血清稀釋倍數(shù)的倒數(shù)，該抗血清稀釋液給出具有未保護(hù)的(殺滅的)細(xì)胞的對照信號兩倍以上的信號。典型地，這些滴度是50％保護(hù)條件下計(jì)算值的兩倍。
結(jié)果受感染者的免疫歷史。自1992以來，有499個成年人接受了MN-rgp120免疫處理，第Ⅰ期試驗(yàn)中是在低等或中等風(fēng)險(xiǎn)個體中進(jìn)行，第Ⅱ期臨床試驗(yàn)中包括中等到高等危險(xiǎn)個體。這里談及的研究過程，即對九個個體中頭七個的遺傳和免疫特征的跟蹤，這些個體在試驗(yàn)期間通過高危行為成為HIV-1的感染體。表2A給出了一系列試驗(yàn)處理結(jié)果和概括了被接種者狀態(tài)，表2B給出了一系列疫苗分析結(jié)果。
表2A對用MN-rgp120免疫后受HIV-1感染的被接種者的描述研究號病例號*風(fēng)險(xiǎn)群體 #抗原劑量/佐劑016 C6M/H300/QS21016 C8M/H600/QS21016 C15 M/H300/QS21201 C7M/H600/Alum201 C11 M/H600/Alum201 C10 M/IDU 600/Alum201 C17 M/IDU 600/Alum*M/H表示男性同性戀者；M/IDU表示男性靜脈內(nèi)用毒者。
#數(shù)字表示每次免疫處理注入的MN-rgp120以微克表示的劑量QS21表示配制在QS21佐劑中的抗原；Alum表示配制在氫氧化鋁中的MN-rgp120。
表2B對用MN-rgp120免疫后受HIV-1感染的被接種者的描述病例號 注射進(jìn)程HIV-1+前HIV-1+的□HIV-1+(月) 注射次數(shù)時間(月)間隔(月)C6 0,1,10,5 24.002.00C8 0,124.003.00C150,1,2 36.254.00C7 0,1,6,12 39.253.00C110,1,6,12 419.50 6.75C100,1,6,19 319.50 13.50C170,1,6,18 424.75 6.25表示最后一次免疫與檢測到HIV-1感染之間的間隔
3個感染出現(xiàn)在Ⅰ期試驗(yàn)(NIH方案AVEG201)中，該試驗(yàn)比較配制在兩種不同佐劑(alum和QS21)中的MN-rgp120的安全性與免疫原性。4個感染出現(xiàn)在Ⅱ期試驗(yàn)中，該試驗(yàn)的目的是在各種高危人群(靜脈用藥者，同性戀者和兩性男性者，HIV-1感染個體的性伙伴)中建立MN-rgp120安全性和免疫原性。
在研究的7個感染(表3)中，兩個(C4和C8)在兩次注射之后出現(xiàn)，三個(C7,C10和C15)在三次注射之后出現(xiàn)，兩個(C11和C17)在接受四次預(yù)定的注射之后出現(xiàn)。接受最后一次免疫與形成感染之間的間隔在2到13.5個月之間。
表3感染HIV-1的被接種者中峰值后加強(qiáng)MN-rgp120抗體滴度注射C6C8C15C7C11C10C171 ＜50 2185 79 ＜50 1890 na na2 21539 10125 na 413 32696 7771 70563 # # 4460 9707 34728 11627 184134 R # # # # # 11340#因受HIV-1感染未分析樣本。
na表示不能用于檢測的樣品。
黑體表示非常低的抗體滴度。
免疫個體中對gp120的抗體反應(yīng)。對所有接受了免疫過程的感染個體，通過ELISA法測定其對MN-rgp120的抗體反應(yīng)的大小與特異性(圖1)。七個受測驗(yàn)者中五個顯示正常的抗體反應(yīng)動力學(xué)過程，其包括一種小的但具有再現(xiàn)性的初次反應(yīng)(1∶100-1∶2,000)和一種強(qiáng)烈的二次反應(yīng)(加強(qiáng))效應(yīng)(滴度范圍1∶7,000-1∶32,000)，以及跟隨第三和第四次注射的抗體反應(yīng)，這兩次反應(yīng)類似于或者稍強(qiáng)于第二次免疫后的反應(yīng)(圖1，表3)。
在C7(圖1C)中觀察到的抗體反應(yīng)不尋常，因?yàn)樵诘谝淮巫⑸浜髾z測不到抗體，且在第二次注射后檢測的滴度僅為1∶350。這樣似乎是C7對首次免疫無反應(yīng)，在第二次注射之后獲得的抗體反應(yīng)代表初次免疫反應(yīng)。與這一假說相一致，第三次注射僅僅產(chǎn)生1∶9,707的滴度，例外于那些通常在兩次免疫之后觀察到的滴度。
從受測驗(yàn)者C15(圖1G)也觀察到一種非典型抗體反應(yīng)，其接受免疫過程根據(jù)一個加速的免疫進(jìn)度表(0、1以及2個月)來進(jìn)行。正如所料，從該受試驗(yàn)者觀察到的抗體滴度(1∶4,460)，位于兩次免疫后觀察到的滴度范圍的低值一端，并且遠(yuǎn)低于三次免疫后的正常值。C15的第三次免疫后缺乏有效的加強(qiáng)反應(yīng)并不奇怪，因?yàn)榍懊娴难芯窟^程在狒狒體內(nèi)進(jìn)行加速的0、1、2月免疫進(jìn)程[Anderson等，211傳染病學(xué)雜志.160:960-9(1989)]，類似于延長了第二次反應(yīng)以及不能引起有效的第三次加強(qiáng)反應(yīng)。
后來對受試驗(yàn)者C6(圖1A)和C8(圖1B)的血清和血漿分析結(jié)果表明，他們在第二次和第三次免疫之間的某點(diǎn)被HIV-1感染。gp120抗體測定中HIV-1感染的血清證明十分明顯，在第二次注射后兩周滴度未減少，相反形成一非特性的高滴度峰值(圖1A和1B)。在第三次注射后MN-rgp120滴度中出現(xiàn)類似的峰值，表明受試驗(yàn)者C7在36周左右，即大約在接受第三次注射(圖1C)后16周受到感染。在gp120滴度中，受試驗(yàn)者C10(圖1E)、C11(圖1D)、C15(圖1G)以及C17(圖1F)在第三次或者第四次免疫后均出現(xiàn)未預(yù)料到的增值，以HIV-1感染最為典型。所獲得的數(shù)據(jù)顯示用于MN-rgp120抗體反應(yīng)的免疫引發(fā)不足以提供免受HIV-1感染的普遍保護(hù)。
抗V3結(jié)構(gòu)域的抗體滴度。為了進(jìn)一步說明抗gp120的抗體反應(yīng)的特征，對一包含主要中和決定簇(PND)的MN-rgp120，測定其合成V3結(jié)構(gòu)域肽的抗體滴度。七個受試驗(yàn)者中五個在第二次免疫后呈現(xiàn)好的V3滴度(1∶400到1∶4000)，然而另外兩個受試驗(yàn)者(C7和C15)卻在出現(xiàn)有效滴度(圖1C和1G)前需要進(jìn)行第三次免疫。如前面所觀察到的(C11)，一些個體(例如C11、C10、C17)的V3滴度峰值似乎隨著每一連續(xù)的免疫過程而下降(圖1D、1E和1F)。在HIV-1感染之后，可觀察到兩種V3反應(yīng)模式。三個受試驗(yàn)者(C6、C7和C10)的V3結(jié)構(gòu)域肽(圖1A、1C和1E)滴度呈現(xiàn)較大幅度的增加，而C8(圖1B)顯示較大幅度的減少。在分析的時候，數(shù)據(jù)不足以得出結(jié)論認(rèn)為對受試驗(yàn)者C11、C15和C17的試驗(yàn)中，V3滴度的變化是對HIV-1感染的反應(yīng)。
所獲得的結(jié)果表明先于HIV-1感染在各時間點(diǎn)以V3結(jié)構(gòu)域形成抗體反應(yīng)的能力，不是一種有效相關(guān)的抵抗所有HIV-1菌株的保護(hù)免疫性。
CD4阻斷滴度。阻斷gp120結(jié)合CD4的抗體代表異種類別病毒中和抗體。已知其中一些能夠結(jié)合gp120的C4結(jié)構(gòu)域[Nakamura等；病毒學(xué)雜志67:6179-91(1993)；Anderson等；傳染病雜志.160:960-9((1983)]，另外一些能夠識別不連續(xù)表位決定的構(gòu)象[Berman等；病毒學(xué)雜志7:4464-9(1932)；Nakamura等；病毒學(xué)雜志67:6179-91(1933)；McKeating等；愛滋病研究和人類逆轉(zhuǎn)錄病毒8:451-9(1992)；Ho等；病毒學(xué)雜志。65:489-93(1991)；Barbas等；美國國家科學(xué)院院報(bào)91:3809-13(1994)]。
檢測兩種類型表位抗體的方法，是測定被接種者血清阻斷[125I]-標(biāo)記gp120結(jié)合到細(xì)胞表面CD4的能力[[Nakamura等；愛滋病和人逆轉(zhuǎn)錄病毒81875-85(1992)；Nakamura等；病毒學(xué)雜志67:6179-91(1993)]。在感染(圖2)之前，測定所有七位被接種者的CD4阻斷滴度，其滴度峰值范圍為1∶10-1∶300。在感染前的最后一時間點(diǎn)上，七個被接種者中五個的CD4滴度很低(1∶30或者更少)。然而一個被接種者(C17)卻在感染(圖2F)之前具有大約1∶300的CD4阻斷滴度。這樣，缺乏阻斷MN-rgp120結(jié)合到CD4的抗體不能說明所有感染過程。在受HIV-1感染之后，從七個受試驗(yàn)者的五個中可觀察到其CD4阻斷滴度(1∶100-1∶1,000)有較大幅度的增加。這五個被接種者包括C6、C7、C8、C10和C11。這些結(jié)果顯示由MN-rgp120引起的CD4阻斷滴度低于天然感染引起的CD4阻斷滴度。]病毒中和活性。利用一種比色分析法測定MN-rgp120免疫受試驗(yàn)者抗血清的病毒中和活性，即測定抗體(用病毒處理過)培育后的MT4細(xì)胞的生存力。由于不知道任一逃逸感染者的實(shí)際受感染日期，同時也不經(jīng)常地收集血清樣品，所以也不知道任一被接種者在受感染時抗體中和效應(yīng)的大小。
在檢驗(yàn)的七個受感染體中，最靠近感染時間的血清樣品是從C7獲得的樣品，在檢測到受HIV-1感染之前，其對HIV-1MN的中和滴度1∶15存在了三周(表4)。然而，在其它情況中，最后一次注射和感染時間之間的間隔是10到25周。
表4HIV-1感染的被接種者的血清的中和活性周 C6 C8 C15 C7 C11 C10C170 ＜10＊＜10＊＜10＊＜10＊＜10＊＜10＊＜10＊2 ＜10＜10＜10 -- - -4 ＜10＊＜10＊nd＊＜10＊＜10＊＜10＊＜10＊6 10 80 - ＜10 30 150 1508 - - nd＊- ---10- - 35 - ---15- - - ＜10 ---16 150#250#-- 30 10 ＜1024 150# ＜10＊20＊＜10＊＜10＊2670 500 200 40030- - 40 1003315 -- -35- 100 - -3630# - 10 405230＊＜10＜1054250 --57100 --6390 --64- - ＜107740# --78 500# 10＊80 10084 60
90 150104 150#*表示免疫。
#表示HIV-1陽性。
nd表示未試驗(yàn)。
-表示不能獲得樣品。
檢驗(yàn)兩個早期受感染個體的血清(表4)，發(fā)現(xiàn)一個個體(C6)在先于HIV-1感染檢測的十周內(nèi)具有的中和滴度峰值為1∶10，而另一個體(C8)在先于HIV-1感染檢測的十周內(nèi)具有的中和滴度峰值為1∶80。受試驗(yàn)者C15，其接受的免疫過程是根據(jù)一個加速進(jìn)度表進(jìn)行的，在先于HIV-1感染檢測的14周，第三次注射后其中和滴度為1∶35。受試驗(yàn)者C10，其在第三次免疫后(第24周)具有的中和滴度峰值為1∶200，在52周，先于第一次指示HIV-1感染的六個月時(第78周)沒有任何可檢測的滴度。
受試驗(yàn)者C11在先于HIV-1的檢測的十四周具有中和滴度1∶90，和第三次免疫后的滴度峰值1∶500。類似地，被接種者C17在先于HIV-1的檢測的十四周具有中和滴度1∶150，和第三次免疫后兩周的滴度峰值1∶400。
基于gp120效應(yīng)的衰變速率約兩個月時間[Belshe等；JAMA272(6):475-80(1994)]，十周內(nèi)從被接種者C10和C17觀察的中和滴度從1∶150衰減為1∶10，在最后的感染前血清樣品和HIV-1檢測時間間隔之間，在C8、C15、C11以及C17中的滴度似乎可以衰減到1∶10或更少。
這些研究結(jié)果顯示了所有的被接種者均在先于HIV-1感染的某些時間發(fā)展一定水平的病毒中和抗體，并且中和效應(yīng)在感染時有可能很小。一般來說，在受HIV-1感染個體中觀察到的病毒中和效應(yīng)的大小，與未受感染個體中觀察到的病毒中和效應(yīng)大小相差不大(Belshe等，JAMA272(6):475-80(1994))。
病毒的序列。確定來自所有七種逃逸病毒的gp120核苷酸序列，是為了評價(jià)具有疫苗抗原的逃逸病毒的相似性。利用聚合酶鏈?zhǔn)椒磻?yīng)，擴(kuò)增來自原病毒DNA的包膜糖蛋白基因。通過直接擴(kuò)增梯度純化的淋巴細(xì)胞溶菌產(chǎn)物的DNA來獲得序列，這些淋巴細(xì)胞直接從患者血液中獲得，沒有任何中間的培養(yǎng)或者擴(kuò)增步驟。
圖3中列出了完整的gp120序列(每個樣品兩個克隆)。所有七種包膜糖蛋白都具有亞類(clade)B病毒的典型序列。與MN-rgp120的全部同源性的大小為69％-80％(表5)。
表5MN-rgp120序列與得自感染的被接種者的序列的比較MNC6.1 C8.3 C7.2 C11.5 C10.5 C17.1 C15.2MN1007978 7075698072C6.110078 7081759079C8.3 1006880768483C7.2 100 80737673C11.5 100 757080C10.5 100 7072C17.1 100 87C15.1 100*數(shù)據(jù)表示百分等同性。
有趣的是，逃逸病毒不同于MN-rgp120的高百分比(七個中四個)為25-30％[Myers等；逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，Los Alamos國家實(shí)驗(yàn)室(1992和1995)]。歷史上這種程度的序列變異是亞型間(intra-clade)而非亞型內(nèi)變異的典型，預(yù)期亞型內(nèi)變異程度在10-20％內(nèi)[Myers等；逆轉(zhuǎn)錄病毒和艾滋病數(shù)據(jù)庫，Los Alamos國家實(shí)驗(yàn)室(1992和1995)]。與MN-rgp120有最大相似性的病毒中，兩個(c6和C8)在完全免疫之前作為早期感染發(fā)生，一個(C17)作為一種后期感染發(fā)生。
在V3結(jié)構(gòu)域中的多態(tài)性。特別令人感興趣的是，在已知結(jié)構(gòu)域多形態(tài)中包含有由病毒中和抗體識別的表位。最具有特征的中和表位，即主要中和決定簇(PND)，發(fā)生在V3環(huán)的尖端。在亞型8病毒中，以與HIV-1原型MN菌株等同性為基礎(chǔ)，大約60％具有MN血清型定義的特征序列，IGPGRAF(SEQ.ID.NO.39)[Berman等；病毒學(xué)雜志7:4464-9(1992)；Myers等；逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，Los Alamos國家實(shí)驗(yàn)室(1992和1995)；La Rosa等；科學(xué)249:932-5(1990)]。
三種病毒(C6、C8和C17)具有MN血清型特征序列(圖3)。相對地，四種病毒在PND中具有帶氨基酸取代基的序列[IGPGRAW(C7),LGPGSTF(C11),IGPGRVL(C10)以及IGPGSAF(C15)](分別為SEQ.ID.NO.40-43)，因此將其分類為＂無MN類似＂病毒。要注意的是，四種＂無MN類似＂序列的每一種都屬稀有序列(表6)，并且其不是亞型B病毒最普通＂無MN＂變體的典型[Myers等；逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，Los Alamos國家實(shí)驗(yàn)室(1992和1995)]。
表6以MN-rgp120免疫的HIV-1感染個體中主要中和決定簇的形態(tài)變化頻率V3序列 觀察值數(shù)據(jù)集合頻率GNE LANL LANL.1 La Rosa序列 n頻率 (n=52)(n=519)(n=160)(n=245)GPGRAF 30.42 0.67 0.57 0.66 0.60GPGRAW 10.14 0.03 0.013 0.06 0.010GPGRVL 10.14 ＜0.020.004 ＜0.006＜0.008GPGSTF**10.14 ＜0.02＜0.002＜0.006＜0.004GPGSAF 10.14 0.02 0.011 ＜0.006＜0.004*數(shù)據(jù)集合GNE指一組在1992年收集的52個不相關(guān)的分離物；數(shù)據(jù)集合LANL指Myers等(逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，LosAlamos國家實(shí)驗(yàn)室1992和1995)報(bào)道的一組519種序列；LANL.1指B.Korber(個人通訊)提供的一組160個流行病學(xué)非相關(guān)個體；數(shù)據(jù)集合La Rosa指由La Rosa等(科學(xué)249:932-5(1990))報(bào)道的序列數(shù)據(jù)。
**在所檢驗(yàn)的數(shù)據(jù)組中不存在的序列。
在Los Alamos國家實(shí)驗(yàn)室搜集的519種亞型B序列中，與逃逸病毒匹配的具有PND序列的病毒傳播范圍從最高的1.3％(C7)到最低的0.2％(C11)[Myers等；逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，Los Alamos國家實(shí)驗(yàn)室(1992和1995)]。同樣地，從其它三個獨(dú)立衍生數(shù)據(jù)組中可觀察到低頻(表6)。這些序列的出現(xiàn)與1985年以前收集的數(shù)據(jù)組間和1992收集的數(shù)據(jù)組間無大的差別[La Rosa；科學(xué)，249:932-5(1990)]，或者不顯著差別于160個流行病學(xué)非相關(guān)個體組(B.Korber，私人通信)。所有四組數(shù)據(jù)都說明具類似MN PND序列病毒的傳播范圍為60％?；谶@一數(shù)據(jù)，確定七種逃逸感染中四種由位于病毒光譜范圍以外的病毒引起，所說的疫苗預(yù)期抑制該病毒。
逃逸病毒V3結(jié)構(gòu)域的其它特性。象MN-rgp120一樣，所有逃逸病毒的V3結(jié)構(gòu)域長為36個氨基酸。然而，所有七種病毒在糖基化位點(diǎn)數(shù)量方面和合胞體誘導(dǎo)(SI)特征序列方面不同于MN-rgp120。
MN-rgp120的序列因在V3結(jié)構(gòu)域中306位置處缺乏N-連接糖基化位點(diǎn)而有些不尋常[Myers等；逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，LosAlamos國家實(shí)驗(yàn)室(1932和1935)]。已知MN-rgp120的抗血清能中和各種在該位置上包含糖基化位點(diǎn)的病毒(例如SF-2,DU6587-5,DU4489-5,CC)，故缺乏這一糖基化位點(diǎn)似乎并不是抗原性上重要的[Berman等；病毒學(xué)雜志7:4464-9(1992)]。
此外，MN-rgp120的V3結(jié)構(gòu)域具有合胞體誘導(dǎo)病毒的典型序列多形態(tài)(R在位置311處，K在位置324處，K在位置328處)[Fouchier等；病毒學(xué)雜志66:3183-87(1992)]，而所有七種逃逸病毒都具有與非合胞體誘導(dǎo)相關(guān)的序列。
合胞體誘導(dǎo)病毒與急性疾病的發(fā)展[Tersmette等；病毒學(xué)雜志62:2026-32(1988)]和T細(xì)胞嗜性有關(guān)[O′Brien等；自然(倫敦)348:69-73(1990)；Shioda等；自然(倫敦)349:167-9(1991)]。迄今，具有這些性質(zhì)的病毒沒有從MN-rgp120免疫的志愿者回收到。
在V1,V2和C4結(jié)構(gòu)域中的多態(tài)性。前面的研究確定了gp120的V1,V2以及C4結(jié)構(gòu)域中的附加中和表位[Nakamura等；病毒學(xué)雜志67:6179-91(1993)；McKeating等；愛滋病研究和人逆轉(zhuǎn)錄病毒8:451-9(1992)；Ho等；病毒學(xué)雜志65:489-93(1991)；Barbas等；美國國家科學(xué)院院報(bào)91:3809-13(1994)；McKeating等；病毒學(xué)雜志67:433244(1993)；Moore等；病毒學(xué)雜志67:6136-6151(1993)；Davis等；J.Gen.Virol.74:2609-17(1993)]。
這些中和表位最具有特色部分在C4結(jié)構(gòu)域內(nèi)，它引起人們的特別關(guān)注，因?yàn)橐蚜私獾浇Y(jié)合到這一區(qū)域的抗體要阻斷gp120結(jié)合到CD4上[Moore等；愛滋病3:155-63(1989)；McKeating等；愛滋病研究和人逆轉(zhuǎn)錄病毒8:451-9(1992)]。因?yàn)楸砦晃挥诒Ｊ?C)結(jié)構(gòu)域內(nèi)，故天然存在的多種形態(tài)，在該區(qū)域中的遠(yuǎn)比其它中和表位中的有限。Nakamura等(病毒學(xué)雜志67:6179-91(1993))報(bào)道一些中和MAbs的結(jié)合依賴于位置429處的K。
比較MN-rgp120序列和HIV-1的其它菌株，顯示在該位置發(fā)生一次普通的同質(zhì)異晶過程，包括E取代為K。確實(shí)，相同分離病毒的附屬菌株在這一位置上經(jīng)常顯示同質(zhì)異晶現(xiàn)象。HIV-1LAI的HXB2附屬菌株在位置429處含有K，而HIV-1LAI的BH10,HIB和LAV附屬菌株在該位置處含有E[Nakamura等；病毒學(xué)雜志67:6179-91(1993)]。同樣地，HIV-1的1984分離菌株在這一位置顯示E，而用以產(chǎn)生MN-rgp120的HIV-1的1990分離菌株，在這一位置具有K。
