專利名稱：基于il－1受體的拮抗劑和制造與使用的方法
本申請(qǐng)是2001年3月22日提交的美國申請(qǐng)09/787,835的部份延續(xù)，后者是1999年9月22日提交的國際申請(qǐng)PCT/US99/22045的美國國家階段申請(qǐng)，所述國際申請(qǐng)要求1999年5月19日提交的美國申請(qǐng)09/313,942(已授權(quán)的)的優(yōu)先權(quán)，所述美國申請(qǐng)09/313,942要求1998年9月25日提交的美國臨時(shí)申請(qǐng)60/101,858(現(xiàn)已放棄)的優(yōu)先權(quán)。本申請(qǐng)通篇參考了各種出版物。這些完全公開的出版物此處引用作為參考。
背景技術(shù)：
細(xì)胞因子CNTF家族在眾多的生理過程中起重要作用，這為拮抗劑和激動(dòng)劑都提供了潛在的治療應(yīng)用。
發(fā)明概述本發(fā)明目的是生產(chǎn)用于治療涉及細(xì)胞因子的疾病或病癥的細(xì)胞因子拮抗劑。
本發(fā)明另一個(gè)目的是已公開的所述細(xì)胞因子拮抗劑在治療涉及細(xì)胞因子的疾病或病癥中的應(yīng)用。
本發(fā)明的另一個(gè)目的是構(gòu)建幾個(gè)特異性的IL-1細(xì)胞因子拮抗劑，稱為IL-1捕獲者(Trap)，各具有不同的序列但都能夠阻止IL-1和它的受體結(jié)合，因此用作IL-1的拮抗劑。
附圖簡(jiǎn)述

圖1人IL-1捕獲者阻斷外源施用的huIL-1的體內(nèi)效應(yīng)。在時(shí)間0時(shí)給BALB/c小鼠皮下注射huIL-1(0.3μg/kg)。在注射huIL-1之前24小時(shí)，用載體或150倍摩爾過量的huI L-1捕獲者預(yù)先處理動(dòng)物。在宰殺動(dòng)物之前2小時(shí)(26小時(shí))，第二次對(duì)小鼠注射huIL-1(0.3μg/kg)以再次刺激免疫反應(yīng)。在各個(gè)時(shí)間點(diǎn)收集血液樣品并檢測(cè)血清中IL-1的水平(用平均值+/-SEM表示；n＝5每組)。
圖2人IL-1捕獲者阻斷外源施用的人IL-1的體內(nèi)效應(yīng)。
圖3人IL-1捕獲者阻斷發(fā)炎的關(guān)節(jié)中的IL-1的效應(yīng)。
圖4-5鼠科動(dòng)物IL-1捕獲者在酵母聚糖加速的膠原蛋白誘導(dǎo)的關(guān)節(jié)炎(Zymosan-Accelerated Collagen-Induced Arthritis)(CIA)模型中減輕關(guān)節(jié)炎癥的嚴(yán)重程度。
圖6在5pM濃度人IL-b存在的情況下，室溫下孵育各種濃度的IL-1捕獲者1649過夜。
發(fā)明詳述本發(fā)明提供了編碼能和細(xì)胞因子結(jié)合生成無功能復(fù)合物的融合多肽的分離的核酸分子，所述核酸分子包含(a)編碼第一個(gè)融合多肽組件的核苷酸序列，所述多肽組件包含細(xì)胞因子受體特異性決定組件的細(xì)胞外結(jié)構(gòu)域中的細(xì)胞因子結(jié)合部位的氨基酸序列；(b)編碼第二個(gè)融合多肽組件的核苷酸序列，所述多肽組件包含細(xì)胞因子受體信號(hào)傳導(dǎo)組件的細(xì)胞外結(jié)構(gòu)域中的細(xì)胞因子結(jié)合部位的氨基酸序列；和(c)編碼第三個(gè)融合多肽組件的核苷酸序列，所述組件包含多聚化組件的氨基酸序列。
“細(xì)胞因子結(jié)合部位”指的是結(jié)合細(xì)胞因子所必需的細(xì)胞外結(jié)構(gòu)域的最小組成部分。本領(lǐng)域技術(shù)人員所接受的細(xì)胞因子受體的一個(gè)明確特征是存在兩個(gè)包含經(jīng)典的半胱氨酸的纖連蛋白樣結(jié)構(gòu)域和存在WSXWS盒(SEQ ID NO26)(Bazan(1990)PNAS 876934-6938)。編碼細(xì)胞因子受體結(jié)合組件的細(xì)胞外結(jié)構(gòu)域和信號(hào)傳導(dǎo)組件的細(xì)胞外結(jié)構(gòu)域的序列也可以用來創(chuàng)建本發(fā)明的融合多肽。類似地，可以使用更長的編碼更大的細(xì)胞因子受體組件部位的序列。然而，由于考慮到比細(xì)胞外結(jié)構(gòu)域更小的片段能行使與細(xì)胞因子結(jié)合的功能，因而本發(fā)明包含的融合多肽包含作為細(xì)胞因子結(jié)合部位的結(jié)合細(xì)胞因子所必需的細(xì)胞外結(jié)構(gòu)域最小組成部分。
本發(fā)明包含細(xì)胞因子受體的“特異性決定組件”和細(xì)胞因子受體的“信號(hào)傳導(dǎo)組件”。不考慮用來稱呼細(xì)胞因子受體特定的組件或亞基的命名系統(tǒng)，本專業(yè)技術(shù)人員能識(shí)別出受體的哪個(gè)組件或亞基負(fù)責(zé)決定細(xì)胞因子的細(xì)胞靶向，于是可以知道哪個(gè)組件構(gòu)成了“特異性決定組件”。類似地，不考慮所用的命名系統(tǒng)，本領(lǐng)域技術(shù)人員能夠知道哪個(gè)組件或亞基可以構(gòu)成“信號(hào)傳導(dǎo)組件”。如此處所使用的，“信號(hào)傳導(dǎo)組件”是天然存在的受體的組件，所述信號(hào)傳導(dǎo)組件不是特異性決定組件并且在缺少特異性決定組件的情況下不與或微弱地與細(xì)胞因子結(jié)合。在天然存在的受體中，“信號(hào)傳導(dǎo)組件”可參與信號(hào)傳導(dǎo)。
在制備編碼本發(fā)明的融合多肽的核酸序列中，所述融合多肽的第一，第二和第三組件編碼在單鏈核酸上，當(dāng)通過宿主載體系統(tǒng)表達(dá)時(shí)便產(chǎn)生單體類型的融合多肽。這樣表達(dá)的單體，接著在多聚化組件(第三個(gè)融合多肽組件)間的相互作用下多聚化。用這種方式產(chǎn)生融合多肽避免了純化異二聚體混合物(如果第一和第二組件由分開的分子產(chǎn)生而且接著多聚化將導(dǎo)致該異二聚體混合物)的需要。例如，于1995年11月28日頒發(fā)的美國專利5,470,952描述了作為CNTF或IL-6拮抗劑的異二聚體蛋白的生產(chǎn)。從由合適的alpha(a)和beta(b)組件共轉(zhuǎn)染的細(xì)胞系中純化異二聚體。接著使用諸如從制備型非變性聚丙酰胺凝膠中被動(dòng)洗脫或使用高壓陽離子交換層析等方法將異二聚體從同二聚體中分離出來。通過本發(fā)明方法可以避免對(duì)這個(gè)純化步驟的需求。
此外，1996年4月18日公布的標(biāo)題為二聚體IL-4抑制劑的PCT國際申請(qǐng)WO96/11213聲稱其申請(qǐng)者已經(jīng)制備出同二聚體，其中二個(gè)IL-4受體通過一個(gè)多聚間隔子相連，和異二聚體，其中一個(gè)IL-4受體通過一個(gè)多聚間隔子與一個(gè)IL-2受體γ鏈相連。所描述的多聚間隔子是聚乙二醇。兩個(gè)受體組件，IL-4R和IL-2Rγ是分開表達(dá)和純化的。通過使用雙功能的PEG試劑將組件連接在一起便產(chǎn)生了聚乙二醇化的(Pegylated)同二聚體和異二聚體。本發(fā)明的優(yōu)勢(shì)是它避免了對(duì)這種費(fèi)時(shí)且昂貴的純化和聚乙二醇化步驟的需要。
在本發(fā)明的一個(gè)實(shí)施方案中，編碼第一個(gè)組件的核酸序列在編碼第二個(gè)組件的核酸序列上游。在另一個(gè)本發(fā)明實(shí)施方案中，編碼第一個(gè)組件的核酸序列在編碼第二個(gè)組件的核酸序列下游。本發(fā)明更近一步的實(shí)施方案可以重新安排融合多肽的第一，第二和第三組件的順序。例如，如果編碼第一個(gè)組件的核酸序列稱作1，編碼第二個(gè)組件的核酸序列稱作2，編碼第三個(gè)組件的核酸序列稱作3，那么組件在本發(fā)明的分離的核酸上從5’到3’讀的順序可以是下列六種組合中的任一種1，2，3；1，3，2；2，1，3；2，3，1；3，1，2；或3，2，1。
在本發(fā)明特定的實(shí)施方案中，被融合多肽結(jié)合的細(xì)胞因子是白細(xì)胞介素-1(IL-1)。
在本發(fā)明優(yōu)選的實(shí)施方案中，多聚化組件包括免疫球蛋白衍生的結(jié)構(gòu)域。更特別的是，免疫球蛋白衍生的結(jié)構(gòu)域可能選自IgG的Fc結(jié)構(gòu)域、IgG的重鏈和IgG的輕鏈。甚至更特別的是，免疫球蛋白的結(jié)構(gòu)域可能選自IgG1或IgG4的Fc結(jié)構(gòu)域、IgG1或IgG4的重鏈和IgG1或IgG4的輕鏈。在另一個(gè)實(shí)施方案中，多聚化組件可能是一個(gè)Fc結(jié)構(gòu)域，其中去除了前五個(gè)氨基酸(包括一個(gè)半胱氨酸)便產(chǎn)生被稱作Fc(DC1)的多聚化組件?；蛘?，多聚化組件可能是其前五個(gè)氨基酸中的半胱氨酸已被另外的氨基酸(例如絲氨酸或丙氨酸)取代的Fc結(jié)構(gòu)域。
本發(fā)明也提供了由本發(fā)明的分離的核苷酸編碼的融合多肽。優(yōu)選地，由于第三個(gè)組件(多聚化組件)的功能，融合多肽以多聚體的形式存在。在優(yōu)選的實(shí)施方案中，所述多聚體是二聚體。合適的多聚化組件是編碼免疫球蛋白重鏈鉸鏈區(qū)的序列(Takahashi等人，(1982)Ce1129671-679)；免疫球蛋白基因序列和其中的部分序列。在優(yōu)選的本發(fā)明實(shí)施方案中，免疫球蛋白基因序列，尤其是編碼Fc結(jié)構(gòu)域的序列，用來編碼多聚化組件。
本發(fā)明也包括包含有此處所述的核酸分子的載體。
本發(fā)明的優(yōu)選實(shí)施方案是編碼具有SEQ ID NO2序列融合多肽的分離的具有SEQ ID NO1序列的核酸分子，其中融合多肽形成能與細(xì)胞因子結(jié)合形成無功能復(fù)合物的多聚體；編碼具有SEQ ID NO4序列的融合多肽的分離的核酸分子，其具有SEQ ID NO3所列序列，其中融合多肽形成能結(jié)合細(xì)胞因子形成無功能復(fù)合物的多聚體；編碼具有SEQ ID NO6序列的融合多肽的分離的核酸分子，其具有SEQ ID NO5所列序列，其中融合多肽形成能結(jié)合細(xì)胞因子形成無功能復(fù)合物的多聚體；以及由上述核酸分子編碼的融合多肽。
其它本發(fā)明優(yōu)選的實(shí)施方案是具有SEQ ID NO7，9，11，13，15，17，19，21和23所列序列的分離的核酸分子，它們分別編碼具有SEQ IDNO8，10，12，14，16，18，20和22所列序列的融合多肽，其中各融合多肽形成能結(jié)合IL-1形成無功能復(fù)合物的多聚體。
本發(fā)明也提供了表達(dá)載體，所述載體包含此處描述的本發(fā)明核酸分子，其中該核酸分子有效地連接表達(dá)控制序列。本發(fā)明也提供了用來生產(chǎn)融合多肽的宿主-載體系統(tǒng)，所述宿主-載體系統(tǒng)包含本發(fā)明的表達(dá)載體，該載體被導(dǎo)入適合用來表達(dá)融合多肽的宿主細(xì)胞。合適的宿主細(xì)胞可以是細(xì)菌細(xì)胞，如大腸桿菌，酵母細(xì)胞，如巴斯德比赤氏酵母(pichiapastoris)，昆蟲細(xì)胞，如草地夜蛾(Spodoptera frug iperda)、或哺乳動(dòng)物細(xì)胞，如COS，CHO，293，BHK或NSO細(xì)胞。
本發(fā)明還提供了生產(chǎn)本發(fā)明融合多肽的方法通過在允許生產(chǎn)融合多肽的條件下培養(yǎng)此處描述的宿主-載體系統(tǒng)中的細(xì)胞和回收所產(chǎn)生的融合多肽以生產(chǎn)本發(fā)明的融合多肽。
根據(jù)本發(fā)明制備的sRα:β1異二聚體為它們的配基提供了有效的捕獲者，它們?cè)谄つ柤?jí)范圍能親合結(jié)合這些配體而不產(chǎn)生有功能的中間體。此處描述的技術(shù)可用于為任何一種細(xì)胞因子開發(fā)細(xì)胞因子捕獲物，其利用α組件(賦予特異性)和β組件(當(dāng)與α特異性組件結(jié)合后，有著比單獨(dú)使用其中任何一種組件時(shí)更高的對(duì)細(xì)胞因子的親合力)。因而，根據(jù)本發(fā)明的拮抗劑包括白細(xì)胞介素-1(IL-1)的拮抗劑。
利用對(duì)本領(lǐng)域技術(shù)人員已知的方法可以制備α和β受體的細(xì)胞外結(jié)構(gòu)域?？梢酝ㄟ^克隆和在原核或真核表達(dá)系統(tǒng)中的表達(dá)來制備用于實(shí)現(xiàn)本發(fā)明的受體分子?？梢允褂萌魏畏N類的方法來表達(dá)和純化重組受體基因。將編碼該因子的基因亞克隆入細(xì)菌表達(dá)載體，諸如例如但不局限于PC110。
可以使用任何允許隨后形成穩(wěn)定的具有生物活性的蛋白技術(shù)純化重組因子。例如但不局限于，所述因子可以以可溶性蛋白或包含體的形式從細(xì)胞中回收，然后可用8M鹽酸胍和透析將蛋白從包含體中定量提取出來。為了進(jìn)一步純化因子，可使用傳統(tǒng)的離子交換層析，疏水相互作用層析，反向?qū)游龌蚰z過濾。
正如公開的PCT申請(qǐng)WO93/10151(描述了β受體異二聚體的產(chǎn)生)所描述的那樣，可使用已知的融合區(qū)域進(jìn)行基因工程制備sRα:β異二聚體受體，或通過化學(xué)方法交聯(lián)細(xì)胞外結(jié)構(gòu)域來制備它們。所使用的結(jié)構(gòu)域可以由α和β組件的完整細(xì)胞外結(jié)構(gòu)域構(gòu)成，或者它們可以由保持有與其配體形成復(fù)合體能力的其突變體或片段和sRα:β復(fù)合物上的其它組件形成。
在本發(fā)明的一個(gè)實(shí)施方案中，細(xì)胞外結(jié)構(gòu)域可用亮氨酸拉鏈進(jìn)行基因工程制備。已經(jīng)顯示人轉(zhuǎn)錄因子c-jun和c-fos的亮氨酸拉鏈結(jié)構(gòu)域可以形成穩(wěn)定的1∶1的化學(xué)計(jì)量的異二聚體(Bush等人，(1990)TrendsGenetics 636-40；Gentz等人，(1989)Science 2431695-1699)。盡管已經(jīng)顯示jun-jun同二聚體能夠形成，但它們的穩(wěn)定性比jun-fos異二聚體的弱1000倍。從未檢測(cè)到Fos-fos同二聚體。
通過遺傳工程形成嵌合基因?qū)-jun或c-fos的亮氨酸拉鏈結(jié)構(gòu)域融合入上述受體組件的可溶性或細(xì)胞外結(jié)構(gòu)域的C末端讀碼框內(nèi)。融合可以是直接的或可以應(yīng)用一個(gè)柔性的連接子結(jié)構(gòu)域如人IgG的鉸鏈區(qū)、或由小氨基酸(諸如甘氨酸，絲氨酸，蘇氨酸或丙氨酸)以各種長度和組合構(gòu)成的多肽連接子。另外，可使用His-His-His-His-His-His(His6)(SEQ ID NO25)標(biāo)記嵌合蛋白，這樣可通過金屬螯合層析快速純化該蛋白，和/或通過可獲得抗體的抗原決定基標(biāo)記，而允許通過蛋白質(zhì)印跡法、免疫共沉淀或活性損耗/阻斷生物鑒定對(duì)該蛋白進(jìn)行檢測(cè)。
