專利名稱：非典型性肺炎病毒特異蛋白質(zhì)和臨床檢測的方法及試劑盒的制作方法
技術領域：
本發(fā)明涉及人類醫(yī)學衛(wèi)生領域，具體地說是一種非典型性肺炎病毒特異蛋白質(zhì)和臨床檢測的方法及試劑盒。
背景技術：
非典型性肺炎病毒，即SARS病毒是一種引起嚴重急性呼吸道疾病綜合癥(Severe Acute Respiratory Syndrome，SARS)，又稱非典型性肺炎的病毒，屬冠狀病毒(coronavirus)類病毒，基因組共含29,737個核苷酸。造成感染嚴重時可致人于死，病死率高達4.9％。主要通過近距離空氣飛沫和密切接觸傳播。該病毒目前還沒有有效的免疫化學檢測方法，只能根據(jù)臨床癥狀(高熱、干咳、肺部X光檢測有陰影等)來診斷，受到很大的局限，只能在受感染者發(fā)病后才能作出診斷，且不易與其它肺炎和一般流感區(qū)別。本發(fā)明根據(jù)該病毒的基因組序列分析結果，選取該病毒特有蛋白質(zhì)和多肽片斷，建立一種檢測非典型性肺炎病毒抗體的免疫學方法和含有它們的試劑盒。

發(fā)明內(nèi)容
本發(fā)明的目的是提供一種非典型性肺炎病毒特異蛋白質(zhì)和臨床檢測的方法及試劑盒。
本發(fā)明采用的技術方案如下1、一種非典型性肺炎病毒特異蛋白質(zhì)，它包括非典型性肺炎病毒的蛋白質(zhì)和經(jīng)修飾的重組非典型性肺炎病毒蛋白質(zhì)，或其具有結合人免疫球蛋白之免疫活性的片段或衍生物，具有通式Seq.ID No.--Seq.ID No.118的氨基酸序列的蛋白質(zhì)。
所說的蛋白質(zhì)相對應的抗體；是單克隆抗體，或是多克隆抗體。
2、一種非典型性肺炎病毒特異蛋白質(zhì)臨床檢測方法，其具體步驟如下1)制備上述的蛋白質(zhì)；通過本領域技術人員周知的PCR擴增技術和重組DNA技術，流程見附圖1，具體如下A、設計相應的PCR引物，同時在所用PCR引物中添加編碼相應數(shù)目氨基酸殘基對應的堿基，分別從臨床非典型性肺炎病毒標準株中擴增編碼上述蛋白質(zhì)的DNA片段；B、將所得PCR產(chǎn)物導入合適的表達載體，構建重組DNA；C、用所得重組DNA轉化合適的宿主細胞，在適于目標蛋白質(zhì)表達的條件下培養(yǎng)如此轉化的宿主細胞；D、回收并純化目標蛋白質(zhì)；2)用純化上述的蛋白質(zhì)或抗體包被一種固相支持物；所說的固相支持物包括本領域周知的有機和無機多聚物，為葡聚糖、天然或經(jīng)修飾的纖維素、聚乙烯、聚苯乙烯；或用膠體金的方法包被到試紙上；3)用比色法、分光光度法或熒光法測定的酶標記的蛋白質(zhì)或抗體，或抗體的第二抗體(IgG、IgM、IgA)；標記酶包括但不局限于氧化還原酶類，為催化酶、過氧化物酶、辣根過氧化物酶、葡萄糖氧化酶、β-半乳糖苷酶；堿性磷酸酶；乙酰膽堿酯酶；使用膠體金，或放射性同位素作為標記；4)將待檢測個體的生物學樣本及由步驟3)標記的蛋白質(zhì)加入經(jīng)包被的固相支持物上，在適合形成抗原抗體免疫結合復合物的條件下孵育一段時間；所述的“適合形成抗原抗體免疫復合物的條件”是指在適當?shù)臏囟燃皶r間條件下孵育抗原與含抗體的樣品的混合物，例如在約0-50攝氏度，標記約30分鐘至約48小時。
5)適度清洗去除任何未結合的非典型性肺炎病毒抗體；所用清洗液一般為pH約5-8的磷酸鹽緩沖溶液。
6)定量檢測孵育混合物中抗原抗體免疫復合物的存在?？刹捎枚喾N本領域周知的方法定量檢測抗原抗體復合物的形成，如以過氧化物溶液為A液及聯(lián)苯胺類物質(zhì)為B液的底物與帶有含標記的辣根過氧化物酶的抗原抗體免疫復合物作用，顯色后在波長為450nm(參比波長630nm)酶標儀讀取吸光值；將所得讀數(shù)與陰性對照的讀數(shù)加以比較，判斷所測樣本的結果。