檢驗(yàn)受感染疫苗接受者的序列時(圖3)，發(fā)現(xiàn)來自受試驗(yàn)者C17的病毒，如MN-rgp120一樣在位置429處含有K，而不同于疫苗免疫原的六種其它病毒在這一位置具有E。這些結(jié)果顯示七種逃逸病毒中的六種，在gp120的C4結(jié)構(gòu)域中的CD4-阻斷的中和表位方面不同于疫苗免疫原。
用單克隆抗體進(jìn)行的研究限定了gp120的V1與V2結(jié)構(gòu)域中的中和表位[McKeating等；病毒學(xué)雜志67:4932-44(1993)；Moore等；病毒學(xué)雜志67:613-6151(1993)；Davis等；J.Gen.Virol.74:2609-17(1993)]。如發(fā)生在C4結(jié)構(gòu)域的多態(tài)性一樣，V2結(jié)構(gòu)域顯示七種普通的多態(tài)性，其影響到病毒中和抗體的結(jié)合。發(fā)生在位置171處的多態(tài)性對鼠科動物MAb 1025的結(jié)合至關(guān)重要，而殘基187對幾個由1088表現(xiàn)的MAbs的MAb結(jié)合來說很重要。
檢驗(yàn)V2結(jié)構(gòu)域序列(圖3)，發(fā)現(xiàn)所有受感染的被接種者病毒均不同于MN-rgp120，因在其位置171處R取代G和在位置187處I或V取代E。在MN-rgp120的V2結(jié)構(gòu)域中識別這些鄰近位點(diǎn)的抗體，不希望它們中和在這些位置上具有附加氨基酸取代基的病毒。這樣，所有七種逃逸病毒在gp120的V2結(jié)構(gòu)域中的中和表位上區(qū)別于MN-rgp120。
有關(guān)人士報(bào)道了gp120V1結(jié)構(gòu)域中的其它中和表位[O′Brien等；自然(倫敦)348:69-73(1990)；McKeating等；病毒學(xué)雜志67:4932-44(1993)]。雖然未鑒定在MN-rgp120的V1結(jié)構(gòu)域中的中和表位，但在逃逸病毒該區(qū)域觀察到的多態(tài)性令人感興趣。特別驚人的是(圖3)該區(qū)域長度范圍從20氨基酸(C17)到45個氨基酸(C6),N-連接糖基化位點(diǎn)的數(shù)量范圍2至6。相對地，MN-rgp120的V1結(jié)構(gòu)域長度為31個氨基酸并編碼三個N-連接糖基化位點(diǎn)。
盡管對序列數(shù)據(jù)庫的檢驗(yàn)結(jié)果表明，在V2結(jié)構(gòu)域中的變異可與V1結(jié)構(gòu)域中的變異相比較，但逃逸病毒的V2結(jié)構(gòu)域仍顯示了比預(yù)期更少的變異。具體地說，V2結(jié)構(gòu)域的長度從36個氨基酸(C7)到39個氨基酸，同時七種病毒的六種在該結(jié)構(gòu)域中包含三個N-連接糖基化位點(diǎn)。在V4結(jié)構(gòu)域中發(fā)現(xiàn)了高度多態(tài)性，其中序列長度范圍從26(C10)到33(C15,C7)個氨基酸，并包含4個或者5個N-連接糖基化位點(diǎn)。
來自逃逸病毒的包膜糖蛋白抗原性。為了確定序列變異對糖蛋白抗原性的重要性，從所有七個受感染的被接種者的病毒制備重組gp120(圖4)。在這一研究過程中，集合了一系列MAbs，并且其與MN-rgp120的結(jié)合，可與通過ELTSA方法從被接種者分離物獲得的rgp120相比較(表7)。
表7與對MN-rgp120的結(jié)合比較，MAb對來自受感染試驗(yàn)者的rgp120的結(jié)合的相對反應(yīng)性V3不連續(xù) C8 V2gp120 1034 50.1 1.5E102510241088MN 1.0 1.00 1.001.001.001.00C6.1 0.37 0.37 0.170.000.000.00C6.5 0.33 0.33 0.750.000.000.00C8.3 0.11 0.37 0.380.000.000.00C8.6 0.14 0.34 0.290.000.000.00C7.2 0.47 0.60 0.710.000.000.00C11.5 0.00 0.00 0.170.000.000.00C11.7 0.00 0.00 0.170.000.000.00C10.5 0.33 0.40 0.460.240.030.04C10.7 0.42 0.48 0.500.290.070.09C17.1 0.33 0.52 0.330.000.300.07C17.3 0.37 0.56 0.330.000.380.06C15.2 0.00 0.47 0.920.000.000.00C15.3 0.00 0.37 0.630.000.000.00*相對反應(yīng)性值表示MAb濃度2μg/ml時，患者分離物的rgp120光密度值與MN-rgp120光密度值的比值。
對照實(shí)驗(yàn)中，MAb5BG的結(jié)合(其特定用于與所有rgp120蛋白質(zhì)N末端融合的HSV gD-1標(biāo)記表位)用來標(biāo)準(zhǔn)化來自每一分離物的gp120數(shù)量(圖5A)。研究結(jié)果顯示該分析測定應(yīng)在微量滴定板孔截留了等量rgp120s的條件下進(jìn)行。
利用1034MAb(分離自用如Nakamura等(病毒學(xué)雜志67:6179-91,1993)描述的MN-rgp120免疫的鼠體)和50.1 MAb(從用如Rini等(美國科學(xué)院學(xué)報(bào)，90:6325-9,1993)描述的合成V3結(jié)構(gòu)域肽免疫的鼠體中制得)檢驗(yàn)V3結(jié)構(gòu)域的抗原結(jié)構(gòu)。已知兩MAbs都顯示出有力的病毒中和活性。當(dāng)檢驗(yàn)MAb與重組體蛋白質(zhì)的結(jié)合時，同與任何逃逸病毒包膜蛋白質(zhì)的結(jié)合相比，MAb與MN-rgp120的結(jié)合至少要大10倍(圖5B和C)。令人驚奇地是，來自三個患者分離物(C8、C6和C17)的rgp120在它們的MAb結(jié)合活性方面是相互變化的，其中這三個患者分離物具有MN血清型限定的序列IGPGRAF(SEQ.ID.NO.39)。因此，Mab1034和MAb50.1與來自C17的rgp120的結(jié)合大大地大于與來自C6和C8的rgp120s的結(jié)合。
在為這些MAbs所識別的表位方面的差異是明顯的，這是由于C6-rgp120和C8-rgp120和50.1的結(jié)合不相上下，而同與C8-衍生的蛋白質(zhì)的結(jié)合相比，1034更好地與C6-衍生的蛋白質(zhì)結(jié)合。最差的MAb反應(yīng)性是與來自C11和C15的rgp120s的反應(yīng)。這一結(jié)果與序列分析的結(jié)果一致，序列分析顯示這兩種病毒都在V3結(jié)構(gòu)域中的第18位置上具有取代R的S自由基。令人驚奇地是，這兩種MAbs都顯示出比預(yù)期更好的與來自C7和C10病毒的rgp120的結(jié)合。如同MN-rgp120一樣，兩種蛋白質(zhì)在V3環(huán)中都包含五-肽、IGPGR序列(SEQ.ID.NO.44)，但它們與MN-rgp120是不同的，在MN-rgp120中V和L在gp120中的第319和320位取代A和F，并且W在來自C7的gp120中的第320位置取代F。這些結(jié)果表明在318位置上的R對這兩種MAb的結(jié)合來說是十分重要的，并且在319和320位置上的疏水取代并不完全破壞1034和50.1所識別的表位。
正如通過序列數(shù)據(jù)所預(yù)測的一樣，如果利用指向MN-rgp120的V2結(jié)構(gòu)域的Mabs(1088和1025)進(jìn)行與逃逸病毒rgp120s的任何結(jié)合，那么幾乎沒有結(jié)合。也與序列數(shù)據(jù)一致的是觀察結(jié)果指向MN-rgp120的C4結(jié)構(gòu)域的MAb1024具有與C17-rgp120(如同在429位置包含K的MN-rgp120一樣)之間的一定反應(yīng)性，但是不具有與其它的分離物(在429殘基上包含E)的任何反應(yīng)性。
同時，這些研究表明所有七種逃逸病毒的抗原結(jié)構(gòu)不同于在三孔上具有中和表位特征的疫苗免疫原。
觀察到一種模式完全不同的與人雜交瘤的反應(yīng)性，正如Ho等(病毒學(xué)雜志65:489-93,1991)描述的，該MAb15e由受HIV-1感染的個體制備。對于這一MAb，可達(dá)到與MN-rgp120和來自C7的rgp120的最大結(jié)合，同時觀測到與來自C11的rgp120的兩個克隆的最差的反應(yīng)性。中等條件下，觀測到與得自C10和C17的rgp120s的不相上下的反應(yīng)性。
這些結(jié)果顯示15e表位是多形的，并且表位在MN-rgp120和得自C7的rgp120上是保守的，但在得自C11的rgp120s上丟失。令人感興趣的是，得自C6的gp120的兩種不同克隆給出極為不同的抗體結(jié)合模式。因此，得自克隆C6.5的rgp120顯示出與這一抗體的強(qiáng)反應(yīng)性，而得自克隆C6.1的rgp120顯示出與這一MAb的極弱的反應(yīng)性。序列數(shù)據(jù)(圖3)的比較表明兩個C6克隆在6個氨基酸位置上是不同的?；诮Y(jié)合到已知序列的其它病毒蛋白質(zhì)上的比較，似乎在Kgp120的C3結(jié)構(gòu)域中的351位取代Ⅰ說明了在結(jié)合活性方面的區(qū)別。這一結(jié)果也與C11的兩克隆的結(jié)果一致，該C11在這一位置同樣包含帶正電的K，并且也具有與這一MAb的弱反應(yīng)性。此外，T在C4結(jié)構(gòu)域中的439位上對Ⅰ的取代能夠說明在C6.1和C6.5之間的15e結(jié)合方面的區(qū)別。雖然在C4結(jié)構(gòu)域中的這一位置上的多態(tài)性不能解釋為什么上述兩種C11克隆不能結(jié)合到15e上，但是可以通過鄰近的T在434位置上對M的取代來影響這一結(jié)合。
討論在這些研究中，在九個疫苗之七個中的病毒和免疫應(yīng)答得到測定，其中的九個疫苗在參與MN-rgp120(一種候選HIV-1疫苗)的1期和2期試驗(yàn)的同時，均已通過高風(fēng)險(xiǎn)的活性感染了HIV-1。期待這樣的感染將因下列兩個理由之一而發(fā)生1)在感染期間缺乏足夠的免疫應(yīng)答；或者2)用落在抗原光譜(期待為疫苗免疫原所覆蓋)之外的病毒進(jìn)行感染。數(shù)據(jù)表明對于所觀測到的感染，兩種解釋都可以包括(表8)。
表8逃逸感染的總結(jié)與MN-rgp120同源病例號 充分免疫 MN-rgp120V3C4V2同源性(％) PND 表位 表位C6- 79+ - -C8- 78+ - -C15- 72- - -C7- 70- - -C11+ 75- - -C10+ 69- - -C17+ 80+ + -在接受沒有最小的三劑量疫苗的個體中發(fā)生兩個感染，其中所謂的最小劑量是用蛋白質(zhì)亞單位疫苗(例如在如由Francis等(Ann.Int.Med.97:302-6,1982)描述的明礬佐劑中的乙型肝炎病毒)誘導(dǎo)保護(hù)免疫性所典型地需要的。兩種附加的逃逸感染發(fā)生于有弱或者不能檢測到的初次(C7)和加強(qiáng)(C15)反應(yīng)的疫苗中。在三個在接收三次或多次生產(chǎn)性免疫(C10、C11和C17)之后而感染HIV-1的個體中，至少兩個，并且有可能所有三個，似乎在接收他們最后免疫之后的六個月以上才受到感染。因?yàn)閷N-rgp120的抗體滴度典型地以2至2.5個月的半衰期衰變[Belshe等；JAMA272(6):475-80(1994)；Berman等；愛滋病，8:591-601(1994)]，所以期待抗體滴度以至少八倍以及在感染期間有可能高達(dá)六十倍來衰變。因此，在感染期間的足夠免疫應(yīng)答的缺乏為七種逃逸感染的至少六個提供了一種可能的解釋。
研究受到HIV-1激發(fā)的gp160免疫黑猩猩[McElrath等；在201方案AVEG第Ⅱ期中的中的縱向疫苗誘導(dǎo)免疫性和研究參與者風(fēng)險(xiǎn)行為。包括于國家愛滋病疫苗協(xié)同發(fā)展組織第八次年度會議文摘。Bethseda,MD1996:216]和受到嵌合SIV/HIV-1病毒(SHIV)激發(fā)的9p120-免疫獼猴短尾猴的疫苗功效，結(jié)果數(shù)據(jù)表明感染時間的中和抗體反應(yīng)的大小與保護(hù)免疫性具有重要關(guān)系。如果證明了保持中和抗體滴度與人類保護(hù)免疫性有效相關(guān)，那么要實(shí)現(xiàn)長期持久的保護(hù)，就要求設(shè)計(jì)有用來最大化抗體反應(yīng)的方式(例如新的佐劑)或免疫制度(頻繁推進(jìn))。每六個月使用一次激發(fā)劑可能有好處。
對于后期感染的其它可能的解釋是，疫苗抗原和逃逸病毒包膜糖蛋白抗原之間的區(qū)別。這一解釋得到觀察結(jié)果的支持，七種逃逸病毒的四種具有在氨基酸水平上有25-30％不同于MN-rgp120的包膜糖蛋白。該區(qū)別的大小歷史性地[Myers等；逆轉(zhuǎn)錄病毒和愛滋病數(shù)據(jù)庫，Los Alamos國家的實(shí)驗(yàn)室(1992和1995)]與內(nèi)亞型變異相關(guān)，并且遠(yuǎn)超過預(yù)期的相同亞型內(nèi)病毒變異的平均值10-20％。盡管對包膜糖蛋白許多結(jié)構(gòu)域中序列變異的生物重要性還不清楚，但在中和表位的多態(tài)性的確是影響疫苗功效的一個重要因子。以前的研究[Salmon-Ceron等；愛滋病研究人類逆轉(zhuǎn)錄病毒11:1479-86(1995)；Javaherian等；科學(xué)250:1590-3(1930)]已經(jīng)證明能由HIV-1包膜衍生疫苗引起的中和活性寬度，關(guān)鍵依賴于在V3結(jié)構(gòu)域中的表位序列(例如；PND)。這樣，基于HIV-1的LAI菌株的供選疫苗(原型＂非MN類似＂亞型B病毒)，顯示極小或沒有與亞型B病毒的交叉中和活性，而包含＂MN類似＂PND序列(IGPGRAF)(SEQ.ID.NO.44)的疫苗顯示廣泛的交叉中和活性。七種逃逸病毒中，具有PND中帶氨基酸取代基的包膜糖蛋白的四種，符合這樣一種解釋，即抗原結(jié)構(gòu)的區(qū)別與某些這樣的感染有關(guān)。
在最近幾年，＂類MN＂病毒發(fā)生在PND表位之外的多態(tài)性逐漸變得清楚。最好的例子是發(fā)生在C4結(jié)構(gòu)域中的多態(tài)性，其中兩個抗原明顯變體因在位置429處存在K或E而有所區(qū)別[Moore等；愛滋病3:155-63(1989)]。因?yàn)槠叻N逃逸病毒的六種不同于疫苗菌株，因此它們在位置429處包含E而非K，對MN-rgp120的C4結(jié)構(gòu)域產(chǎn)生的抗體不可能中和在七個被接種者的六個中感染的病毒。
已知在gp120的V1與V2結(jié)構(gòu)域中存在著其它中和表位。雖然這些結(jié)構(gòu)域因?yàn)椴迦牒腿笔Ф叨瓤勺儯琈cKeating等仍描述了其中和表位(McKeating等；病毒學(xué)雜志67:4932-44(1993)；Moore等；病毒學(xué)雜志67:6136-6151(1993)；和Davis等；J.Gen.Virol.74:2609-17(1993))。如C4表位一樣，已知其序列變異發(fā)生在由相同雙親分離體衍生而來的不同亞菌株之間。因?yàn)樗衅叻N逃逸病毒的疫苗抗原內(nèi)存在RDK序列而非GDK序列，故其不同于MN-rgp120，對MN-rgp120的V2結(jié)構(gòu)域的中和抗體，將不能如預(yù)期的那樣中和任一從以MN-rgp120免疫的被接種者回收的病毒。
雖然中和表位的多態(tài)性可以解釋大多數(shù)感染中的保護(hù)性缺乏，但看來似乎不能解釋被接種者C17的感染，其受到在V3與C4結(jié)構(gòu)域中配合MN-rgp120的病毒的感染。如果序列差異是在這種情況下引起保護(hù)性缺乏的原因，那么關(guān)鍵性的差異就可能與該逃逸病毒gp120的V1結(jié)構(gòu)域中不尋常的序列有關(guān)。幾項(xiàng)研究結(jié)果表明V1結(jié)構(gòu)域具有通過病毒中和單克隆抗體識別的表位[McKeating等；病毒學(xué)雜志67:4932-44(1993)；Davis等；J.Gen.Virol.74:2609-17(1993)；Kayman等；病毒學(xué)雜志68:400-410(1994)]。
雖然我們對V1表位的了解相對于其它中和位點(diǎn)來說還很少，V1表位似乎是依賴于構(gòu)象的，并由來自HIV-1感染個體的抗血清在V1與V2結(jié)構(gòu)域中識別的表位[McKeating等；病毒學(xué)雜志67:4932-44(1993)；Kayman等；病毒學(xué)雜志68:400-410(1994)]。來自C17的病毒的V1序列很值得注意，因?yàn)樗萂N-rgp120或者其它任何逃逸病毒都要小并且含有較少的N-連接糖基化位點(diǎn)?；谕瑯拥睦碛桑瑏碜訡11和C6的包膜糖蛋白也值得注意，因?yàn)樗鼈儽萂N-rgp120或者其它的逃逸病毒更大并含有更多的糖基化位點(diǎn)。
氨基酸序列的差異可以為抗原結(jié)構(gòu)差異提供線索，而這種多態(tài)性的后果只有通過對抗體結(jié)合的研究方可得到證明。為了確定序列差異與抗原結(jié)構(gòu)差異的相關(guān)性，表達(dá)來自所有七種逃逸病毒每一種兩個克隆的gp120，同時用單克隆抗體板檢驗(yàn)克隆的抗原性。正如從序列數(shù)據(jù)預(yù)測的一樣，對MN-rgp120的V2結(jié)構(gòu)域沒有逃逸病毒包膜糖蛋白與中和MAbs進(jìn)行反應(yīng)。檢驗(yàn)對C4結(jié)構(gòu)域的MAbs時，僅有C17包膜糖蛋白(其關(guān)于K429配合MN-rgp120)顯示雖然較弱但很重要的結(jié)合。令人驚奇的是，三種包含亞型B PND共有序列IGPGRAF(SEQ.ID.NO.!!)的逃逸包膜糖蛋白，盡管它們具有的PND序列與疫苗免疫原密切有關(guān)，但其與所有三種PND的指向MAbs反應(yīng)微弱。這樣，所有三個攜帶疫苗分離物似乎都具有受MAb結(jié)合干擾的識別位點(diǎn)之外的變化。
許多年來人們已經(jīng)知道抗體外中和作用的抗性是由于在遠(yuǎn)程序列方面的突變所致，該遠(yuǎn)程序列改變中和表位的構(gòu)象，并且通過病毒中和抗體來干擾識別[Nara等；病毒學(xué)雜志64:3773-91(1990)；cordonnier等；自然，340:571-4(1989)]。同時，這些結(jié)果表明從逃逸病毒回收的包膜糖蛋白的抗原結(jié)構(gòu)極為不同于疫苗抗原的結(jié)構(gòu)。
新結(jié)果是殘基在C3結(jié)構(gòu)域中的定位，該C3結(jié)構(gòu)域似乎影響結(jié)合病毒中和人MAb,15e的結(jié)合。已知這一MAb識別不連續(xù)表位，阻斷CD4結(jié)合，以及中和HIV-1的各種實(shí)驗(yàn)室和初級分離物[Ho等；病毒學(xué)雜志65:489-93(1991)；Thali等；病毒學(xué)雜志66:5635-5641(1992)；Moore等；AIDS研究.人類逆轉(zhuǎn)錄病毒9:1179-1187(1993)]。
逃逸病毒與包膜糖蛋白的結(jié)合的比較表明這一抗體的識別關(guān)鍵取決于gp120的C3或者C4結(jié)構(gòu)域中的殘基。在C3結(jié)構(gòu)域中的351位的帶正電荷的K的唯一存在為HIV-1的C11.5、C11.7和C6.1菌株不能與15e結(jié)合的現(xiàn)象提供了一種普通的解釋。此外，可能的是，不同的氨基酸在不同位置的取代說明15e不能與得自C6和C11克隆的rgp120s結(jié)合。發(fā)生這一類型取代的唯一明顯位置是在C4結(jié)構(gòu)域中，其中T在434(C11)取代M以及T在439取代I。
本研究表明MN-rgp120的當(dāng)前制劑抗HIV-1感染的有效性不到100％?；谝郧暗挠肕N-rgp120進(jìn)行的體外和體內(nèi)研究，期待防止天然HIV-1在人類中感染的保護(hù)取決于病毒中和抗體的域值濃度，以及疫苗免疫原和激發(fā)病毒之間的抗原相似性。
在這一點(diǎn)上，七種逃逸感染(C17)中只有一種是未預(yù)料到的。這一個體接收全程的免疫，也在至少兩個的重要中和表位(V3和C4結(jié)構(gòu)域)上感染有類似于MN-rgp120的病毒。這一感染可能與感染期間抗體應(yīng)答的大小有關(guān)，或者與逃逸病毒和疫苗菌株之間的抗原差別有關(guān)，或者與在這一方案中得到監(jiān)控的感染環(huán)境(例如潰瘍性的損害，由具有急性感染或嚴(yán)重病毒血癥的供體引起的感染)有關(guān)。此外，這一個體可以代表一個真正的疫苗無效，而無需明確的解釋。
在平衡方面，本文所描述的逃逸感染的分析并沒有揭示任何數(shù)據(jù)，該數(shù)據(jù)使作為抑制HIV-1感染的疫苗的MN-rgp120的繼續(xù)發(fā)展受挫。上述結(jié)果支持如下推測提高疫苗免疫原性(如通過附加的加強(qiáng)免疫)可能是需要的，以便保持長期的保護(hù)免疫性，并且得自除MN-rgp120之外的病毒的其它抗原不同的菌株的rgp120的加入對于擴(kuò)展保護(hù)的寬度是有用的。
得自逃逸感染的病毒和病毒糖蛋白的可利用性可以提供一種使鑒定新抗原(包埋在多價(jià)疫苗中)的過程合理化的重要方法。按照定義，由逃逸病毒制備的重組病毒糖蛋白具有與MN-rgp120極為不同的抗原結(jié)構(gòu)，并且當(dāng)前所傳遞的病毒的代表。因此，把逃逸病毒的rgp120與MN-rgp120相結(jié)合是有效的補(bǔ)充方法，并且極大地?cái)U(kuò)展了抗原復(fù)雜度和增加了交叉中和活性的寬度。
序列表(1)一般信息(ⅰ)申請人Berman,Phillip W.
(ⅱ)發(fā)明名稱HIV包膜多肽和疫苗(ⅲ)序列數(shù)44個(ⅳ)通訊地址(A)收信人SKJERVEN,MORRILL,MACPHERSON等.