在另一個(gè)實(shí)施方案中，用人IgG1的Fc結(jié)構(gòu)域來制備sRα:β1異二聚體(Aruffo等人(1991)Cell 6735-44)。與前者相反，由于攜帶有Fc結(jié)構(gòu)域的嵌合分子將表達(dá)為通過二硫鍵連接的同二聚體，所以異二聚體的形成必須通過生物化學(xué)方法獲得。于是在有利于打斷鏈間二硫鍵但不影響鏈內(nèi)二硫鍵的條件下同二聚體將會(huì)減少。然后將具有不同細(xì)胞外組成部分的單體等摩爾量混合并氧化形成同和異二聚體的混合物?？梢酝ㄟ^層析技術(shù)對(duì)這個(gè)混合物的組分進(jìn)行分離?；蛘?，通過遺傳改造和表達(dá)由受體組件的可溶性或細(xì)胞外部位，其后連接hIgG的Fc結(jié)構(gòu)域，再連接上述的c-jun或c-fos亮氨酸拉鏈(Kostelny等人，(1992)J.Immunol.1481547-1553))構(gòu)成的分子，可以偏向性形成這種異二聚體。由于這些亮氨酸拉鏈主要形成異二聚體，它們可以用來驅(qū)動(dòng)形成目的二聚體。對(duì)于描述過的使用亮氨酸拉鏈的嵌合蛋白，它們也可以使用金屬螯合劑或抗原決定基標(biāo)記。通過金屬螯合層析可快速純化這種帶有標(biāo)簽的結(jié)構(gòu)域，和/或通過抗體使用蛋白質(zhì)印跡、免疫共沉淀或活性損耗/阻斷生物鑒定方法來檢測(cè)這種帶有抗原決定基標(biāo)簽的結(jié)構(gòu)域。
在另外的實(shí)施方案中，可使用驅(qū)動(dòng)二聚體形成的其它免疫球蛋白來源的結(jié)構(gòu)域來制備異二聚體。例如這樣的結(jié)構(gòu)域包括IgG的重鏈(Cg1和Cg4)以及人免疫球蛋白的kappa(K)和lambda(λ)輕鏈的恒定區(qū)。Cg和輕鏈的異二聚體化發(fā)生在Cg的CH1結(jié)構(gòu)域和輕鏈(CL)的恒定區(qū)之間并且通過單個(gè)二硫鍵橋共價(jià)連接兩個(gè)結(jié)構(gòu)域而穩(wěn)定。因此，可以使用這些免疫球蛋白的結(jié)構(gòu)域制備構(gòu)建體。或者，免疫球蛋白的結(jié)構(gòu)域包括來源于驅(qū)動(dòng)二聚體化的T細(xì)胞受體組件的結(jié)構(gòu)域。
在本發(fā)明另一個(gè)實(shí)施方案中，通過表達(dá)利用柔性連接環(huán)的嵌合分子可以制備sRα:β1異二聚體。編碼嵌合蛋白的DNA構(gòu)建體設(shè)計(jì)為能表達(dá)通過柔性環(huán)以串連方式(“頭對(duì)頭”)融合在一起的二個(gè)可溶性或細(xì)胞外結(jié)構(gòu)域。這個(gè)環(huán)可以是完全人造的(例如在某些間隔處被絲氨酸或蘇氨酸插入打斷的多聚甘氨酸重復(fù))或從天然存在的蛋白(例如hIgG的鉸鏈區(qū))中“借來”。通過基因工程可以構(gòu)建其中交換可溶性結(jié)構(gòu)域或融合的細(xì)胞外結(jié)構(gòu)域順序的分子(例如sIL6Ra/環(huán)/sgp130或sgp130/環(huán)/sIL-6Ra)和/或其中改變環(huán)的長度和組成允許選擇具有所需特征的分子。
或者，跟據(jù)本發(fā)明制造的異二聚體可以從用合適的α和β組件共轉(zhuǎn)染的細(xì)胞系進(jìn)行純化。本領(lǐng)域技術(shù)人員使用已知方法從同二聚體中分離出異二聚體。例如，通過從制備型的非變性聚丙烯酰胺凝膠中被動(dòng)洗脫可以回收有限量的異二聚體。或者，使用高壓陽離子交換層析可以純化異二聚體。使用Mono S陽離子交換柱可以獲得高質(zhì)量的純化。
使用本領(lǐng)域技術(shù)人員已知的方法(例如參看Fingl等人，(1975)ThePharmacological Basis of Therapeutics，Goodman and Gilman，eds.Macmill an Publishing Co.，New York，pp.1-46)可以確定用來治療IL-1相關(guān)疾病和病癥的有效劑量。根據(jù)本發(fā)明使用的藥物組合物包括上述的拮抗劑，所述拮抗劑在體內(nèi)施用之前存在于藥學(xué)可接受的液體、固體或半固體載體、連接到載體或靶向分子(例如抗體、激素、生長因子等)和/或包裝入脂質(zhì)體、微囊體和受控釋放制劑內(nèi)(包括拮抗劑表達(dá)細(xì)胞)。例如，藥物組合物可能在含水溶液中(如無菌水、鹽水、磷酸緩沖液或葡萄糖溶液)包含一種或更多種拮抗劑。或者，活性藥劑可以包含在固體(例如石蠟)或半固體(例如凝膠狀的)制劑中，所述制劑可以植入到需要這種治療的病人體內(nèi)。給藥途徑可以是任何一種本領(lǐng)域已知的給藥模式，包括但不局限于靜脈內(nèi)地、鞘內(nèi)地、皮下地，通過注射入所涉及的組織，動(dòng)脈內(nèi)地、鼻內(nèi)地、經(jīng)口地，或通過植入設(shè)備。
給藥可以導(dǎo)致本發(fā)明活性藥劑全身或局部分布。例如，在某些涉及神經(jīng)系統(tǒng)長距區(qū)域的情況下，靜脈內(nèi)或皮下施用藥劑可能比較理想。某些情況下，將包含有活性藥劑的植入物植入受損的區(qū)域或附近。合適的植入物包括但不局限于明膠海綿、蠟或基于微顆粒的植入物。
實(shí)施例實(shí)施例1.克隆融合多肽組件通過利用組織cDNA的標(biāo)準(zhǔn)PCR技術(shù)(CLONTECH)進(jìn)行PCR可獲得人細(xì)胞因子受體細(xì)胞外結(jié)構(gòu)域，將其克隆入表達(dá)載體pMT21(GeneticsInstitute，Inc.)并且用ABI 373A DNA測(cè)序列儀和Taq雙脫氧終止子循環(huán)測(cè)序試劑盒(Applied Biosystems，Inc.，F(xiàn)os ter City，CA))對(duì)該序列進(jìn)行測(cè)序。對(duì)于IL-IRAcP，克隆來自Genbank序列AB006357的從1到1074位的核苷酸(對(duì)應(yīng)氨基酸1-358)。對(duì)于IL-1RI，克隆來自Genbank序列X16896的從55到999位的核苷酸(對(duì)應(yīng)氨基酸19-333)。
實(shí)施例2.生產(chǎn)融合多肽(細(xì)胞因子捕獲者)通過標(biāo)準(zhǔn)的克隆和PCR技術(shù)由單個(gè)克隆的DNA(上面描述的)構(gòu)建編碼細(xì)胞因子捕獲者的核酸序列。在每一個(gè)案例中，將所述序列構(gòu)建入讀碼框內(nèi)，使得編碼第一個(gè)融合多肽組件的序列可以與編碼第二個(gè)融合多肽組件的序列融合，其后連接一個(gè)作為多聚化組件的Fc結(jié)構(gòu)域(鉸鏈，人IgG1的CH2和CH3區(qū)域)。在某些情況下，將額外的核苷酸插入編碼第一個(gè)和第二個(gè)融合多肽組件的序列之間的讀碼框內(nèi)，為兩個(gè)組件之間添加一個(gè)連接子區(qū)域。
實(shí)施例3.對(duì)IL-1捕獲子的MRC5生物鑒定MRC5人肺成纖維細(xì)胞通過分泌IL-6對(duì)IL-1作出反應(yīng)，于是可以用MRC5細(xì)胞來檢測(cè)IL-1捕獲者對(duì)IL-1依賴性的IL-6生成的阻斷能力。IL1捕獲子ISC569針對(duì)IL-1-RI.Fc進(jìn)行測(cè)試，所述IL-1-RI.Fc是融入一個(gè)Fc結(jié)構(gòu)域的IL-1 I型受體的細(xì)胞外結(jié)構(gòu)。
將MRC5細(xì)胞以每毫升1×105個(gè)細(xì)胞懸浮在培養(yǎng)基里并將0.1ml細(xì)胞(每孔10,000個(gè)細(xì)胞)鋪到96孔組織培養(yǎng)板的孔里。培養(yǎng)板在加濕的含5％CO2的培養(yǎng)箱中于37℃孵育24小時(shí)。
將IL-1捕獲者和不同劑量的重組人IL-1預(yù)先孵育在96孔培養(yǎng)皿內(nèi)并在37℃孵育2小時(shí)。然后將0.1ml該混合物加入到含有MRC5細(xì)胞的96孔板內(nèi)，這樣IL-1捕獲者的終濃度為10nM而IL-1終濃度的范圍是從2.4pM到5nM。對(duì)照孔只含捕獲者或什么都不加。
然后將培養(yǎng)板放入加濕的含5％CO2的培養(yǎng)箱中于37℃條件下孵育24小時(shí)。收集上清液，根據(jù)產(chǎn)商說明使用R&D Systems QuantikineImmunoassay Kit檢測(cè)IL-6的水平。
實(shí)施例4.在體內(nèi)用IL-1捕獲子阻斷注射的IL-1IL-1是一個(gè)促炎細(xì)胞因子。已經(jīng)顯示在動(dòng)物中系統(tǒng)性地給藥IL-1會(huì)引發(fā)急性反應(yīng)，包括短暫高血糖癥、胰島素分泌過少、發(fā)燒、食欲減退和增加的血清白細(xì)胞介素6(IL-6)水平(Reimers，1998)。因?yàn)樾∈髮?duì)鼠和人的IL-1都會(huì)起反應(yīng)，可以使用人IL-1并且可以估計(jì)人特異性IL-1拮抗劑在體內(nèi)的結(jié)合效果。這個(gè)急性小鼠模型可以用來決定人IL-1捕獲者拮抗外源施用的人IL-1引起的體內(nèi)效應(yīng)的能力。這個(gè)模型提供了人IL-1捕獲者體內(nèi)效應(yīng)的快速指示并且可以用作幫助分子選擇的檢測(cè)方法。
實(shí)驗(yàn)設(shè)計(jì)給雄性C57BL/6小鼠皮下注射重組人IL-1β(rhIL-1β；0.3mg/kg)。在rhIL-1β給藥前24小時(shí)，用載體、人IL-1捕獲者569(50或150倍摩爾過量；分別為0.18或0.54mg/kg)或重組的鼠IL-1受體拮抗劑(rmIL-1ra；150或750倍摩爾過量；分別為45.8或229μg/kg)處理動(dòng)物。施用rhIL-1β2小時(shí)后取血液樣品并用小鼠IL-6ELISA方法檢測(cè)血清中IL-6水平。外源給藥的rhIL-1β顯著地增加了血清中的IL-6水平。用50或150倍摩爾過量的hIL-1捕獲者預(yù)處理阻斷rhIL-1β誘導(dǎo)的IL-6。與此相反，注射150或750倍摩爾過量的rmIL-Ira都不會(huì)阻斷IL-6的誘導(dǎo)。
結(jié)果外源施用人IL-1導(dǎo)致顯著誘導(dǎo)的血清IL-6水平。在150倍摩爾過量時(shí)，人IL-1捕獲者完全阻斷IL-6的增加(圖1)。而且，人IL-1捕獲者的藥效會(huì)持續(xù)至少另外24小時(shí)，甚至當(dāng)重新施用IL-1時(shí)仍能阻止IL-6的增加(圖1)。這種持續(xù)長時(shí)間的藥效暗示在應(yīng)用于長期治療時(shí)沒有必要每天注射IL-1捕獲者。
在分開的實(shí)驗(yàn)中，IL-Ira在150倍或750倍摩爾過量時(shí)都沒有明顯地阻斷IL6的誘導(dǎo)。因而，在這個(gè)例子中，IL-1捕獲者似乎是一個(gè)更好的IL-1活性阻斷劑。
實(shí)施例5.構(gòu)建額外的單鏈IL-1捕獲者此處描述的用于構(gòu)建DNA載體的技術(shù)是本專業(yè)技術(shù)人員熟悉的標(biāo)準(zhǔn)生物學(xué)技術(shù)(參見例如Sambrook，J.，E.F.Fritsch And T.Maniatis.Molecular CloningA Laboratory Manual，第二版，Vols 1，2，和3，1989；Current Protocols in Molecular Biology，Eds.Ausubel等人，Greene Publ.Assoc.，Wiley Interscience，NY)。利用ABI 373ADNA測(cè)序列儀和Taq雙脫氧終止子循環(huán)測(cè)序試劑盒(AppliedBiosystems，Inc.，F(xiàn)oster City，CA))標(biāo)準(zhǔn)技術(shù)對(duì)所有DNA序列進(jìn)行測(cè)序。
(a)IL-1捕獲者823序列-所述IL-1捕獲者823序列由人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO7中的核苷酸1-1077)，之后連接人IL-1RI的細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO7中的核苷酸1078-2013)，之后連接鉸鏈區(qū)，人IgG1的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO7中的核苷酸2014-2703)的一部分構(gòu)成，其中在核苷酸2017-2019含有由半胱氨酸變?yōu)楦拾彼?TGT-＞GGA)的突變。核酸序列編碼的融合多肽序列為SEQ ID NO8。
(b)IL-1捕獲者823-1198-B序列-所述IL-1捕獲者823-1198-B序列由人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于SEQ ID NO9的核苷酸1-1077)，之后連接人IL-1RI細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO9中的核苷酸1078-2013)，之后連接一段氨基酸片段(對(duì)應(yīng)于SEQ ID NO9中的核苷酸2014-2019)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO9中的核苷酸2020-2709)構(gòu)成。核酸序列編碼融合的多肽序列為SEQ ID NO10所列。
(c)IL-1捕獲者823-1267-C序列-所述IL-1捕獲者823-1267-C序列由人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO11中的核苷酸1-1077)，之后連接人IL-1RI細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ IDNO11中的核苷酸1078-2013)，之后連接一段氨基酸片段(對(duì)應(yīng)于SEQID NO11中的核苷酸2014-2019)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO7中的核苷酸2020-2709)構(gòu)成，其中包含在核苷酸2047(T＞C)將絲氨酸改變成脯氨酸的突變。核酸序列編碼的融合多肽序列為SEQ ID NO12所列。
(d)IL-1捕獲者570-FE序列-所述IL-1捕獲者570-FE序列由人IL-1RI細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO1中的核苷酸1-996)，，之后連接人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO1中的核苷酸997-2013)，之后連接鉸鏈區(qū)，人IgG1的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO1中的核苷酸2014-2703)的一部分構(gòu)成，其中包含在核苷酸2017-2019由半胱氨酸變?yōu)楦拾彼?TGT-＞GGA)的突變。核酸序列編碼融合多肽為SEQ ID NO2所列序列。
(e)IL-1捕獲者570-FE-B序列-所述IL-1捕獲者570-FE-B序列由人IL-1RI細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO3中的核苷酸1-996)，之后連接人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO3中的核苷酸997-2013)，之后連接一段氨基酸片段(對(duì)應(yīng)于SEQ ID NO3中的核苷酸2014-2019)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO3中的核苷酸2020-2709)構(gòu)成。核酸序列編碼的融合多肽為SEQ ID NO4所列序列。
(f)IL-1捕獲者570-FE-C序列-所述IL-1捕獲者570-FE-C序列由人IL-1RI的細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO5中的核苷酸1-996)，之后連接人IL-RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO5中的核苷酸997-2013)，之后連接一段氨基酸片段(對(duì)應(yīng)于SEQ ID NO5中的核苷酸2014-2019)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO7中的核苷酸2020-2709)構(gòu)成，其中包含在核苷酸2047(T＞C)將絲氨酸改變成脯氨酸的突變。核酸序列編碼的融合多肽為SEQ ID NO6所列序列。