3、一種非典型性肺炎病毒特異蛋白質(zhì)臨床檢測的試劑盒，包括1)包被有純化的蛋白質(zhì)的固相支持物；相應的經(jīng)標記的的蛋白質(zhì)和使用該試劑盒的說明(雙抗原夾心法)；2)包被有純化的抗體的固相支持物；相應的經(jīng)標記的抗體和使用該試劑盒的說明(雙抗體夾心法)；3)包被有純化的蛋白質(zhì)的固相支持物；相應的第二抗體，該第二抗體是抗IgG抗體和/或抗IgM抗體和使用該試劑盒的說明(Elisa法)。
所說的快速檢測非典型性肺炎病毒的試紙，是由噴涂過的蛋白或抗體，及噴涂有膠體金的試紙基質(zhì)粘合而成；在上述檢測試劑盒中，經(jīng)標記的非典型性肺炎病毒蛋白質(zhì)是液體的形式包裝在容器中，或便于重構的凍干物的形式置于的包裝中。在后一種情況中，檢測試劑盒中還優(yōu)選含有用于重構經(jīng)標記的非典型性肺炎病毒蛋白質(zhì)的緩沖液；在上述檢測試劑盒中，還含有提供判斷檢測結果的相應的范圍。
本發(fā)明與背景技術相比具有的有益效果是它解決了臨床對非典型性肺炎病毒感染者的早期快速診斷問題，具有較高的準確性，為臨床檢測本病提供了快速、準確的診斷方法和試劑盒。


圖1是將PCR產(chǎn)物與載體連接構建重組DNA示意圖；圖2是高壓液相色譜(HPLC)圖。
具體實施例方式
B、將所得PCR產(chǎn)物導入合適的表達載體，如附圖1所示的pET-28質(zhì)粒，構建重組DNA；1μg的質(zhì)粒DNA于37℃下經(jīng)NdeI和XhoI酶完全酶切，酚/氯仿抽提，-20℃下無水乙醇沉淀過夜。離心后沉淀溶于13μl H2O。經(jīng)過NdeI和XhoI酶充分酶切的PCR產(chǎn)物與上述預酶切的質(zhì)粒連接。
C、用所得重組DNA轉化合適的宿主細胞，在適于目標蛋白質(zhì)表達的條件下培養(yǎng)如此轉化的宿主細胞；轉化至大腸桿菌ER2566，涂布含有200μg/ml氨芐青霉素的LB選擇平板。37℃過夜培養(yǎng)。構建過程參見圖1。所述連接及轉化的方法和條件參見，J.薩姆布雷奇等著的《分子克隆實驗指南》(Molecular CloningA LaboratoryManual)P672-849(科學出版社，1998)從轉化平板上挑取抗性菌落，提取質(zhì)粒并電泳檢查，篩選重組子。通過限制性內(nèi)切酶NdeI和XhoI酶切分析，篩選含有正確插入的目的基因的重組子。對所得含有插入片段的重組質(zhì)粒測序，確證在PCR過程中目的基因片段未發(fā)生變異或出現(xiàn)差錯。
D、回收并純化目標蛋白質(zhì)；將選定的重組子菌株接種于含200μg/ml氨芐青霉素的LB培養(yǎng)基中，置于恒溫搖床培養(yǎng)，37℃培養(yǎng)至菌液0D值約0.6后加入1mM IPTG誘導，30℃繼續(xù)培養(yǎng)2小時。然后將發(fā)酵培養(yǎng)物離心收集菌體沉淀。用超聲波將所得細菌菌體破碎后再次離心，收集上清液。利用金屬鎳-Sepharose(Pharmacia公司)的親和層析柱進行蛋白的分離純化(洗脫條件0.02M pH7.4磷酸鹽緩沖溶液(PBS)，洗脫速度1ml/分鐘)。經(jīng)充分洗滌去除非特異性結合的蛋白質(zhì)后，分離并收集與金屬鎳-Sepharose結合的表達蛋白，進行SDS-PAGE電泳和高壓液相色譜分析。
附圖2為經(jīng)純化的蛋白質(zhì)的高壓液相色譜圖。
二、間接法檢測非典型性肺炎病毒抗體1)每次試驗設空白對照、陰性對照和陽性對照各1孔?？瞻卓撞患右后w；陰、陽性對照無需稀釋直接取50μl加入陰、陽性對照孔中；其余檢測孔加入樣品稀釋液，每孔100μl，再加入待測樣品10μl。充分混勻，貼上封口膠，置37℃溫育30分鐘。
2)將50倍濃縮洗板液用雙蒸水50倍稀釋后用于洗滌板孔。