(B)街道25Metro Drive,Suite700(C)城市San Jose(D)州加利福尼亞(E)國家美國(F)ZIP:95110(ⅴ)計(jì)算機(jī)可讀形式(A)介質(zhì)類型3.5英寸，1.44Mb軟盤(B)計(jì)算機(jī)IBMPC兼容機(jī)(C)操作系統(tǒng)PC-DOS/MS-DOS(D)軟件WinPatin(Genentech)(ⅵ)當(dāng)前的申請數(shù)據(jù)(A)申請?zhí)?B)申請日(C)分類號(ⅷ)律師/代理人信息(A)姓名Terlizzi,Laura(B)登記號31,307(C)證書號M-3897US(ⅸ)電訊信息
(A)電話(408)4539200(B)傳真(408)4537979(2)SEQ ID NO:1的信息(ⅰ)序列特征(A)長度1503個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ IDNO:1:
GGG GTA CCT GTG TGG AAG GAA GCA ACC ACC ACT CTA 36Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GAG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val15 20 25CAT AAT GTT TGG GCC ACA CAT GCT TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp30 35CCA AAC CCA CAA GAA ATG GTA TTG GAA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Met Val Leu Glu Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT GAC ATG GTA GAA CAG ATG 192Asp Phe Asn Met Trp Lys Asn Asp Met Val Glu Gln Met55 60CAT GAG GAT ATA ATC AGT TTA TGG GAT CAA AGC CTA AAA 231His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys65 70 75CCA TGT GTA AAA TTA ACC CCA CTC TGT ATT ACT TTA AAT 27CPro Cys Val Lys Leu Thr Pro Leu Cys Ile Thr Leu Asn80 85 90TGC ACC AAT TGG AAG AAG AAT GAT ACT AAA ACT AAT AGT 309Cys Thr Asn Trp Lys Lys Asn Asp Thr Lys Thr Asn Ser95 100AGT AGT ACT ACA ACT AAT AAT AGT AGT GCT ACA GCT AAT 348Ser Ser Thr Thr Thr Asn Asn Ser Ser Ala Thr Ala Asn105 110 115AGT AGT AGT ACT ACA ACT AAT AGT AGT TGG GGA GAG ATA 387Ser Ser ser Thr Thr Thr Asn Ser Ser Trp Gly Glu Ile120 125AAG GAG GGA GAA ATA AAG AAC TGC TCT TTC AAT ATC ACC 426Lys Glu Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr130 135 140ACA AGC ATA AGA GAC AAG GTG AAG AAA GAA TAT GCA CTT 465Thr Ser Ile Arg Asp Lys Val Lys Lys Glu Tyr Ala Leu145 150 155TTT TAT AGC CTT GAT GTA GTA CCA ATA GAA AAT GAT AAT 504Phe Tyr Ser Leu Asp Val Val Pro Ile Glu Asn Asp Asn160 165ACT AGC TAT AGG TTG AGA AGT TGT AAC ACC TCA GTC ATT 543Thr Ser Tyr Arg Leu Arg Ser Cys Asn Thr Ser Val Ile170 175 180ACA CAA GCC TGT CCA AAG GTA ACT TTT GAG CCA ATT CCC 582Thr Gln Ala Cys Pro Lys Val Thr Phe Glu Pro Ile Pro185 190ATA CAT TAT TGT ACC CCG GCT GGT TTT GCG ATT CTG AAG 621Ile His Tyr Cys Thr Pro Ala Gly Phe Ala Ile Leu Lys195 200 205TGT AGA GAT AAA AAG TTC AAT GGA ACA GGA CCA TGC AAA 660Cys Arg Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Lys210 215 220AAT GTT AGC ACA GTA CAA TGT GCA CAT GGA ATT AAG CCA 699Asn Val Ser Thr Val Gln Cys Ala His Gly Ile Lys Pro225 230GTA GTG TCA ACT CAA CTG CTG TTA AAT GGC AGC CTA GCA 738Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala235 240 245GAA GAA GAG GTA ATA ATT AGA TCT GCC AAT TTC TCA AAC 777Glu Glu Glu Val Ile Ile Arg Ser Ala Asn Phe Ser Asn250 255AAT GCT AAA ATC ATA ATA GTA CAG TTG AGG GAA CCT GTA 816Asn Ala Lys Ile Ile Ile Val Gln Leu Arg Glu Pro Val260 265 270GAA ATT AAT TGT ACA AGA CCC AGC AAC AAT ACA ATA AAA 855Glu Ile Asn Cys Thr Arg Pro Ser Asn Asn Thr Ile Lys275 280 285GGT ATA CAC ATA GGA CCA GGG AGA GCA TTT TAT GCA ACA 894Gly Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr290 295GGA GAC ATA CGA GGA GAT ATA AGA CAA GCA CAT TGT AAC 933Gly Asp Ile Arg Gly Asp Ile Arg Gln AlaHis Cys Asn300 305310ATT AGT GGA GCA AAA TGG AAT AAC ACT TTA AAG AAG GTA 972Ile Ser Gly Ala Lys Trp Asn Asn Thr Leu Lys Lys Val315 320GTT AAA AAA TTA AAA GAA CAA TTT CCA AAT AAA ACA ATA 1011Val Lys Lys Leu Lys Glu Gln Phe Pro Asn Lys Thr Ile325 330 335GTC TTT AAC CAT TCC TCA GCA GGG GAC CCA GAA ATT GTA 1050Val Phe Asn His Ser Ser Gly Gly Asp Pro Glu Ile Val340 345 350ATG CAC AGT TTT AAT TGT CAA GGG GAA TTT TTC TAC TGT 1089Met His Ser Phe Asn Cys Gln Gly Glu Phe Phe Tyr Cys355 360AAT ACA ACA AAG CTG TTT AAT AGT ACT TGG AAT GAT ACT 1128Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Asp Thr365 370 375ACA GAG TCA AAT AAC AAT GAT AGT ACT ATT ACA CTC CCA 1167Thr Glu Ser Asn Asn Asn Asp Ser Thr Ile Thr Leu Pro380 385TGC AGA ATA AAA CAA ATT ATA AAC ATG TGG CAG GAA ATA 1206Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Ile390 395 400GGA AAA GCA ATG TAT GCC CCT CCC ACC AGA GGA GAA ATT 1245Gly Lys Ala Met Tyr Ala Pro Pro Thr Arg Gly Glu Ile405 410 415AAA TGT TCA TCA AAT ATT ACA GGA CTA CTG TTA ATA AGA 1284Lys Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Ile Arg420 425GAT GGT GGT ATT AAC ACT AGC GAT GCC ACC GAG ACC TTC 1323Asp Gly Gly Ile Asn Thr Ser Asp Ala Thr Glu Thr Phe430 435 440AGA CCG GGA GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT 1362Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg Ser445 450GAA TTA TAT AAA TAT AAA GTA GTG AAA ATT GAG CCA TTA 1401Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu455 460 465GGA GTA GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG 1440Gly Val Ala Pro Thr Lys Ala Lys Ar9 Ar9 Val Val Gln470 475 480AGA GAA AAA AGA GCA GTA ACA CTA GGA GCT ATG TTC CTT 1479Arg Glu Lys Arg Ala Val Thr Leu Gly Ala Met Phe Leu485 490GGG TTC TTA GGA GCA TAA AGC TTC 1503Gly Phe Leu Gly Ala Xaa Ser Phe495 500 501(2)SEQ ID NO:2的信息(ⅰ)序列特征(A)長度501個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:2:Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala1 5 10 15Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala20 25 30Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Met Val35 40 45Leu Glu Asn Val Thr Glu Asp Phe Asn Met Trp Lys Asn Asp Met50 55 60Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser65 70 75Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Ile Thr Leu Asn80 85 90Cys Thr Asn Trp Lys Lys Asn Asp Thr Lys Thr Asn Ser Ser Ser95 100 105Thr Thr Thr Asn Asn Ser Ser Ala Thr Ala Asn Ser Ser Ser Thr110 115 120Thr Thr Asn Ser Ser Trp Gly Glu Ile Lys Glu Gly Glu Ile Lys125 130 135Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp Lys Val Lys140 145 150Lys Glu Tyr Ala Leu Phe Tyr Ser Leu Asp Val Val Pro Ile Glu155 160 165Asn Asp Asn Thr Ser Tyr Arg Leu Arg Ser Cys Asn Thr Ser Val170 175 180Ile Thr Gln Ala Cys Pro Lys Val Thr Phe Glu Pro Ile Pro Ile185 190 195His Tyr Cys Thr Pro Ala Gly Phe Ala Ile Leu Lys Cys Arg Asp200 205 210Lys Lys Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val215 220 225Gln Cys Ala His Gly Ile Lys Pro Val Val Ser Thr Gln Leu Leu230 235 240Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Ile Ile Arg Ser Ala245 250 255Asn Phe Ser Asn Asn Ala Lys Ile Ile Ile Val Gln Leu Arg Glu260 265 270Pro Val Glu Ile Asn Cys Thr Arg Pro Ser Asn Asn Thr Ile Lys275 280 285Gly Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr Gly Asp290 295 300Ile Arg Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Gly Ala305 310 315Lys Trp Asn Asn Thr Leu Lys Lys Val Val Lys Lys Leu Lys Glu320 325 330Gln Phe Pro Asn Lys Thr Ile Val Phe Asn His Ser Ser Gly Gly335 340 345Asp Pro Glu Ile Val Met His Ser Phe Asn Cys Gln Gly Glu Phe350 355 360Phe Tyr Cys Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Asp365 370 375Thr Thr Glu Ser Asn Asn Asn Asp Ser Thr Ile Thr Leu Pro Cys380 385 390Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Ile Gly Lys Ala395 400 405Met Tyr Ala Pro Pro Thr Arg Gly Glu Ile Lys Cys Ser Ser Asn410 415 420Ile Thr Gly Leu Leu Leu Ile Arg Asp Gly Gly Ile Asn Thr Ser425 430 435Asp Ala Thr Glu Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp440 445 450Asn Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu455 460 465Pro Leu Gly Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln470 475 480Arg Glu Lys Arg Ala Val Thr Leu Gly Ala Met Phe Leu Gly Phe485 490 495Leu Gly Ala Xaa Ser Phe500 501(2)SEQIDNO:3的信息(ⅰ)序列特征(A)長度1503個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:3:
GGG GTA CCT GTA TGG AAA GAA GCA ACC ACC ACT CTA 36Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GAG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val15 20 25CAT AAT GTT TGG GCC ACA CAT GCT TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr Mis Ala Cys Val Pro Thr Asp30 35CCA AAC CCA CAA GAA ATG GTA TTG GAA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Met Val Leu Glu Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT GAC ATG GTA GAA CAG ATG 192Asp Phe Asn Met Trp Lys Asn Asp Met Val Glu Gln Met55 60CAT GAG ANT ATA ATC AGT TTA TGG GAT CAA AGC CTA AAA 231His Glu Xaa Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys65 70 75CCA TGT GTA AAA TTA ACC CCA CTC TGT ATT ACT TTA AAT 270Pro Cys Val Lys Leu Thr Pro Leu Cys Ile Thr Leu Asn80 85 90TGC ACC AAT TGG AAG GAG AAT GAT ACT AAA ACT AAT AGT 309Cys Thr Asn Trp Lys Glu Ash Asp Thr Lys Thr Asn Ser95 100AGT AGT ACT ACA ACT AAT AAT AGT AGT GCT ACA GCT AAT 348Ser Ser Thr Thr Thr Asn Asn Ser Ser Ala Thr Ala Asn105 110 115AGT AGT AGT ACT ACA ACT AAT AGT AGT TGG GGA GAG ATA 387Ser Ser Ser Thr Thr Thr Asn Ser Ser Trp Gly Glu Ile120 125AAG GAG GGA GAA ATA AAG AAC TGC TCT TTC AAT ATC ACC 426Lys Glu Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr130 135 140ACA GGC ATA AGA GAC AAG GTG AAG AAA GAA TAT GCA CTT 465Thr Gly Ile Arg Asp Lys Val Lys Lys Glu Tyr Ala Leu145 150 155TTT TAT AGC CTT GAT GTA GTA CCA ATA GAA AAT GAT AAT 504Phe Tyr Ser Leu Asp Val Val Pro Ile Glu Asn Asp Asn160 165ACT AGC TAT AGG TTG AGA AGT TGT AAC ACC TCA GTC ATT 543Thr Ser Tyr Arg Leu Arg Ser Cys Asn Thr Ser Val Ile170 175 180ACA CAA GCC TGT CCA AAG GTA ACT TTT GAG CCA ATT CCC 582Thr Gln Ala Cys Pro Lys Val Thr Phe Glu Pro Ile Pro185 190ATA CAT TAT TGT ACC CCG GCT GGT TTT GCG ATT CTG AAG 621Ile His Tyr Cys Thr Pro Ala Gly Phe Ala Ile Leu Lys195 200 205TGT AAA GAT AAA AAG TTC AAT GGA ACA GGA CCA TGC AAA 660Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Lys210 215 220AAT GTT AGC ACA GTA CAA TGT ACA CAT GGA ATT AAG CCA 699Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys Pro225 230GTA GTG TCA ACT CAA CTG CTG TTA AAT GGC AGC CTA GCA 738Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala235 240 245GAA GAA GAG GTA ATA ATT AGA TCT GCC AAT TTC TCA AAC 777Glu Glu Glu Val Ile Ile Arg Ser Ala Asn Phe Ser Asn250 255AAT GCT AAA ATC ATA ATA GTA CAG TTG AAG GAA CCT GTA 816Asn Ala Lys Ile Ile Ile Val Gln Leu Lys Glu Pro Val260 265 270GAA ATT AAT TGT ACA AGA CCC AGC AAC AAT ACA ATA AAA 855Glu Ile Asn Cys Thr Arg Pro Ser Asn Asn Thr Ile Lys275 280 285GGT ATA CAC ATA GGA CCA GGG AGA GCA TTT TAT GCA ACA 894Gly Ils His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr290 295GGA GAC ATA CGA GGA GAT ATA AGA CAA GCA CAT TGT AAC 933Gly Asp Ile Arg Gly Asp Ile Arg Gln Ala His Cys Asn300 305 310ATT AGT GGA GCA AAA TGG AAT AAC ACT TTA AAG AAG GTA 972Ile Ser Gly Ala Lys Trp Asn Asn Thr Leu Lys Lys Val315 320CTT ATA AAA TTA AAA GAA CAA TTT CCA AAT AAA ACA ATA 1011Val Ile Lys Leu Lys Glu Gln Phe Pro Asn Lys Thr Ile325 330 335GTC TTT AAC CAT TCC TCA GGA GGG GAC CCA GAA ATT GTA 1050Val Phe Asn His Ser Ser Gly Gly Asp Pro Glu Ile Val340 345 350ATG CAC AGT TTT AAT TGT CAA GGG GAA TTT TTC TAC TGT 1089Met His Ser Phe Asn Cys Gln Gly Glu Phe Phe Tyr Cys355 360AAT ACA ACG AAG CTG TTT AAT AGT ACT TGG AAT GAT ACT 1128Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Asp Thr365 370 375ACA GAG TCA AAT AAC AAT GAT AGT ACT ATT ACA CTC CCA 1167Thr Glu Ser Asn Asn Asn Asp Ser Thr Ile Thr Leu Pro380 385TGC AGA ATA AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA 1206Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val390 395 400GGA AAA GCA ATG TAT GCC CCT CCC ATC AGA GGA GAA ATT 1245Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile405 410 415AAA TGT TCA TCA AAT ATT ACA GGA CTA CTG TTA ACA AGA 1284Lys Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg420 425GAT GGT GGT ATT AAC ACT AGC GAT GCC ACC GAG ACC TTC 1323Asp Gly Gly Ile Asn Thr Ser Asp Ala Thr Glu Thr Phe430 435 440AGA CCG GGA GGA GGA GAT ATG AGG GAC AAT TG6 AGA AGT 1362Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg Ser445 450GAA TTA TAT AAA TAT AAA GTA GTG AAA ATT GAG CCA TTA 1401Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu455 460 465GGA GTA GCA CCC ACC AAG GCA AAG AGA AGA GTC GTG CAG 1440Gly Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln470 475 480AGA GAA AAA AGA GCA GTA ACA CTA GGA GCT ATG TTC CTT 1479Arg Glu Lys Arg Ala Val Thr Leu Gly Ala Met Phe Leu485 490GGG TTC TTG GGA GCA TAA AGC TTC 1503Gly Phe Leu Gly Ala Xaa Ser Phe495 500 501(2)SEQ ID NO:4的信息(ⅰ)序列特征(A)長度501個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:4:Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala1 5 10 15Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala20 25 30Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Met Val35 40 45Leu Glu Asn Val Thr Glu Asp Phe Asn Met Trp Lys Asn Asp Met50 55 60Val Glu Gln Met His Glu Xaa Ile Ile Ser Leu Trp Asp Gln Ser65 70 75Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Ile Thr Leu Asn80 85 90Cys Thr Asn Trp Lys Glu Asn Asp Thr Lys Thr Asn Ser Ser Ser95 100 105Thr Thr Thr Asn Asn Ser Ser Ala Thr Ala Asn Ser Ser Ser Thr110 115 120Thr Thr Asn Ser Ser Trp Gly Glu Ile Lys Glu Gly Glu Ile Lys125 130 135Asn Cys Ser Phe Asn Ile Thr Thr Gly Ile Arg Asp Lys Val Lys140 145 150Lys Glu Tyr Ala Leu Phe Tyr Ser Leu Asp Val Val Pro Ile Glu155 160 165Asn Asp Asn Thr Ser Tyr Arg Leu Arg Ser Cys Asn Thr Ser Val170 175 180Ile Thr Gln Ala Cys Pro Lys Val Thr Phe Glu Pro Ile Pro Ile185 190 195His Tyr Cys Thr Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp200 205 210Lys Lys Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val215 220 225Gln Cys Thr Mis Gly Ile Lys Pro Val Val Ser Thr Gln Leu Leu230 235 240Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Ile Ile Arg Ser Ala245 250 255Asn Phe Ser Asn Asn Ala Lys Ile Ile Ile Val Gln Leu Lys Glu260 265 270Pro Val Glu Ile Asn Cys Thr Arg Pro Ser Asn Asn Thr Ile Lys275 280 285Gly Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr Gly Asp290 295 300Ile Arg Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Gly Ala305 310 315Lys Trp Asn Asn Thr Leu Lys Lys Val Val Ile Lys Leu Lys Glu320 325 330Gln Phe Pro Asn Lys Thr Ile Val Phe Asn His Ser Ser Gly Gly335 340 345Asp Pro Glu Ile Val Met His Ser Phe Asn Cys Gln Gly Glu Phe350 355 360Phe Tyr Cys Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Asp365 370 375Thr Thr Glu Ser Asn Asn Asn Asp Ser Thr Ile Thr Leu Pro Cys380 385 390Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala395 400 405Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile Lys Cys Ser Ser Asn410 415 420Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Ile Asn Thr Ser425 430 435Asp Ala Thr Glu Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp440 445 450Asn Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu455 460 465Pro Leu Gly Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln470 475 480Arg Glu Lys Arg Ala Val Thr Leu Gly Ala Met Phe Leu Gly Phe485 490 495Leu Gly Ala Xaa Ser Phe500 501(2)SEQ ID NO:5的信息(ⅰ)序列特征(A)長度1461個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:5:G GTA CCT GTA TGG AAA GAA GCA ACC ACC ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT GAT ACA GAG GTA CAT 76Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His15 20 25AAT GTT TGG GCT ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AAC CCA CAA GAA GTA GTA TTG GAA AAT GTA ACA GAA AAT 154Asn Pro Gln Glu Val Val Leu Glu Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His55 60GAG GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA AAG CCA 232Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA AAT TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90ACT AAT TTG GAG AAT GCT AAT AAT ACC GAG AAT GCT AAT 310Thr Asn Leu Glu Asn Ala Asn Asn Thr Glu Asn Ala Asn95 100AAT ACC AAT AAT TAT ACC TTG GGG ATG GAG AGA GGT GAA 349Asn Thr Asn Asn Tyr Thr Leu Gly Met Glu Arg Gly Glu105 110 115ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA AGC TTA AGA 388Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Leu Arg120 125GAT AAG GTG AAA AAA GAA TAT GCA TTG TTT TAT AAA CTT 427Asp Lys Val Lys Lys Glu Tyr Ala Leu Phe Tyr Lys Leu130 135 140GAT GTA GTA CAA ATA GAT AAT AGT ACC AAC TAT AGG CTG 466Asp Val Val Gln Ile Asp Asn Ser Thr Asn Tyr Arg Leu145 150 155ATA AGT TGT AAT ACC TCA GTC ATT ACA CAG GCC TGT CCA 505Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro160 165AAG GTA TCC TTT GAG CTA ATT CCC ATA CAT TAT TGT GCC 544Lys Val Ser Phe Glu Leu Ile Pro Ile His Tyr Cys Ala170 175 180CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG AAG 583Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys185 190TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGC ACA GTA 622Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val195 200 205CAA TGT ACA CAT GGA ATT AGA CCA GTA GTA TCA ACT CAA 661Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln210 215 220CTA CTG TTA AAT GGC AGT CTA GCA GAA GAA GAG ATA GTA 700Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Ile Val225 230ATT AGA TCT GAA AAT ATC ACA GAC AAT GCT AAA ACC ATA 739Ile Arg Ser Glu Asn Ile Thr Asp Asn Ala Lys Thr Ile235 240 245ATA GTG CAG CTA AAT GAA TCT ATA GTG ATT AAT TGT ACA 778Ile Val Gln Leu Asn Glu Ser Ile Val Ile Asn Cys Thr250 255AGA CCC AAT AAC AAC ACA AGA AAA AGT ATA AAT ATA GGA 817Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Asn Ile Gly260 265 270CCA GGG AGA GCA TTC TAT ACA ACA GGA GAC ATA ATA GGA 856Pro Gly Arg Ala Phe Tyr Thr Thr Gly Asp Ile Ile Gly275 280 285GAT ATA AGA CAA GCA CAT TGT AAC CTT AGT AAA ACA CAA 895Asp Ile Arg Gln Ala His Cys Asn Leu Ser Lys Thr Gln290 295TGG GAA AAA ACG TTA AGA CAG ATA GCT ATA AAA TTA GAA 934Trp Glu Lys Thr Leu Arg Gln Ile Ala Ile Lys Leu Glu300 305 310GAA AAA TTT AAG AAT AAA ACA ATA GCC TTT AAT AAA TCC 973Glu Lys Phe Lys Asn Lys Thr Ile Ala Phe Asn Lys Ser315 320TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT AAT 1012Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe Asn325 330 335TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA AAA CTG 1051Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Lys Leu340 345 350TTT AAT AGT ACC TGG AAT TTA ACA CAA CCG TTT AGT AAT 1090Phe Asn Ser Thr Trp Asn Leu Thr Gln Pro Phe Ser Asn355 360ACC GGG AAT CGT ACT GAA GAG TTA AAT ATT ACA CTC CCA 1129Thr Gly Asn Arg Thr Glu Glu Leu Asn Ile Thr Leu Pro365 370 375TGC AGA ATA AAA CAA ATC ATA AAC TTG TGG CAG GAA GTA 1168Cys Arg Ile Lys Gln Ile Ile Asn Leu Trp Gln Glu Val380 385GGC AAA GCA ATG TAT GCC CCT CCC ATC AGA GGA CAA ATT 1207Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile390 395 400AGA TGT TCA TCA AAT ATT ACA GGG CTA CTA TTA ACA AGA 1246Arg Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg405 410 415GAT GGT GGA AGT AAC ACC GGT GAC AAC AGG ACT GAG ACC 1285Asp Gly Gly Ser Asn Thr Gly Asp Asn Arg Thr Glu Thr420 425TTT AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG AGA 1324Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg430 435 440AGT GAA TTA TAT AAA TAT AAA GTA GTA AGA ATT GAA CCA 1363Ser Glu Leu Tyr Lys Tyr Lys Val Val Arg Ile Glu Pro445 450TTA GGA GTA GCA CCC ACC CAG GCA AAG AGA AGA GTG GTG 1402Leu Gly Val Ala Pro Thr Gln Ala Lys Arg Arg Val Val45 460 465CAA AGA GAA AAA AGA GCA GTG GGG ATA GGA GCT ATG TTC 1441Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Met Phe470 475 480CTT GGG TTC TTG GGA GAT AA 1461Leu Gly Phe Leu Gly Asp485 486(2)SEQ ID NO:6的信息(ⅰ)序列特征(A)長度486個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:6:Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala Ser1 5 10 15Asp Ala Lys Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala Thr20 25 30His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Val Val Leu35 40 45Glu Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val50 55 60Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90Thr Asn Leu Glu Asn Ala Asn Asn Thr Glu Asn Ala Asn Asn Thr95 100 105Asn Asn Tyr Thr Leu Gly Met Glu Arg Gly Glu Ile Lys Asn Cys110 115 120Ser Phe Asn Ile Thr Thr Ser Leu Arg Asp Lys Val Lys Lys Glu125 130 135Tyr Ala Leu Phe Tyr Lys Leu Asp Val Val Gln Ile Asp Asn Ser140 145 150Thr Asn Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln155 160 165Ala Cys Pro Lys Val Ser Phe Glu Leu Ile Pro Ile His Tyr Cys170 175 180Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys Phe185 190 195Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val Gln Cys Thr200 205 210His Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly215 220 225Ser