(g)IL-1捕獲者1647-CtF序列-所述IL-1捕獲者1647-CtF序列由人IL-1RII細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO13中的核苷酸1-1044)，之后連接人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO13中的核苷酸1045-2058)，之后連接鉸鏈區(qū)，人IgG1的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO13中的核苷酸2059-2748)的一部分構(gòu)成，其中包含在核苷酸2017-2019由半胱氨酸變?yōu)楦拾彼?TGT-＞GGA)的突變。核酸序列編碼的融合多肽為SEQ ID NO14所列序列。
(h)IL-1捕獲者1647-CtF-B序列-所述IL-1捕獲者1647-CtF-B序列由人IL-1RII細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO15中的核苷酸1-1044)，之后連接人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ IDNO15中的核苷酸1045-2058)，之后連接一段氨基酸片段(對(duì)應(yīng)于EQID NO15中的核苷酸2059-2064)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO15中的核苷酸2065-2754)構(gòu)成。核酸序列編碼的融合多肽為SEQ ID NO16所列序列。
(i)IL-1捕獲者1647-CtF-C序列-所述IL-1捕獲者1647-CtF-C序列由人IL-1RII細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO17中的核苷酸1-1044)，之后連接人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ IDNO17中的核苷酸1045-2058)，之后連接一段氨基酸片段(對(duì)應(yīng)于SEQID NO17中的核苷酸2059-2064)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO17中的核苷酸2065-2754)構(gòu)成，其中包含在核苷酸2092(T＞C)將絲氨酸改變成脯氨酸的突變。核酸序列編碼的融合多肽為SEQ ID NO17所列序列。
(j)IL-1捕獲者1649序列-所述IL-1捕獲者1649序列由人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO19中的核苷酸1-1074)，之后連接人IL-1RII細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO19中的核苷酸1075-2058)，之后連接鉸鏈區(qū)，人IgG1的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO19中的核苷酸2059-2748)的一部分構(gòu)成，其中包含在核苷酸2062-2064位點(diǎn)由半胱氨酸變?yōu)楦拾彼?TGT-＞GGA)的突變。核酸序列編碼的融合多肽為SEQ ID NO20所列序列。
(k)IL-1捕獲者1649-B序列-所述IL-1捕獲者1649-B序列由人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO21中的核苷酸1-1074)，之后連接人IL-1RII細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO21中的核苷酸1075-2058)，之后連接一段氨基酸片段(對(duì)應(yīng)于核苷酸2059-2064)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO21中的核苷酸2065-2754)構(gòu)成。核酸序列編碼的融合多肽為SEQ ID NO22所列序列。
(1)IL-1捕獲者1649-C序列-所述IL-1捕獲者1649-C序列由人IL-1RAcP細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO23中的核苷酸1-1074)，之后連接人IL-1RII細(xì)胞外結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO23中的核苷酸1075-2058)，之后連接一段氨基酸片段(對(duì)應(yīng)于SEQ ID NO23中的核苷酸2059-2064)，之后連接鉸鏈區(qū)，人IgG4的CH2和CH3結(jié)構(gòu)域(對(duì)應(yīng)于序列SEQ ID NO23中的核苷酸2065-2754)構(gòu)成，其中包含在核苷酸2092(T＞C)將絲氨酸改變成脯氨酸的突變。核酸序列編碼的融合多肽為SEQ ID NO24所列序列。
除了上面所描述的序列外，下列對(duì)這些序列的修飾也是本主題發(fā)明所包括的。
1.對(duì)于IL1捕獲者823，823-1198.B和823-1267.CAcP替換在核苷酸1043位從A到C的改變導(dǎo)致從賴氨酸到蘇氨酸的氨基酸改變。SG插入在核苷酸1077和1078位之間插入核苷酸TCC GGA將在捕獲者的兩個(gè)受體結(jié)構(gòu)域之間加上絲氨酸甘氨酸氨基酸片段。
2.對(duì)于IL1捕獲者570-FE，570-FE.B和570-FE.CAcP替換在核苷酸1979位從A到C的改變導(dǎo)致從賴氨酸到蘇氨酸的氨基酸改變。SG插入在核苷酸996和977位之間插入核苷酸TCC GGA將在捕獲者的兩個(gè)受體結(jié)構(gòu)域之間加上絲氨酸甘氨酸氨基酸片段。
3.對(duì)于IL1捕獲者1647-CtF，1647-CtF.B和1647-CtF.CAcP替換在核苷酸2027位從A到C的改變導(dǎo)致從賴氨酸到蘇氨酸氨基酸的改變。SG插入在核苷酸1044和1045位之間插入核苷酸TCC GGA將在捕獲者的兩個(gè)受體結(jié)構(gòu)域之間加上絲氨酸甘氨酸氨基酸片段。
4.對(duì)于IL1捕獲者1649，1649-B和1649-CAcP替換在核苷酸1043位從A到C的改變導(dǎo)致從賴氨酸到蘇氨酸的氨基酸改變。SG插入在核苷酸1074和1075位之間插入核苷酸TCC GGA將在捕獲者的兩個(gè)受體結(jié)構(gòu)域之間加上絲氨酸甘氨酸氨基酸片段。
此外，本領(lǐng)域技術(shù)人員將認(rèn)可構(gòu)建其中Fc結(jié)構(gòu)域源于不同的同種異型免疫球蛋白的IL-1捕獲者是理想的。上面描述的修飾不會(huì)改變捕獲者對(duì)IL1的結(jié)合能力。
例6.人IL-1捕獲者阻止IL-1在發(fā)炎關(guān)節(jié)中的作用背景酵母聚糖(zymosan)是-種酵母細(xì)胞壁提取物，當(dāng)它注射入膝蓋時(shí)會(huì)在關(guān)節(jié)處引發(fā)急性炎癥和上調(diào)IL-1β的水平(Joosten等人(1994)Clin Exp Immunol 97204-211.)。軟骨細(xì)胞通過下調(diào)蛋白聚糖的合成對(duì)炎癥和局部IL-1β作出反應(yīng)，這種反應(yīng)是促成關(guān)節(jié)處軟骨漸近損壞的人關(guān)節(jié)炎的一個(gè)特征(Van den Berg等人(1982)Rheum Intl1165-169)。IL-1β的拮抗劑可以用來評(píng)估它們對(duì)酵母聚糖誘導(dǎo)的IL-1β上升的效應(yīng)的阻斷能力。
材料和方法通過經(jīng)髕韌帶麻痹的雄性C57BL/6小鼠右膝關(guān)節(jié)關(guān)節(jié)內(nèi)(i.a.)注射酵母聚糖A(Sigma；300μg每10μ1)。通過髕韌帶向其左膝關(guān)節(jié)關(guān)節(jié)內(nèi)(i.a.)注射無菌PBS。在關(guān)節(jié)內(nèi)注射之前24小時(shí)，用載體或hIL-1捕獲子569(19mg/kg，皮下注射)處理動(dòng)物。根據(jù)上面van den Berg等人(1982)描述的方法在注射酵母聚糖24小時(shí)之后去除小鼠髕骨以測(cè)量蛋白聚糖的合成。簡(jiǎn)而言之，將每個(gè)髕骨及其相連系的韌帶放入含有10μCi/ml35S-硫酸鹽(NEN DuPont)的培養(yǎng)基(含HEPES、HCO3、谷氨酰胺和青霉素/鏈霉素的RPMI)中于37℃，5％CO2條件下孵育3小時(shí)。孵育后，將組織清洗并用10％的福爾馬琳(VWR)固定過夜。然后在將髕骨從周圍組織分離出來之前將組織放入脫鈣溶液(J.T.Baker)中放置4小時(shí)。然后將每個(gè)膝關(guān)節(jié)放在Solvable(Packard)中于50℃下孵育過夜。加入U(xiǎn)tima Gold液體閃爍液(Packard)并且將樣品用液體閃爍計(jì)數(shù)器計(jì)數(shù)。對(duì)每只動(dòng)物測(cè)量的值為酵母聚糖關(guān)節(jié)的cpm/載體關(guān)節(jié)的cpm比值。
結(jié)果與載體注射相比，關(guān)節(jié)內(nèi)注射酵母聚糖減少了近50％的蛋白聚糖的合成(圖3)。在酵母聚糖注射之前施用hIL-1捕獲者阻斷了IL-1β的局部效應(yīng)并且蛋白聚糖的合成恢復(fù)到對(duì)照的大約90％。這些數(shù)據(jù)顯示hIL-1捕獲者569皮下注射后能滲入關(guān)節(jié)中有效地中和IL-1在這些關(guān)節(jié)中的生物效應(yīng)。
實(shí)施例7.在一個(gè)酵母聚糖加速的膠原蛋白誘導(dǎo)的關(guān)節(jié)炎(CIA)模型中，鼠IL-1捕獲者減輕關(guān)節(jié)炎癥的嚴(yán)重度背景已經(jīng)顯示IL-1涉及到類風(fēng)濕性關(guān)節(jié)炎中炎癥和軟骨破壞的發(fā)展(Dinarello(1996)Blood 87(6)2095-2147；Wooley等人(1993)Arthritis & Rheumatism 36(9)1305-1314)。膠原蛋白誘導(dǎo)的關(guān)節(jié)炎(CIA)是與類風(fēng)濕性關(guān)節(jié)炎相似的受到廣泛研究的炎性多關(guān)節(jié)炎動(dòng)物模型。普通組織病理特征包括關(guān)節(jié)的炎癥和侵蝕、滑液超常增生和炎性細(xì)胞浸潤(Joe等人(1999)Mol Med Today 5367-369)。由于前人的研究已經(jīng)顯示各種抗IL-1處理對(duì)減輕CIA動(dòng)物中的關(guān)節(jié)炎癥狀有積極作用(van den Berg等人(1994)Clin Exp Immunol 95237-243；Joosten等人(1999)J Immunol 1635049-5055.；van de Loo等人(1992)J Rheumatol 19348-356.)，申請(qǐng)者在這種動(dòng)物模型中檢查了鼠IL-1捕獲者(mIL-1捕獲者)對(duì)關(guān)節(jié)炎癥狀發(fā)展進(jìn)程的影響。人IL-1捕獲者與嚙齒動(dòng)物的IL-1很少有交叉反應(yīng)。mIL-1捕獲者由鼠IL-1RAcP細(xì)胞外結(jié)構(gòu)域之后連接鼠IL-IRI細(xì)胞外結(jié)構(gòu)域，之后連接鉸鏈，鼠科動(dòng)物IgG2a的CH2和CH3結(jié)構(gòu)域而構(gòu)成。
材料和方法用100μg/50μl經(jīng)完全和不完全弗式佐劑(2∶1∶1比例；Chondrex)乳化過的牛II型膠原蛋白(CII；Chondrex)在尾巴基部對(duì)雄性DBA-1小鼠進(jìn)行皮內(nèi)免疫并在第21天用經(jīng)不完全弗式佐劑乳化過的II型膠原蛋白(100μg/50μl)進(jìn)行皮內(nèi)免疫加強(qiáng)。由于CIA在DBA-1小鼠體內(nèi)以較低發(fā)生率經(jīng)過較長的時(shí)期逐漸發(fā)生(Joosten等人，(1994)Clin Exp Immunol 97204-211)，申請(qǐng)者通過在第30天用3mg酵母聚糖(Sigma)腹膜內(nèi)注射動(dòng)物以同步化關(guān)節(jié)炎癥狀的起始。在注射酵母聚糖之前2小時(shí)，小鼠被隨機(jī)分配到幾個(gè)處理組并用載體或mIL-1捕獲者(31或10mg/kg，3X/周，8次皮下注射)對(duì)其進(jìn)行注射。由不知道處理組的人每周3次估量爪子的關(guān)節(jié)炎癥狀(ASI分值，如Wooley等人(1993)Arthritis & Rheumatism 36(9)1305-1314中所描述)。在第8次注射后24小時(shí)，殺死動(dòng)物，測(cè)量此時(shí)的爪子寬度連同ASI分值。
結(jié)果相對(duì)那些接受mIL-1捕獲者處理(圖4)的在酵母聚糖注射10到15后癥狀達(dá)到最大值的動(dòng)物，用載體處理的動(dòng)物在皮下注射酵母聚糖后5天內(nèi)ASI分值有了顯著的增加。鼠IL-1捕獲者以劑量依賴的方式起作用，因此接受10mg/kg捕獲者處理的動(dòng)物具有比那些接受31mg/kg捕獲者處理的動(dòng)物更嚴(yán)重的關(guān)節(jié)炎癥狀(更大的ASI分值)。然而，兩個(gè)mIL-1捕獲者處理組都具有顯著低于載體處理組的關(guān)節(jié)炎癥狀。這種ASI分值的不同也反應(yīng)在宰殺時(shí)動(dòng)物爪子的寬度(圖5)。接受mIL-1捕獲者處理的動(dòng)物有著與那些天然的沒有用膠原蛋白免疫過的動(dòng)物相似寬度的爪子。這些數(shù)據(jù)顯示mIL-1捕獲者能有效地中和IL-1并阻止發(fā)炎關(guān)節(jié)的發(fā)展。
實(shí)施例8.IL-1捕獲者1649能阻斷IL-1β的活性在室溫下，將各種濃度的IL-1捕獲者1649與5pM的人IL-1b共同孵育過夜。然后在37℃、5％CO2的條件下將該混合物加入到二個(gè)293-NFkB細(xì)胞(20,000細(xì)胞/孔)孔里孵育5小時(shí)。293-NFkB細(xì)胞含有一個(gè)穩(wěn)定整合的具有熒光素酶基因(該基因被一個(gè)包含5個(gè)NFkB位點(diǎn)的啟動(dòng)子驅(qū)動(dòng))的報(bào)告質(zhì)粒。IL-1b的加入導(dǎo)致熒光素酶基因表達(dá)的增加。在室溫下將Steady-Glo Reagent(Promega)加入到細(xì)胞中過15分鐘并且用照度計(jì)將熒光素酶基因表達(dá)定量為相對(duì)光單位(RLU)。IL-1捕獲子1649顯示IC50為32pM，該值暗示了約30pM的Kd(圖6)。這些數(shù)據(jù)顯示IL-1捕獲子1649潛在地阻斷IL-1。
本發(fā)明不局限于此處描述的具體實(shí)施方案的范圍內(nèi)。事實(shí)上，從前面的描述和附圖來看，除了此處描述的修改，本發(fā)明的各種修改對(duì)本領(lǐng)域技術(shù)人員來說是明顯的。這樣的修改屬于權(quán)利要求的范圍內(nèi)。
序列表<110>REGENERON PHARMACEUTICALS，INC.
<120>基于IL-1受體的拮抗劑和制造與使用的方法<130>REG 203C-WO<140>to be assigned<141>2003-10-24<150>10/282,162<151>2002-10-28<150>09/787,835<151>2001-03-22<150>PCT/US99/22045<151>1999-09-22<160>26<170>FastSEQ for Windows Version 3.0<210>1<211>2703<212>DNA<213>人<400>1atggtgttac tcagacttat ttgtttcata gctctactga tttcttctct ggaggctgat60aaatgcaagg aacgtgaaga aaaaataatt ttagtgtcat ctgcaaatga aattgatgtt120cgtccctgtc ctcttaaccc aaatgaacac aaaggcacta taacttggta taaggatgac180agcaagacac ctgtatctac agaacaagcc tccaggattc atcaacacaa agagaaactt240tggtttgttc ctgctaaggt ggaggattca ggacattact attgcgtggt aagaaattca300tcttactgcc tcagaattaa aataagtgca aaatttgtgg agaatgagcc taacttatgt360tataatgcac aagccatatt taagcagaaa ctacccgttg caggagacgg aggacttgtg420tgcccttata tggagttttt taaaaatgaa aataatgagt tacctaaatt acagtggtat480aaggattgca aacctctact tcttgacaat atacacttta gtggagtcaa agataggctc540atcgtgatga atgtggctga aaagcataga gggaactata cttgtcatgc atcctacaca600tacttgggca agcaatatcc tattacccgg gtaatagaat ttattactct agaggaaaac660aaacccacaa ggcctgtgat tgtgagccca gctaatgaga caatggaagt agacttggga720tcccagatac aattgatctg taatgtcacc ggccagttga gtgacattgc ttactggaag780tggaatgggt cagtaattga tgaagatgac ccagtgctag gggaagacta ttacagtgtg840gaaaatcctg caaacaaaag aaggagtacc ctcatcacag tgcttaatat atcggaaatt900gagagtagat tttataaaca tccatttacc tgttttgcca agaatacaca tggtatagat960gcagcatata tccagttaat atatccagtc actaattcag aacgctgcga tgactgggga1020ctagacacca tgaggcaaat ccaagtgttt gaagatgagc cagctcgcat caagtgccca1080ctctttgaac acttcttgaa attcaactac agcacagccc attcagctgg ccttactctg1140atctggtatt ggactaggca ggaccgggac cttgaggagc caattaactt ccgcctcccc1200gagaaccgca ttagtaagga gaaagatgtg ctgtggttcc ggcccactct cctcaatgac1260actggcaact atacctgcat gttaaggaac actacatatt gcagcaaagt tgcatttccc1320ttggaagttg ttcaaaaaga cagctgtttc aattccccca tgaaactccc agtgcataaa1380ctgtatatag aatatggcat tcagaggatc acttgtccaa atgtagatgg atattttcct1440tccagtgtca aaccgactat cacttggtat atgggctgtt ataaaataca gaattttaat1500
aatgtaatac ccgaaggtat gaacttgagt ttcctcattg ccttaatttc aaataatgga1560aattacacat gtgttgttac atatccagaa aatggacgta cgtttcatct caccaggact1620ctgactgtaa aggtagtagg ctctccaaaa aatgcagtgc cccctgtgat ccattcacct1680aatgatcatg tggtctatga gaaagaacca ggagaggagc tactcattcc ctgtacggtc1740tattttagtt ttctgatgga ttctcgcaat gaggtttggt ggaccattga tggaaaaaaa1800cctgatgaca tcactattga tgtcaccatt aacgaaagta taagtcatag tagaacagaa1860gatgaaacaa gaactcagat tttgagcatc aagaaagtta cctctgagga tctcaagcgc1920agctatgtct gtcatgctag aagtgccaaa ggcgaagttg ccaaagcagc caaggtgaag1980cagaaagtgc cagctccaag atacacagtg gaatccggag acaaaactca cacatgccca2040ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc2100aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc2160cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc2220aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc2280gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc2340ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag2400gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag cctgacctgc2460ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg2520gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctat2580agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg2640atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa2700tga 2703<210>2<211>900<212>PRT<213>人<400>2Met Val Leu Leu Arg Leu Ile Cys Phe Ile Ala Leu Leu Ile Ser Ser1 5 10 15Leu Glu Ala Asp Lys Cys Lys Glu Arg Glu Glu Lys Ile Ile Leu Val20 25 30Ser Ser Ala Asn Glu Ile Asp Val Arg Pro Cys Pro Leu Asn Pro Asn35 40 45Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser Lys Thr Pro50 55 60Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys Glu Lys Leu65 70 75 80Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr Tyr Cys Val85 90 95Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys Phe100 105 110Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala Ile Phe Lys115 120 125Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys Pro Tyr Met130 135 140Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu Gln Trp Tyr145 150 155 160Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe Ser Gly Val165 170 175Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His Arg Gly Asn180 185 190Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln Tyr Pro Ile195 200 205Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys Pro Thr Arg210 215 220Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val Asp Leu Gly
225 230 235 240Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu Ser Asp Ile245 250 255Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp Asp Pro Val260 265 270Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn Lys Arg Arg275 280 285Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu Ser Arg Phe290 295 300Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His Gly Ile Asp305 310 315 320Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn Ser Glu Arg Cys325 330335Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp340 345 350Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Phe355 360 365Asn Tyr Ser Thr Ala His Ser Ala Gly Leu Thr Leu Ile Trp Tyr Trp370 375 380Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro385 390 395 400Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr405 410 415Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr420 425 430Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser435 440 445Cys Phe Asn Ser Pro Met Lys Leu Pro Val His Lys Leu Tyr Ile Glu450 455 460Tyr Gly Ile Gln Arg Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro465 470 475 480Ser Ser Val Lys Pro Thr Ile Thr Trp Tyr Met Gly Cys Tyr Lys Ile485 490 495Gln Asn Phe Asn Asn Val Ile Pro Glu Gly Met Asn Leu Ser Phe Leu500 505 510Ile Ala Leu Ile Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr515 520 525Pro Glu Asn Gly Arg Thr Phe His Leu Thr Arg Thr Leu Thr Val Lys530 535 540Val Val Gly Ser Pro Lys Asn Ala Val Pro Pro Val Ile His Ser Pro545 550 555 560Asn Asp His Val Val Tyr Glu Lys Glu Pro Gly Glu Glu Leu Leu Ile565 570 575Pro Cys Thr Val Tyr Phe Ser Phe Leu Met Asp Ser Arg Asn Glu Val580 585 590Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Ile Thr Ile Asp Val595 600 605Thr Ile Asn Glu Ser Ile Ser His Ser Arg Thr Glu Asp Glu Thr Arg610 615 620Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Ser Glu Asp Leu Lys Arg625 630 635 640Ser Tyr Val Cys His Ala Arg Ser Ala Lys Gly Glu Val Ala Lys Ala645 650 655Ala Lys Val Lys Gln Lys Val Pro Ala Pro Arg Tyr Thr Val Glu Ser660 665 670Gly Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu675 680 685
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu690 695 700Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser705 710 715 720His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu725 730 735Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr740 745 750Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn755 760 765Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro770 775 780Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln785 790 795 800Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val805 810 815Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val820 825 830Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro835 840 845Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr850 855 860Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val865 870 875 880Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu885 890 895Ser Pro Gly Lys900<210>3<211>2709<212>DNA<213>人<400>3atggtgttac tcagacttat ttgtttcata gctctactga tttcttctct ggaggctgat60aaatgcaagg aacgtgaaga aaaaataatt ttagtgtcat ctgcaaatga aattgatgtt120cgtccctgtc ctcttaaccc aaatgaacac aaaggcacta taacttggta taaggatgac180agcaagacac ctgtatctac agaacaagcc tccaggattc atcaacacaa agagaaactt240tggtttgttc ctgctaaggt ggaggattca ggacattact attgcgtggt aagaaattca300tcttactgcc tcagaattaa aataagtgca aaatttgtgg agaatgagcc taacttatgt360tataatgcac aagccatatt taagcagaaa ctacccgttg caggagacgg aggacttgtg420tgcccttata tggagttttt taaaaatgaa aataatgagt tacctaaatt acagtggtat480aaggattgca aacctctact tcttgacaat atacacttta gtggagtcaa agataggctc540atcgtgatga atgtggctga aaagcataga gggaactata cttgtcatgc atcctacaca600tacttgggca agcaatatcc tattacccgg gtaatagaat ttattactct agaggaaaac660aaacccacaa ggcctgtgat tgtgagccca gctaatgaga caatggaagt agacttggga720tcccagatac aattgatctg taatgtcacc ggccagttga gtgacattgc ttactggaag780tggaatgggt cagtaattga tgaagatgac ccagtgctag gggaagacta ttacagtgtg840gaaaatcctg caaacaaaag aaggagtacc ctcatcacag tgcttaatat atcggaaatt900gagagtagat tttataaaca tccatttacc tgttttgcca agaatacaca tggtatagat960gcagcatata tccagttaat atatccagtc actaattcag aacgctgcga tgactgggga1020ctagacacca tgaggcaaat ccaagtgttt gaagatgagc cagctcgcat caagtgccca1080ctctttgaac acttcttgaa attcaactac agcacagccc attcagctgg ccttactctg1140atctggtatt ggactaggca ggaccgggac cttgaggagc caattaactt ccgcctcccc1200gagaaccgca ttagtaagga gaaagatgtg ctgtggttcc ggcccactct cctcaatgac1260actggcaact atacctgcat gttaaggaac actacatatt gcagcaaagt tgcatttccc1320
ttggaagttg ttcaaaaaga cagctgtttc aattccccca tgaaactccc agtgcataaa1380ctgtatatag aatatggcat tcagaggatc acttgtccaa atgtagatgg atattttcct1440tccagtgtca aaccgactat cacttggtat atgggctgtt ataaaataca gaattttaat1500aatgtaatac ccgaaggtat gaacttgagt ttcctcattg ccttaatttc aaataatgga1560aattacacat gtgttgttac atatccagaa aatggacgta cgtttcatct caccaggact1620ctgactgtaa aggtagtagg ctctccaaaa aatgcagtgc cccctgtgat ccattcacct1680aatgatcatg tggtctatga gaaagaacca ggagaggagc tactcattcc ctgtacggtc1740tattttagtt ttctgatgga ttctcgcaat gaggtttggt ggaccattga tggaaaaaaa1800cctgatgaca tcactattga tgtcaccatt aacgaaagta taagtcatag tagaacagaa1860gatgaaacaa gaactcagat tttgagcatc aagaaagtta cctctgagga tctcaagcgc1920agctatgtct gtcatgctag aagtgccaaa ggcgaagttg ccaaagcagc caaggtgaag1980cagaaagtgc cagctccaag atacacagtg gaatccggag agtccaaata cggtccgcca2040tgcccatcat gcccagcacc tgagttcctg gggggaccat cagtcttcct gttcccccca2100aaacccaagg acactctcat gatctcccgg acccctgagg tcacgtgcgt ggtggtggac2160gtgagccagg aagaccccga ggtccagttc aactggtacg tggatggcgt ggaggtgcat2220aatgccaaga caaagccgcg ggaggagcag ttcaacagca cgtaccgtgt ggtcagcgtc2280ctcaccgtcc tgcaccagga ctggctgaac ggcaaggagt acaagtgcaa ggtctccaac2340aaaggcctcc cgtcctccat cgagaaaacc atctccaaag ccaaagggca gccccgagag2400ccacaggtgt acaccctgcc cccatcccag gaggagatga ccaagaacca ggtcagcctg2460acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga gagcaatggg2520cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc2580ctctacagca ggctaaccgt ggacaagagc aggtggcagg aggggaatgt cttctcatgc2640tccgtgatgc atgaggctct gcacaaccac tacacacaga agagcctctc cctgtctctg2700ggtaaatga2709<210>4<211>902<212>PRT<213>人<400>4Met Val Leu Leu Arg Leu Ile Cys Phe Ile Ala Leu Leu Ile Ser Ser1 5 10 15Leu Glu Ala Asp Lys Cys Lys Glu Arg Glu Glu Lys Ile Ile Leu Val20 25 30Ser Ser Ala Asn GluIle Asp Val Arg Pro Cys Pro Leu Asn Pro Asn35 40 45Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser Lys Thr Pro50 55 60Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys Glu Lys Leu65 70 75 80Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr Tyr Cys Val85 90 95Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys Phe100 105 110Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala Ile Phe Lys115 120 125Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys Pro Tyr Met130 135 140Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu Gln Trp Tyr145 150 155 160Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe Ser Gly Val165 170 175Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His Arg Gly Asn180 185 190Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln Tyr Pro Ile195 200 205
Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys Pro Thr Arg210 215 220Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val Asp Leu Gly225 230 235 240Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu Ser Asp Ile245 250 255Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp Asp Pro Val260265 270Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn Lys Arg Arg275 280 285Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu Ser Arg Phe290 295 300Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His Gly Ile Asp305 310 315 320Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn Ser Glu Arg Cys325 330 335Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp340 345 350Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Phe355 360 365Asn Tyr Ser Thr Ala His Ser Ala Gly Leu Thr Leu Ile Trp Tyr Trp370 375 380Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro385 390 395 400Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr405 410 415Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr420 425 430Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser435 440 445Cys Phe Asn Ser Pro Met Lys Leu Pro Val His Lys Leu Tyr Ile Glu450 455 460Tyr Gly Ile Gln Arg Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro465 470 475 480Ser Ser Val Lys Pro Thr Ile Thr Trp Tyr Met Gly Cys Tyr Lys Ile485 490 495Gln Asn Phe Asn Asn Val Ile Pro Glu Gly Met Asn Leu Ser Phe Leu500 505 510Ile Ala Leu Ile Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr515 520 525Pro Glu Asn Gly Arg Thr Phe His Leu Thr Arg Thr Leu Thr Val Lys530 535 540Val Val Gly Ser Pro Lys Asn Ala Val Pro Pro Val Ile His Ser Pro545 550 555 560Asn Asp His Val Val Tyr Glu Lys Glu Pro Gly Glu Glu Leu Leu Ile565 570 575Pro Cys Thr Val Tyr Phe Ser Phe Leu Met Asp Ser Arg Asn Glu Val580 585 590Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Ile Thr Ile Asp Val595 600 605Thr Ile Asn Glu Ser Ile Ser His Ser Arg Thr Glu Asp Glu Thr Arg610 615 620Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Ser Glu Asp Leu Lys Arg625 630 635 640Ser Tyr Val Cys His Ala Arg Ser Ala Lys Gly Glu Val Ala Lys Ala645 650 655Ala Lys Val Lys Gln Lys Val Pro Ala Pro Arg Tyr Thr Val Glu Ser
660 665 670Gly Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu675 680 685Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp690 695 700Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp705 710 715 720Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly725 730 735Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn740 745 750Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp755 760 765Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro770 775 780Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu785 790 795 800Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn805 810 815Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile820 825 830Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr835 840 845Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg850 855 860Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys865 870 875 880Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu885 890 895Ser Leu Ser Leu Gly Lys900<210>5<211>2709<212>DNA<213>人<400>5atggtgttac tcagacttat ttgtttcata gctctactga tttcttctct ggaggctgat60aaatgcaagg aacgtgaaga aaaaataatt ttagtgtcat ctgcaaatga aattgatgtt120cgtccctgtc ctcttaaccc aaatgaacac aaaggcacta taacttggta taaggatgac180agcaagacac ctgtatctac agaacaagcc tccaggattc atcaacacaa agagaaactt240tggtttgttc ctgctaaggt ggaggattca ggacattact attgcgtggt aagaaattca300tcttactgcc tcagaattaa aataagtgca aaatttgtgg agaatgagcc taacttatgt360tataatgcac aagccatatt taagcagaaa ctacccgttg caggagacgg aggacttgtg420tgcccttata tggagttttt taaaaatgaa aataatgagt tacctaaatt acagtggtat480aaggattgca aacctctact tcttgacaat atacacttta gtggagtcaa agataggctc540atcgtgatga atgtggctga aaagcataga gggaactata cttgtcatgc atcctacaca600tacttgggca agcaatatcc tattacccgg gtaatagaat ttattactct agaggaaaac660aaacccacaa ggcctgtgat tgtgagccca gctaatgaga caatggaagt agacttggga720tcccagatac aattgatctg taatgtcacc ggccagttga gtgacattgc ttactggaag780tggaatgggt cagtaattga tgaagatgac ccagtgctag gggaagacta ttacagtgtg840gaaaatcctg caaacaaaag aaggagtacc ctcatcacag tgcttaatat atcggaaatt900gagagtagat tttataaaca tccatttacc tgttttgcca agaatacaca tggtatagat960gcagcatata tccagttaat atatccagtc actaattcag aacgctgcga tgactgggga1020ctagacacca tgaggcaaat ccaagtgttt gaagatgagc cagctcgcat caagtgccca1080ctctttgaac acttcttgaa attcaactac agcacagccc attcagctgg ccttactctg1140
atctggtatt ggactaggca ggaccgggac cttgaggagc caattaactt ccgcctcccc1200gagaaccgca ttagtaagga gaaagatgtg ctgtggttcc ggcccactct cctcaatgac1260actggcaact atacctgcat gttaaggaac actacatatt gcagcaaagt tgcatttccc1320ttggaagttg ttcaaaaaga cagctgtttc aattccccca tgaaactccc agtgcataaa1380ctgtatatag aatatggcat tcagaggatc acttgtccaa atgtagatgg atattttcct1440tccagtgtca aaccgactat cacttggtat atgggctgtt ataaaataca gaattttaat1500aatgtaatac ccgaaggtat gaacttgagt ttcctcattg ccttaatttc aaataatgga1560aattacacat gtgttgttac atatccagaa aatggacgta cgtttcatct caccaggact1620ctgactgtaa aggtagtagg ctctccaaaa aatgcagtgc cccctgtgat ccattcacct1680aatgatcatg tggtctatga gaaagaacca ggagaggagc tactcattcc ctgtacggtc1740tattttagtt ttctgatgga ttctcgcaat gaggtttggt ggaccattga tggaaaaaaa1800cctgatgaca tcactattga tgtcaccatt aacgaaagta taagtcatag tagaacagaa1860gatgaaacaa gaactcagat tttgagcatc aagaaagtta cctctgagga tctcaagcgc1920agctatgtct gtcatgctag aagtgccaaa ggcgaagttg ccaaagcagc caaggtgaag1980cagaaagtgc cagctccaag atacacagtg gaatccggag agtccaaata cggtccgcca2040tgcccaccat gcccagcacc tgagttcctg gggggaccat cagtcttcct gttcccccca2100aaacccaagg acactctcat gatctcccgg acccctgagg tcacgtgcgt ggtggtggac2160gtgagccagg aagaccccga ggtccagttc aactggtacg tggatggcgt ggaggtgcat2220aatgccaaga caaagccgcg ggaggagcag ttcaacagca cgtaccgtgt ggtcagcgtc2280ctcaccgtcc tgcaccagga ctggctgaac ggcaaggagt acaagtgcaa ggtctccaac2340aaaggcctcc cgtcctccat cgagaaaacc atctccaaag ccaaagggca gccccgagag2400ccacaggtgt acaccctgcc cccatcccag gaggagatga ccaagaacca ggtcagcctg2460acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga gagcaatggg2520cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc2580ctctacagca ggctaaccgt ggacaagagc aggtggcagg aggggaatgt cttctcatgc2640tccgtgatgc atgaggctct gcacaaccac tacacacaga agagcctctc cctgtctctg2700ggtaaatga2709<210>6<211>902<212>PRT<213>人<400>6Met Val Leu Leu Arg Leu Ile Cys Phe Ile Ala Leu Leu Ile Ser Ser1 5 10 15Leu Glu Ala Asp Lys Cys Lys Glu Arg Glu Glu Lys Ile Ile Leu Val20 25 30Ser Ser Ala Asn Glu Ile Asp Val Arg Pro Cys Pro Leu Asn Pro Asn35 40 45Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser Lys Thr Pro50 55 60Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys Glu Lys Leu65 70 75 80Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr Tyr Cys Val85 90 95Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys Phe100 105 110Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala Ile Phe Lys115 120 125Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys Pro Tyr Met130 135 140Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu Gln Trp Tyr145 150 155 160Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe Ser Gly Val165 170 175Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His Arg Gly Asn