Leu Ala Clu Glu Glu Ile Val Ile Arg Ser Glu Asn Ile Thr230 235 240Asp Asn Ala Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Ile Val245 250 255Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Asn260 265 270Ile Gly Pro Gly Arg Ala Phe Tyr Thr Thr Gly Asp Ile Ile Gly275 280 285Asp Ile Arg Gln Ala His Cys Asn Leu Ser Lys Thr Gln Trp Glu290 295 300Lys Thr Leu Arg Gln Ile Ala Ile Lys Leu Glu Glu Lys Phe Lys305 310 315Asn Lys Thr Ile Ala Phe Asn Lys Ser Ser Gly Gly Asp Pro Glu320 325 330Ile Val Met His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys335 340 345Asn Thr Thr Lys Leu Phe Asn Ser Thr Trp Asn Leu Thr Gln Pro350 355 360Phe Ser Asn Thr Gly Asn Arg Thr Glu Glu Leu Asn Ile Thr Leu365 370 375Pro Cys Arg Ile Lys Gln Ile lle Asn Leu Trp Gln Glu Val Gly380 385 390Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile Arg Cys Ser395 400 405Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Ser Asn410 415 420Thr Gly Asp Asn Arg Thr Glu Thr Phe Arg Pro Gly Gly Gly Asp425 430 435Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val440 445 450Arg Ile Glu Pro Leu Gly Val Ala Pro Thr Gln Ala Lys Arg Arg455 460 465Val Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Met Phe470 475 480Leu Gly Phe Leu Gly Asp485 486(2)SEQ ID NO:7的信息(ⅰ)序列特征(A)長度1474個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:7G GTA CCT GTG TGG AAA GAA GCA ACC ACC ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT GAT ACA GAG GTA CAT 76Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His15 20 25AAT GTT TGG GCT ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AAC CCA CAA GAA GTA GTA TTG GAA AAT GTA ACA GAA AAT 154Asn Pro Gln Glu Val Val Leu Glu Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His55 60GAG GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA AAG CCA 232Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA AAT TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90ACT AAT TTG GAG AAT GCT AAT AAT ACC GAG AAT GCT AAT 310Thr Asn Leu Glu Asn Ala Asn Asn Thr Glu Asn Ala Asn95 100AAT ACC AAT AAT TAT ACC TTG GGG ATG GAG AGA GGT GAA 349Asn Thr Asn Asn Tyr Thr Leu Gly Met Glu Arg Gly Glu105 110 115AGA AAA AAC TGC TCT TTC AAT ATC ACC ACA AGC TTA AGA 388Arg Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Leu Arg120 125GAT AAG GGG AAA AAA GAA TAT GCA TTG TTT TAT AAA CTT 427Asp Lys Gly Lys Lys Glu Tyr Ala Leu Phe Tyr Lys Leu130 135 140GAT GTA GTA CAA ATA GAT AAT AGT ACC AAC TAT AGG CTG 466Asp Val Val Gln Ile Asp Asn Ser Thr Asn Tyr Arg Leu145 150 155ATA AGT TGT AAT ACC TCA GTC ATT ACA CAG GCC TGT CCA 505Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro160 165AAG GTA TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT GCC 544Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala170 175 180CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG AAG 583Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys185 190TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGG ACA GTA 622Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Arg Thr Val195 200 205CAA TGT ACA CAT GGA ATT AGA CCA GTA GTA TCA ACT CAA 661Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln210 215 220CTA CTG TTA AAT GGC AGT CTA GCA GAA GAA GAG ATA GTA 700Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Ile Val225 230ATT AGA TCT GAA AAT ATC ACA GAC AAT GCT AAA ACC ATA 739Ile Arg Ser Glu Asn Ile Thr Asp Asn Ala Lys Thr Ile235 240 245ATA GTG CAG CTA AAT GAA TCT ATA GTG ATT AAT TGT ACA 778Ile Val Gln Leu Asn Glu Ser Ile Val Ile Asn Cys Thr250 255AGA CCC AAT AAC AAC ACA AGA AAA AGT ATA AAT ATA GGA 817Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Asn Ile Gly260 265 270CCA GGG AGA GCA TTC TAT ACA ACA GGA GAC ATA ATA GGA 856Pro Gly Arg Ala Phe Tyr Thr Thr Gly Asp Ile Ile Gly275 280 285GAT ATA AGA CAA GCA CAT TGT AAC CTT AGT AAA ACA CAA 895Asp Ile Arg Gln Ala His Cys Asn Leu Ser Lys Thr Gln290 295TGG GAA AAA ACG TTA AGA CAG ATA GCT ATA AAA TTA GAA 934Trp Glu Lys Thr Leu Arg Gln Ile Ala Ile Lys Leu Glu300 305 310GAA AAA TTT AAG AAT AAA ACA ATA GCC TTT AAT AAA TCC 973Glu Lys Phe Lys Asn Lys Thr Ile Ala Phe Asn Lys Ser315 320TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT AAT 1012Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe Asn325 330 335TGT GGA GGG GGA TTT TTC TAC TGT AGT ACG AGA AAA CTG 1051Cys Gly Gly Gly Phe Phe Tyr Cys Ser Thr Arg Lys Leu340 345 350TTT AAT AGT ACC TGG AAT TTA ACA CAA CCG TTT AGT AAT 1090Phe Asn Ser Thr Trp Asn Leu Thr Gln Pro Phe Ser Asn355 360ACC GGG GAT CGT ACT GAA GAG TTA AAT ATT ACA CTC CCA 1129Thr Gly Asp Arg Thr Glu Glu Leu Asn Ile Thr Leu Pro365 370 375TGC AGA ATA AAA CAA ATC ATA AAC TTG TGG CAG GAA GTA 1168Cys Arg Ile Lys Gln Ile Ile Asn Leu Trp Gln Glu Val380 385GGC AAA GCA ATG TAT GCC CCT CCC ATC AGA GGA CAA ATT 1207Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile390 395 400AGA TGT TCA TCA AAT ATT ACA GGG CTA CTA TTA AGG AGA 1246Arg Cys Ser Ser Asn Ile Thr Gly Leu Leu Leu Arg Arg405 410 415GAT GGT GGA AGT AAC ACC AGT GAC AAC CAG ACT GAG ACC 1285Asp Gly Gly Ser Asn Thr Ser Asp Asn Gln Thr Glu Thr420 425TTT AGA CCT GGG GGA GGA GAT ATG AGG GAC AAG TGG AGA 1324Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Lys Trp Arg430 435 440AGT GAA TTA TAT AAA TAT AAA GTA GTA AGA ATT GAA CCA 1363Ser Glu Leu Tyr Lys Tyr Lys Val Val Arg Ile Glu Pro445 450TTA GGA GTA GCA CCC ACC CAG GCA AAG AGA AGA GTG GTG 1402Leu Gly Val Ala Pro Thr Gln Ala Lys Arg Arg Val Val455 460 465CAA AGA GAA AAA AGA GCA GTG GGG ATA GGA GCT ATG TTC 1441Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Met Phe470 475 480CTT AGG TTC TTA GGA GAT AAA GCT TCT AGA GTC 1474Leu Arg Phe Leu Gly Asp Lys Ala Ser Arg Val485 490 491(2)SEQ ID NO:8的信息(ⅰ)序列特征(A)長度491個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:8:Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala Ser1 5 10 15Asp Ala Lys Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala Thr20 25 30His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Val Val Leu35 40 45Glu Asn Val Thr Glu Asn Phe Asn Met Trp Ly5 Asn Asn Met Val50 55 60Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90Thr Asn Leu Glu Asn Ala Asn Asn Thr Glu Asn Ala Asn Asn Thr95 100 105Asn Asn Tyr Thr Leu Gly Met Glu Arg Gly Glu Arg Lys Asn Cys110 115 120Ser Phe Asn Ile Thr Thr Ser Leu Arg Asp Lys Gly Lys Lys Glu125 130 135Tyr Ala Leu Phe Tyr Lys Leu Asp Val Val Gln lle Asp Asn Ser140 145 150Thr Asn Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln155 160 165Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys170 175 180Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys Phe185 190 195Asn Gly Thr Gly Pro Cys Lys Asn Val Arg Thr Val Gln Cys Thr200 205 210His Gly Ile Ar9 Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly215 220 225Ser Leu Ala Glu Glu Glu Ile Val Ile Arg Ser Glu Asn Ile Thr230 235 240Asp Asn Ala Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Ile Val245 250 255Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Asn260 265 270Ile Gly Pro Gly Arg Ala Phe Tyr Thr Thr Gly Asp Ile Ile Gly275 280 285Asp Ile Arg Gln Ala His Cys Asn Leu Ser Lys Thr Gln Trp Glu290 295 300Lys Thr Leu Arg Gln Ile Ala Ile Lys Leu Glu Glu Lys Phe Lys305 310 315Asn Lys Thr Ile Ala Phe Asn Lys Ser Ser Gly Gly Asp Pro Glu320 325 330Ile Val Met His Ser Phe Asn Cys Gly Gly Gly Phe Phe Tyr Cys335 340 345Ser Thr Arg Lys Leu Phe Asn Ser Thr Trp Asn Leu Thr Gln Pro350 355 360Phe Ser Asn Thr Gly Asp Arg Thr Glu Glu Leu Asn Ile Thr Leu365 370 375Pro Cys Arg Ile Lys Gln Ile Ile Asn Leu Trp Gln Glu Val Gly380 385 390Lys Ala Met Tyr Ala Pro Pro Ile Ar9 Gly Gln Ile Arg Cys Ser395 400 405Ser Asn Ile Thr Gly Leu Leu Leu Arg Arg Asp Gly Gly Ser Asn410 415 420Thr Ser Asp Asn Gln Thr Glu Thr Phe Arg Pro Gly Gly Gly Asp425 430 435Met Arg Asp Lys Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val440 445 450Arg Ile Glu Pro Leu Gly Val Ala Pro Thr Gln Ala Lys Arg Arg455 460 465Val Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Met Phe470 475 480Leu Arg Phe Leu Gly Asp Lys Ala Ser Arg Val485 490 491(2)SEQ ID NO:9的信息(ⅰ)序列特征(A)長度1512個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:9:
CTC GAG GTA CCT GTA TGG AAA GAA GCA ACT ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT AAT ACA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asn Thr Glu15 20 25AAA CAT AAT GTT TGG GCC ACA CAC GCC TGT GTA CCC ACA 114Lys His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr30 35GAT CCC AAC CCA CAA GAA GTA GTA TTG GGA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Val Leu Gly Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA 192Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln55 60ATG CAT GAA GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA 231Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAT GAT TTA GGG AAT GCT ACT AAT ACC AAT 309Asn Cys Thr Asp Asp Leu Gly Asn Ala Thr Asn Thr Asn95 100AGT AGT GCC ACT ACC AAT AGT AGT AGT TGG GAA GAA ATG 348Ser Ser Ala Thr Thr Asn Ser Ser Ser Trp Glu Glu Met105 110 115AAG GGG GAA ATG AAA AGA TGC TCT TTC AAT ATC ACC ACA 387Lys Gly Glu Met Lys Arg Cy5 Ser Phe Asn Ile Thr Thr120 125AGC ATA AGA GAT AAG ATT AAG AAA GAA CAT GCA CTT TTC 426Ser Ile Arg Asp Lys Ile Lys Lys Glu His Ala Leu Phe130 135 140TAT AGA CTT GAT GTA GTA CCA ATA GAT AAT GAT AAT ACC 465Tyr Arg Leu Asp Val Val Pro Ile Asp Asn Asp Asn Thr145 150 155ACA TAT AGG TTG ATA AAT TGT AAT ACC TCA GTC ATT ACA 504Thr Tyr Arg Leu Ile Asn Cys Asn Thr Ser Val Ile Thr160 165CAG GCC TGT CCA AAG GTA TCA TTT GAG CCA ATT CCC ATA 543Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile170 175 180CAT TTT TGT GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT 582His Phe Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys185 190AAT AAT AAG ACG TTC GAG GGA AAA GGA CCA TGT AAA AAT 621Asn Asn Lys Thr Phe Glu Gly Lys Gly Pro Cys Lys Asn195 200 205GTC AGT ACA GTA CAA TGC ACA CAT GGA ATT AGG CCA GTA 660Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro Val210 215 220GTG TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA GAA 699Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu225 230GAA GAG GTA ATA ATT AGA TCT GAC AAT ATC ACA GAC AAT 738Glu Glu Val Ile Ile Arg Ser Asp Asn Ile Thr Asp Asn235 240 245ACT AAA ACC ATT ATA GTA CAG CTA AAC GAA TCT GTA GTA 777Thr Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Val Val250 255ATT AAT TGT ACA AGA CCC AAC AAC AAT ACA AGA AAA AGT 816Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser260 265 270ATA CAT ATA GGA CCA GGG AGT GCA TTT TTT GCA ACA GGA 855Ile His Ile Gly Pro Gly Ser Ala Phe Phe Ala Thr Gly275 280 285GAA ATA ATA GGA GAT ATA AGA CAA GCA CAC TGT AAC CTT 894Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Leu290 295AGT AGA ACA CAA TGG AAT AAC ACT TTA GGA AAG ATA GTC 933Ser Arg Thr Gln Trp Asn Asn Thr Leu Gly Lys Ile Val300 305 310ATA AAA TTA AGA GAA CAA TTT AGA AAA CAA TTT GGA GAA 972Ile Lys Leu Arg Glu Gln Phe Arg Lys Gln Phe Gly Glu315 320AAA ACA ATA GTC TTT AAT CGA TCC TCA GGA GGG GAC CCG 1011Lys Thr Ile Val Phe Asn Arg Ser Ser Gly Gly Asp Pro325 330 335GAA ATT GCA ATG CAC AGT TTT AAT TGT GGA GGG GAA TTT 1050Glu Ile Ala Met His Ser Phe Asn Cys Gly Gly Glu Phe340 345 350TTC TAC TGT AAC ACA ACA GCA CTG TTT AAT AGT ACC TGG 1089Phe Tyr Cys Asn Thr Thr Ala Leu Phe Asn Ser Thr Trp355 360AAT GTT ACT AAA GGG TTG AAT AAC ACT GAA GGA AAT AGC 1128Asn Val Thr Lys Gly Leu Asn Asn Thr Glu Gly Asn Ser365 370 375ACA GGA GAT GAA AAT ATC ATA CTC CCA TGT AGA ATA AAA 1167Thr Gly Asp Glu Asn Ile Ile Leu Pro Cys Arg Ile Lys380 385CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA ATG 1206Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met390 395 400TAT GCC CCT CCC ATC AGT GGA CAA ATT AGA TGT TCA TCA 1245Tyr Ala Pro Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser405 410 415AAC ATT ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT AGT 1284Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Ser420 425AAG AAC GAG AGC ATC ACC ACC GAG GTC TTC AGA CCT GGA 1323Lys Asn Glu Ser Ile Thr Thr Glu Val Phe Ar9 Pro Gly430 435 440GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT GAA TTA TAT 1362Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr445 450AAA TAT AAA GTA GTA AAA ATT GAA CCA TTA GGA GTA GCG 1401Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala455 460 465CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG AGA GAA AAA 1440Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys470 475 480AGA GCA GTG GGA ACA ATA GGA GCT ATG TTC CTT GGG TTC 1479Arg Ala Val Gly Thr Ile Gly Ala Met Phe Leu Gly Phe485 490TTG GGA GCA TAA AGC TTC TAG AGT CGA CCT GCA 1512Leu Gly Ala Xaa Ser Phe Xaa Ser Arg Pro Ala495 500 504(2)SEQ ID NO:10的信息(ⅰ)序列特征(A)長度504個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:10:Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys1 5 10 15Ala Ser Asp Ala Lys Ala Tyr Asn Thr Glu Lys His Asn Val Trp20 25 30Ala Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Val35 40 45Val Leu Gly Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn50 55 60Met Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln65 70 75Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90Asn Cys Thr Asp Asp Leu Gly Asn Ala Thr Asn Thr Asn Ser Ser95 100 105Ala Thr Thr Asn Ser Ser Ser Trp Glu Glu Met Lys Gly Glu Met110 115 120Lys Arg Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp Lys Ile125 130 135Lys Lys Glu His Ala Leu Phe Tyr Arg Leu Asp Val Val Pro Ile140 145 150Asp Asn Asp Asn Thr Thr Tyr Arg Leu Ile Asn Cys Asn Thr Ser155 160 165Val Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro170 175 180Ile His Phe Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn185 190 195Asn Lys Thr Phe Glu Gly Lys Gly Pro Cys Lys Asn Val Ser Thr200 205 210Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln Leu215 220 225Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Ile Ile Arg Ser230 235 240Asp Asn Ile Thr Asp Asn Thr Lys Thr Ile Ile Val Gln Leu Asn245 250 255Glu Ser Val Val Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg260 265 270Lys Ser Ile His Ile Gly Pro Gly Ser Ala Phe Phe Ala Thr Gly275 280 285Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Leu Ser Arg290 295 300Thr Gln Trp Asn Asn Thr Leu Gly Lys Ile Val Ile Lys Leu Arg305 310 315Glu Gln Phe Arg Lys Gln Phe Gly Glu Lys Thr Ile Val Phe Asn320 325 330Arg Ser Ser Gly Gly Asp Pro Glu Ile Ala Met His Ser Phe Asn335 340 345Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Ala Leu Phe Asn350 355 360Ser Thr Trp Asn Val Thr Lys Gly Leu Asn Asn Thr Glu Gly Asn365 370 375Ser Thr Gly Asp Glu Asn Ile Ile Leu Pro Cys Arg Ile Lys Gln380 385 390Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met Tyr Ala Pro395 400 405Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser Asn Ile Thr Gly Leu410 415 420Leu Leu Thr Arg Asp Gly Gly Ser Lys Asn Glu Ser Ile Thr Thr425 430 435Glu Val Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg440 445 450Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly455 460 465Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys470 475 480Arg Ala Val Gly Thr Ile Gly Ala Met Phe Leu Gly Phe Leu Gly485 490 495Ala Xaa Ser Phe Xaa Ser Arg Pro Ala500 504(2)SEQ ID NO:11的信息(ⅰ)序列特征(A)長度1501個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQIDNO:11:
CTC GAG GTA CCT GTG TGG AAA GAA GCA ACT ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT AAT ACA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asn Thr Glu15 20 25AAA CAT AAT GTT TGG GCC ACA CAC GCC TGT GTA CCC ACA 114Lys His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr30 35GAT CCC AAC CCA CAA GAA GTA GTA TTG GGA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Val Leu Gly Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA 192Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln55 60ATG CAT GAA GAT ATA ATC AGT TTA TGG GAT CAA AGT CTA 231Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAT GAT TTA GGG AAT GCT ACT AAT ACC AAT 309Asn Cys Thr Asp Asp Leu Gly Asn Ala Thr Asn Thr Asn95 100AGC AGT GCC ACT ACC AAT AGT AGT AGT TGG GAA GAA ATG 348Ser Ser Ala Thr Thr Asn Ser Ser Ser Trp Glu Glu Met105 110 115AAG GGG GAA ATG AAA AGG TGC TCT TTC AAT ATC ACC ACA 387Lys Gly Glu Met Lys Arg Cys Ser Phe Asn Ile Thr Thr120 125AGC ATA AGA GAT AAG ATT AAG AAA GAA CAT GCA CTT TTC 426Ser Ile Arg Asp Lys Ile Lys Lys Glu His Ala Leu Phe130 135 140TAT AGA CTT GAT GTA GTA CCA ATA GAT AAT GAT AAT ACC 465Tyr Arg Leu Asp Val Val Pro Ile Asp Asn Asp Asn Thr145 150 155ACA TAT AGG TTG ATA AAT TGT AAT ACC TCA GTC ATT ACA 504Thr Tyr Arg Leu Ile Asn Cys Asn Thr Ser Val Ile Thr160 165CAG GCC TGT CCA AAG GTA TCA TTT GAG CCA ATT CCC ATA 543Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile170 175 180CAT TTT TGT GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT 582His Phe Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys185 190AAT AAT AAG ACG TTC GAG GGA AAA GGA CCA TGT AAA AAT 621Asn Asn Lys Thr Phe Glu Gly Lys Gly Pro Cys Lys Asn195 200 205GTC AGT ACA GTA CAA TGC ACA CAT GGA ATT AGG CCA GTA 650Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro Val210 215 220GTG TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA GAA 699Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu225 230GAA GAG GTA ATA ATT AGA TCT GGC AAT ATC ACA GAC AAT 738Glu Glu Val Ile Ile Arg Ser Gly Asn Ile Thr Asp Asn235 240 245ACT AAA ACC ATT ATA GTA CAG CTA AAC GAA TCT GTA GTA 777Thr Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Val Val250 255ATT AAT TGT ACA AGA TCC AAC AAC AAT ACA AGA AAA AGT 816Ile Asn Cys Thr Arg Ser Asn Asn Asn Thr Arg Lys Ser260 265 270ATA CAT ATA GGA CCA GGG AGT GCA TTT TTT GCA ACA GGA 855Ile His Ile Gly Pro Gly Ser Ala Phe Phe Ala Thr Gly275 280 285GAA ATA ATA GGA GAT ATA AGA CAA GCA CAC TGT AAC CTT 894Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Leu290 295AGT AGA ACA CAA TGG AAT AAC ACT TTA GGA AAG ATA GTC 933Ser Arg Thr Gln Trp Asn Asn Thr Leu Gly Lys Ile Val300 305 310ATA AAA TTA AGA GAA CAA TTT AGA AAA CAA TTT GGA GAA 972Ile Lys Leu Arg Glu Gln Phe Arg Lys Gln Phe Gly Glu315 320AAA ACA ATA GTC TTT AAT CGA TCC TCA GGA GGG GAC CCG 1011Lys Thr Ile Val Phe Asn Arg Ser Ser Gly Gly Asp Pro325 330 335GAA ATT GCA ATG CAC AGT TTT AAT TGT GGA GGG GAA TTT 1050Glu Ile Ala Met His Ser Phe Asn Cys Gly Gly Glu Phe340 345 350TTC TAC TGT AAC ACA ACA GCA CTG TTT AAT AGT ACC TGG 1089Phe Tyr Cys Asn Thr Thr Ala Leu Phe Asn Ser Thr Trp355 360AAT GTT ACT AAA GGG TTG AAT AAC ACT GAA GGA AAT AGC 1128Asn Val Thr Lys Gly Leu Asn Asn Thr Glu Gly Asn Ser365 370 375ACA GGG GAT GAA AAT ATC ATA CTC CCA TGT AGA ATA AAA 1167Thr Gly Asp Glu Asn Ile Ile Leu Pro Cys Arg Ile Lys380 385CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA ATG 1206Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met390 395 400TAT GCC CCT CCC ATC AGT GGA CAA ATT AGA TGT TCA TCA 1245Tyr Ala Pro Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser405 410 415AAT ATT ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT AGT 1284Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly ser420 425AAG AAC GAG AGC ATC ACC ACC GAG GTC TTC AGA CCT GGA 1323Lys Asn Glu Ser Ile Thr Thr Glu Val Phe Arg Pro Gly430 435 440GGA GGA GAT ATG AGG GAC AAT TGG AGA AGT GAA TTA TAT 1362Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr445 450AAA TAT AAA GTA GTA AAA ATT GAA CCA TTA GGA GTA GCG 1401Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala455 460 465CCC ACC AAG GCA AAG AGA AGA GTG GTG CAG AGA GAA AAA 1440Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys470 475 480AGA GCA GTG GGA ACA ATA GGA GCT ATG TTC CTT GGG TTC 1479Arg Ala Val Gly Thr Ile Gly Ala Met Phe Leu Gly Phe485 490TTA GGA GCA TAA AGC TTC TAG A 1501Leu Gly Ala Xaa Ser Phe Xaa495 500(2)SEQ ID NO:12的信息(ⅰ)序列特征(A)長度500個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:12:Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys1 5 10 15Ala Ser Asp Ala Lys Ala Tyr Ash Thr Glu Lys His Ash Val Trp20 25 30Ala Thr His Ala Cys Val Pro Thr Asp Pro Ash Pro Gln Glu Val35 40 45Val Leu Gly Ash Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn50 55 60Met Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln65 70 75Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90Asn Cys Thr Asp Asp Leu Gly Asn Ala Thr Asn Thr Asn Ser Ser95 100 105Ala Thr Thr Asn Ser Ser Ser Trp Glu Glu Met Lys Gly Glu Met110 115 120Lys Arg Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp Lys Ile125 130 135Lys Lys Glu His Ala Leu Phe Tyr Arg Leu Asp Val Val Pro Ile140 145 150Asp Asn Asp Asn Thr Thr Tyr Arg Leu Ile Asn Cys Asn Thr Ser155 160 165Val Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro170 175 180Ile His Phe Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn185 190 195Asn Lys Thr Phe Glu Gly Lys Gly Pro Cys Lys Asn Val Ser Thr200 205 210Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln Leu215 220 225Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Ile Ile Arg Ser230 235 240Gly Asn Ile Thr Asp Asn Thr Lys Thr Ile Ile Val Gln Leu Asn245 250 255Glu Ser Val Val Ile Asn Cys Thr Arg Ser Asn Asn Asn Thr Arg260 265 270Lys Ser Ile His Ile Gly Pro Gly Ser Ala Phe Phe Ala Thr Gly275 280 285Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Leu Ser Arg290 295 300Thr Gln Trp Asn Asn Thr Leu Gly Lys Ile Val Ile Lys Leu Arg305 310 315Glu Gln Phe Arg Lys Gln Phe Gly Glu Lys Thr Ile Val Phe Asn320 325 330Arg Ser Ser Gly Gly Asp Pro Glu Ile Ala Met His Ser Phe Asn335 340 345Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Ala Leu Phe Asn350 355 360Ser Thr Trp Asn Val Thr Lys Gly Leu Asn Asn Thr Glu Gly Asn365 370 375Ser Thr Gly Asp Glu Asn Ile Ile Leu Pro Cys Arg Ile Lys Gln380 385 390Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met Tyr Ala Pro395 400 405Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser Asn Ile Thr Gly Leu410 415 420Leu Leu Thr Arg Asp Gly Gly Ser Lys Asn Glu Ser Ile Thr Thr425 430 435Glu Val Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg440 445 450Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly455 460 465Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys470 475 480Arg Ala Val Gly Thr Ile Gly Ala Met Phe Leu Gly Phe Leu Gly485 490 495Ala Xaa Ser Phe Xaa500(2)SEQ ID NO:13的信息(ⅰ)序列特征(A)長度1514個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:13:GG GAA TTC GGA TCC GGG GTA CCT GTG TGG AAG GAA GCA 38Glu Phe Gly Ser Gly Val Pro Val Trp Lys Glu Ala1 5 10ACC ACC ACT CTA TTC TGT GCA TCA GAT GCT AGA GCA TAT 77Thr Thr Thr Leu Phe Cys Ala Ser Asp Ala Ar9 Ala Tyr15 20 25GAC ACA GAG GTA CAT AAT GTT TGG GCC ACA CAT GCC TGT 116Asp Thr Glu Val His Asn Val Trp Ala Thr His Ala Cys30 35GTA CCC ACA GAC CCT AGT CCA CAA GAA GTA GTT TTG GAA 155Val Pro Thr Asp Pro Ser Pro Gln Glu Val Val Leu Glu40 45 50AAT GTG ACA GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG 194Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met55 60GTA GAA CAA ATG CAT GAG GAT ATA ATT AGT TTA TGG GAT 233Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp65 70 75CAA AGC TTA AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT 272Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys80 85 90GTT ACT TTA AAT TGC AGT GAT TAT AGG AAT GCT ACT GAT 311Val Thr Leu Asn Cys Ser Asp Tyr Arg Asn Ala Thr Asp95 100TAT AAG AAT GCT ACT GAT ACC ACT AGT AGT AAC GAG GGA 350Tyr Lys Asn Ala Thr Asp Thr Thr Ser Ser Asn Glu Gly105 110 115AAG ATG GAG AGA GGA GAA ATA AAA AAC TGC TCT TTC AAT 389Lys Met Glu Arg Gly Glu Ile Lys Asn Cys Ser Phe Asn120 125ATT ACC ACA AGC ATA AAA AAT AAG ATG CAG AAA GAA TAT 428Ile Thr Thr Ser Ile Lys Asn Lys Met Gln Lys Glu Tyr130 135 140GCA CTT TTC TAT AAA CTT GAT ATA GTA CCA ATA GAT AAT 467Ala Leu Phe Tyr Lys Leu Asp Ile Val Pro Ile Asp Asn145 150 155ACA AGC TAT ACA TTG ATA AGT TGT AAC ACC TCA GTC ATT 506Thr Ser Tyr Thr Leu Ile Ser Cys Asn Thr Ser Val Ile160 165ACA CAG GCC TGT CCA AAG GTA TCC TTT GAA CCA ACT CCC 545Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Thr Pro170 175 180ATA CAT TAT TGT GCT CCG GCT GGT TTT GCG ATT CTA AAG 584Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys185 190TGT AAT GAT AAG AAG TTC AGT GGA AAA GGA GAA TGT AAA 623Cys Asn Asp Lys Lys Phe Ser GIy Lys Gly Glu Cys Lys195 200 205AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AGG CCA 662Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro210 215 220GTA GTA TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA 701Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala225 230GAA GAA GAG GTG GTA ATT AGA TCT GAC AAT TTC ATA GAC 740Glu Glu Glu Val Val Ile Arg Ser Asp Asn Phe Ile Asp235 240 245AAT ACT AAA ACC ATA ATA GTA CAG CTG AAA GAA TCT GTA 779Asn Thr Lys Thr Ile Ile Val Gln Leu Lys Glu Ser Val250 255GAA ATT AAT TGT ATA AGA CCC AAC AAT AAT ACA AGA AAA 818Glu Ile Asn Cys Ile Arg Pro Asn Asn Asn Thr Arg Lys260 265 270GGT ATA CAT ATA GGA CCA GGG AGA GCA TGG TAT GCA ACA 857Gly Ile His Ile Gly Pro Gly Arg Ala Trp Tyr Ala Thr275 280 285GGA GAA ATA GTA GGA GAT ATA AGA AAG GCA TAT TGT AAC 896Gly Glu Ile Val Gly Asp Ile Arg Lys Ala Tyr Cys Asn290 295ATT AGT AGA ACA AAA TGG AAT AAC ACT TTA ATA CAG ATA 935Ile Ser Arg Thr Lys Trp Asn Asn Thr Leu Ile Gln Ile300 305 310GCT AAC AAA TTA AAA GAA AAA TAT AAT ACA ACA ATA AGC 974Ala Asn Lys Leu Lys Glu Lys Tyr Asn Thr Thr Ile Ser315 320TTT AAT CGA TCC TCA GGA GGG GAC CCA GAA ATT GTA ACG 1013Phe Asn Arg Ser Ser Gly Gly Asp Pro Glu Ile Val Thr325 330 335CAT AGT TTT AAT TGT GGA GGG GAG TTT TTC TAC TGT GAT 1052His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asp340 345 350TCA ACA CAA CTG TTT AAT AGT ACT TGG AAT TTA AAT GGT 1091Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Leu Asn Gly355 360ACT TGG AAT TTT ACT GCA GGG TCA AAT GAA ACT GAA GGC 1130Thr Trp Asn Phe Thr Ala Gly Ser Asn Glu Thr Glu GIy365 370 375AAT ATC ACA CTC CCA TGC AGA ATA AAA CAA ATT ATA AAC 1169Asn Ile Thr Leu Pro Cys Arg Ile Lys Glln Ile Ile Asn380 385AGG TGG CAG GAA GTA GGG AAA GCA ATG TAT GCC CCT CCC 1208Arg Trp Gln Glu Val Gly Lys Ala Met Tyr Ala Pro Pro390 395 400ATC AGT GGA CAA ATA AAA TGC TCA TCA AAC ATT ACA GGG 1247Ile Ser Gly Gln Ile Lys Cys Ser Ser Asn Ile Thr Gly405 410 415ATG ATA TTA ACA AGG GAT GGT GGT AAC GAG AAC AAT AAT 1286Met Ile Leu Thr Arg Asp Gly Gly Asn Glu Asn Asn Asn420 425GAG AGC AGT ACT ACT GAG ACC TTC AGA CCG GGA GGA GGA 1325Glu Ser Ser Thr Thr Glu Thr Phe Arg Pro Gly Gly Gly430 435 440GAT ATG AGG AAC AAT TGG AGA AGT GAA TTA TAT AAA TAT 1364Asp Met Arg Asn Asn Trp Arg Ser Glu Leu Tyr Lys Tyr445 450AAA GTA GTA AAA ATT GAA CCA TTA GGA GTA GCA CCC ACC 1403Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala Pro Thr455 460 465AAG GCA AAG AGA AGA GTG GTG CAG AGA GAA AAA AGA GCA 1442Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys Arg Ala470 475 480GTG GGA GCG CTA GGA GCT ATG TTC CTT GGG TTC TTA GGA 1481Val Gly Ala Leu Gly Ala Met Phe Leu Gly Phe Leu Gly485 490GCA TAA AGC TTC TAG ACC GAC TCT AGA GGA TCC 1514Ala Xaa Ser Phe Xaa Thr Asp Ser Arg Gly Ser495 500 504(2)SEQ ID NO:14的信息(ⅰ)序列特征(A)長度504個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:14:Glu Phe Gly Ser Gly Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10 15Leu Phe Cys Ala Ser Asp Ala Arg Ala Tyr Asp Thr Glu Val His20 25 30Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro Ser Pro35 40 45Gln Glu Val Val Leu Glu Asn Val Thr Glu Asn Phe Asn Met Trp50 55 60Lys Asn Asn Met Val Glu Gln Met His Glu Asp Ile Ile Ser Leu65 70 75Trp Asp Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys80 85 90Val Thr Leu Asn Cys Ser Asp Tyr Arg Asn Ala Thr Asp Tyr Lys95 100 105Asn Ala Thr Asp Thr Thr Ser Ser Asn Glu Gly Lys Met Glu Arg110 115 120Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Lys125 130 135Asn Lys Met Gln Lys GLu Tyr Ala Leu Phe Tyr Lys Leu Asp Ile140 145 150Val Pro Ile Asp Asn Thr Ser Tyr Thr Leu Ile Ser Cys Asn Thr155 160 165Ser Val Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Thr170 175 180Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys185 190 195Asn Asp Lys Lys Phe Ser Gly Lys Gly Glu Cys Lys Asn Val Ser200 205 210Thr Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln215 220 225Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Val Ile Arg230 235 240Ser Asp Asn Phe Ile Asp Asn Thr Lys Thr Ile Ile Val Gln Leu245 250 255Lys Glu Ser Val Glu Ile Asn Cys Ile Arg Pro Asn Asn Asn Thr260 265 270Arg Lys Gly Ile His Ile Gly Pro Gly Arg Ala Trp Tyr Ala Thr275 280 285Gly Glu Ile Val Gly Asp Ile Arg Lys Ala Tyr Cys Asn Ile Ser290 295 300Arg Thr Lys Trp Asn Asn Thr Leu Ile Gln Ile Ala Asn Lys Leu305 310 315Lys Glu Lys Tyr Asn Thr Thr Ile Ser Phe Asn Arg Ser Ser Gly320 325 330Gly Asp Pro Glu Ile Val Thr His Ser Phe Asn Cys Gly Gly Glu335 340 345Phe Phe Tyr Cys Asp Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn350 355 360Leu Asn Gly Thr Trp Asn Phe Thr Ala Gly Ser Asn Glu Thr Glu365 370 375Gly Asn Ile Thr Leu Pro Cys Arg Ile Lys Gln Ile Ile Asn Arg380 385 390Trp Gln Glu Val Gly Lys Ala Met Tyr Ala Pro Pro Ile Ser Gly395 400 405Gln Ile Lys Cys Ser Ser Asn Ile Thr Gly Met Ile Leu Thr Arg410 415 420Asp Gly Gly Asn Glu Asn Asn Asn Glu Ser Ser Thr Thr Glu Thr425 430 435Phe Arg Pro Gly Gly Gly Asp Met Arg Asn Asn Trp Arg Ser Glu440 445 450Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala455 460 465Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys Arg Ala470 475 480Val Gly Ala Leu Gly Ala Met Phe Leu Gly Phe Leu Gly Ala Xaa485 490 495Ser Phe Xaa Thr Asp Ser Arg Gly Ser500 504(2)SEQ ID NO:15的信息(ⅰ)序列特征(A)長度1408個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:15:G GTA CCT GTG TGG AAG GAA GCA ACC ACC ACT CTA TTC 37Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AGA GCA TAT GAC ACA GAG GTA CAT 76Cys Ala Ser Asp Ala Arg Ala Tyr Asp Thr Glu Val His15 20 25AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC CCT 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AGT CCA CAA GAA GTA TTT TTG GGA AAT GTG ACA GAA AAT 154Ser Pro Gln Glu Val Phe Leu Gly Asn Val Thr Glu Asn40 45 50TTT AAT ATG TGG AAA AAT AAC ATG GTA GAA CAA ATG TAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met Tyr55 60GAG GAT ATA ATT AGT TTA TGG GAT CAA AGC TTA AAG CCA 232Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAA TTA ACC CCA CTC TGT GTT ACT TTA AAT TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90AGT GAT TAT AGG AAT GCT ACT GAT TAT AAG AAT GCT ACT 310Ser Asp Tyr Arg Asn Ala Thr Asp Tyr Lys Asn Ala Thr95 100GAT ACC ACT AGT AGT AAC GAG GGA AAG ATG GAG AGA GGA 349Asp Thr Thr Ser Ser Asn Glu Gly Lys Met Glu Arg Gly105 110 115GAA ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA AGC ATA 388Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile120 125AAA AAT AAG ATG CAG AAA GAA TAT GCA CTT TTC TAT AAA 427Lys Asn Lys Met Gln Lys Glu Tyr Ala Leu Phe Tyr Lys130 135 140CTT AAT ATA GTA CCA ATA GAT AAT ACA AGC TAT ACA TTG 466Leu Asn Ile Val Pro Ile Asp Asn Thr Ser Tyr Thr Leu145 150 155ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCC TGT CCA 505Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro160 165AAG GTA TCC TTT GAA CCA ATT CCC ATA CAT TAT TGT GCT 544Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala170 175 180CCG GCT GGT TTT GCG ATT CTA AAG TGT AAT GAT AAG AAG 583Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys Lys185 190TTC AGT GGA AAA GGA GAA TGT AAA AAT GTC AGC ACA GTA 622Phe Ser Gly Lys Gly Glu Cys Lys Asn Val Ser Thr Val195 200 205CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT CAA 661Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln210 215 220CTG CTG TTA AAT GGC AGT CTA GCA GAA GAA GAG GTG GTA 700Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Val225 230ATT AGA TCT GAC AAT TTC ACA GAC AAT ACT AAA ACC ATA 739Ile Arg Ser Asp Asn Phe Thr Asp Asn Thr Lys Thr Ile235 240 245ATA GTA CAG CTG AAA GAA TCT GTA GAA ATT AAT TGT ATA 778Ile Val Gln Leu Lys Glu Set Val Glu Ile Asn Cys Iie250 255AGA CCC AAC AAT AAT ACA AGA AAA GGT ATA CAT ATA GGA 817Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Ile Gly260 265 270CCA GGG AGA GCA TGG TAT GCA ACA GGA GAA ATA GTA GGA 856Pro Gly Arg Ala Trp Tyr Ala Thr Gly Glu Ile Val Gly275 280 285GAT ATA AGA CAG GCA TAT TGT AAC ATT AGT AGA ACA AAA 895Asp Ile Arg Gln Ala Tyr Cys Asn Ile Ser Arg Thr Lys290 295TGG AAT AAC ACT TTA ATA CAG ATA GCT AAC AAA TTA AAA 934Trp Asn Asn Thr Leu Ile Gln Ile Ala Asn Lys Leu Lys300 305 310GAA AAA TAT AAT ACA ACA ATA AGC TTT AAT CGA TCC TCA 973Glu Lys Tyr Asn Thr Thr Ile Ser Phe Asn Arg Ser Ser315 320GGA GGG GAC CCA GAA ATT GTA ACC CAT AGT TTT AAT TGT 1012Gly Gly Asp Pro Glu Ile Val Thr His Ser Phe Asn Cys325 330 335GGA GGG GAA TTT TTC TAC TGT AAT TCA ACA CAA CTG TTT 1051Gly Gly Glu Phe Phe Tyr Cys Asn Ser Thr Gln Leu Phe340 345 350AAT AGT ACT TGG AAT TTA AAT GGT ACT TGG AAT TTT ACT 1090Asn Ser Thr Trp Asn Leu Asn Gly Thr Trp Asn Phe Thr355 360GCA GGG TCA AAT GAA ACT GAA GGC AAT ATC ACA CTC CCA 1129Ala Gly Ser Asn Glu Thr Glu Gly Asn Ile Thr Leu Pro365 370 375TGC AGA ATA AAA CAA ATT ATA AAC AGG TGG CAG GAA GTA 1165Cys Arg Ile Lys Gln Ile Ile Asn Arg Trp Gln Glu Val380 385GGA AAA GCA ATG TAT GCC CCT CCC ATC AGT GGA CAA ATA 1207Gly Lys Ala Met Tyr Ala Pro Pro Ile Ser Gly Gln Ile390 395 400AGA TGC TCA TCA AAC ATT ACA GGG ATG ATA TTA ACA AGG 1246Arg Cys Ser Ser Asn Ile Thr Gly Met Ile Leu Thr Arg405 410415GAT GGT GGT AAC GAG AAC AAT AAT GAG AGC AGT ACT ACT 12/5Asp Gly Gly Asn Glu Asn Asn Asn Glu Ser Ser Thr Thr420 425GAG ACC TTC AGA CCG GGA GGA GGA GAT ATG AGG AAC AAT 1324Glu Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asn Asn430 435 440TGG AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA AAA ATT 1363Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile445 450GAG CCA TTA GGA GTA GCA CCC ACC GAC TCT AGA GGA TCC 1402Glu Pro Leu Gly Val Ala Pro Thr Asp Ser Arg Gly Ser455 460 465TCT AGA 1408Ser Arg469(2)SEQ ID NO:16的信息(ⅰ)序列特征(A)長度469個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:16:Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala Ser1 5 10 15Asp Ala Arg Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala Thr2 25 30His Ala Cys Val Pro Thr Asp Pro Ser Pro Gln Glu Val Phe Leu35 40 45Gly Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val50 55 60Glu Gln Met Tyr Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys80 85 90Ser Asp Tyr Arg Asn Ala Thr Asp Tyr Lys Asn Ala Thr Asp Thr95 100 105Thr Ser Ser Asn Glu Gly Lys Met Glu Arg Gly Glu Ile Lys Asn110 115 120Cys Ser Phe Asn Ile Thr Thr Ser Ile Lys Asn Lys Met Gln Lys125 130 135Glu Tyr Ala Leu Phe Tyr Lys Leu Asn Ile Val Pro Ile Asp Asn140 145 150Thr Ser Tyr Thr Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln155 160 165Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys170 175 180Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys Lys Phe185 190 195Ser Gly Lys Gly Glu Cys Lys Asn Val Ser Thr Val Gln Cys Thr200 205 210His Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly215 220 225Ser Leu Ala Glu Glu Glu Val Val Ile Arg Ser Asp Asn Phe Thr230 235 240Asp Asn Thr Lys Thr Ile Ile Val Gln Leu Lys Glu Ser Val Glu245 250 255Ile Asn Cys Ile Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His260 265 270Ile Gly Pro Gly Arg Ala Trp Tyr Ala Thr Gly Glu Ile Val Gly275 280 285Asp Ile Arg Gln Ala Tyr Cys Asn Ile Ser Ar9 Thr Lys Trp Asn290 295 300Asn Thr Leu Ile Gln Ile Ala Asn Lys Leu Lys Glu Lys Tyr Asn305 310 315Thr Thr Ile Ser Phe Asn Arg Ser ser Gly Gly Asp Pro Glu Ile320 325 330Val Thr His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn335 340 345Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Leu Asn Gly Thr Trp350 355 360Asn Phe Thr Ala Gly Ser Asn Glu Thr Glu Gly Asn Ile Thr Leu365 370 375Pro Cys Ar9 Ile Lys Gln Ile Ile Asn Arg Trp Gln Glu Val Gly380 385 390Lys Ala Met Tyr Ala Pro Pro Ile Ser Gly Gln Ile Arg Cys Ser395 400 405Ser Asn Ile Thr Gly Met Ile Leu Thr Arg Asp Gly Gly Asn Glu410 415 420Asn Asn Asn Glu Ser Ser Thr Thr Glu Thr Phe Arg Pro Gly Gly425 430 435Gly Asp Met Arg Asn Asn Trp Arg Ser Glu Leu Tyr Lys Tyr Lys440 445 450Val Val Lys Ile Glu Pro Leu Gly Val Ala Pro Thr Asp Ser Arg455 460 465Gly ser Ser Arg469(2)SEQ ID NO:17的信息(ⅰ)序列特征(A)長度1499個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:17:
GAG GTA CCT GTG TGG AAA GAA GCA ACC ACT ACT CTA 36Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GGG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Gly Val15 20 25CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp30 35CCC AAC CCA CAA GAA ATA GAA TTG GTA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Ile Glu Leu Val Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT AAA ATG GTA GAC CAG ATG 192Asp Phe Asn Met Trp Lys Asn Lya Met Val Asp Gln Met55 60CAT GAG GAT ATA ATC AGT TTA TGG GAT GAA AGC CTA AAG 231His Glu Asp Ile Ile Ser Leu Trp Asp Glu Ser Leu Lys65 70 75CCA TGT GTA AAG TTA ACC CCA CTT TGT GTT ACT CTA AAC 270Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn80 85 90TGC AGT GAT GTG AAC AAT TCC ACA AAT CCT AAT GAT ACT 309Cys Ser Asp Val Asn Asn Ser Thr Asn Pro Asn Asp Thr95 100AAT ACT AAT TCC ACT AAT ACT ACT TCC TCT ACT CCT ACG 348Asn Thr Asn Ser Thr Asn Thr Thr Ser Ser Thr Pro Thr105 110 115GCC ACT ACT AGT AGC GAG GAA AAG ATG GAG AAG GGA GAA 387Ala Thr Thr Ser Ser Glu Glu Lys Met Glu Lys Gly Glu120 125ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA CAC ATG AAA 426Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr His Met Lys130 135 140GAT AAG GCA CAG AAA GAA TAT GCA CTT TTT TAT AAA CTT 465Asp Lys Ala Gln Lys Glu Tyr Ala Leu Phe Tyr Lys Leu145 150 155GAT ATA GTA CCA ATA GAT GAT AAT AAT GCC AGC TAT AGG 504Asp Ile Val Pro Ile Asp Asp Asn Asn Ala Ser Tyr Arg160 165TTG ATA AGT TGT AAT ACC TCA GAC ATT ACA CAG GCC TGT 543Leu Ile Ser Cys Asn Thr Ser Asp Ile Thr Gln Ala Cys170 175 180CCA AAG GTG ACC TTT GAG CCA ATT CCC ATA CAT TAT TGT 582Pro Lys Val Thr Phe Glu Pro Ile Pro Ile His Tyr Cys185 190GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG 621Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys195 200 205AAG TTC AAT GGA ACA GGA CCA TGT TCA AAG GTC AGC ACA 660Lys Phe Asn Gly Thr Gly Pro Cys Ser Lys Val Ser Thr210 215 220GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 699Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr225 230CAA CTG TTG TTA AAT GGC AGT CTT GCA GAA GAA GAA GTA 738Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val235 240 245GTA ATT AGA TCT GTC AAT TTC ACA GAC AAT GCT AAA ATC 777Val Ile Arg Ser Val Asn Phe Thr Asp Asn Ala Lys Ile250 255ATA ATA GTA CAG CTG AAA GAA CCT GTA GCA ATT AAT TGT 816Ile Ile Val Gln Leu Lys Glu Pro Val Ala Ile Asn Cys260 265 270ACA AGA CCC AAC AAC AAT ACA AGA AAA GGT ATA CAT CTA 855Thr Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Leu275 280 285GGA CCA GGG AgC ACA TTT TAT ACA ACA GGA GAA ATA ATA 894Gly Pro Gly Ser Thr Phe Tyr Thr Thr Gly Glu Ile Ile290 295GGA GAC ATA AGA AAA GCA TAT TGC AAG ATT AGT AAA GAA 933Gly Asp Ile Arg Lys Ala Tyr Cys Lys Ile Ser Lys Glu300 305 310AAA TGG AAT AAC ACT TTA AGA CAG GTA GTT AAA AAA TTA 972Lys Trp Asn Asn Thr Leu Arg Gln Val Val Lys Lys Leu315 320AGA GAA CAA TTT GGG AAT AAA ACA ATA ATT TTT AAT CGA 1011Arg Glu Gln Phe Gly Asn Lys Thr Ile Ile Phe Asn Arg325 330 335TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT 1050Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe340 345350AAC TGT GGA GGG GAG TTT TTC TAC TGT AAT ACA ACA CAA 1089Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Gln355 360CTG TTT AAT AGT ACT TGG AAT AAT ACT GAA GGG ACA AAT 1128Leu Phe Asn Ser Thr Trp Asn Asn Thr Glu Gly Thr Asn365 370 375AGC ACT GAA GGA AAT AGC ACA ATC ACA CTC CCA TGC AGA 1167Ser Thr Glu Gly Asn Ser Thr Ile Thr Leu Pro Cys Arg380 385ATA AAA CAA ATT ATA AAT ATG TGG CAG GAA GTA GGA AAA 1206Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys390 395 400GCA ACG TAT GCC CCT CCC ATC AGA GGA CGA ATT AGA TGC 1245Ala Thr Tyr Ala Pro Pro Ile Arg Gly Arg Ile Arg Cys405 410 415ATA TCA AAT ATT ACA GGA CTG CTA TTA ACA AGA GAT GGT 1284Ile Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly420 425GGT AGG AAT GTC ACA AAC AAT ACCGAA ACC TTC AGA CCT 1323Gly Arg Asn Val Thr Asn Asn Thr Glu Thr Phe Arg Pro430 435 440GGA GGA GGA GAC ATG AGG GAC AAT TGG AGA AGT GAA TTA 1362Gly Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu445 450TAT AAA TAT AAA GTA GTA AAA GTT GAA CCA TTA GGA ATA 1401Tyr Lys Tyr Lys Val Val Lys Val Glu Pro Leu Gly Ile455 460 465GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAC AGA GAC 1440Ala Pro Thr Lys Ala Lys Arg Arg Val Val His Arg Asp470 475 480AAA AGA GCA GCA CTA GGA GCC TTG TTC CTT GGG TTC TTA 1479Lys Arg Ala Ala Leu Gly Ala Leu Phe Leu Gly Phe Leu485 490GGA GCA TAA AAG CTT CTA GA 1499Gly Ala Xaa Lys Leu Leu495 499(2)SEQ ID NO:18的信息(ⅰ)序列特征(A)長度499個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:18:Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala1 5 10 15Ser Asp Ala Lys Ala Tyr Asp Thr Gly Val His Asn Val Trp Ala20 25 30Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Ile Glu35 40 45Leu Val Asn Val Thr Glu Asp Phe Asn Met Trp Lys Asn Lys Met50 55 60Val Asp Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Glu Ser65 70 75Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn80 85 90Cys Ser Asp Val Asn Asn Ser Thr Asn Pro Asn Asp Thr Asn Thr95 100 105Asn Ser Thr Asn Thr Thr Ser Ser Thr Pro Thr Ala Thr Thr Ser110 115 120Ser Glu Glu Lys Met Glu Lys Gly Glu Ile Lys Asn Cys Ser Phe125 130 135Asn Ile Thr Thr His Met Lys Asp Lys Ala Gln Lys Glu Tyr AIa140 145 150Leu Phe Tyr Lys Leu Asp Ile Val Pro Ile Asp Asp Asn Asn Ala155 160 165Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Asp Ile Thr Gln Ala170 175 180Cys Pro Lys Val Thr Phe Glu Pro Ile Pro Ile His Tyr Cys Ala185 190 195Pro Ala Gly Phe Ale Ile Leu Lys Cys Lys Asp Lys Lys Phe Asn200 205 210Gly Thr Gly Pro Cys Ser Lys Val Ser Thr Val Gln Cys Thr His215 220 225Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser230 235 240Leu Ala Glu Glu Glu Val Val Ile Arg Ser Val Asn Phe Thr Asp245 250 255Asn Ala Lys Ile Ile Ile Val Gln Leu Lys Glu Pro Val Ala I1e260 265 270Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Leu275 280 285Gly Pro Gly Ser Thr Phe Tyr Thr Thr Gly Glu Ile Ile Gly Asp290 295 300Ile Arg Lys Ala Tyr Cys Lys Ile Ser Lys Glu Lys Trp Asn Asn305 310 315Thr Leu Arg Gln Val Val Lys Lys Leu Arg Glu Gln Phe Gly Asn320 325 330Lys Thr Ile Ile Phe Asn Arg Ser Ser Gly Gly Asp Pro Glu Ile335 340 345Val Met His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn350 355 360Thr Thr Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr Glu Gly Thr365 370 375Asn Ser Thr Glu GIy Asn Ser Thr Ile Thr Leu Pro Cys Arg Ile380 385 390Lys Gln Ile Ile Asn Mer Trp Gln Glu Val Gly Lys Ala Thr Tyr395 400 405Ala Pro Pro Ile Arg Gly Arg Ile Arg Cys Ile Ser Asn Ile Thr410 415 420Gly Leu Leu Leu Thr Arg Asp Gly Gly Arg Asn Val Thr Asn Asn425 430 435Thr Glu Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp440 445 450Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Val Glu Pro Leu455 460 465Gly Ile Ala Pro Thr Lys Ala Lys Arg Arg Val Val His Arg Asp470 475 480Lys Arg Ala Ala Leu Gly Ala Leu Phe Leu Gly Phe Leu Gly Ala485 490 495Xaa Lys Leu Leu499(2)SEQ ID NO:19的信息(ⅰ)序列特征(A)長度1499個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:19:
GAG GTA CCT GTA TGG AAA GAA GCA ACC ACT ACT CTA 36Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu1 5 10TTT TGT GCA TCA GAT GCT AAA GCA TAT GAC ACA GAG GTG 75Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val15 20 25CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC 114His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp30 35CCC AAC CCA CAA GAA ATA GAA TTG GTA AAT GTG ACA GAA 153Pro Asn Pro Gln Glu Ile Glu Leu Val Asn Val Thr Glu40 45 50GAT TTT AAC ATG TGG AAA AAT AAA ATG GTA GAC CAG ATG 192Asp Phe Asn Met Trp Lys Asn Lys Met Val Asp Gln Met55 60CAT GAG GAT ATA ATC AGT TTA TGG GAT GAA AGC CTA AAG 231His Glu Asp Ile Ile Ser Leu Trp Asp Glu Ser Leu Lys65 70 75CCA TGT GTA AAG TTA ACC CCA CTT TGT GTT ACT CTA AAC 270Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn80 85 90TGC AGT GAT GTG AAC AAT TCC ACA AAT CCT AAT GAT ACT 309Cys Ser Asp Val Asn Asn Ser Thr Asn Pro Asn Asp Thr95 100AAT ACT AAT TCC ACT AAT ACT ACT TCC TCT ACT CCT ACG 348Asn Thr Asn Ser Thr Asn Thr Thr Ser Ser Thr Pro Thr105 110 115GCC ACT ACT AGT AGC GAG GAA AAG ATG GAG AAG GGA GAA 387Ala Thr Thr Ser Ser Glu Glu Lys Met Glu Lys Gly Glu120 125ATA AAA AAC TGC TCT TTC AAT ATC ACC ACA CAC ATG AAA 426Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr His Met Lys130 135 140GAT AAG GTA CAG AAA GAA TAT GCA CTT TTT TAT AAA CTT 465Asp Lys Val Gln Lys Glu Tyr Ala Leu Phe Tyr Lys Leu145 150 155GAT ATA GTA CCA ATA GAT GAT AAT AAT ACC AGC TAT AGG 504Asp Ile Val Pro Ile Asp Asp Asn Asn Thr Ser Tyr Arg160 165TTG ATA AGT TGT AAT ACC TCA GTC ATT ACA CAG GCC TGT 543Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys170 175 180CCA ATG GTG ACC TTT GAG CCA ATT CCC ATA CAT TAT TGT 582Pro Met Val Thr Phe Glu Pro Ile Pro Ile His Tyr Cys185 190GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAA GAT AAG 621Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys195 200 205AAG TTC AAT GGA ACA GGA CCA TGT TCA AAG GTC AGC ACA 660Lys Phe Asn Gly Thr Gly Pro Cys Ser Lys Val Ser Thr210 215 220GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 699Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr225 230CAA CTG TTG TTA AAT GGC AGT CTT GCA GAA GAA GAA GTA 738Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val235 240 245GTA ATT AGA TCT GTC AAT TTC ACA GAC AAT GCT AAA ATC 777Val Ile Arg Ser Val Asn Phe Thr Asp Asn Ala Lys Ile250 255ATA ATA GTA CAG CTG AAA GAA CCT GTA GCA ATT AAT TGT 816Ile Ile Val Gln Leu Lys Glu Pro Val Ala Ile Asn Cys260 265 270ACA AGA CCC AAC AAC AAT ACA AGA AAA GGT ATA CAT CTA 855Thr Ar9 Pro Asn Asn Asn Thr Arg Lys Gly Ile His Leu275 280 285GGA CCA GGG AGC ACA TTT TAT ACA ACA GGA GAA ATA ATA 894Gly Pro Gly Ser Thr Phe Tyr Thr Thr Gly Glu Ile Ile290 295GGA GAC ATA AGA AAA GCA TAT TGC AAG ATT AGT AAA GAA 933Gly Asp Ile Arg Lys Ala Tyr Cys Lys Ile Ser Lys Glu300 305 310AAA TGG AAT AAC ACT TTA AGA CAG GTA GTT AAA AAA TTA 972Lys Trp Asn Asn Thr Leu Arg Gln Val Val Lys Lys Leu315 320AGA GAA CAA TTT GGG AAT AAA ACA ATA ATT TTT AAT CGA 1011Arg Glu Gln Phe Gly Asn Lys Thr Ile Ile Phe Asn Arg325 330 335TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT 1050Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe340 345 350AAC TGT GGA GGG GAG TTT TTC TAC TGT AAT ACA ACA CAA 1089Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Gln355 360CTG TTT AAT AGT ACT TGG AAT AAT ACT GAA GGG ACA AAT 1128Leu Phe Asn Ser Thr Trp Asn Asn Thr Glu Gly Thr Asn365 370 375AGC ACT GAA GGA AAT AGC ACA ATC ACA CTC CCA TGC AGA 1167Ser Thr Glu Gly Asn Ser Thr Ile Thr Leu Pro Cys Arg380 385ATA AAA CAA ATT ATA AAT ATG TGG CAG GAA GTA GGA AAA 1206Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys390 395 400GCA ACG TAT GCC CCT CCC ATC AGA GGA CGA ATT AGA TGC 1245Ala Thr Tyr Ala Pro Pro Ile Arg Gly Arg Ile Arg Cys405 410 415ATA TCA AAT ATT ACA GGA CTG CTA TTA ACA AGA GAT GGT 1284Ile Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly420 425GGT AGG AAT GTC ACA AAC AAT ACC GAN NCC TTC AGA CCT 1323Gly Arg Asn Val Thr Asn Asn Thr Xaa Xaa Phe Arg Pro430 435 440GGA GGA GGA GAC ATG AGG GAC AAT TGG AGA AGT GAA TTA 1362Gly Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu445 450TAT AAA TAT AAA GTA GTA AAA GTT GAA CCA TTA GGA ATA 1401Tyr Lys Tyr Lys Val Val Lys Val Glu Pro Leu Gly Ile455 460 465GCA CCC ACC AAG GCA AAG AGA AGA GTG GTG CAC AGA GAC 1440Ala Pro Thr Lys Ala Lys Arg Arg Val Val His Arg Asp470 475 480AAA AGA GCA GCA CTA GGA GCT TTG TTC CTT GGG TTC TTA 1479Lys Arg Ala Ala Leu Gly Ala Leu Phe Leu Gly Phe Leu485 490GGA GCA TAA AAG CTT CTA GA 1499Gly Ala Xaa Lys Leu Leu495 499(2)SEQ ID NO:20的信息(ⅰ)序列特征(A)長度499個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:20:Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala1 5 10 15Ser Asp Ala Lys Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala20 25 30Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Ile Glu35 40 45Leu Val Asn Val Thr Glu Asp Phe Asn Met Trp Lys Asn Lys Met50 55 60Val Asp Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Glu Ser65 70 75Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn80 85 90Cys Ser Asp Val Asn Asn Ser Thr Asn Pro Asn Asp Thr Asn Thr95 100 105Asn Ser Thr Asn Thr Thr Ser Ser Thr Pro Thr Ala Thr Thr Ser110 115 120Ser Glu Glu Lys Met Glu Lys Gly Glu Ile Lys Asn Cys Ser Phe125 130 135Asn Ile Thr Thr His Met Lys Asp Lys Val Gln Lys Glu Tyr Ala140 145 150Leu Phe Tyr Lys Leu Asp Ile Val Pro Ile Asp Asp Asn Asn Thr155 160 165Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala170 175 180Cys Pro Met Val Thr Phe Glu Pro Ile Pro Ile His Tyr Cys Ala185 190 195Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys Phe Asn200 205 210Gly Thr Gly Pro Cys Ser Lys Val Ser Thr Val Gln Cys Thr His215 220 225Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser230 235 240Leu Ala Glu Glu Glu Val Val Ile Arg Ser Val Asn Phe Thr Asp245 250 255Asn Ala Lys Ile Ile Ile Val Gln Leu Lys Glu pro Val Ala Ile260 265 270Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Gly Ile His Leu275 280 285Gly Pro Gly Ser Thr Phe Tyr Thr Thr Gly Glu Ile Ile Gly Asp290 295 300Ile Arg Lys Ala Tyr Cys Lys Ile Ser Lys Glu Lys Trp Asn Asn305 310 315Thr Leu Arg Gln Val Val Lys Lys Leu Arg Glu Gln Phe Gly Asn320 325 330Lys Thr Ile Ile Phe Asn Arg Ser Ser Gly Gly Asp Pro Glu Ile335 340 345Val Met His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn350 355 360Thr Thr Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr Glu Gly Thr365 370 375Asn Ser Thr GLu Gly Asn Ser Thr Ile Thr Leu Pro Cys Arg Ile380 385 390Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Thr Tyr395 400 405Ala Pro Pro Ile Arg Gly Arg Ile Arg Cys Ile Ser Asn Ile Thr410 415 420Gly Leu Leu Leu Thr Arg Asp Gly Gly Arg Asn Val Thr Asn Asn425 430 435Thr Xaa Xaa Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp440 445 450Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Val Glu Pro Leu455 460 465Gly Ile Ala Pro Thr Lys Ala Lys Arg Arg Val Val His Arg Asp470 475 480Lys Arg Ala Ala Leu Gly Ala Leu Phe Leu Gly Phe Leu Gly Ala485 490 495Xaa Lys Leu Leu499(2)SEQ ID NO:21的信息(ⅰ)序列特征(A)長度1475個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:21:
GTA CCT GTG TGG AAA GAA GCA AAC ACA ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT GAT AGA GAA GTA CAT 76Cys Ala Ser Asp Ala Lys Ala Tyr Asp Arg Glu Val His15 20 25AAT GTT TGG GCA ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AAC CCA CAA GAA ATA GTA TTG GGA AAT GTG ACA GAA AAT 154Asn Pro Gln Glu Ile Val Leu Gly Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His55 60GAG GAT ATA ATC AAT TTA TGG GAT CAA AGC TTA AAG CCA 232Glu Asp Ile Ile Asn Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAG TTA ACT CCA CTC TGT GTT ACT TTA AAG TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Lys Cys80 85 90AAG GAT CTG GAG AGG AAT ACT ACC TAT AAT AGC ACT ATT 310Lys Asp Leu Glu Arg Asn Thr Thr Tyr Asn Ser Thr Ile95 100ACC AAT AAT AGT AGT TTG GAG GGA CTA AGA GAA CAA ATG 349Thr Asn Asn Ser Ser Leu Glu Gly Leu Arg Glu Gln Met105 110 115ACA AAC TGC TCT TTC AAC ATC ACC ACA AGT ATA AGA GAT 388Thr Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp120 125AAG GTG CAG AAA GAA TAT GCA CTT TTG TAT AAA CTT GAT 427Lys Val Gln Lys Glu Tyr Ala Leu Leu Tyr Lys Leu Asp130 135 140GTA GTA CCA ATA GAA GAA GAT GAC AAT ACT AGC TAT AGA 466Val Val Pro Ile Glu Glu Asp Asp Asn Thr Ser Tyr Arg145 150 155TTG ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCT TGT 505Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys160 165CCA AAG ACA TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT 544Pro Lys Thr Ser Phe Glu Pro Ile Pro Ile His Tyr Cys170 175 180GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAT GAT AAG 583Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys185 190AAG TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGC ACA 622Lys Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr195 200 205GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 661Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr210 215 220CAA CTG TTG TTA AAT GGC AGT CTA GCA GAA GAA GAG GTA 700Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val225 230GTA ATC AGA TCT GCC AAT TTC ACA GAC AAT GCT AAA ACC 739Val Ile Arg Ser Ala Asn Phe Thr Asp Asn Ala Lys Thr235 240 245ATA ATA GTA CAT CTA AAT GAA ACT GTA AAA ATT AAT TGT 778Ile Ile Val His Leu Asn Glu Thr Val Lys Ile Asn Cys250 255ACA AGA CTT GGC AAC AAT ACA AGA AAA AGT ATA AAT ATA 817Thr Arg Leu Gly Asn Asn Thr Arg Lys Ser Ile Asn Ile260 265 270GGA CCA GGG AGA GTA CTC TAT GCA ACA GGA GAA ATA ATA 856Gly Pro Gly Arg Val Leu Tyr Ala Thr Gly Glu Ile Ile275 280 285GGA GAC ATA AGA CAA GCA CAT TGT AAC ATT AGT AGA GCA 895Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Arg Ala290 295CAA TGG AAT AAG ACT TTA GAA AAG GTA GTT GAC AAA TTA 934Gln Trp Asn Lys Thr Leu Glu Lys Val Val Asp Lys Leu300 305 310AGA AAA CAA TTT GGG GAT AAT ACA ACA ATA GCT TTT AAT 973Arg Lys Gln Phe Gly Asp Asn Thr Thr Ile Ala Phe Asn315 320CGA TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC ACT 1012Arg Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Thr325 330 335TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA 1051Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr340 345 350CAA CTG TTT AAT AGT ACT TGG AAT AAT ACT TGG AAG GAT 1090Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr Trp Lys Asp355 360CCT AAC AGG AGT GAC AAT ATC ACA CTC CCA TGC AGA ATA 1129Pro Asn Arg Ser Asp Asn Ile Thr Leu Pro Cys Arg Ile365 370 375AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA 1168Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala380 385ATG TAC GCC CCT CCC ATC AGA GGG GAA ATT AGA TGT TCA 1207Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile Arg Cys Ser390 395 400TCA AAT ATC ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT 1246Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly405 410 415AAT GAC GAT GGT AAT GAC ACG ACC ACA AAC AGG ACC GAG 1285Asn Asp Asp Gly Asn Asp Thr Thr Thr Asn Arg Thr Glu420 425ATC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1324Ile Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp430 435 440AGA AGT GAA TTA TAT AGA TAT AAA GTA GTA AAA ATT GAA 1363Arg Ser Glu Leu Tyr Arg Tyr Lys Val Val Lys Ile Glu445 450CCA TTA GGA ATA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1402Pro Leu Gly Ile Ala Pro Thr Arg Ala Lys Arg Arg Val455 460 465GTG CAG AGA GAA AAA AGA GCA GTA GGA CTA GGA GCT TTG 1441Val Gln Arg Glu Lys Arg Ala Val Gly Leu Gly Ala Leu47 475 480TTC CTT GGG T TCTTAGGAG CATAAAGCTT CTAGA 1475Phe Leu Gly483(2)SEQ ID NO:22的信息(ⅰ)序列特征(A)長度491個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:22:Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe Cys Ala Ser1 5 10 15Asp Ala Lys Ala Tyr Asp Arg Glu Val His Asn Val Trp Ala Thr20 25 30His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Ile Val Leu35 40 45Gly Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val50 55 60Glu Gln Met His Glu Asp Ile Ile Asn Leu Trp Asp Gln Ser Leu65 70 75Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Lys Cys80 85 90Lys Asp Leu Glu Arg Asn Thr Thr Tyr Asn Ser Thr Ile Thr Asn95 100 105Asn Ser Ser Leu Glu Gly Leu Arg Glu Gln Met Thr Asn Cys Ser110 115 120Phe Asn Ile Thr Thr Ser Ile Arg Asp Lys Val Gln Lys Glu Tyr125 130 135Ala Leu Leu Tyr Lys Leu Asp Val Val Pro Ile Glu Glu Asp Asp140 145 150Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr155 160 165Gln Ala Cys Pro Lys Thr Ser Phe Glu Pro Ile Pro Ile His Tyr170 175 180Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys Lys185 190 195Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val Gln Cys200 205 210Thr His Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn215 220 225Gly Ser Leu Ala Glu Glu Glu Val Val Ile Arg Ser Ala Asn Phe230 235 240Thr Asp Asn Ala Lys Thr Ile Ile Val His Leu Asn Glu Thr Val245 250 255Lys Ile Asn Cys Thr Arg Leu Gly Asn Asn Thr Arg Lys Ser Ile260 265 270Asn Ile Gly Pro Gly Arg Val Leu Tyr Ala Thr Gly Glu Ile Ile275 280 285Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Arg Ala Gln Trp290 295 300Asn Lys Thr Leu Glu Lys Val Val Asp Lys Leu Arg Lys Gln Phe305 310 315Gly Asp Asn Thr Thr Ile Ala Phe Asn Arg Ser Ser Gly Gly Asp320 325 330Pro Glu Ile Val Met His Thr Phe Asn Cys Gly Gly Glu Phe Phe335 340 345Tyr Cys Asn Thr Thr Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr350 355 360Trp Lys Asp Pro Asn Arg Ser Asp Asn Ile Thr Leu Pro Cys Arg365 370 375Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met380 385 390Tyr Ala Pro Pro Ile Arg Gly Glu Ile Arg Cys Ser Ser Asn Ile395 400 405Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Asn Asp Asp Gly Asn410 415 420Asp Thr Thr Thr Asn Arg Thr Glu Ile Phe Arg Pro Gly Gly Gly425 430 435Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr Arg Tyr Lys Val440 445 450Val Lys Ile Glu Pro Leu Gly Ile Ala Pro Thr Arg Ala Lys Arg455 460 465Arg Val Val Gln Arg Glu Lys Arg Ala Val Gly Leu Gly Ala Leu470 475 480Phe Leu Gly Phe Leu Gly Ala Leu Phe Leu Gly485 490 491(2)SEQ ID NO:23的信息(ⅰ)序列特征(A)長度1475個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:23:G GTA CCT GTG TGG AAA GAA GCA AAC ACA ACT CTA TTT 37Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe1 5 10TGT GCA TCA GAT GCT AAA GCA TAT GAT AGA GAA GTA CAT 76Cys Ala Ser Asp Ala Lys Ala Tyr Asp Arg Glu Val His15 20 25AAT GTT TGG GCA ACA CAT GCC TGT GTA CCC ACA GAC CCC 115Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro30 35AAC CCA CAA GAA ATA GTA TTG GGA AAT GTG ACA GAA AAT 154Asn Pro Gln Glu Ile Val Leu Gly Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAA ATG CAT 193Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His55 60GAG GAT ATA ATC AAT TTA TGG GAT CAA AGC TTA AAG CCA 232Glu Asp Ile Ile Asn Leu Trp Asp Gln Ser Leu Lys Pro65 70 75TGT GTA AAG TTA ACT CCA CTC TGT GTT ACT TTA AAG TGC 271Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Lys Cys80 85 90AAG GAT CTG GAG AGG AAT ACT ACC TAT AAT AGC ACT ATT 310Lys Asp Leu Glu Arg Asn Thr Thr Tyr Asn Ser Thr Ile95 100ACC AAT AAT AGT AGT TTG GAG GGA CTA AGA GAA CAA ATG 349Thr Asn Asn Ser Ser Leu Glu Gly Leu Arg Glu Gln Met105 110 115ACA AAC TGC TCT TTC AAC ATC ACC ACA AGT ATA AGA GAT 388Thr Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Arg Asp120 125AAG GTG CAG AAA GAA TAT GCA CTT TTG TAT AAA CTT GAT 427Lys Val Gln Lys Glu Tyr Ala Leu Leu Tyr Lys Leu Asp130 135 140GTA GTA CCA ATA GAA GAA GAT GAC AAT ACT AGC TAT AGA 466Val Val Pro Ile Glu Glu Asp Asp Asn Thr Ser Tyr Arg145 150 155TTG ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCT TGT 505Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys160 165CCA AAG ACA TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT 544Pro Lys Thr Ser Phe Glu Pro Ile Pro Ile His Tyr Cys170 175 180GCC CCG GCT GGT TTT GCG ATT CTA AAG TGT AAT GAT AAG 583Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys185 190AAG TTC AAT GGA ACA GGA CCA TGT AAA AAT GTC AGC ACA 622Lys Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr195 200 205GTA CAA TGT ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT 661Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser Thr210 215 220CAA CTG TTG TTA AAT GGC AGT CTA GCA GAA GAA GAG GTA 700Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val225 230GTA ATC AGA TCT GCC AAT TTC ACA GAC AAT GCT AAA ACC 739Val Ile Arg Ser Ala Asn Phe Thr Asp Asn Ala Lys Thr235 240 245ATA ATA GTA CAT CTA AAT GAA ACT GTA AAA ATT AAT TGT 778Ile Ile Val His Leu Asn Glu Thr Val Lys Ile Asn Cys250 255ACA AGA CTT GGC AAC AAT ACA AGA AAA AGT ATA AAT ATA 817Thr Arg Leu Gly Asn Asn Thr Arg Lys Ser Ile Asn Ile260 265 270GGA CCA GGG AGA GTA CTC TAT GCA ACA GGA GAA ATA ATA 856Gly Pro Gly Arg Val Leu Tyr Ala Thr Gly Glu Ile Ile275 280 285GGA GAC ATA AGA CAA GCA CAT TGT AAC ATT AGT AGA GCA 895Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Arg Ala290 295CAA TGG AAT AAG ACT TTA GAA AAG GTA GTT GAC AAG TTA 934Gln Trp Asn Lys Thr Leu Glu Lys Val Val Asp Lys Leu300 305 310AGA AAA CAA TTT GGG GAT AAT ACA ACA ATA GCT TTT AAT 973Arg Lys Gln Phe Gly Asp Asn Thr Thr Ile Ala Phe Asn315 320CGA TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC ACT 1012Arg Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Thr325 330 335TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA 1051Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr340 345 350CAA CTG TTT AAT AGT ACT TGG AAT AAT ACT TGG AAG GAT 1090Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr Trp Lys Asp355 360CCT AAC AGG AGT GAC AAT ATC ACA CTC CCA TGC AGA ATA 1129Pro Asn Arg Ser Asp Asn Ile Thr Leu Pro Cys Arg Ile365 370 375AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA GCA 1168Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala380 385ATG TAC GCC CCT CCC ATC AGA GGG GAA ATT AGA TGT TCA 1207Met Tyr Ala Pro Pro Ile Arg Gly Glu Ile Arg Cys Ser390 395 400TCA AAT ATC ACA GGG CTG CTA CTA ACA AGA GAT GGT GGT 1246Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly405 410 415AAT GAC GAT GGT AAT GAC ACG ACC ACA AAC AGG ACC GAG 1285Asn Asp Asp Gly Asn Asp Thr Thr Thr Asn Arg Thr Glu420 425ATC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1324Ile Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp430 435 440AGA AGT GAA TTA TAT AGA TAT AAA GTA GTA AAA ATT GAA 1363Arg Ser Glu Leu Tyr Arg Tyr Lys Val Val Lys Ile Glu445 450CCA TTA GGA ATA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1402Pro Leu Gly Ile Ala Pro Thr Arg Ala Lys Arg Arg Val455 460 465GTG CAG AGA GAA AAA AGA GCA GTA GGA CTA GGA GCT TTG 1441Val Gln Arg Glu Lys Arg Ala Val Gly Leu Gly Ala Leu470 475 480TTC CTT GGG TTC TTG GGA GCA TAA AGC TTC TAG A 1475Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa485 490 491(2)SEQ ID NO:24的信息(ⅰ)序列特征(A)長度491個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹi)序列描述SEQ ID NO:24:Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe Cys Ala Ser1 5 10 15Asp Ala Lys Ala Tyr Asp Arg Glu Val His Asn Val Trp Ala Thr20 25 30His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Ile Val Leu35 40 45Gly Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val50 55 60Glu Gln Met His Glu Asp Ile Ile Asn Leu Trp Asp Gln Ser Leu65 70 75Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Lys Cys80 85 90Lys Asp Leu Glu Arg Asn Thr Thr Tyr Asn Ser Thr Ile Thr Asn95 100 105Asn Ser Ser Leu Glu Gly Leu Arg Glu Gln Met Thr Asn Cys Ser110 115 120Phe Asn Ile Thr Thr Ser Ile Arg Asp Lys Val Gln Lys Glu Tyr125 130 135Ala Leu Leu Tyr Lys Leu Asp Val Val Pro Ile Glu Glu Asp Asp140 145 150Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr155 160 165Gln Ala Cys Pro Lys Thr Ser Phe Glu Pro Ile Pro Ile His Tyr170 175 180Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys Lys185 190 195Phe Asn Gly Thr Gly Pro Cys Lys Asn Val Ser Thr Val Gln Cys200 205 210Thr His Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn215 220 225Gly Ser Leu Ala Glu Glu Glu Val Val Ile Arg Ser Ala Asn Phe230 235 240Thr Asp Asn Ala Lys Thr Ile Ile Val His Leu Asn Glu Thr Val245 250 255Lys Ile Asn Cys Thr Arg Leu Gly Asn Asn Thr Arg Lys Ser Ile260 265 270Asn Ile Gly Pro Gly Arg Val Leu Tyr Ala Thr Gly Glu Ile Ile275 280 285Gly Asp Ile Arg Gln Ala His Cys Asn Ile Ser Arg Ala Gln Trp290 295 300Asn Lys Thr Leu Glu Lys Val Val Asp Lys Leu Arg Lys Gln Phe305 310 315Gly Asp Asn Thr Thr Ile Ala Phe Asn Arg Ser Ser Gly Gly Asp320 325 330Pro Glu Ile Val Met His Thr Phe Asn Cys Gly Gly Glu Phe Phs335 340 345Tyr Cys Asn Thr Thr Gln Leu Phe Asn Ser Thr Trp Asn Asn Thr350 355 360Trp Lys Asp Pro Asn Arg Ser Asp Asn Ile Thr Leu Pro Cys Arg365 370 375Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met380 385 390Tyr Ala Pro Pro Ile Arg Gly Glu Ile Arg Cys Ser Ser Asn Ile395 400 405Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Asn Asp Asp Gly Asn410 415 420Asp Thr Thr Thr Asn Arg Thr Glu Ile Phe Arg Pro Gly Gly Gly425 430 435Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr Arg Tyr Lys Val440 445 450Val Lys Ile Glu Pro Leu Gly Ile Ala Pro Thr Arg Ala Lys Arg455 460 465Arg Val Val Gln Arg Glu Lys Arg Ala Val Gly Leu Gly Ala Leu470 475 480Phe Leu Gly Phe Leu Gly ALa Xaa Ser Phe Xaa485 490 491(2)SEQ ID NO:25的信息(ⅰ)序列特征(A)長度1435個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:25:
CTC GAG GTA CCT GTG TGG AAA GAA GCA ACC ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT GAT TCA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Ser Glu15 20 25GCA CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA 114Ala His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr30 35GAC CCC AAC CCA CAA GAA GTA GAA TTG GAA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Glu Leu Glu Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG 192Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln55 60ATG CAT GGG GAT ATA ATT AGT TTA TGG GAT CAA AGC CTA 231Met His Gly Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACG TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAC CCA AAT GTT ACT AAT AGC GAG AGA ACG 309Asn Cys Thr Asp Pro Asn Val Thr Asn Ser Glu Arg Thr95 100ATA GAG GGG GGA GAA ATA AAA AAT TGC TCT TTC AAT ATC 348Ile Glu Gly Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile105 110 115ACC ACA AAC ATA AGA GAT AGG TTT CAG AAA GAA TAT GCA 387Thr Thr Asn Ile Arg Asp Arg Phe Gln Lys Glu Tyr Ala120 125CTT TTT TAT AAA CTT GAT GTA ATA CCA TTA GGT AAT GAT 426Leu Phe Tyr Lys Leu Asp Val Ile Pro Leu Gly Asn Asp130 135 140AAT ACT AGC TAT AGG TTG ATA AGT TGT AAC ACC TCA GTC 465Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val145 150 155ATT ACA CAG GCC TGT CCA AAG GTA TCC TTT GAG CCA ATT 504Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile160 165CCC ATA CAT TAT TGT GCC CCG GCT GGT TTT GCG ATT CTA 543Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu170 175 180AAG TGT AAA GAT AAG AAG TTC AAT GGA ACA GGA CCA TGT 582Lys Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys185 190ACA AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AAG 621Thr Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys195 200 205CCA GTA GTA TCA ACT CAA CTG TTG TTA AAT GGC AGT CTA 660Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu210 215 220GCA GAA GAA GAC ATA GTA ATT AGA TCC GCC AAT CTC ACA 699Ala Glu Glu Asp Ile Val Ile Arg Ser Ala Asn Leu Thr225 230GAC AAT GCT AAA AAC ATA ATA GTA CAG CTG AAT GAA TCT 738Asp Asn Ala Lys Asn Ile Ile Val Gln Leu Asn Glu Ser235 240 245GTA ACA ATG AAT TGT ACA AGA CCC AAC AAC AAT ACA ATG 777Val Thr Met Asn Cys Thr Arg Pro Asn Asn Asn Thr Met250 255AAA AGT ATA CAT ATA GGA CCA GGC ACA GCA TTT TAT GCA 816Lys Ser Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala260 265 270ACA GGA AAC ATA ATA GGA GAT ATA AGA CAA GCA CAT TGT 855Thr Gly Asn Ile Ile Gly Asp Ile Arg Gln Ala His Cys275 280 285AAC ATT AGT GGA ACA AAA TGG AAT GAC ACT TTG AAA AAG 894Asn Ile Ser Gly Thr Lys Trp Asn Asp Thr Leu Lys Lys290 295ATA GCT ATA AAA TTA AGA GAA CAA TTT AAT AAG ACA ATA 933Ile Ala Ile Lys Leu Arg Glu Gln Phe Asn Lys Thr Ile300 305 310GTC TTT AAT CAA TCC TCA GGA GGG GAC CCA GAA ATT GCA 972Val Phe Asn Gln Ser Ser Gly Gly Asp Pro Glu Ile Ala315 320ACG CTC AGT TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT 1011Thr Leu Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys325 330 335AAT TCA ACA CAA CTG TTT AAT AGT ACT TGG AAT AGT ACT 1050Asn Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Ser Thr340 345 350GGG TCA AAT AAC ACT AAA GGA AAT GAC ACA ATC ACA CTC 1089Gly Ser Asn Asn Thr Lys Gly Asn Asp Thr Ile Thr Leu355 360CCA TGC AGA ATA AGA CAA ATT ATA AAC ATG TGG CAG AAA 1128Pro Cys Arg Ile Arg Gln Ile Ile Asn Met Trp Gln Lys365 370 375ATA GGA AAA GCA ATG TAT GCC CCT CCC ATC AAA GGG CAA 1167Ile Gly Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln380 385ATT AGA TGT TCA TCA AAT ATT ACA GGG CTA ATA TTA ACA 1206Ile Arg Cys Ser Ser Asn Ile Thr Gly Leu Ile Leu Thr390 395 400AGA GAT GGT GGT AAC AAC AAC ATG AGC AAG ACC ACC GAG 1245Arg Asp Gly Gly Asn Asn Asn Met Ser Lys Thr Thr Glu405 410 415ACC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1284Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp420 425AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA AAA ATT GAA 1323Arg Ser Glu Leu Tyr Lys Tyr Lys val Val Lys Ile Glu430 435 440CCA TTA GGA GTA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1362Pro Leu Gly Val Ala Pro Thr Arg Ala Lys Arg Arg Val445 450GTG CAG AGA GAA AAA AGA GCA GTG GGA ATA GGA GCT GTG 1401Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Val455 460 465TTC CTT GGG TTC TTG GGA GCA TAA AGC TTC TAG A 1435Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa470 475 478(2)SEQ ID NO:26的信息(ⅰ)序列特征(A)長度478個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:26:Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys1 5 10 15Ala Ser Asp Ala Lys Ala Tyr Asp Ser Glu Ala His Asn Val Trp20 25 30Ala Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Val35 40 45Glu Leu Glu Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn50 55 60Met Val Glu Gln Met His Gly Asp lle lle Ser Leu Trp Asp Gln65 70 75Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90Asn Cys Thr Asp Pro Asn Val Thr Asn Ser Glu Arg Thr Ile Glu95 100 105Gly Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Asn Ile110 115 120Arg Asp Arg Phe Gln Lys Glu Tyr Ala Leu Phe Tyr Lys Leu Asp125 130 135Val Ile Pro Leu Gly Asn Asp Asn Thr Ser Tyr Arg Leu Ile Ser140 145 150Cys ASn Thr Ser Val Ile Thr Gln Ala Cys Pro Lys Val Ser Phe155 160 165Glu Pro Ile Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile170 175 180Leu Lys Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Thr185 190 195Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys Pro Val Val200 205 210Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Asp Ile215 220 225Val Ile Arg Ser Ala Asn Leu Thr Asp Asn Ala Lys Asn Ile Ile230 235 240Val Gln Leu Asn Glu Ser Val Thr Met Asn Cys Thr Arg Pro Asn245 250 255Asn Asn Thr Met Lys Ser Ile His Ile Gly Pro Gly Arg Ala Phe260 265 270Tyr Ala Thr Gly Asn Ile Ile Gly Asp Ile Arg Gln Ala His Cys275 280 285Asn Ile Ser Gly Thr Lys Trp Asn Asp Thr Leu Lys Lys Ile Ala290 295 300Ile Lys Leu Arg Glu Gln Phe Asn Lys Thr Ile Val Phe Asn Gln305 310 315Ser Ser Gly Gly Asp Pro Glu Ile Ala Thr Leu Ser Phe Asn Cys320 325 330Gly Gly Glu Phe Phe Tyr Cys Asn Ser Thr Gln Leu Phe Asn Ser335 340 345Thr Trp Asn Ser Thr Gly Ser Asn Asn Thr Lys Gly Asn Asp Thr350 355 360Ile Thr Leu Pro Cys Arg Ile Arg Gln Ile Ile Asn Met Trp Gln365 370 375Lys Ile Gly Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln Ile380 385 390Arg Cys Ser Ser Asn Ile Thr Gly Leu Ile Leu Thr Arg Asp Gly395 400 405Gly Asn Asn Asn Met Ser Lys Thr Thr Glu Thr Phe Arg Pro Gly410 415 420Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr Lys Tyr425 430 435Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala Pro Thr Arg Ala440 445 450Lys Arg Ar9 Val Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly455 460 465Ala VaI Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa470 475 478(2)SEQ ID NO:27的信息(ⅰ)序列特征(A)長度1435個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:27:
CTC GAG GTA CCT GTG TGG AAA GAA GCA ACC ACC ACT 36Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr1 5 10CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT GAT TCA GAG 75Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Ser Glu15 20 25GCA CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA 114Ala His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr30 35GAC CCC AAC CCA CAA GAA GTA GAA TTG GAA AAT GTG ACA 153Asp Pro Asn Pro Gln Glu Val Glu Leu Glu Asn Val Thr40 45 50GAA AAT TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG 192Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln55 50ATG CAT GGG GAT ATA ATT AGT TTA TGG GAT CAA AGC CTA 231Met His Gly Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu65 70 75AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT ACG TTA 270Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90AAT TGC ACT GAC CCA AAT GTT ACT AAT AGC GAG AGA ACG 309Asn Cys Thr Asp Pro Asn Val Thr Asn Ser Glu Arg Thr95 100ATA GAG GGG GGA GAA ATA AAA AAT TGC TCT TTC AAT ATC 348Ile Glu Gly Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile105 110 115ACC ACA AAC ATA AGA GAT AGG TTT CAG AAA GAA TAT GCA 387Thr Thr Asn Ile Arg Asp Arg Phe Gln Lys Glu Tyr Ala120 125CTT TTT TAT AAA CTT GAT GTA ATA CCA TTA GGT AAT GAT 426Leu Phe Tyr Lys Leu Asp Val Ile Pro Leu Gly Asn Asp130 135 140AAT ACT AGC TAT AGG TTG ATA AGT TGT AAC ACC TCA GTC 465Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val145 150 155ATT ACA CAG GCC TGT CCA AAG GTA TCC TTT GAG CCA ATT 504Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile160 165CCC ATA CAT TAT TGT GCC CCG GCT GGT TTT GCG ATT CTA 543Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu170 175 180AAG TGT AAA GAT AAG AAG TTC AAT GGA ACA GGA CCA TGT 582Lys Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys185 190ACA AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AAG 621Thr Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys195 200 205CCA GTA GTA TCA ACT CAA CTG TTG TTA AAT GGC AGT CTA 660Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu210 215 220GCA GAA GAA GAC ATA GTA ATT AGA TCC GCC AAT CTC ACA 699Ala Glu Glu Asp Ile Val Ile Arg Ser Ala Asn Leu Thr225 230GAC AAT GCT AAA AAC ATA ATA GTA CAG CTG AAT GAA TCT 738Asp Asn Ala Lys Asn Ile Ile Val Gln Leu Asn Glu Ser235 240 245GTA ACA ATG AAT TGT ACA AGA CCC AAC AAC AAT ACA ATG 777Val Thr Met Asn Cys Thr Arg Pro Asn Asn Asn Thr Met250 255AAA AGT ATA CAT ATA GGA CCA GGC AGA GCA TTT TAT GCA 816Lys Ser Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala260 265 270ACA GGA AAC ATA ATA GGA GAT ATA AGA CAA GCA CAT TGT 855Thr Gly Asn Ile Ile Gly Asp Ile Arg Gln Ala His Cys275 280 285AAC ATT AGT GGA ACA AAA TGG AAT GAC ACT TTG AAA AAG 894Asn Ile Ser Gly Thr Lys Trp Asn Asp Thr Leu Lys Lys290 295ATA GCT ATA AAA TTA AGA GAA CAA TTT AAT AAG ACA ATA 933Ile Ala Ile Lys Leu Arg Glu Gln PheAsn Lys Thr Ile300 305310GTC TTT AAT CAA TCC TCA GGA GGG GAC CCA GAA ATT GCA 972Val Phe Asn Gln Ser Ser Gly Gly Asp Pro Glu Ile Ala315 320ACG CTC AGT TTT AAT TGT GGA GGG GAA TTT TTC TAC TGT 1011Thr Leu Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys325 330 335AAT TCA ACA CAA CTG TTT AAT AGT ACT TGG AAT AGT ACT 1050Asn Ser Thr Gln Leu Phe Asn Ser Thr Trp Asn Ser Thr340 345 350GGG TCA AAT AAC ACT AAA GGA AAT GAC ACA ATC ACA CTC 1089Gly Ser Asn Asn Thr Lys Gly Asn Asp Thr Ile Thr Leu355 360CCA TGC AGA ATA AGA CAA ATT ATA AAC ATG TGG CAG AAA 1128Pro Cys Arg Ile Arg Gln Ile Ile Asn Met Trp Gln Lys365 370 375ATA GGA AAA GCA ATG TAT GCC CCT CCC ATC AAA GGG CAA 1167Ile Gly Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln380 385ATT AGA TGT TCA TCA AAT ATT ACA GGG CTA ATA TTA ACA 1206Ile Arg Cys Ser Ser Asn Ile Thr Gly Leu Ile Leu Thr390 395 400AGA GAT GGT GGT AAC AAC AAC ATG AGC AAG ACC ACC GAG 1245Arg Asp Gly Gly Asn Asn Asn Met Ser Lys Thr Thr Glu405 410 415ACC TTC AGA CCT GGA GGA GGA GAT ATG AGG GAC AAT TGG 1284Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp420 425AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA AAA ATT GAA 1323Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu430 435 440CCA TTA GGA GTA GCA CCC ACC AGG GCA AAG AGA AGA GTG 1362Pro Leu Gly Val Ala Pro Thr Arg Ala Lys Arg Arg Val445 450GTG CAG AGA GAA AAA AGA GCA GTG GGA ATA GGA GCT GTG 1401Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly Ala Val455 460 465TTC CTT GGG TTC TTG GGA GCA TAA AGC TTC TAG A 1435Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa470 475 478(2)SEQ ID NO:28的信息(ⅰ)序列特征(A)長度478個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:28:Leu Glu Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys1 5 10 15Ala Ser Asp Ala Lys Ala Tyr Asp Ser Glu Ala His Asn Val Trp20 25 30Ala Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Val35 40 45Glu Leu Glu Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn50 55 60Met Val Glu Gln Met His Gly Asp Ile Ile Ser Leu Trp Asp Gln65 70 75Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu80 85 90Asn Cys Thr Asp Pro Asn Val Thr Asn Ser Glu Arg Thr Ile Glu95 100 105Gly Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Asn Ile110 115 120Arg Asp Arg Phe Gln Lys Glu Tyr Ala Leu Phe Tyr Lys Leu Asp125 130 135Val Ile Pro Leu Gly Asn Asp Asn Thr Ser Tyr Arg Leu Ile Ser140 145 150Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro Lys Val Ser Phe155 160 165Glu Pro Ile Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile170 175 180Leu Lys Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Thr185 190 195Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Lys Pro Val Val200 205 210Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Asp Ile215 220 225Val Ile Arg Ser Ala Asn Leu Thr Asp Asn Ala Lys Asn Ile Ile230 235 240Val Gln Leu Asn Glu Ser Val Thr Met Asn Cys Thr Arg Pro Asn245 250 255Asn Asn Thr Met Lys Ser Ile His Ile Gly Pro Gly Arg Ala Phe260 265 270Tyr Ala Thr Gly Asn Ile Ile Gly Asp Ile Arg Gln Ala His Cys275 280 285Asn Ile Ser Gly Thr Lys Trp Asn Asp Thr Leu Lys Lys Ile Ala290 295 300Ile Lys Leu Arg Glu Gln Phe Asn Lys Thr Ile Val Phe Asn Gln305 310 315Ser Ser Gly Gly Asp Pro Glu Ile Ala Thr Leu Ser Phe Asn Cys320 325 330Gly Gly Glu Phe Phe Tyr Cys Asn Ser Thr Gln Leu Phe Asn Ser335 340 345Thr Trp Asn Ser Thr Gly Ser Asn Asn Thr Lys Gly Asn Asp Thr350 355 360Ile Thr Leu Pro Cys Arg Ile Arg Gln Ile Ile Asn Met Trp Gln365 370 375Lys Ile Gly Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln Ile380 385 390Arg Cys Ser Ser Asn Ile Thr Gly Leu Ile Leu Thr Arg Asp Gly395 400 405Gly Asn Asn Asn Met Ser Lys Thr Thr Glu Thr Phe Arg Pro Gly410 415 420Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr Lys Tyr425 430 435Lys Val Val Lys Ile Glu Pro Leu Gly Val Ala Pro Thr Arg Ala440 445 450Lys Arg Arg Val Val Gln Arg Glu Lys Arg Ala Val Gly Ile Gly455 460 465Ala Val Phe Leu Gly Phe Leu Gly Ala Xaa Ser Phe Xaa470 475 478(2)SEQ ID NO:29的信息(ⅰ)序列特征(A)長度511個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:29:Met Arg Val Lys Gly Ile Arg Arg Asn Tyr Gln His Trp Trp Gly Arg1 5 10 15Gly Thr Met Leu Leu Gly Leu Leu Met Ile Cys Ser Ala Thr Glu Lys20 25 30Leu Trp Val Thr Val Tyr Tyr Gly Val Pro Val Trp Lys Glu Ala Thr35 40 45Thr Thr Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu Ala50 55 60His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro Asn Pro65 70 75 80Gln Glu Val Glu Leu Val Asn Val Thr Glu Asn Phe Asn Met Trp Lys85 90 95Asn Asn Met Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asn100 105 110Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu115 120 125Asn Cys Thr Asp Leu Arg Asn Thr Thr Asn Thr Asn Asn Ser Thr Asp130 135 140Asn Asn Asn Ser Lys Ser Glu Gly Thr Ile Lys Gly Gly Glu Met Lys145 150 155 160Asn Cys ser Phe Asn Ile Thr Thr Ser Ile Gly Asp Lys Met Gln Lys165 170 175Glu Tyr Ala Leu Leu Tyr Lys Leu Asp Ile Glu Pro Ile Asp Asn Asp180 185 190Ser Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln195 200 205Ala Cys Pro Lys Ile Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala210 215 220Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp Lys Lys Phe Ser Gly225 230 235 240Lys Gly Ser Cys Lys Asn Val Ser Thr Val Gln Cys Thr His Gly Ile245 250 255Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu260 265 270Glu Glu Val Val Ile Arg Ser Glu Asp Phe Thr Asp Asn Ala Lys Thr275 280 285Ile Ile Val His Leu Lys Glu Ser Val Gln Ile Asn Cys Thr Arg Pro290 295 300Asn Tyr Asn Lys Arg Lys Arg Ile His Ile Gly Pro Gly Arg Ala Phe305 310 315 320Tyr Thr Thr Lys Asn Ile Lys Gly Thr Ile Arg Gln Ala His Cys Ile325 330 335Ile Ser Arg Ala Lys Trp Asn Asp Thr Leu Arg Gln Ile Val Ser Lys340 345 350Leu Lys Glu Gln Phe Lys Asn Lys Thr Ile Val Phe Asn Pro Ser Ser355 360 365Gly Gly Asp Pro Glu Ile Val Met His Ser Phe Asn Cys Gly Gly Glu370 375 380Phe Phe Tyr Cys Asn Thr Ser Pro Leu Phe Asn Ser Ile Trp Asn Gly385 390 395 400Asn Asn Thr Trp Asn Asn Thr Thr Gly Ser Asn Asn Asn Ile Thr Leu405 410 415Gln Cys Lys Ile Lys Gln Ile Ile Asn Met Trp Gln Lys Val Gly Lys420 425 430Ala Met Tyr Ala Pro Pro Ile Glu Gly Gln Ile Arg Cys Ser Ser Asn435 440 445Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Glu Asp Thr Asp Thr450 455 460Asn Asp Thr Glu Ile Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn465 470 475 480Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Thr Ile Glu Pro Leu485 490 495Gly Val Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu500 