180 185 190Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln Tyr Pro Ile195 200 205Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys Pro Thr Arg210 215 220Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val Asp Leu Gly225 230 235 240Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu Ser Asp Ile245 250 255Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp Asp Pro Val260 265 270Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn Lys Arg Arg275 280 285Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu Ser Arg Phe290 295 300Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His Gly Ile Asp305 310 315 320Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn Ser Glu Arg Cys325 330 335Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp340 345 350Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Phe355 360 365Asn Tyr Ser Thr Ala His Ser Ala Gly Leu Thr Leu Ile Trp Tyr Trp370 375 380Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro385 390 395 400Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr405 410 415Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr420 425 430Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser435 440 445Cys Phe Asn Ser Pro Met Lys Leu Pro Val His Lys Leu Tyr Ile Glu450 455 460Tyr Gly Ile Gln Arg Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro465 470 475 480Ser Ser Val Lys Pro Thr Ile Thr Trp Tyr Met Gly Cys Tyr Lys Ile485 490 495Gln Asn Phe Asn Asn Val Ile Pro Glu Gly Met Asn Leu Ser Phe Leu500 505 510Ile Ala Leu Ile Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr515 520 525Pro Glu Asn Gly Arg Thr Phe His Leu Thr Arg Thr Leu Thr Val Lys530 535 540Val Val Gly Ser Pro Lys Asn Ala Val Pro Pro Val Ile His Ser Pro545 550 555 560Asn Asp His Val Val Tyr Glu Lys Glu Pro Gly Glu Glu Leu Leu Ile565 570 575Pro Cys Thr Val Tyr Phe Ser Phe Leu Met Asp Ser Arg Asn Glu Val580 585 590Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Ile Thr Ile Asp Val595 600 605Thr Ile Asn Glu Ser Ile Ser His Ser Arg Thr Glu Asp Glu Thr Arg610 615 620Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Ser Glu Asp Leu Lys Arg625 630 635 640
Ser Tyr Val Cys His Ala Arg Ser Ala Lys Gly Glu Val Ala Lys Ala645 650 655Ala Lys Val Lys Gln Lys Val Pro Ala Pro Arg Tyr Thr Val Glu Ser660 665 670Gly Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu675 680 685Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp690 695 700Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp705 710 715 720Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly725 730 735Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn740 745 750Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp755 760 765Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro770 775 780Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu785 790 795 800Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn805 810 815Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile820 825 830Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr835 840 845Thr Pr0 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg850 855 860Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys865 870 875 880Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu885 890 895Ser Leu Ser Leu Gly Lys900<210>7<211>2703<212>DNA<213>人<400>7atggtgcttc tgtggtgtgt agtgagtctc tacttttatg gaatcctgca aagtgatgcc60tcagaacgct gcgatgactg gggactagac accatgaggc aaatccaagt gtttgaagat120gagccagctc gcatcaagtg cccactcttt gaacacttct tgaaattcaa ctacagcaca180gcccattcag ctggccttac tctgatctgg tattggacta ggcaggaccg ggaccttgag240gagccaatta acttccgcct ccccgagaac cgcattagta aggagaaaga tgtgctgtgg300ttccggccca ctctcctcaa tgacactggc aactatacct gcatgttaag gaacactaca360tattgcagca aagttgcatt tcccttggaa gttgttcaaa aagacagctg tttcaattcc420cccatgaaac tcccagtgca taaactgtat atagaatatg gcattcagag gatcacttgt480ccaaatgtag atggatattt tccttccagt gtcaaaccga ctatcacttg gtatatgggc540tgttataaaa tacagaattt taataatgta atacccgaag gtatgaactt gagtttcctc600attgccttaa tttcaaataa tggaaattac acatgtgttg ttacatatcc agaaaatgga660cgtacgtttc atctcaccag gactctgact gtaaaggtag taggctctcc aaaaaatgca720gtgccccctg tgatccattc acctaatgat catgtggtct atgagaaaga accaggagag780gagctactca ttccctgtac ggtctatttt agttttctga tggattctcg caatgaggtt840tggtggacca ttgatggaaa aaaacctgat gacatcacta ttgatgtcac cattaacgaa900agtataagtc atagtagaac agaagatgaa acaagaactc agattttgag catcaagaaa960
gttacctctg aggatctcaa gcgcagctat gtctgtcatg ctagaagtgc caaaggcgaa1020gttgccaaag cagccaaggt gaagcagaaa gtgccagctc caagatacac agtggaaaaa1080tgcaaggaac gtgaagaaaa aataatttta gtgagctcag caaatgaaat cgatgttcgt1140ccctgtcctc ttaacccaaa tgaacacaaa ggcactataa cttggtataa ggatgacagc1200aagacacctg tatctacaga acaagcctcc aggattcatc aacacaaaga gaaactttgg1260tttgttcctg ctaaggtgga ggattcagga cattactatt gcgtggtaag aaattcatct1320tactgcctca gaattaaaat aagtgcaaaa tttgtggaga atgagcctaa cttatgttat13 8 0aatgcacaag ccatatttaa gcagaaacta cccgttgcag gagacggagg acttgtgtgc1440ccttatatgg agttttttaa aaatgaaaat aatgagttac ctaaattaca gtggtataag1500gattgcaaac ctctacttct tgacaatata cactttagtg gagtcaaaga taggctcatc1560gtgatgaatg tggctgaaaa gcatagaggg aactatactt gtcatgcatc ctacacatac1620ttgggcaagc aatatcctat tacccgggta atagaattta ttactctaga ggaaaacaaa1680cccacaaggc ctgtgattgt gagcccagct aatgagacaa tggaagtaga cttgggatcc1740cagatacaat tgatctgtaa tgtcaccggc cagttgagtg acattgctta ctggaagtgg1800aatgggtcag taattgatga agatgaccca gtgctagggg aagactatta cagtgtggaa1860aatcctgcaa acaaaagaag gagtaccctc atcacagtgc ttaatatatc ggaaattgag1920agtagatttt ataaacatcc atttacctgt tttgccaaga atacacatgg tatagatgca1980gcatatatcc agttaatata tccagtcact aattccggag acaaaactca cacatgccca2040ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc2100aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc2160cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc2220aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc2280gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc2340ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag2400gtgtacaccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag cctgacctgc2460ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg2520gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac2580agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg2640atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa2700tga 2703<210>8<211>900<212>PRT<213>人<400>8Met Val Leu Leu Trp Cys Val Val Ser Leu Tyr Phe Tyr Gly Ile Leu1 5 10 15Gln Ser Asp Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met20 25 30Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro35 40 45Leu Phe Glu His Phe Leu Lys Phe Asn Tyr Ser Thr Ala His Ser Ala50 55 60Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu65 70 75 80Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys85 90 95Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr100 105 110Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro115 120 125Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Pro Met Lys Leu130 135 140Pro Val His Lys Leu Tyr Ile Glu Tyr Gly Ile Gln Arg Ile Thr Cys145 150 155 160
Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Thr Ile Thr165 170 175Trp Tyr Met Gly Cys Tyr Lys Ile Gln Asn Phe Asn Asn Val Ile Pro180 185 190Glu Gly Met Asn Leu Ser Phe Leu Ile Ala Leu Ile Ser Asn Asn Gly195 200 205Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Thr Phe His210 215 220Leu Thr Arg Thr Leu Thr Val Lys Val Val Gly Ser Pro Lys Asn Ala225 230 235 240Val Pro Pro Val Ile His Ser Pro Asn Asp His Val Val Tyr Glu Lys245 250 255Glu Pro Gly Glu Glu Leu Leu Ile Pro Cys Thr Val Tyr Phe Ser Phe260 265 270Leu Met Asp Ser Arg Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys275 280 285Pro Asp Asp Ile Thr Ile Asp Val Thr Ile Asn Glu Ser Ile Ser His290 295 300Ser Arg Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys305 310 315 320Val Thr Ser Glu Asp Leu Lys Arg Ser Tyr Val Cys His Ala Arg Ser325 330 335Ala Lys Gly Glu Val Ala Lys Ala Ala Lys Val Lys Gln Lys Val Pro340 345 350Ala Pro Arg Tyr Thr Val Glu Lys Cys Lys Glu Arg Glu Glu Lys Ile355 360 365Ile Leu Val Ser Ser Ala Asn Glu Ile Asp Val Arg Pro Cys Pro Leu370 375 380Asn Pro Asn Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser385 390 395 400Lys Thr Pro Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys405 410 415Glu Lys Leu Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr420 425 430Tyr Cys Val Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser435 440 445Ala Lys Phe Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala450 455 460Ile Phe Lys Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys465 470 475 480Pro Tyr Met Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu485 490 495Gln Trp Tyr Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe500 505 510Ser Gly Val Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His515 520 525Arg Gly Asn Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln530 535 540Tyr Pro Ile Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys545 550 555 560Pro Thr Arg Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val565 570 575Asp Leu Gly Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu580 585 590Ser Asp Ile Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp595 600 605Asp Pro Val Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn
610 615 620Lys Arg Arg Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu625 630 635 640Ser Arg Phe Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His645 650 655Gly Ile Asp Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn Ser660 665 670Gly Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu675 680 685Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu690 695 700Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser705 710 715 720His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu725 730 735Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr740 745 750Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn755 760 765Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro770 775 780Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln785 790 795 800Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val805 810 815Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val820 825 830Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro835 840 845Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr850 855 860Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val865 870 875 880Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu885 890 895Ser Pro Gly Lys900<210>9<211>2709<212>DNA<213>人<400>9atggtgcttc tgtggtgtgt agtgagtctc tacttttatg gaatcctgca aagtgatgcc60tcagaacgct gcgatgactg gggactagac accatgaggc aaatccaagt gtttgaagat120gagccagctc gcatcaagtg cccactcttt gaacacttct tgaaattcaa ctacagcaca180gcccattcag ctggccttac tctgatctgg tattggacta ggcaggaccg ggaccttgag240gagccaatta acttccgcct ccccgagaac cgcattagta aggagaaaga tgtgctgtgg300ttccggccca ctctcctcaa tgacactggc aactatacct gcatgttaag gaacactaca360tattgcagca aagttgcatt tcccttggaa gttgttcaaa aagacagctg tttcaattcc420cccatgaaac tcccagtgca taaactgtat atagaatatg gcattcagag gatcacttgt480ccaaatgtag atggatattt tccttccagt gtcaaaccga ctatcacttg gtatatgggc540tgttataaaa tacagaattt taataatgta atacccgaag gtatgaactt gagtttcctc600attgccttaa tttcaaataa tggaaattac acatgtgttg ttacatatcc agaaaatgga660cgtacgtttc atctcaccag gactctgact gtaaaggtag taggctctcc aaaaaatgca720gtgccccctg tgatccattc acctaatgat catgtggtct atgagaaaga accaggagag780
gagctactca ttccctgtac ggtctatttt agttttctga tggattctcg caatgaggtt840tggtggacca ttgatggaaa aaaacctgat gacatcacta ttgatgtcac cattaacgaa900agtataagtc atagtagaac agaagatgaa acaagaactc agattttgag catcaagaaa960gttacctctg aggatctcaa gcgcagctat gtctgtcatg ctagaagtgc caaaggcgaa1020gttgccaaag cagccaaggt gaagcagaaa gtgccagctc caagatacac agtggaaaaa1080tgcaaggaac gtgaagaaaa aataatttta gtgagctcag caaatgaaat cgatgttcgt1140ccctgtcctc ttaacccaaa tgaacacaaa ggcactataa cttggtataa ggatgacagc1200aagacacctg tatctacaga acaagcctcc aggattcatc aacacaaaga gaaactttgg1260tttgttcctg ctaaggtgga ggattcagga cattactatt gcgtggtaag aaattcatct1320tactgcctca gaattaaaat aagtgcaaaa tttgtggaga atgagcctaa cttatgttat1380aatgcacaag ccatatttaa gcagaaacta cccgttgcag gagacggagg acttgtgtgc1440ccttatatgg agttttttaa aaatgaaaat aatgagttac ctaaattaca gtggtataag1500gattgcaaac ctctacttct tgacaatata cactttagtg gagtcaaaga taggctcatc1560gtgatgaatg tggctgaaaa gcatagaggg aactatactt gtcatgcatc ctacacatac1620ttgggcaagc aatatcctat tacccgggta atagaattta ttactctaga ggaaaacaaa1680cccacaaggc ctgtgattgt gagcccagct aatgagacaa tggaagtaga cttgggatcc1740cagatacaat tgatctgtaa tgtcaccggc cagttgagtg acattgctta ctggaagtgg1800aatgggtcag taattgatga agatgaccca gtgctagggg aagactatta cagtgtggaa1860aatcctgcaa acaaaagaag gagtaccctc atcacagtgc ttaatatatc ggaaattgag1920agtagatttt ataaacatcc atttacctgt tttgccaaga atacacatgg tatagatgca1980gcatatatcc agttaatata tccagtcact aattccggag agtccaaata cggtccgcca2040tgcccatcat gcccagcacc tgagttcctg gggggaccat cagtcttcct gttcccccca2100aaacccaagg acactctcat gatctcccgg acccctgagg tcacgtgcgt ggtggtggac2160gtgagccagg aagaccccga ggtccagttc aactggtacg tggatggcgt ggaggtgcat2220aatgccaaga caaagccgcg ggaggagcag ttcaacagca cgtaccgtgt ggtcagcgtc2280ctcaccgtcc tgcaccagga ctggctgaac ggcaaggagt acaagtgcaa ggtctccaac2340aaaggcctcc cgtcctccat cgagaaaacc atctccaaag ccaaagggca gccccgagag2400ccacaggtgt acaccctgcc cccatcccag gaggagatga ccaagaacca ggtcagcctg2460acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga gagcaatggg2520cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc2580ctctacagca ggctaaccgt ggacaagagc aggtggcagg aggggaatgt cttctcatgc2640tccgtgatgc atgaggctct gcacaaccac tacacacaga agagcctctc cctgtctctg2700ggtaaatga2709<210>10<211>902<212>PRT<213>人<400>10Met Val Leu Leu Trp Cys Val Val Ser Leu Tyr Phe Tyr Gly Ile Leu1 5 10 15Gln Ser Asp Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met20 25 30Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro35 40 45Leu Phe Glu His Phe Leu Lys Phe Asn Tyr Ser Thr Ala His Ser Ala50 55 60Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu65 70 75 80Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys85 90 95Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr100 105 110Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro115 120 125Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Pro Met Lys Leu
130 135 140Pro Val His Lys Leu Tyr Ile Glu Tyr Gly Ile Gln Arg Ile Thr Cys145 150 155 160Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Thr Ile Thr165 170 175Trp Tyr Met Gly Cys Tyr Lys Ile Gln Asn Phe Asn Asn Val Ile Pro180 185 190Glu Gly Met Asn Leu Ser Phe Leu Ile Ala Leu Ile Ser Asn Asn Gly195 200 205Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Thr Phe His210 215220Leu Thr Arg Thr Leu Thr Val Lys Val Val Gly Ser Pro Lys Asn Ala225 230 235 240Val Pro Pro Val Ile His Ser Pro Asn Asp His Val Val Tyr Glu Lys245 250 255Glu Pro Gly Glu Glu Leu Leu Ile Pro Cys Thr Val Tyr Phe Ser Phe260 265 270Leu Met Asp Ser Arg Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys275 280 285Pro Asp Asp Ile Thr Ile Asp Val Thr Ile Asn Glu Ser Ile Ser His290 295 300Ser Arg Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys305 310 315 320Val Thr Ser Glu Asp Leu Lys Arg Ser Tyr Val Cys His Ala Arg Ser325 330 335Ala Lys Gly Glu Val Ala Lys Ala Ala Lys Val Lys Gln Lys Val Pro340 345 350Ala Pro Arg Tyr Thr Val Glu Lys Cys Lys Glu Arg Glu Glu Lys Ile355 360 365Ile Leu Val Ser Ser Ala Asn Glu Ile Asp Val Arg Pro Cys Pro Leu370 375 380Asn Pro Asn Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser385 390 395 400Lys Thr Pro Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys405 410 415Glu Lys Leu Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr420 425 430Tyr Cys Val Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser435 440 445Ala Lys Phe Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala450 455 460Ile Phe Lys Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys465 470 475 480Pro Tyr Met Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu485 490 495Gln Trp Tyr Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe500 505 510Ser Gly Val Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His515 520 525Arg Gly Asn Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln530 535 540Tyr Pro Ile Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys545 550 555 560Pro Thr Arg Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val565 570 575Asp Leu Gly Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu580 585 590
Ser Asp Ile Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp595 600 605Asp Pro Val Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn610 615 620Lys Arg Arg Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu625 630 635 640Ser Arg Phe Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His645 650 655Gly Ile Asp Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn Ser660 665 670Gly Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu675 680 685Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp690 695 700Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp705 710 715 720Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly725 730 735Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn740 745 750Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp755 760 765Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro770 775 780Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu785 790 795 800Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn805 810 815Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile820 825 830Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr835 840 845Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg850 855 860Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys865 870 875 880Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu885 890 895Ser Leu Ser Leu Gly Lys900<210>11<211>2709<212>DNA<213>人<400>11atggtgcttc tgtggtgtgt agtgagtctc tacttttatg gaatcctgca aagtgatgcc60tcagaacgct gcgatgactg gggactagac accatgaggc aaatccaagt gtttgaagat120gagccagctc gcatcaagtg cccactcttt gaacacttct tgaaattcaa ctacagcaca180gcccattcag ctggccttac tctgatctgg tattggacta ggcaggaccg ggaccttgag240gagccaatta acttccgcct ccccgagaac cgcattagta aggagaaaga tgtgctgtgg300ttccggccca ctctcctcaa tgacactggc aactatacct gcatgttaag gaacactaca360tattgcagca aagttgcatt tcccttggaa gttgttcaaa aagacagctg tttcaattcc420cccatgaaac tcccagtgca taaactgtat atagaatatg gcattcagag gatcacttgt480ccaaatgtag atggatattt tccttccagt gtcaaaccga ctatcacttg gtatatgggc540tgttataaaa tacagaattt taataatgta atacccgaag gtatgaactt gagtttcctc600
attgccttaa tttcaaataa tggaaattac acatgtgttg ttacatatcc agaaaatgga660cgtacgtttc atctcaccag gactctgact gtaaaggtag taggctctcc aaaaaatgca720gtgccccctg tgatccattc acctaatgat catgtggtct atgagaaaga accaggagag780gagctactca ttccctgtac ggtctatttt agttttctga tggattctcg caatgaggtt840tggtggacca ttgatggaaa aaaacctgat gacatcacta ttgatgtcac cattaacgaa900agtataagtc atagtagaac agaagatgaa acaagaactc agattttgag catcaagaaa960gttacctctg aggatctcaa gcgcagctat gtctgtcatg ctagaagtgc caaaggcgaa1020gttgccaaag cagccaaggt gaagcagaaa gtgccagctc caagatacac agtggaaaaa1080tgcaaggaac gtgaagaaaa aataatttta gtgagctcag caaatgaaat cgatgttcgt1140ccctgtcctc ttaacccaaa tgaacacaaa ggcactataa cttggtataa ggatgacagc1200aagacacctg tatctacaga acaagcctcc aggattcatc aacacaaaga gaaactttgg1260tttgttcctg ctaaggtgga ggattcagga cattactatt gcgtggtaag aaattcatct1320tactgcctca gaattaaaat aagtgcaaaa tttgtggaga atgagcctaa cttatgttat1380aatgcacaag ccatatttaa gcagaaacta cccgttgcag gagacggagg acttgtgtgc1440ccttatatgg agttttttaa aaatgaaaat aatgagttac ctaaattaca gtggtataag1500gattgcaaac ctctacttct tgacaatata cactttagtg gagtcaaaga taggctcatc1560gtgatgaatg tggctgaaaa gcatagaggg aactatactt gtcatgcatc ctacacatac1620ttgggcaagc aatatcctat tacccgggta atagaattta ttactctaga ggaaaacaaa1680cccacaaggc ctgtgattgt gagcccagct aatgagacaa tggaagtaga cttgggatcc1740cagatacaat tgatctgtaa tgtcaccggc cagttgagtg acattgctta ctggaagtggl800aatgggtcag taattgatga agatgaccca gtgctagggg aagactatta cagtgtggaa1860aatcctgcaa acaaaagaag gagtaccctc atcacagtgc ttaatatatc ggaaattgag1920agtagatttt ataaacatcc atttacctgt tttgccaaga atacacatgg tatagatgca1980gcatatatcc agttaatata tccagtcact aattccggag agtccaaata cggtccgcca2040tgcccaccat gcccagcacc tgagttcctg gggggaccat cagtcttcct gttcccccca2100aaacccaagg acactctcat gatctcccgg acccctgagg tcacgtgcgt ggtggtggac2160gtgagccagg aagaccccga ggtccagttc aactggtacg tggatggcgt ggaggtgcat2220aatgccaaga caaagccgcg ggaggagcag ttcaacagca cgtaccgtgt ggtcagcgtc2280ctcaccgtcc tgcaccagga ctggctgaac ggcaaggagt acaagtgcaa ggtctccaac2340aaaggcctcc cgtcctccat cgagaaaacc atctccaaag ccaaagggca gccccgagag2400ccacaggtgt acaccctgcc cccatcccag gaggagatga ccaagaacca ggtcagcctg2460acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga gagcaatggg2520cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc2580ctctacagca ggctaaccgt ggacaagagc aggtggcagg aggggaatgt cttctcatgc2640tccgtgatgc atgaggctct gcacaaccac tacacacaga agagcctctc cctgtctctg2700ggtaaatga2709<210>12<211>902<212>PRT<213>人<400>12Met Val Leu Leu Trp Cys Val Val Ser Leu Tyr Phe Tyr Gly Ile Leu1 5 10 15Gln Ser Asp Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met20 25 30Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro35 40 45Leu Phe Glu His Phe Leu Lys Phe Asn Tyr Ser Thr Ala His Ser Ala50 55 60Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu65 70 75 80Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys85 90 95Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr100 105 110
Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro115 120 125Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Pro Met Lys Leu130 135 140Pro Val His Lys Leu Tyr Ile Glu Tyr Gly Ile Gln Arg Ile Thr Cys145 150 155 160Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Thr Ile Thr165 170 175Trp Tyr Met Gly Cys Tyr Lys Ile Gln Asn Phe Asn Asn Val Ile Pro180 185 190Glu Gly Met Asn Leu Ser Phe Leu Ile Ala Leu Ile Ser Asn Asn Gly195 200 205Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Thr Phe His210 215 220Leu Thr Arg Thr Leu Thr Val Lys Val Val Gly Ser Pro Lys Asn Ala225 230 235 240Val Pro Pro Val Ile His Ser Pro Asn Asp His Val Val Tyr Glu Lys245 250 255Glu Pro Gly Glu Glu Leu Leu Ile Pro Cys Thr Val Tyr Phe Ser Phe260 265 270Leu Met Asp Ser Arg Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys275 280 285Pro Asp Asp Ile Thr Ile Asp Val Thr Ile Asn Glu Ser Ile Ser His290 295 300Ser Arg Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys305 310 315 320Val Thr Ser Glu Asp Leu Lys Arg Ser Tyr Val Cys His Ala Arg Ser325 330 335Ala Lys Gly Glu Val Ala Lys Ala Ala Lys Val Lys Gln Lys Val Pro340 345 350Ala Pro Arg Tyr Thr Val Glu Lys Cys Lys Glu Arg Glu Glu Lys Ile355 360 365Ile Leu Val Ser Ser Ala Asn Glu Ile Asp Val Arg Pro Cys Pro Leu370 375 380Asn Pro Asn Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser385 390 395 400Lys Thr Pro Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys405 410 415Glu Lys Leu Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr420 425 430Tyr Cys Val Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser435 440 445Ala Lys Phe Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala450 455 460Ile Phe Lys Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys465 470 475 480Pro Tyr Met Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu485 490 495Gln Trp Tyr Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe500 505 510Ser Gly Val Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His515 520 525Arg Gly Asn Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln530 535 540Tyr Pro Ile Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys545 550 555 560Pro Thr Arg Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val
565 570 575Asp Leu Gly Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu580 585 590Ser Asp Ile Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp595 600 605Asp Pro Val Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn610 615 620Lys Arg Arg Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu625 630 635 640Ser Arg Phe Tyr Lys Hig Pro Phe Thr Cys Phe Ala Lys Asn Thr His645 650 655Gly Ile Asp Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn Ser660 665 670Gly Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu675 680 685Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp690 695 700Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp705 710 715 720Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly725 730 735Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn740 745 750Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp755 760 765Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro770 775 780Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu785 790 795 800Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn805 810 815Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile820 825 830Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr835 840 845Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg850 855 860Leu Thr val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys865 870 875 880Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu885 890 895Ser Leu Ser Leu Gly Lys900<210>13<211>2748<212>DNA<213>人<400>13atggtgcgct tgtacgtgtt ggtaatggga gtttctgcct tcacccttca gcctgcggca60cacacagggg ctgccagaag ctgccggttt cgtgggaggc attacaagcg ggagttcagg120ctggaagggg agcctgtagc cctgaggtgc ccccaggtgc cctactggtt gtgggcctct180gtcagccccc gcatcaacct gacatggcat aaaaatgact ctgctaggac ggtcccagga240gaagaagaga cacggatgtg ggcccaggac ggtgctctgt ggcttctgcc agccttgcag300gaggactctg gcacctacgt ctgcactact agaaatgctt cttactgtga caaaatgtcc360attgagctca gagtttttga gaatacagat gctttcctgc cgttcatctc atacccgcaa420
attttaacct tgtcaacctc tggggtatta gtatgccctg acctgagtga attcacccgt480gacaaaactg acgtgaagat tcaatggtac aaggattctc ttcttttgga taaagacaat540gagaaatttc taagtgtgag ggggaccact cacttactcg tacacgatgt ggccctggaa600gatgctggct attaccgctg tgtcctgaca tttgcccatg aaggccagca atacaacatc660actaggagta ttgagctacg catcaagaaa aaaaaagaag agaccattcc tgtgatcatt720tcccccctca agaccatatc agcttctctg gggtcaagac tgacaatccc atgtaaggtg780tttctgggaa ccggcacacc cttaaccacc atgctgtggt ggacggccaa tgacacccac840atagagagcg cctacccggg aggccgcgtg accgaggggc cacgccagga atattcagaa900aataatgaga actacattga agtgccattg atttttgatc ctgtcacaag agaggatttg960cacatggatt ttaaatgtgt tgtccataat accctgagtt ttcagacact acgcaccaca1020gtcaaggaag cctcctccac gttctcagaa cgctgcgatg actggggact agacaccatg1080aggcaaatcc aagtgtttga agatgagcca gctcgcatca agtgcccact ctttgaacac1140ttcttgaaat tcaactacag cacagcccat tcagctggcc ttactctgat ctggtattgg1200actaggcagg accgggacct tgaggagcca attaacttcc gcctccccga gaaccgcatt1260agtaaggaga aagatgtgct gtggttccgg cccactctcc tcaatgacac tggcaactat1320acctgcatgt taaggaacac tacatattgc agcaaagttg catttccctt ggaagttgtt1380caaaaagaca gctgtttcaa ttcccccatg aaactcccag tgcataaact gtatatagaa1440tatggcattc agaggatcac ttgtccaaat gtagatggat attttccttc cagtgtcaaa1500ccgactatca cttggtatat gggctgttat aaaatacaga attttaataa tgtaataccc1560gaaggtatga acttgagttt cctcattgcc ttaatttcaa ataatggaaa ttacacatgt1620gttgttacat atccagaaaa tggacgtacg tttcatctca ccaggactct gactgtaaag1680gtagtaggct ctccaaaaaa tgcagtgccc cctgtgatcc attcacctaa tgatcatgtg1740gtctatgaga aagaaccagg agaggagcta ctcattccct gtacggtcta ttttagtttt1800ctgatggatt ctcgcaatga ggtttggtgg accattgatg gaaaaaaacc tgatgacatc1860actattgatg tcaccattaa cgaaagtata agtcatagta gaacagaaga tgaaacaaga1920actcagattt tgagcatcaa gaaagttacc tctgaggatc tcaagcgcag ctatgtctgt1980catgctagaa gtgccaaagg cgaagttgcc aaagcagcca aggtgaagca gaaagtgcca2040gctccaagat acacagtgtc cggagacaaa actcacacat gcccaccgtg cccagcacct2100gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga caccctcatg2160atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag2220gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccgcgg2280gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac2340tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc2400gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta caccctgccc2460ccatcccggg atgagctgac caagaaccag gtcagcctga cctgcctggt caaaggcttc2520tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag2580accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctatagcaa gctcaccgtg2640gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg2700cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatga 2748<210>14<211>915<212>PRT<213>人<400>14Met Val Arg Leu Tyr Val Leu Val Met Gly Val Ser Ala Phe Thr Leu1 5 10 15Gln Pro Ala Ala His Thr Gly Ala Ala Arg Ser Cys Arg Phe Arg Gly20 25 30Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val Ala Leu35 40 45Arg Cys Pro Gln Val Pro Tyr Trp Leu Trp Ala Ser Val Ser Pro Arg50 55 60Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val Pro Gly65 70 75 80Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp Leu Leu
85 90 95Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr Arg Asn100 105 110Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe Glu Asn115 120 125Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu Thr Leu130 135 140Ser Thr Ser Gly Val Leu Val Cys Pro Asp Leu Ser Glu Phe Thr Arg145 150 155 160Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu Leu Leu165 170 175Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr His Leu180 185 190Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg Cys Va1195 200 205Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn Ile Thr Arg Ser Ile210 215 220Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val Ile Ile225 230 235 240Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu Thr Ile245 250 255Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr Met Leu260 265 270Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro Gly Gly275 280 285Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn Glu Asn290 295 300Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu Asp Leu305 310 315 320His Met Asp Phe Lys Cys Val Val His Asn Thr Leu Ser Phe Gln Thr325 330 335Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Glu Arg Cys340 345 350Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp355 360 365Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Phe370 375 380Asn Tyr Ser Thr Ala His Ser Ala Gly Leu Thr Leu Ile Trp Tyr Trp385 390 395 400Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro405 410 415Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr420 425 430Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr435 440 445Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser450 455 460Cys Phe Asn Ser Pro Met Lys Leu Pro Val His Lys Leu Tyr Ile Glu465 470 475 480Tyr Gly Ile Gln Arg Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro485 490 495Ser Ser Val Lys Pro Thr Ile Thr Trp Tyr Met Gly Cys Tyr Lys Ile500 505 510Gln Asn Phe Asn Asn Val Ile Pro Glu Gly Met Asn Leu Ser Phe Leu515 520 525Ile Ala Leu Ile Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr530 535 540
Pro Glu Asn Gly Arg Thr Phe His Leu Thr Arg Thr Leu Thr Val Lys545 550 555 560Val Val Gly Ser Pro Lys Asn Ala Val Pro Pro Val Ile His Ser Pro565 570 575Asn Asp His Val Val Tyr Glu Lys Glu Pro Gly Glu Glu Leu Leu Ile580 585 590Pro Cys Thr Val Tyr Phe Ser Phe Leu Met Asp Ser Arg Asn Glu Val595 600 605Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Ile Thr Ile Asp Val610 615 620Thr Ile Asn Glu Ser Ile Ser His Ser Arg Thr Glu Asp Glu Thr Arg625 630 635 640Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Ser Glu Asp Leu Lys Arg645 650 655Ser Tyr Val Cys His Ala Arg Ser Ala Lys Gly Glu Val Ala Lys Ala660 665 670Ala Lys Val Lys Gln Lys Val Pro Ala Pro Arg Tyr Thr Val Ser Gly675 680 685Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly690 695 700Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met705 710 715 720Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His725 730 735Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val740 745 750His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr755 760 765Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly770 775 780Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile785 790 795 800Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val805 810 815Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser820 825 830Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu835 840 845Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro850 855 860Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val865 870 875 880Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met885 890 895His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser900 905 910Pro Gly Lys915<210>15<211>2754<212>DNA<213>人<400> 15atggtgcgct tgtacgtgtt ggtaatggga gtttctgcct tcacccttca gcctgcggca60cacacagggg ctgccagaag ctgccggttt cgtgggaggc attacaagcg ggagttcagg120
ctggaagggg agcctgtagc cctgaggtgc ccccaggtgc cctactggtt gtgggcctct180gtcagccccc gcatcaacct gacatggcat aaaaatgact ctgctaggac ggtcccagga240gaagaagaga cacggatgtg ggcccaggac ggtgctctgt ggcttctgcc agccttgcag300gaggactctg gcacctacgt ctgcactact agaaatgctt cttactgtga caaaatgtcc360attgagctca gagtttttga gaatacagat gctttcctgc cgttcatctc atacccgcaa420attttaacct tgtcaacctc tggggtatta gtatgccctg acctgagtga attcacccgt480gacaaaactg acgtgaagat tcaatggtac aaggattctc ttcttttgga taaagacaat540gagaaatttc taagtgtgag ggggaccact cacttactcg tacacgatgt ggccctggaa600gatgctggct attaccgctg tgtcctgaca tttgcccatg aaggccagca atacaacatc660actaggagta ttgagctacg catcaagaaa aaaaaagaag agaccattcc tgtgatcatt720tcccccctca agaccatatc agcttctctg gggtcaagac tgacaatccc atgtaaggtg780tttctgggaa ccggcacacc cttaaccacc atgctgtggt ggacggccaa tgacacccac840atagagagcg cctacccggg aggccgcgtg accgaggggc cacgccagga atattcagaa900aataatgaga actacattga agtgccattg atttttgatc ctgtcacaag agaggatttg960cacatggatt ttaaatgtgt tgtccataat accctgagtt ttcagacact acgcaccaca1020gtcaaggaag cctcctccac gttctcagaa cgctgcgatg actggggact agacaccatg1080aggcaaatcc aagtgtttga agatgagcca gctcgcatca agtgcccact ctttgaacac1140ttcttgaaat tcaactacag cacagcccat tcagctggcc ttactctgat ctggtattgg1200actaggcagg accgggacct tgaggagcca attaacttcc gcctccccga gaaccgcatt1260agtaaggaga aagatgtgct gtggttccgg cccactctcc tcaatgacac tggcaactat1320acctgcatgt taaggaacac tacatattgc agcaaagttg catttccctt ggaagttgtt1380caaaaagaca gctgtttcaa ttcccccatg aaactcccag tgcataaact gtatatagaa1440tatggcattc agaggatcac ttgtccaaat gtagatggat attttccttc cagtgtcaaa1500ccgactatca cttggtatat gggctgttat aaaatacaga attttaataa tgtaataccc1560gaaggtatga acttgagttt cctcattgcc ttaatttcaa ataatggaaa ttacacatgt1620gttgttacat atccagaaaa tggacgtacg tttcatctca ccaggactct gactgtaaag1680gtagtaggct ctccaaaaaa tgcagtgccc cctgtgatcc attcacctaa tgatcatgtg1740gtctatgaga aagaaccagg agaggagcta ctcattccct gtacggtcta ttttagtttt1800ctgatggatt ctcgcaatga ggtttggtgg accattgatg gaaaaaaacc tgatgacatc1860actattgatg tcaccattaa cgaaagtata agtcatagta gaacagaaga tgaaacaaga1920actcagattt tgagcatcaa gaaagttacc tctgaggatc tcaagcgcag ctatgtctgt1980catgctagaa gtgccaaagg cgaagttgcc aaagcagcca aggtgaagca gaaagtgcca2040gctccaagat acacagtgtc cggagagtcc aaatacggtc cgccatgccc atcatgccca2100gcacctgagt tcctgggggg accatcagtc ttcctgttcc ccccaaaacc caaggacact2160ctcatgatct cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccaggaagac2220cccgaggtcc agttcaactg gtacgtggat ggcgtggagg tgcataatgc caagacaaag2280ccgcgggagg agcagttcaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac2340caggactggc tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccgtcc2400tccatcgaga aaaccatctc caaagccaaa gggcagcccc gagagccaca ggtgtacacc2460ctgcccccat cccaggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa2520ggcttctacc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac2580tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaggcta2640accgtggaca agagcaggtg gcaggagggg aatgtcttct catgctccgt gatgcatgag2700gctctgcaca accactacac acagaagagc ctctccctgt ctctgggtaa atga 2754<210>16<211>917<212>PRT<213>人<400>16Met Val Arg Leu Tyr Val Leu Val Met Gly Val Ser Ala Phe Thr Leu1 5 10 15Gln Pro Ala Ala His Thr Gly Ala Ala Arg Ser Cys Arg Phe Arg Gly20 25 30Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val Ala Leu35 40 45