505 510(2)SEQ ID NO:30的信息(ⅰ)序列特征(A)長度2800個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:30:TTCGAGCTCG CCCGACATTG ATTATTGACT AGAGTCGATC GACAGCTGTG 50GAATGTGTGT CAGTTAGGGT GTGGAAAGTC CCCAGGCTCC CCAGCAGGCA 100GAAGTATGCA AAGCATGCAT CTCAATTAGT CAGCAACCAG GTGTGGAAAG 150TCCCCAGGCT CCCCAGCAGG CAGAAGTATG CAAAGCATGC ATCTCAATTA 200GTCAGCAACC ATAGTCCCGC CCCTAACTCC GCCCATCCCG CCCCTAACTC 250CGCCCAGTTC CGCCCATTCT CCGCCCCATG GCTGACTAAT TTTTTTTATT 300TATGCAGAGG CCGAGGCCGC CTCGGCCTCT GAGCTATTCC AGAAGTAGTG 350AGGAGGCTTT TTTGGAGGCC TAGGCTTTTG CAAAAAGCTA GCTTATCCGG 400CCGGGAACGG TGCATTGGAA CGCGGATTCC CCGTGCCAAG AGTCAGGTAA 450GTACCGCCTA TAGAGTCTAT AGGCCCACCC CCTTGGCTTC GTTAGAACGC 500GGCTACAATT AATACATAAC CTTTTGGATC GATCCTACTG ACACTGACAT 550CCACTTTTTC TTTTTCTCCA CAGGTGTCCA CTCCCAGGTC CAACTGCACC 600TCGGTTCGCG AAGCTAGCTT GGGCTGCATC GATTGAATTC CACTGCCTTC 650CACCAAGCTC TGCAGGATCC CAGAGTCAGG GG TCT GTA TCT TCC TGC 697Ser Val Ser Ser Cys1 5TGG TGG CTC CAG TTC AGG AAC AGT AAA CCC TGC TCC GAA TAT 739Trp Trp Leu Gln Phe Arg Asn Ser Lys Pro Cys Ser Glu Tyr10 15TGC CTC TCA CAT CTC GTC AAT CTC CGC GAG GAC TGG GGA CCC 781Cys Leu Ser His Leu Val Asn Leu Arg Glu Asp Trp Gly Pro20 25 30TCT GAC AAG CTT CAG CGC GAA CGA CCA ACT ACC CCG ATC ATC 823Cys Asp Lys Leu Gln Arg Glu Arg Pro Thr Thr Pro Ile Ile35 40 45AGT TAT CCT TAA GGT CTC TTT TGT GTG GTG CGT TCC GGT ATG 865Ser Tyr Pro ＊ Gly Leu Phe Cys Val Val Arg Ser Gly Met50 55 60GGG GGG ACT GCC GCC AGG TTG GGG GCC GTG ATT TTG TTT GTC 907Gly Gly Thr Ala Ala Arg Leu Gly Ala Val Ile Leu Phe Val62 65 70 75GTC ATA GTG GGC CTC CAT GGG GTC CGC GGC AAA TAT GCC TTG 949Val Ile Val Gly Leu His Gly Val Arg Gly Lys Tyr Ala Leu80 85GCG GAT GCC TCT CTC AAG ATG GCC GAC CCC AAT CGA TTT CGC 991Ala Asp Ala Ser Leu Lys Met Ala Asp Pro Asn Arg Phe Arg90 95 100GGC AAA GAC CTT CCG GTC CTG GAC CAG CTG CTC GAG GTA CCT 1033Gly Lys Asp Leu Pro Val Leu Asp Gln Leu Leu Glu Val Pro105 110 115GTG TGG AAA GAA GCA AAC ACC ACT CTA TTT TGT GCA TCA GAT 1075Val Trp Lys Glu Ala Asn Thr Thr Leu Phe Cys Ala Ser Asp120 125 130GCT AAA GCA TAT AAG ACA GAG GCA CAT AAT GTT TGG GCC ACA 1117Ala Lys Ala Tyr Lys Thr Glu Ala His Asn Val Trp Ala Thr135 140 145CAT GCC TGT GTA CCC ACA GAC CCC AAA CCA CAA GAA ATA AAA 1159His Ala Cys Val Pro Thr Asp Pro Lys Pro Gln Glu Ile Lys150 155TTG GAA AAT GTG ACA GAA AAT TTT AAC ATG TGG AAA AAT AAC 1201Leu Glu Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn160 165 170ATG GTA GAA CAG ATG CAT GAG GAT ATA ATC AGT TTA TGG GAT 1243Met Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp175 180 185CAA AGC CTA AAG CCA TGT GTA AAA TTA ACC CCA CTC TGT GTT 1285Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val190 195 200ACT TTA AAT TGC ACT GAT TTG AGG AAT AAT ACT AAT ACC AAT 1327Thr Leu Asn Cys Thr Asp Leu Arg Asn Asn Thr Asn Thr Asn205 210 215AGT ACC TAC GGA AAA ATA ATG GAG GGA GGA GAG ATA AAA AAC 1369Ser Thr Tyr Gly Lys Ile Met Glu Gly Gly Glu Ile Lys Asn220 225TGC TCT TTC AAT ATC ACC ACA AGC ATA AAA GAT AAG CTG AAA 1411Cys Ser Phe Asn Ile Thr Thr Ser Ile Lys Asp Lys Leu Lys230 235 240GAT ATG TCA CTT TTT TAT AAA CTT GAT GTA GTA CCA ATA GGT 1453Asp Met Ser Leu Phe Tyr Lys Leu Asp Val Val Pro Ile Gly245 250 255AAT AAT AGT AAT ACT ACT AGT TAT AGG TTG ATA AGT TGT AAC 1495Asn Asn Ser Asn Thr Thr Ser Tyr Arg Leu Ile Ser Cys Asn260 265 270ACC TCA GTC ATT ACA CAA GCC TGT CCA AAG ACA TCC TTT GAG 1537Thr Ser Val Ile Thr Gln Ala Cys Pro Lys Thr Ser Phe Glu275 280 285CCA ATT CCC ATA CAT TAT TGT GCC CCG GCT GGT TTT GCG ATT 1579Pro Ile Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile290 295CTC AAG TGT AAT GAT AAT AAG TTC AAT GGA ACA GGA CCA TGT 1621Leu Lys Cys Asn Asp Asn Lys Phe Asn Gly Thr Gly Pro Cys300 305 310CCA AAT GTC AGC ACA GTA CAA TGT ACA CAT GGA ATT AGG CCA 1663Pro Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro315 320 325GTA GTA TCA ACT CAA CTG CTG TTA AAT GGC AGT CTA GCA GAA 1705Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu330 335 340AAA GAG GTA GTC CTT AGA TCT GAA AAT TTC ACG GAC AAT GCT 1747Lys Glu Val Val Leu Arg Ser Glu Asn Phe Thr Asp Asn Ala345 350 355AAA ACC ATA ATA GTA CAG CTG AAC GAA TCT GTA ATA ATT GAT 1789Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Val Ile Ile Asp360 365TGT ATG AGA CCC AAC AAC AAT ACA AGA ACA AGT ATA CCT ATG 1831Cys Met Arg Pro Asn Asn Asn Thr Arg Thr Ser Ile Pro Met370 375 380GGA CCA GGG AAA GCA TTT TAT GCA ACA GGA GAT GTA ATA GGA 1873Gly Pro Gly Lys Ala Phe Tyr Ala Thr Gly Asp Val Ile Gly385 390 395GAT ATA AGA CGA GCA CAT TGT AAC ATT AGT AGA GCA GGA TGG 1915Asp Ile Arg Arg Ala His Cys Asn Ile Ser Arg Ala Gly Trp400 405 410AAT ACC ACT TTA CAA CAG ATA GCT AAA AAA TTA AGA GAA AAA 1957Asn Thr Thr Leu Gln Gln Ile Ala Lys Lys Leu Arg Glu Lys415 420 425TTT GAG AAC AAA ACA ATA GTT TTT AAT CAC TCC TCA GGA GGG 1999Phe Glu Asn Lys Thr Ile Val Phe Asn His Ser Ser Gly Gly430 435GAC CCA GAA ATT GTA ATG CAC ACT TTT AAT TGT GGA GGG GAA 2041Asp Pro Glu Ile Val Met His Thr Phe Asn Cys Gly Gly Glu440 445 450TTT TTC TGC TGT AAT TCA ACA CCA CTG TTT AAT AGT ACT TGG 2083Phe Phe Cys Cys Asn Ser Thr Pro Leu Phe Asn Ser Thr Trp455 460 465AAT GAT GCA CAA CTG TTT AAT AGT ACT TGG GAT GAT ACT AAA 2125Asn Asp Ala Gln Leu Phe Asn Ser Thr Trp Asp Asp Thr Lys470 475 480TGG TCA AAA GGC ACT AAC GAA AAT GAC ACA ATC ACC CTC CAT 2167Trp Ser Lys Gly Thr Asn Glu Asn Asp Thr Ile Thr Leu His485 490 495TGC AGA ATA AAA CAA ATT ATA AAT ATG TGG CAG GAA GTA GGA 2209Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly500 505AAA GCA ATG TAT GCC CCT CCC ATC AAA GGA CAA ATT AGA TGT 2251Lys Ala Met Tyr Ala Pro Pro Ile Lys Gly Gln Ile Arg Cys510 515 520GAA TCA AAT ATT ACA GGG CTG CTA TTA ACA AGA GAT GGT GGT 2293Glu Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly525 530 535AAC GAC ACG AGC AAG AAT AAC ACT GAG ATT TTC AGA CCT GGA 2335Asn Asp Thr Ser Lys Asn Asn Thr Glu Ile Phe Arg Pro Gly540 545 550GGA GGA AAT ATG AAG GAC AAT TGG AGA AGT GAA TTA TAT AAA 2377Gly Gly Asn Met Lys Asp Asn Trp Arg Ser Glu Leu Tyr Lys555 560 565TAT AAA GTA ATA AAA ATT GAA CCA TTA GGA GTA GCA CCC ATC 2419Tyr Lys Val Ile Lys Ile Glu Pro Leu Gly Val Ala Pro Ile570 575 579TAGGCAAAGA GAAGAGTGGT GCAGAGAGAA AAAAGAGCAG TGACACTAGG 2469AGCTATGTTC CTTGGGTTCT TGGGAGCAGC AGGAAGCACT ATGGGCGATA 2519AGCTTTAATG CGGTAGTTTA TCACAGTTAA ATTCGTAACG CAGTCAGGCA 2569CCGTGTATGA AATCTAACAA TGCGACCTGC AGAAGCTTAG AACCGAGGAA 2619CTTGTTTATT GCAGCTTATA ATGGTTACAA ATAAAGCAAT AGCATCACAA 2669ATTTCACAAA TAAAGCATTT TTTTCACTGC ATTCTAGTTG TGGTTTGTCC 2719AAACTCATCA ATGTATCTTA TCATGTCTGG ATCGGGAATT AATTCGGCGC 2769AGCACCATGG CCTGAAATAA CCTCTGAAAG A 2800(2)SEQ ID NO:31的信息(ⅰ)序列特征(A)長度579個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:31:Ser Val Ser Ser Cys Trp Trp Leu Gln Phe Arg Asn Ser Lys Pro Cys1 5 10 15Ser Glu Tyr Cys Leu Ser His Leu Val Asn Leu Arg Glu Asp Trp Gly20 25 30Pro Cys Asp Lys Leu Gln Arg Glu Arg Pro Thr Thr Pro Ile Ile Ser35 40 45Tyr Pro ＊ Gly Leu Phe Cys Val Val Arg Ser Gly Met Gly Gly Thr50 55 60Ala Ala Arg Leu Gly Ala Val Ile Leu Phe Val Val Ile Val Gly Leu65 70 75 80His Gly Val Arg Gly Lys Tyr Ala Leu Ala Asp Ala Ser Leu Lys Met85 90 95Ala Asp Pro Asn Arg Phe Arg Gly Lys Asp Leu Pro Val Leu Asp Gln100 105 110Leu Leu Glu Val Pro Val Trp Lys Glu Ala Asn Thr Thr Leu Phe Cys115 120 125Ala Ser Asp Ala Lys Ala Tyr Lys Thr Glu Ala His Asn Val Trp Ala130 135 140Thr His Ala Cys Val Pro Thr Asp Pro Lys Pro Gln Glu Ile Lys Leu145 150 155 160Glu Asn Val Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu165 170 175Gln Met His Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro180 185 190Cys Val Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys Thr Asp Leu195 200 205Arg Asn Asn Thr Asn Thr Asn Ser Thr Tyr Gly Lys Ile Met Glu Gly210 215 220Gly Glu Ile Lys Asn Cys Ser Phe Asn Ile Thr Thr Ser Ile Lys Asp225 230 235 240Lys Leu Lys Asp Met Ser Leu Phe Tyr Lys Leu Asp Val Val Pro Ile245 250 255Gly Asn Asn Ser Asn Thr Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr260 265 270Ser Val Ile Thr Gln Ala Cys Pro Lys Thr Ser Phe Glu Pro Ile Pro275 280 285Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Asn Asp290 295 300Asn Lys Phe Asn Gly Thr Gly Pro Cys Pro Asn Val Ser Thr Val Gln305 310 315 320Cys Thr His Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn325 330 335Gly Ser Leu Ala Glu Lys Glu Val Val Leu Arg Ser Glu Asn Phe Thr340345350Asp Asn Ala Lys Thr Ile Ile Val Gln Leu Asn Glu Ser Val Ile Ile355 360 365Asp Cys Met Arg Pro Asn Asn Asn Thr Arg Thr Ser Ile Pro Met Gly370 375 380Pro Gly Lys Ala Phe Tyr Ala Thr Gly Asp Val Ile Gly Asp Ile Arg385 390 395 400Arg Ala His Cys Asn Ile Ser Arg Ala Gly Trp Asn Thr Thr Leu Gln405 410 415Gln Ile Ala Lys Lys Leu Arg Glu Lys Phe Glu Asn Lys Thr Ile Val420 425 430Phe Asn His Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Thr Phe435 440 445Asn Cys Gly Gly Glu Phe Phe Cys Cys Asn Ser Thr Pro Leu Phe Asn450 455 460Ser Thr Trp Asn Asp Ala Gln Leu Phe Asn Ser Thr Trp Asp Asp Thr465 470 475 480Lys Trp Ser Lys Gly Thr Asn Glu Asn Asp Thr Ile Thr Leu His Cys485 490 495Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys Ala Met500 505 510Tyr Ala Pro Pro Ile Lys Gly Gln Ile Arg Cys Glu Set Asn Ile Thr515 520 525Gly Leu Leu Leu Thr Arg Asp Gly Gly Asn Asp Thr Ser Lys Asn Asn530 535 540Thr Glu Ile Phe Arg Pro Gly Gly Gly Asn Met Lys Asp Asn Trp Arg545 550 555 560Ser Glu Leu Tyr Lys Tyr Lys Val Ile Lys Ile Glu Pro Leu Gly Val565 570 575Ala Pro Ile(2)SEQ ID NO:32的信息(ⅰ)序列特征(A)長度1533個堿基對(B)類型核酸(C)鏈型單鏈D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:32:ATGGGGGGGA CTGCCGCCAG GTTGGGGGCC GTGATTTTGT TTGTCGTCAT 50AGTGGGCCTC CATGGGGTCC GCGGCAAATA TGCCTTGGCG GATGCCTCTC 100TCAAGATGGC CGACCCCAAT CGATTTCGCG GCAAAGACCT TCCGGTCCTG 150GACCAGCTGC TCGAG GTA CCT GTG TGG AAA GAA GCA ACC ACC 192Val Pro Val Trp Lys Glu Ala Thr Thr1 5ACT CTA TTT TGT GCA TCA GAT GCT AAA GCA TAT GAT ACA GAG 234Thr Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu10 15 20GTA CAT AAT GTT TGG GCC ACA CAT GCC TGT GTA CCC ACA GAC 276Val His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp25 30 35CCC AAC CCA CAA GAA ATA GGA TTG GAA AAT GTA ACA GAA AAT 318Pro Asn Pro Gln Glu Ile Gly Leu Glu Asn Val Thr Glu Asn40 45 50TTT AAC ATG TGG AAA AAT AAC ATG GTA GAA CAG ATG CAT GAG 360Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His Glu55 60 65GAT ATA ATC AGT TTA TGG GAT CAA AGC TTA AAG CCA TGT GTA 402Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro Cys Val70 75AAA TTA ACC CCA CTA TGT GTT ACT TTA AAT TGC ACT GAT TTG 444Lys Leu Thr Pro Leu Cys Val Thr Leu Asn Cys Thr Asp Leu80 85 90AAA AAT GCT ACT AAT ACC ACT AGT AGC AGC TGG GGA AAG ATG 486Lys Asn Ala Thr Asn Thr Thr Ser Ser Ser Trp Gly Lys Met95 100 105GAG AGA GGA GAA ATA AAA AAC TGC TCT TTC AAT GTC ACC ACA 528Glu Arg Gly Glu Ile Lys Asn Cys Ser Phe Asn Val Thr Thr110 115 120AGT ATA AGA GAT AAG ATG AAG AAT GAA TAT GCA CTT TTT TAT 570Ser Ile Arg Asp Lys Met Lys Asn Glu Tyr Ala Leu Phe Tyr125 130 135AAA CTT GAT GTA GTA CCA ATA GAT AAT GAT AAT ACT AGC TAT 612Lys Leu Asp Val Val Pro Ile Asp Asn Asp Asn Thr Ser Tyr140 145AGG TTG ATA AGT TGT AAC ACC TCA GTC ATT ACA CAG GCC TGT 654Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys150 155 160CCA AAG GTG TCC TTT GAG CCA ATT CCC ATA CAT TAT TGT GCC 696Pro Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala165 170 175CCG GCT GGT TTT GCG ATT CTA AAG TGT AGA GAT AAA AAG TTC 738Pro Ala Gly Phe Ala Ile Leu Lys Cys Arg Asp Lys Lys Phe180 185 190AAC GGA ACA GGA CCA TGT ACA AAT GTC AGC ACA GTA CAA TGT 780Asn Gly Thr Gly Pro Cys Thr Asn Val Ser Thr Val Gln Cys195 200 205ACA CAT GGA ATT AGG CCA GTA GTA TCA ACT CAA CTG CTG TTA 822Thr His Gly Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu210 215AAT GGC AGT TTA GCA GAA GAA GAA GTA GTA ATT AGA TCT GCC 864Asn Gly Ser Leu Ala Glu Glu Glu Val Val Ile Arg Ser Ala220 225 230AAT TTC TCG GAC AAT GCT AAA ACC ATA ATA GTA CAG CTG AAC 906Asn Phe Ser Asp Asn Ala Lys Thr Ile Ile Val Gln Leu Asn235 240 245GAA TCT GTA GAA ATT AAT TGT ACA AGA CCC AAC AAC AAT ACA 948Glu Ser Val Glu Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr250 255 260AGA AGA AGT ATA CAT ATA GGA CCA GGG AGA GCA TTT TAT GCA 990Arg Ar9 Ser Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala265 270 275ACA GGA GAA ATA ATA GGA GAC ATA AGA CAA GCA CAT TGT AAC 1032Thr Gly Glu Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn280 285CTT AGT AGC ACA AAA TGG AAT AAT ACT TTA AAA CAG ATA GTT 1074Leu Ser Ser Thr Lys Trp Asn Asn Thr Leu Lys Gln Ile Val290 295 300ACA AAA TTA AGA GAA CAT TTT AAT AAA ACA ATA GTC TTT AAT 1116Thr Lys Leu Arg Glu His Phe Asn Lys Thr Ile Val Phe Asn305 310 315CAC TCC TCA GGA GGG GAC CCA GAA ATT GTA ATG CAC AGT TTT 1158His Ser Ser Gly Gly Asp Pro Glu Ile Val Met His Ser Phe320 325 330AAT TGT GGA GGG GAA TTT TTC TAC TGT AAT ACA ACA CCA CTG 1200Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asn Thr Thr Pro Leu335 340 345TTT AAT AGT ACT TGG AAT TAT ACT TAT ACT TGG AAT AAT ACT 1242Phe Asn Ser Thr Trp Asn Tyr Thr Tyr Thr Trp Asn Asn Thr350 355GAA GGG TCA AAT GAC ACT GGA AGA AAT ATC ACA CTC CAA TGC 1284Glu Gly Ser Asn Asp Thr Gly Arg Asn Ile Thr Leu Gln Cys360 365 370AGA ATA AAA CAA ATT ATA AAC ATG TGG CAG GAA GTA GGA AAA 1326Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu Val Gly Lys375 380 385GCA ATG TAT GCC CCT CCC ATA AGA GGA CAA ATT AGA TGC TCA 1368Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile Arg Cys Ser390 395 400TCA AAT ATT ACA GGG CTG CTA TTA ACA AGA GAT GGT GGT AAT 1410Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Asn405 410 415AAC AGC GAA ACC GAG ATC TTC AGA CCT GGA GGA GGA GAT ATG 1452Asn Ser Glu Thr Glu Ile Phe Arg Pro Gly Gly Gly Asp Met420 425AGG GAC AAT TGG AGA AGT GAA TTA TAT AAA TAT AAA GTA GTA 1494Arg Asp Asn Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val430 435 440AAA ATT GAA CCA TTA GGA GTA GCA CCC ACC AAG GCA TAA 1533Lys Ile Glu Pro Leu Gly Val Ala Pro Thr Lys Ala ＊445 450 455(2)SEQ ID NO:33的信息(ⅰ)序列特征(A)長度456個氨基酸(B)類型氨基酸(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:33:Val Pro Val Trp Lys Glu Ala Thr Thr Thr Leu Phe Cys Ala Ser Asp1 5 10 15Ala Lys Ala Tyr Asp Thr Glu Val His Asn Val Trp Ala Thr His Ala20 25 30Cys Val Pro Thr Asp Pro Asn Pro Gln Glu Ile Gly Leu Glu Asn Val35 40 45Thr Glu Asn Phe Asn Met Trp Lys Asn Asn Met Val Glu Gln Met His50 55 60Glu Asp Ile Ile Ser Leu Trp Asp Gln Ser Leu Lys Pro Cys Val Lys65 70 75 80Leu Thr Pro Leu Cys Val Thr Leu Asn Cys Thr Asp Leu Lys Asn Ala85 90 95Thr Asn Thr Thr Ser Ser Ser Trp Gly Lys Met Glu Arg Gly Glu Ile100 105 110Lys Asn Cys Ser Phe Asn Val Thr Thr Ser Ile Arg Asp Lys Met Lys115 120125Asn Glu Tyr Ala Leu Phe Tyr Lys Leu Asp Val Val Pro Ile Asp Asn130 135 140Asp Asn Thr Ser Tyr Arg Leu Ile Ser Cys Asn Thr Ser Val Ile Thr145 150 155 160Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys165170175Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Arg Asp Lys Lys Phe Asn180 185 190Gly Thr Gly Pro Cys Thr Asn Val Ser Thr Val Gln Cys Thr His Gly195 200205Ile Arg Pro Val Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala210 215 220Glu Glu Glu Val Val Ile Arg Ser Ala Asn Phe Ser Asp Asn Ala Lys225 230 235240Thr Ile Ile Val Gln Leu Asn Glu Ser Val Glu Ile Asn Cys Thr Arg245 250 255Pro Asn Asn Asn Thr Arg Arg Ser Ile His Ile Gly Pro Gly Arg Ala260 265 270Phe Tyr Ala Thr Gly Glu Ile Ile G1y Asp Ile Arg Gln Ala His Cys275 280 285Asn Leu Ser Ser Thr Lys Trp Asn Asn Thr Leu Lys Gln Ile Val Thr290 295 300Lys Leu Arg Glu His Phe Asn Lys Thr Ile Val Phe Asn His Ser Ser305 310 315 320Gly Gly Asp Pro Glu Ile Val Met His Ser Phe Asn Cys Gly Gly Glu325 330 335Phe Phe Tyr Cys Asn Thr Thr Pro Leu Phe Asn Ser Thr Trp Asn Tyr340 345 350Thr Tyr Thr Trp Asn Asn Thr Glu Gly Ser Asn Asp Thr Gly Arg Asn355 360 365Ile Thr Leu Gln Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Glu370 375 380Val Gly Lys Ala Met Tyr Ala Pro Pro Ile Arg Gly Gln Ile Arg Cys385 390 395 400Ser Ser Asn Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Asn Asn405 410 415Ser Glu Thr Glu Ile Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn420 425 430Trp Arg Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu435 440 445Gly Val Ala Pro Thr Lys Ala ＊450 455(2)SEQ ID NO:34的信息(ⅰ)序列特征(A)長度37個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:34:
GGGAATTCGG ATCCAGAGCA GAAGACAGTG GCAATGA 37(2)SEQ IDNO:35的信息(ⅰ)序列特征(A)長度33個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ IDNO:35:
CTCGAGCTCC TGAACACAGT CAGACTCATC AAG 33(2)SEQ ID NO:36的信息(ⅰ)序列特征(A)長度39個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:36:
GGTCTAGAAG CTTTAGCCCA TAGTGCTTCC TGCTGCTCC 39(2)SEQ ID NO:37的信息(ⅰ)序列特征(A)長度36個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:37:
GGGCGGATCC TCGAGGTACC TGTRTGGAAA GAAGCA 36(2)SEQ ID NO:38的信息(ⅰ)序列特征(A)長度38個堿基對(B)類型核酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQID NO:38:
GGTCTAGAAG CTTTATGCTC CYAAGAACCC AAGGAACA38(2)SEQ ID NO:39的信息(ⅰ)序列特征(A)長度7個氨基酸(B)類型氨基酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:39:
Ile GlyPro GlyArgAlaPhel5(2)SEQID NO:40的信息(ⅰ)序列特征(A)長度7個氨基酸(B)類型氨基酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:40:
Ile Gly Pro Gly Arg Ala Trp1 5(2)SEQ IDNO41的信息(ⅰ)序列特征(A)長度7個氨基酸(B)類型氨基酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:41:
Leu Gly Pro Gly Ser Thr Phel 5(2)SEQIDNO:42的信息(ⅰ)序列特征(A)長度7個氨基酸
(B)類型氨基酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:42:
Ile Gly Pro Gly Arg Val Leul 5(2)SEQ ID NO:43的信息(ⅰ)序列特征(A)長度7個氨基酸(B)類型氨基酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:43:
Ile Gly Pro Gly Ser Ala Phe1 5(2)SEQ ID NO:44的信息(ⅰ)序列特征(A)長度5個氨基酸(B)類型氨基酸(C)鏈型單鏈(D)拓?fù)浣Y(jié)構(gòu)線型(ⅹⅰ)序列描述SEQ ID NO:44:
Ile Gly Pro Gly Arg1 權(quán)利要求
1．一種分離的多肽，該多肽包含HIV gp120的氨基酸序列，所說的氨基酸序列選自由SEQ IDNo.2,4,6,8,10,12,14,16,18,20,22,24,26和28，以及它們的片段組成的組。
2．權(quán)利要求1的多肽，其中所說的多肽還包含標(biāo)記表位序列。
3．權(quán)利要求2的多肽，其中所說的標(biāo)記表位序列是HSV gD-1標(biāo)記表位序列。
4．權(quán)利要求2的多肽，其中所說的標(biāo)記表位序列是融合到HIVgp120氨基酸序列中的。
5．一種不超過五千堿基的寡核苷酸序列，其編碼HIV gp120多肽序列，該多肽序列包含選自由SEQ ID No.2,4,6,8,10,12,14,16,18,20,22,24,26和28，以及它們的片段組成的組的氨基酸序列。
6．權(quán)利要求5的寡核苷酸，其中所說的寡核苷酸包含選自由SEQ ID No.1,3,5,7,9,11,13,15,17,19,21,23,25和27，以及它們的片段組成的組的核苷酸序列。
7．權(quán)利要求5的寡核苷酸，其中由該寡核苷酸編碼的氨基酸序列還包含一個標(biāo)記表位。
8．權(quán)利要求5的寡核苷酸，其中所說的標(biāo)記表位是HSV gD-1標(biāo)記表位。
9．權(quán)利要求7的寡核苷酸，其中所說的標(biāo)記表位是融合到HIVgp120氨基酸序列中的。
10．一種疫苗，該疫苗包含在合適載體中的gp120 MN和HIVgp120多肽序列，所說的多肽序列選自由SEQ ID No.2,4,6,8,10,12,14,16,18,20,22,24,26和28，以及它們的片段組成的組。
11．一種疫苗，該疫苗包含a．第一gp120多肽序列或其片段；和b．一種逃逸分離物HIV gp120多肽序列或其片段，其來自用所說的第一HIV gp120多肽序列接種的被接種者；其中所說的HIV gp120多肽序列在合適的載體中。
12．權(quán)利要求11的疫苗，其中所說的第一HIV gp120多肽序列包括gp120MN,gp120 A244,gp120MN-GNE6(SEQ ID No.31)，或gp120MN-GNE8(SEQ ID No.33)。
13．權(quán)利要求12的疫苗，其中所說的疫苗還包含第二gp120多肽序列或其片段，所說的第二gp120多肽序列包括gp120MN,gp120A244,gp120MN-GNE6(SEQ ID No.31)，或gp120MN-GNE8(SEQ IDNo.33)，其中所說的第二HIV gp120多肽序列不同于第一HIV gp120多肽序列。
14．權(quán)利要求13的疫苗，其中所說的第一gp120多肽序列包括gp120MN，并且所說的第二gp120多肽序列包括gp120.A244。
15．權(quán)利要求14的疫苗，其中所說的逃逸分離物包含在合適載體中的HIV gp120多肽序列，該序列選自由SEQID No.2,4,6,8,10,12,14,16,18,20,22,24,26和28，以及它們的片段組成的組。
16．權(quán)利要求13的疫苗，其中所說的第一gp120多肽序列包括gp120MN，并且所說的第二gp120多肽序列包括gp120 MN-GNE8(SEQ ID No.33)。
17．權(quán)利要求16的疫苗，其中所說的逃逸分離物HIV gp120多肽序列是在合適載體中的HIV gp120多肽序列，該序列選自由SEQ IDNo.2,4,6,8,10,12,14,16,18,20,22,24,26和28，以及它們的片段組成的組。
18．權(quán)利要求13的疫苗，其中所說的逃逸分離物HIV gp120多肽來自用所說的第一和第二HIV gp120多肽序列接種的被接種者。
19．一種制備HIV疫苗的方法，該方法包括將來自被接種者的逃逸分離物HIV gp120多肽序列或其片段添加至用于接種到被接種者的疫苗中。
20．權(quán)利要求11的疫苗，其中所說的第一gp120多肽序列來自親巨噬細(xì)胞的HIV-1菌株。
21．權(quán)利要求11的疫苗，其中所說的第一gp120多肽序列來自親T-細(xì)胞的HIV-1菌株。
22．權(quán)利要求21的疫苗，其中所說的疫苗還包含來自親巨噬細(xì)胞的HIV-1菌株的第二gp120多肽序列或其片段。
23．權(quán)利要求22的疫苗，其中所說的第一和第二gp120多肽序列結(jié)合在不同的趨化因子受體上。
24．權(quán)利要求23的疫苗，其中所說的第一gp120多肽序列結(jié)合在CC-CKR-5上，而所說的第二gp120多肽序列結(jié)合在CXC-CKR-4上。
25．權(quán)利要求11的疫苗，其中所說的疫苗還包含人工改造的用于誘導(dǎo)細(xì)胞毒性T-細(xì)胞反應(yīng)的病毒。
全文摘要
本發(fā)明提供了編碼gp120多肽(其來自用MN－rgp120的疫苗試驗(yàn)的逃逸分離物)的寡核苷酸序列和所編碼的gp120多肽。利用亞單位疫苗中的一種或多種分離物的gp120多肽(通常與MN－rgp120一起),可提供抗一些HIV菌株的保護(hù)作用,這些HIV菌株與疫苗菌株(例如;MN－rgp120)有足夠的差異,使得所說的疫苗不提供抗這些菌株的保護(hù)作用。本發(fā)明也提供了所述多肽誘導(dǎo)的抗體。
文檔編號A61K39/21GK1225132SQ97196244
公開日1999年8月4日 申請日期1997年7月3日 優(yōu)先權(quán)日1997年7月3日
發(fā)明者菲利普·W·伯曼 申請人:遺傳技術(shù)研究公司