Arg Cys Pro Gln Val Pro Tyr Trp Leu Trp Ala Ser Val Ser Pro Arg50 55 60Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val Pro Gly65 70 75 80Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp Leu Leu85 90 95Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr Arg Asn100 105 110Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe Glu Asn115 120 125Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu Thr Leu130 135 140Ser Thr Ser Gly Val Leu Val Cys Pro Asp Leu Ser Glu Phe Thr Arg145 150 155 160Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu Leu Leu165 170 175Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr His Leu180 185 190Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg Cys Val195 200 205Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn Ile Thr Arg Ser Ile210 215 220Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val Ile Ile225 230 235 240Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu Thr Ile245 250 255Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr Met Leu260 2652 70Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro Gly Gly275 280 285Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn Glu Asn290 295 300Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu Asp Leu305 310 315 320His Met Asp Phe Lys Cys Val Val His Asn Thr Leu Ser Phe Gln Thr325 330 335Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Glu Arg Cys340 345 350Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp355 360 365Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Phe370 375 380Asn Tyr Ser Thr Ala His Ser Ala Gly Leu Thr Leu Ile Trp Tyr Trp385 390 395 400Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro405 410 415Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr420 425 430Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr435 440 445Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser450 455 460Cys Phe Asn Ser Pro Met Lys Leu Pro Val His Lys Leu Tyr Ile Glu465 470 475 480Tyr Gly Ile Gln Arg Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro485 490 495Ser Ser Val Lys Pro Thr Ile Thr Trp Tyr Met Gly Cys Tyr Lys Ile
500 505 510Gln Asn Phe Asn Asn Val Ile Pro Glu Gly Met Asn Leu Ser Phe Leu515 520 525Ile Ala Leu Ile Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr530 535 540Pro Glu Asn Gly Arg Thr Phe His Leu Thr Arg Thr Leu Thr Val Lys545 550 555 560Val Val Gly Ser Pro Lys Asn Ala Val Pro Pro Val Ile His Ser Pro565 570 575Asn Asp His Val Val Tyr Glu Lys Glu Pro Gly Glu Glu Leu Leu Ile580 585 590Pro Cys Thr Val Tyr Phe Ser Phe Leu Met Asp Ser Arg Asn Glu Val595 600 605Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Ile Thr Ile Asp Val610 615 620Thr Ile Asn Glu Ser Ile Ser His Ser Arg Thr Glu Asp Glu Thr Arg625 630 635 640Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Ser Glu Asp Leu Lys Arg645 650 655Ser Tyr Val Cys His Ala Arg Ser Ala Lys Gly Glu Val Ala Lys Ala660 665 670Ala Lys Val Lys Gln Lys Val Pro Ala pro Arg Tyr Thr Val Ser Gly675 680 685Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe690 695 700Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr705 710 715 720Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val725 730 735Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val740 745 750Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser755 760 765Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu770 775 780Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser785 790 795 800Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro805 810 815Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln820 825 830Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala835 840 845Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr850 855 860Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu865 870 875 880Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser885 890 895Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser900 905 910Leu Ser Leu Gly Lys915<210>17<211>2754<212>DNA
<213>人<400>17atggtgcgct tgtacgtgtt ggtaatggga gtttctgcct tcacccttca gcctgcggca60cacacagggg ctgccagaag ctgccggttt cgtgggaggc attacaagcg ggagttcagg120ctggaagggg agcctgtagc cctgaggtgc ccccaggtgc cctactggtt gtgggcctct180gtcagccccc gcatcaacct gacatggcat aaaaatgact ctgctaggac ggtcccagga240gaagaagaga cacggatgtg ggcccaggac ggtgctctgt ggcttctgcc agccttgcag300gaggactctg gcacctacgt ctgcactact agaaatgctt cttactgtga caaaatgtcc360attgagctca gagtttttga gaatacagat gctttcctgc cgttcatctc atacccgcaa420attttaacct tgtcaacctc tggggtatta gtatgccctg acctgagtga attcacccgt480gacaaaactg acgtgaagat tcaatggtac aaggattctc ttcttttgga taaagacaat540gagaaatttc taagtgtgag ggggaccact cacttactcg tacacgatgt ggccctggaa600gatgctggct attaccgctg tgtcctgaca tttgcccatg aaggccagca atacaacatc660actaggagta ttgagctacg catcaagaaa aaaaaagaag agaccattcc tgtgatcatt720tcccccctca agaccatatc agcttctctg gggtcaagac tgacaatccc atgtaaggtg780tttctgggaa ccggcacacc cttaaccacc atgctgtggt ggacggccaa tgacacccac840atagagagcg cctacccggg aggccgcgtg accgaggggc cacgccagga atattcagaa900aataatgaga actacattga agtgccattg atttttgatc ctgtcacaag agaggatttg960cacatggatt ttaaatgtgt tgtccataat accctgagtt ttcagacact acgcaccaca1020gtcaaggaag cctcctccac gttctcagaa cgctgcgatg actggggact agacaccatg1080aggcaaatcc aagtgtttga agatgagcca gctcgcatca agtgcccact ctttgaacac1140ttcttgaaat tcaactacag cacagcccat tcagctggcc ttactctgat ctggtattgg1200actaggcagg accgggacct tgaggagcca attaacttcc gcctccccga gaaccgcatt1260agtaaggaga aagatgtgct gtggttccgg cccactctcc tcaatgacac tggcaactat1320acctgcatgt taaggaacac tacatattgc agcaaagttg catttccctt ggaagttgtt1380caaaaagaca gctgtttcaa ttcccccatg aaactcccag tgcataaact gtatatagaa1440tatggcattc agaggatcac ttgtccaaat gtagatggat attttccttc cagtgtcaaa1500ccgactatca cttggtatat gggctgttat aaaatacaga attttaataa tgtaataccc1560gaaggtatga acttgagttt cctcattgcc ttaatttcaa ataatggaaa ttacacatgt1620gttgttacat atccagaaaa tggacgtacg tttcatctca ccaggactct gactgtaaag1680gtagtaggct ctccaaaaaa tgcagtgccc cctgtgatcc attcacctaa tgatcatgtg1740gtctatgaga aagaaccagg agaggagcta ctcattccct gtacggtcta ttttagtttt1800ctgatggatt ctcgcaatga ggtttggtgg accattgatg gaaaaaaacc tgatgacatc1860actattgatg tcaccattaa cgaaagtata agtcatagta gaacagaaga tgaaacaaga1920actcagattt tgagcatcaa gaaagttacc tctgaggatc tcaagcgcag ctatgtctgt1980catgctagaa gtgccaaagg cgaagttgcc aaagcagcca aggtgaagca gaaagtgcca2040gctccaagat acacagtgtc cggagagtcc aaatacggtc cgccatgccc accatgccca2100gcacctgagt tcctgggggg accatcagtc ttcctgttcc ccccaaaacc caaggacact2160ctcatgatct cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccaggaagac2220cccgaggtcc agttcaactg gtacgtggat ggcgtggagg tgcataatgc caagacaaag2280ccgcgggagg agcagttcaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac2340caggactggc tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccgtcc2400tccatcgaga aaaccatctc caaagccaaa gggcagcccc gagagccaca ggtgtacacc2460ctgcccccat cccaggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa2520ggcttctacc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac2580tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaggcta2640accgtggaca agagcaggtg gcaggagggg aatgtcttct catgctccgt gatgcatgag2700gctctgcaca accactacac acagaagagc ctctccctgt ctctgggtaa atga 2754<210>18<211>917<212>PRT<213>人<400>18Met Val Arg Leu Tyr Val Leu Val Met Gly Val Ser Ala Phe Thr Leu
1 5 10 15Gln Pro Ala Ala His Thr Gly Ala Ala Arg Ser Cys Arg Phe Arg Gly20 25 30Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val Ala Leu35 40 45Arg Cys Pro Gln Val Pro Tyr Trp Leu Trp Ala Ser Val Ser Pro Arg50 55 60Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val Pro Gly65 70 75 80Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp Leu Leu85 90 95Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr Arg Asn100 105 110Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe Glu Asn115 120 125Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu Thr Leu130 135 140Ser Thr Ser Gly Val Leu Val Cys Pro Asp Leu Ser Glu Phe Thr Arg145 150 155 160Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu Leu Leu165 170 175Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr His Leu180 185 190Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg Cys Val195 200 205Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn Ile Thr Arg Ser Ile210 215 220Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val Ile Ile225 230 235 240Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu Thr Ile245 250 255pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr Met Leu260 265 270Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro Gly Gly275 280 285Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn Glu Asn290 295 300Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu Asp Leu305 310 315 320His Met Asp Phe Lys Cys Val Val His Asn Thr Leu Ser Phe Gln Thr325 330 335Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Glu Arg Cys340 345 350Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp355 360 365Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Phe370 375 380Asn Tyr Ser Thr Ala His Ser Ala Gly Leu Thr Leu Ile Trp Tyr Trp385 390 395 400Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro405 410 415Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr420 425 430Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr435 440 445Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser450 455 460
Cys Phe Asn Ser Pro Met Lys Leu Pro Val His Lys Leu Tyr Ile Glu465 470 475 480Tyr Gly Ile Gln Arg Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro485 490 495Ser Ser Val Lys Pro Thr Ile Thr Trp Tyr Met Gly Cys Tyr Lys Ile500 505 510Gln Asn Phe Asn Asn Val Ile Pro Glu Gly Met Asn Leu Ser Phe Leu515 520 525Ile Ala Leu Ile Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr530 535 540Pro Glu Asn Gly Arg Thr Phe His Leu Thr Arg Thr Leu Thr Val Lys545 550 555 560Val Val Gly Ser Pro Lys Asn Ala Val Pro Pro Val Ile His Ser Pro565 570 575Asn Asp His Val Val Tyr Glu Lys Glu Pro Gly Glu Glu Leu Leu Ile580 585 590Pro Cys Thr Val Tyr Phe Ser Phe Leu Met Asp Ser Arg Asn Glu Val595 600 605Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Ile Thr Ile Asp Val610 615 620Thr Ile Asn Glu Ser Ile Ser His Ser Arg Thr Glu Asp Glu Thr Arg625 630 635 640Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Ser Glu Asp Leu Lys Arg645 650 655Ser Tyr Val Cys His Ala Arg Ser Ala Lys Gly Glu Val Ala Lys Ala660 665 670Ala Lys Val Lys Gln Lys Val Pro Ala Pro Arg Tyr Thr Val Ser Gly675 680 685Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe690 695 700Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr705 710 715 720Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val725 730 735Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val740 745 750Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser755 760 765Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu770 775 780Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser785 790 795 800Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro805 810 815Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln820 825 830Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala835 840 845Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr850 855 860Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu865 870 875 880Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser885 890 895Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser900 905 910Leu Ser Leu Gly Lys
915<210>19<211>2748<212>DNA<213>人<400>19atggtgcttc tgtggtgtgt agtgagtctc tacttttatg gaatcctgca aagtgatgcc60tcagaacgct gcgatgactg gggactagac accatgaggc aaatccaagt gtttgaagat120gagccagctc gcatcaagtg cccactcttt gaacacttct tgaaattcaa ctacagcaca180gcccattcag ctggccttac tctgatctgg tattggacta ggcaggaccg ggaccttgag240gagccaatta acttccgcct ccccgagaac cgcattagta aggagaaaga tgtgctgtgg300ttccggccca ctctcctcaa tgacactggc aactatacct gcatgttaag gaacactaca360tattgcagca aagttgcatt tcccttggaa gttgttcaaa aagacagctg tttcaattcc420cccatgaaac tcccagtgca taaactgtat atagaatatg gcattcagag gatcacttgt480ccaaatgtag atggatattt tccttccagt gtcaaaccga ctatcacttg gtatatgggc540tgttataaaa tacagaattt taataatgta atacccgaag gtatgaactt gagtttcctc600attgccttaa tttcaaataa tggaaattac acatgtgttg ttacatatcc agaaaatgga660cgtacgtttc atctcaccag gactctgact gtaaaggtag taggctctcc aaaaaatgca720gtgccccctg tgatccattc acctaatgat catgtggtct atgagaaaga accaggagag780gagctactca ttccctgtac ggtctatttt agttttctga tggattctcg caatgaggtt840tggtggacca ttgatggaaa aaaacctgat gacatcacta ttgatgtcac cattaacgaa900agtataagtc atagtagaac agaagatgaa acaagaactc agattttgag catcaagaaa960gttacctctg aggatctcaa gcgcagctat gtctgtcatg ctagaagtgc caaaggcgaa1020gttgccaaag cagccaaggt gaagcagaaa gtgccagctc caagatacac agtgcacaca1080ggggctgcca gaagctgccg gtttcgtggg aggcattaca agcgggagtt caggctggaa1140ggggagcctg tagccctgag gtgcccccag gtgccctact ggttgtgggc ctctgtcagc1200ccccgcatca acctgacatg gcataaaaat gactctgcta ggacggtccc aggagaagaa1260gagacacgga tgtgggccca ggacggtgct ctgtggcttc tgccagcctt gcaggaggac1320tctggcacct acgtctgcac tactagaaat gcttcttact gtgacaaaat gtccattgag1380ctcagagttt ttgagaatac agatgctttc ctgccgttca tctcataccc gcaaatttta1440accttgtcaa cctctggggt attagtatgc cctgacctga gtgaattcac ccgtgacaaa1500actgacgtga agattcaatg gtacaaggat tctcttcttt tggataaaga caatgagaaa1560tttctaagtg tgagggggac cactcactta ctcgtacacg atgtggccct ggaagatgct1620ggctattacc gctgtgtcct gacatttgcc catgaaggcc agcaatacaa catcactagg1680agtattgagc tacgcatcaa gaaaaaaaaa gaagagacca ttcctgtgat catttccccc1740ctcaagacca tatcagcttc tctggggtca agactgacaa tcccatgtaa ggtgtttctg1800ggaaccggca cacccttaac caccatgctg tggtggacgg ccaatgacac ccacatagag1860agcgcctacc cgggaggccg cgtgaccgag gggccacgcc aggaatattc agaaaataat1920gagaactaca ttgaagtgcc attgattttt gatcctgtca caagagagga tttgcacatg1980gattttaaat gtgttgtcca taataccctg agttttcaga cactacgcac cacagtcaag2040gaagcctcct ccacgttctc cggagacaaa actcacacat gcccaccgtg cccagcacct2100gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga caccctcatg2160atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag2220gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccgcgg2280gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac2340tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc2400gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta caccctgccc2460ccatcccggg atgagctgac caagaaccag gtcagcctga cctgcctggt caaaggcttc2520tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag2580accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctatagcaa gctcaccgtg2640gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg2700cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatga 2748<210>20<211>915
<212>PRT<213>人<400>20Met Val Leu Leu Trp Cys Val Val Ser Leu Tyr Phe Tyr Gly Ile Leu1 5 10 15Gln Ser Asp Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met20 25 30Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro35 40 45Leu Phe Glu His Phe Leu Lys Phe Asn Tyr Ser Thr Ala His Ser Ala50 55 60Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu65 70 75 80Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys85 90 95Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr100 105 110Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro115 120 125Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Pro Met Lys Leu130 135 140Pro Val His Lys Leu Tyr Ile Glu Tyr Gly Ile Gln Arg Ile Thr Cys145 150 155 160Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Thr Ile Thr165 170 175Trp Tyr Met Gly Cys Tyr Lys Ile Gln Asn Phe Asn Asn Val Ile Pro180 185 190Glu Gly Met Asn Leu Ser Phe Leu Ile Ala Leu Ile Ser Asn Asn Gly195 200 205Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Thr Phe His210 215 220Leu Thr Arg Thr Leu Thr Val Lys Val Val Gly Ser Pro Lys Asn Ala225 230 235 240Val Pro Pro Val Ile His Ser Pro Asn Asp His Val Val Tyr Glu Lys245 250 255Glu Pro Gly Glu Glu Leu Leu Ile Pro Cys Thr Val Tyr Phe Ser Phe260 265 270Leu Met Asp Ser Arg Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys275 280 285Pro Asp Asp Ile Thr Ile Asp Val Thr Ile Asn Glu Ser Ile Ser His290 295 300Ser Arg Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys305 310 315 320Val Thr Ser Glu Asp Leu Lys Arg Ser Tyr Val Cys His Ala Arg Ser325 330 335Ala Lys Gly Glu Val Ala Lys Ala Ala Lys Val Lys Gln Lys Val Pro340 345 350Ala Pro Arg Tyr Thr Val His Thr Gly Ala Ala Arg Ser Cys Arg Phe355 360 365Arg Gly Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val370 375 380Ala Leu Arg Cys Pro Gln val Pro Tyr Trp Leu Trp Ala Ser Val Ser385 390 395 400Pro Arg Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val405 410 415Pro Gly Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp
420 425 430Leu Leu Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr435 440 445Arg Asn Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe450 455 460Glu Asn Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu465 470 475 480Thr Leu Ser Thr Ser Gly Val Leu Val Cys Pro Asp Leu Ser Glu Phe485 490 495Thr Arg Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu500 505 510Leu Leu Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr515 520 525His Leu Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg530 535 540Cys Val Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn Ile Thr Arg545 550 555 560Ser Ile Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val565 570 575Ile Ile Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu580 585 590Thr Ile Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr595 600 605Met Leu Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro610 615 620Gly Gly Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn625 630 635 640Glu Asn Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu645 650 655Asp Leu His Met Asp Phe Lys cys Val Val His Asn Thr Leu Ser Phe660 665 670Gln Thr Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Gly675 680 685Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly690 695 700Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met705 710 715 720Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His725 730 735Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val740 745 750His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr755 760 765Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly770 775 780Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile785 790 795 800Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val805 810 815Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser820 825 830Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu835 840 845Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro850 855 860Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val865 870 875 880
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met885 890 895His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser900 905 910Pro Gly Lys915<210>21<211>2754<212>DNA<213>人<400>21atggtgcttc tgtggtgtgt agtgagtctc tacttttatg gaatcctgca aagtgatgcc60tcagaacgct gcgatgactg gggactagac accatgaggc aaatccaagt gtttgaagat120gagccagctc gcatcaagtg cccactcttt gaacacttct tgaaattcaa ctacagcaca180gcccattcag ctggccttac tctgatctgg tattggacta ggcaggaccg ggaccttgag240gagccaatta acttccgcct ccccgagaac cgcattagta aggagaaaga tgtgctgtgg300ttccggccca ctctcctcaa tgacactggc aactatacct gcatgttaag gaacactaca360tattgcagca aagttgcatt tcccttggaa gttgttcaaa aagacagctg tttcaattcc420cccatgaaac tcccagtgca taaactgtat atagaatatg gcattcagag gatcacttgt480ccaaatgtag atggatattt tccttccagt gtcaaaccga ctatcacttg gtatatgggc540tgttataaaa tacagaattt taataatgta atacccgaag gtatgaactt gagtttcctc600attgccttaa tttcaaataa tggaaattac acatgtgttg ttacatatcc agaaaatgga660cgtacgtttc atctcaccag gactctgact gtaaaggtag taggctctcc aaaaaatgca720gtgccccctg tgatccattc acctaatgat catgtggtct atgagaaaga accaggagag780gagctactca ttccctgtac ggtctatttt agttttctga tggattctcg caatgaggtt840tggtggacca ttgatggaaa aaaacctgat gacatcacta ttgatgtcac cattaacgaa900agtataagtc atagtagaac agaagatgaa acaagaactc agattttgag catcaagaaa960gttacctctg aggatctcaa gcgcagctat gtctgtcatg ctagaagtgc caaaggcgaa1020gttgccaaag cagccaaggt gaagcagaaa gtgccagctc caagatacac agtgcacaca1080ggggctgcca gaagctgccg gtttcgtggg aggcattaca agcgggagtt caggctggaa1140ggggagcctg tagccctgag gtgcccccag gtgccctact ggttgtgggc ctctgtcagc1200ccccgcatca acctgacatg gcataaaaat gactctgcta ggacggtccc aggagaagaa1260gagacacgga tgtgggccca ggacggtgct ctgtggcttc tgccagcctt gcaggaggac1320tctggcacct acgtctgcac tactagaaat gcttcttact gtgacaaaat gtccattgag1380ctcagagttt ttgagaatac agatgctttc ctgccgttca tctcataccc gcaaatttta1440accttgtcaa cctctggggt attagtatgc cctgacctga gtgaattcac ccgtgacaaa1500actgacgtga agattcaatg gtacaaggat tctcttcttt tggataaaga caatgagaaa1560tttctaagtg tgagggggac cactcactta ctcgtacacg atgtggccct ggaagatgct1620ggctattacc gctgtgtcct gacatttgcc catgaaggcc agcaatacaa catcactagg1680agtattgagc tacgcatcaa gaaaaaaaaa gaagagacca ttcctgtgat catttccccc1740ctcaagacca tatcagcttc tctggggtca agactgacaa tcccatgtaa ggtgtttctg1800ggaaccggca cacccttaac caccatgctg tggtggacgg ccaatgacac ccacatagag1860agcgcctacc cgggaggccg cgtgaccgag gggccacgcc aggaatattc agaaaataat1920gagaactaca ttgaagtgcc attgattttt gatcctgtca caagagagga tttgcacatg1980gattttaaat gtgttgtcca taataccctg agttttcaga cactacgcac cacagtcaag2040gaagcctcct ccacgttctc cggagagtcc aaatacggtc cgccatgccc atcatgccca2100gcacctgagt tcctgggggg accatcagtc ttcctgttcc ccccaaaacc caaggacact2160ctcatgatct cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccaggaagac2220cccgaggtcc agttcaactg gtacgtggat ggcgtggagg tgcataatgc caagacaaag2280ccgcgggagg agcagttcaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac2340caggactggc tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccgtcc2400tccatcgaga aaaccatctc caaagccaaa gggcagcccc gagagccaca ggtgtacacc2460ctgcccccat cccaggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa2520ggcttctacc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac2580tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaggcta2640
accgtggaca agagcaggtg gcaggagggg aatgtcttct catgctccgt gatgcatgag2700gctctgcaca accactacac acagaagagc ctctccctgt ctctgggtaa atga 2754<210>22<211>917<212>PRT<213>人<400>22Met Val Leu Leu Trp Cys Val Val Ser Leu Tyr Phe Tyr Gly Ile Leu1 5 10 15Gln Ser Asp Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met20 25 30Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro35 40 45Leu Phe Glu His Phe Leu Lys Phe Asn Tyr Ser Thr Ala His Ser Ala50 55 60Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu65 70 75 80Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys85 90 95Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr100 105 110Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro115 120 125Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Pro Met Lys Leu130 135 140Pro Val His Lys Leu Tyr Ile Glu Tyr Gly Ile Gln Arg Ile Thr Cys145 150 155 160Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Thr Ile Thr165 170 175Trp Tyr Met Gly Cys Tyr Lys Ile Gln Asn Phe Asn Asn Val Ile Pro180 185 190Glu Gly Met Asn Leu Ser Phe Leu Ile Ala Leu Ile Ser Asn Asn Gly195 200 205Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Thr Phe His210 215 220Leu Thr Arg Thr Leu Thr Val Lys Val Val Gly Ser Pro Lys Asn Ala225 230 235 240Val Pro Pro Val Ile His Ser Pro Asn Asp His Val Val Tyr Glu Lys245 250 255Glu Pro Gly Glu Glu Leu LeuIle Pro Cys Thr Val Tyr Phe Ser Phe260 265 270Leu Met Asp Ser Arg Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys275 280 285Pro Asp Asp Ile Thr Ile Asp Val Thr Ile Asn Glu Ser Ile Ser His290 295 300Ser Arg Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys305 310 315 320Val Thr Ser Glu Asp Leu Lys Arg Ser Tyr Val Cys His Ala Arg Ser325 330 335Ala Lys Gly Glu Val Ala Lys Ala Ala Lys Val Lys Gln Lys Val Pro340 345 350Ala Pro Arg Tyr Thr Val His Thr Gly Ala Ala Arg Ser Cys Arg Phe355 360 365Arg Gly Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val370 375 380
Ala Leu Arg Cys Pro Gln Val Pro Tyr Trp Leu Trp Ala Ser Val Ser385 390 395 400Pro Arg Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val405 4l0 4l5Pro Gly Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp420 425 430Leu Leu Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr435 440 445Arg Asn Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe450 455 460Glu Asn Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu465 470 475 480Thr Leu Ser Thr Ser Gly Val Leu val Cys Pro Asp Leu Ser Glu Phe485 490 495Thr Arg Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu500 505 510Leu Leu Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr515 520 525His Leu Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg530 535 540Cys Val Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn Ile Thr Arg545 550 555 560Ser Ile Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val565 570 575Ile Ile Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu580 585 590Thr Ile Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr595 600 605Met Leu Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro610 615 620Gly Gly Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn625 630 635 640Glu Asn Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu645 650 655Asp Leu His Met Asp Phe Lys Cys Val Val His Asn Thr Leu Ser Phe660 665 670Gln Thr Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Gly675 680 685Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe690 695 700Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr705 710 715 720Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val725 730 735Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val740 745 750Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser755 760 765Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu770 775 780Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser785 790 795 800Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro805 810 815Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln820 825 830Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
835 840 845Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr850 855 860Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu865 870 875 880Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser885 890 895Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser900 905 910Leu Ser Leu Gly Lys915<210>23<211>2754<212>DNA<213>人<400>23atggtgcttc tgtggtgtgt agtgagtctc tacttttatg gaatcctgca aagtgatgcc60tcagaacgct gcgatgactg gggactagac accatgaggc aaatccaagt gtttgaagat120gagccagctc gcatcaagtg cccactcttt gaacacttct tgaaattcaa ctacagcaca180gcccattcag ctggccttac tctgatctgg tattggacta ggcaggaccg ggaccttgag240gagccaatta acttccgcct ccccgagaac cgcattagta aggagaaaga tgtgctgtgg300ttccggccca ctctcctcaa tgacactggc aactatacct gcatgttaag gaacactaca360tattgcagca aagttgcatt tcccttggaa gttgttcaaa aagacagctg tttcaattcc420cccatgaaac tcccagtgca taaactgtat atagaatatg gcattcagag gatcacttgt480ccaaatgtag atggatattt tccttccagt gtcaaaccga ctatcacttg gtatatgggc540tgttataaaa tacagaattt taataatgta atacccgaag gtatgaactt gagtttcctc600attgccttaa tttcaaataa tggaaattac acatgtgttg ttacatatcc agaaaatgga660cgtacgtttc atctcaccag gactctgact gtaaaggtag taggctctcc aaaaaatgca720gtgccccctg tgatccattc acctaatgat catgtggtct atgagaaaga accaggagag780gagctactca ttccctgtac ggtctatttt agttttctga tggattctcg caatgaggtt840tggtggacca ttgatggaaa aaaacctgat gacatcacta ttgatgtcac cattaacgaa900agtataagtc atagtagaac agaagatgaa acaagaactc agattttgag catcaagaaa960gttacctctg aggatctcaa gcgcagctat gtctgtcatg ctagaagtgc caaaggcgaa1020gttgccaaag cagccaaggt gaagcagaaa gtgccagctc caagatacac agtgcacaca1080ggggctgcca gaagctgccg gtttcgtggg aggcattaca agcgggagtt caggctggaa1140ggggagcctg tagccctgag gtgcccccag gtgccctact ggttgtgggc ctctgtcagc1200ccccgcatca acctgacatg gcataaaaat gactctgcta ggacggtccc aggagaagaa1260gagacacgga tgtgggccca ggacggtgct ctgtggcttc tgccagcctt gcaggaggac1320tctggcacct acgtctgcac tactagaaat gcttcttact gtgacaaaat gtccattgag1380ctcagagttt ttgagaatac agatgctttc ctgccgttca tctcataccc gcaaatttta1440accttgtcaa cctctggggt attagtatgc cctgacctga gtgaattcac ccgtgacaaa1500actgacgtga agattcaatg gtacaaggat tctcttcttt tggataaaga caatgagaaa1560tttctaagtg tgagggggac cactcactta ctcgtacacg atgtggccct ggaagatgct1620ggctattacc gctgtgtcct gacatttgcc catgaaggcc agcaatacaa catcactagg1680agtattgagc tacgcatcaa gaaaaaaaaa gaagagacca ttcctgtgat catttccccc1740ctcaagacca tatcagcttc tctggggtca agactgacaa tcccatgtaa ggtgtttctg1800ggaaccggca cacccttaac caccatgctg tggtggacgg ccaatgacac ccacatagag1860agcgcctacc cgggaggccg cgtgaccgag gggccacgcc aggaatattc agaaaataat1920gagaactaca ttgaagtgcc attgattttt gatcctgtca caagagagga tttgcacatg1980gattttaaat gtgttgtcca taataccctg agttttcaga cactacgcac cacagtcaag2040gaagcctcct ccacgttctc cggagagtcc aaatacggtc cgccatgccc accatgccca2100gcacctgagt tcctgggggg accatcagtc ttcctgttcc ccccaaaacc caaggacact2160ctcatgatct cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccaggaagac2220cccgaggtcc agttcaactg gtacgtggat ggcgtggagg tgcataatgc caagacaaag2280ccgcgggagg agcagttcaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac2340
caggactggc tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccgtcc2400tccatcgaga aaaccatctc caaagccaaa gggcagcccc gagagccaca ggtgtacacc2460ctgcccccat cccaggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa2520ggcttctacc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac2580tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaggcta2640accgtggaca agagcaggtg gcaggagggg aatgtcttct catgctccgt gatgcatgag2700gctctgcaca accactacac acagaagagc ctctccctgt ctctgggtaa atga 2754<210>24<211>917<212>PRT<213>人<400>24Met Val Leu Leu Trp Cys Val Val Ser Leu Tyr Phe Tyr Gly Ile Leu1 5 10 15Gln Ser Asp Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met20 25 30Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro35 40 45Leu Phe Glu His Phe Leu Lys Phe Asn Tyr Ser Thr Ala His Ser Ala50 55 60Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu65 70 75 80Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys85 90 95Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr100 105 110Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro115 120 125Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Pro Met Lys Leu130 135 140Pro Val His Lys Leu Tyr Ile Glu Tyr Gly Ile Gln Arg Ile Thr Cys145 150 155 160Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Thr Ile Thr165 170 175Trp Tyr Met Gly Cys Tyr Lys Ile Gln Asn Phe Asn Asn Val Ile Pro180 185 190Glu Gly Met Asn Leu Ser Phe Leu Ile Ala Leu Ile Ser Asn Asn Gly195 200 205Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Thr Phe His210 215 220Leu Thr Arg Thr Leu Thr Val Lys Val Val Gly Ser Pro Lys Asn Ala225 230 235 240Val Pro Pro Val Ile His Ser Pro Asn Asp His Val Val Tyr Glu Lys245 250 255Glu Pro Gly Glu Glu Leu Leu Ile Pro Cys Thr Val Tyr Phe Ser Phe260 265 270Leu Met Asp Ser Arg Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys275 280 285Pro Asp Asp Ile Thr Ile Asp Val Thr Ile Asn Glu Ser Ile Ser His290 295 300Ser Arg Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys305 310 315 320Val Thr Ser Glu Asp Leu Lys Arg Ser Tyr Val Cys His Ala Arg Ser325 330 335Ala Lys Gly Glu Val Ala Lys Ala Ala Lys Val Lys Gln Lys Val Pro
340 345 350Ala Pro Arg Tyr Thr Val His Thr Gly Ala Ala Arg Ser Cys Arg Phe355 360 365Arg Gly Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val370 375 380Ala Leu Arg Cys Pro Gln Val Pro Tyr Trp Leu Trp Ala Ser Val Ser385 390 395 400Pro Arg Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val405 410 415Pro Gly Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp420 425 430Leu Leu Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr435 440 445Arg Asn Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe450 455 460Glu Asn Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu465 470 475 480Thr Leu Ser Thr Ser Gly Val Leu Val Cys Pro Asp Leu Ser Glu Phe485 490 495Thr Arg Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu500 505 510Leu Leu Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr515 520 525His Leu Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg530 535 540Cys Val Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn Ile Thr Arg545 550 555 560Ser Ile Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val565 570 575Ile Ile Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu580 585 590Thr Ile Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr595 600 605Met Leu Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro610 615 620Gly Gly Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn625 630 635 640Glu Asn Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu645 650 655Asp Leu His Met Asp Phe Lys Cys Val Val His Asn Thr Leu Ser Phe660 665 670Gln Thr Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Gly675 680 685Glu Ser Lys Tyr Gly Pro Pro cys Pro Pro Cys Pro Ala Pro Glu Phe690 695 700Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr705 710 715 720Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val725 730 735Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val740 745 750Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser755 760 765Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu770 775 780Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser785 790 795 800
Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro805 810 815Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln820 825 830Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala835 840 845Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr850 855 860Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu865 870 875 880Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser885 890 895Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser900 905 910Leu Ser Leu Gly Lys915<210>25<211>6<212>PRT<213>人工序列<220>
<223>合成的<400>25His His His His His His1 5<210>26<211>5<212>PRT<213>人<220>
<221>變體<222>3<223>Xaa＝任何氨基酸<400>26Trp Ser Xaa Trp Ser1 權(quán)利要求
1.分離的核酸分子，其包含選自下列的序列SEQ ID NO1，3，5，7，9，11，13，15，17，19，21和23，其中所述分離的核酸分子編碼的融合多肽形成能結(jié)合白細(xì)胞介素1(IL-1)而形成無功能復(fù)合物的多聚體。
2.權(quán)利要求1的分離的核酸分子，其編碼分別選自下列的序列SEQ ID NO2，4，6，8，10，12，14，16，18，20，22，和24。
3.權(quán)利要求1的分離的核酸分子，其中該序列為SEQ ID NO7。
4.權(quán)利要求3的分離的核酸分子，其中該核酸分子編碼包含序列SEQ ID NO8的融合多肽。
5.包含權(quán)利要求1到4中任一項(xiàng)的分離的核酸分子的載體。
6.包含有效地連接表達(dá)控制序列的權(quán)利要求1到4中任一項(xiàng)的分離的核酸分子的表達(dá)載體。
7.用于在合適的宿主細(xì)胞內(nèi)生產(chǎn)融合多肽的宿主-載體系統(tǒng)，其包含權(quán)利要求6的表達(dá)載體。
8.權(quán)利要求7的宿主-載體系統(tǒng)，其中宿主細(xì)胞是細(xì)菌細(xì)胞、酵母細(xì)胞、昆蟲細(xì)胞或哺乳動(dòng)物細(xì)胞。
9.權(quán)利要求8的宿主-載體系統(tǒng)，其中宿主細(xì)胞是CHO細(xì)胞。
10.產(chǎn)生融合多肽的方法，包括在允許產(chǎn)生融合多肽的條件下培養(yǎng)權(quán)利要求8的宿主-載體系統(tǒng)的細(xì)胞，和回收這樣產(chǎn)生的融合多肽。
11.權(quán)利要求1的分離的核酸，其中SEQ ID NO1，3或5的核酸包含一或多個(gè)的如下改變(a)核苷酸1979位由A變成C；并且(b)將核苷酸TCC GGA插入到核苷酸996和997位之間；
12.權(quán)利要求1的分離的核酸，其中SEQ ID NO7，9，或11的核酸包含一或多個(gè)如下改變(a)SEQ ID NO7，9，或11的核苷酸1043位由A變成C；并且(b)將核苷酸TCC GGA插入到核苷酸1077和1078位點(diǎn)之間。
13.權(quán)利要求1的分離的核酸，其中SEQ ID NO13，15或17的核酸包含一或多個(gè)的如下改變(a)核苷酸2027位由A變成C；(b)將核苷酸TCC GGA插入到核苷酸1044和1045位之間。
14.權(quán)利要求1的分離的核酸，其中SEQ ID NO19，21或23的核酸包含一或多個(gè)的如下改變(a)核苷酸1043位點(diǎn)由A變成C；且(b)將核苷酸TCC GGA插入到核苷酸1074和1075位點(diǎn)之間。
15.由權(quán)利要求1或11到14中的任一項(xiàng)的核酸分子編碼的融合多肽。
16.融合多肽，其包含氨基酸序列SEQ ID NO2，4，6，8，10，12，14，16，18，20，22和24。
17.包含權(quán)利要求6的融合多肽的多聚體，其能與IL-1結(jié)合形成無功能復(fù)合物。
18.權(quán)利要求17的多聚體，其為二聚體。
19.權(quán)利要求18的多聚體，其中各融合蛋白具有SEQ ID NO8的序列。
全文摘要
本發(fā)明提供了能與白細(xì)胞介素－1(IL－1)結(jié)合而形成無功能復(fù)合物的融合多肽。本發(fā)明也提供了編碼該融合多肽的核酸序列和制造與使用該融合多肽的方法。
文檔編號(hào)A61K38/00GK1878871SQ200380102337
公開日2006年12月13日 申請(qǐng)日期2003年10月24日 優(yōu)先權(quán)日2002年10月28日
發(fā)明者N·斯塔爾, G·D·揚(yáng)科波洛斯 申請(qǐng)人:瑞澤恩制藥公司