3)棄去各孔中樣品，每孔加滿洗液，靜置數(shù)秒后棄去，重復5次，拍干。
4)每孔加入酶標抗人IgG工作液100μl，貼上封口膠。置37℃溫育20分鐘。
5)棄去各孔中液體，用洗板液反復洗滌5次，拍干。
6)每孔加入底物液A 50μl，再加入底物液B 50μl，輕拍混勻，置37℃避光溫育10分鐘。
7)顯色完畢后，每孔加入終止液50μl，輕拍混勻，立即以空白孔調(diào)零，置酶標儀450nm波長下測定OD值(參考波長為630nm)。實施例2、雙抗原夾心法檢測非典型性肺炎病毒抗體在96孔微孔反應板上加入預先用0.02M pH9.6的碳酸鹽緩沖液(CB)稀釋抗原至最適濃度的包被液，4℃靜置24小時，加入含0.1％TW-20的0.02M pH7.4 PBS洗滌液(PBS-T)加滿各孔，靜置數(shù)秒后棄去，重復清洗兩次。每孔加入200μl入1％牛血清白蛋白(BSA)和1％脫脂奶粉的0.02M pH7.4磷酸鹽緩沖液(PBS)，4℃下封閉2小時。棄去封閉液，室溫下晾干，用鋁薄袋封板，4℃保藏待用。在微孔反應板孔中加入預先用酶標抗原稀釋液(20％山羊血清，1％酪蛋白，0.02MPBS，pH7.4)以優(yōu)化好的濃度稀釋的SARS冠狀病毒抗原相應的酶標抗原，每孔50μl，再加入15個來自不同待檢測的受試者血清，37℃溫育60分鐘。然后用含0.1％TW-20的0.02M PBS-T洗滌液加滿各孔，靜置數(shù)秒后棄去，重復清洗5次。每孔中加入50μl由0.06％過氧化脲(Sigma)0.01M pH4.5的醋酸鹽緩沖液組成的酶作用底物A液以及50μl由0.06％TMB(Sigma)0.01M pH4.5的檸檬酸鹽緩沖液組成的酶作用底物B液，37℃溫育15分鐘。每孔中加入50μl的2MH2SO4終止反應。在酶標儀(Dynatech MR 4100)上讀取450nm處的吸光值(以630為參比波長)。其中陽性對照和陰性對照均為經(jīng)過臨床確診及其他可能的方法復核的人血清，根據(jù)實預先確定的閾值判定樣本結果，高于閾值的樣本為非典型性肺炎病毒陽性。
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Ala Arg Asp Gly Cys Val Pro Leu Asn Ile Ile Pro85 90 95Leu Thr Thr Ala Ala Lys Leu Met Val Val Val Pro Asp Tyr Gly Thr100 105110Tyr Lys Asn Thr Cys Asp Gly Asn Thr Phe Thr Tyr Ala Ser Ala Leu115 120 125Trp Glu Ile Gln Gln Val Val Asp Ala Asp Ser Lys Ile Val Gln Leu130 135140Ser Glu Ile145147<210>9<211>153<212>PRT<213>SARS coronavirus<400>9Val Val Asp Ala Asp Ser Lys Ile Val Gln Leu Ser Glu Ile Asn Met1 510 15Asp Asn Ser Pro Asn Leu Ala Trp Pro Leu Ile Val Thr Ala Leu Arg20 25 30Ala Asn Ser Ala Val Lys Leu Gln Asn Asn Glu Leu Ser Pro Val Ala35 4045Leu Arg Gln Met Ser Cys Ala Ala Gly Thr Thr Gln Thr Ala Cys Thr50 55 60Asp Asp Asn Ala Leu Ala Tyr Tyr Asn Asn Ser Lys Gly Gly Arg Phe65 70 75 80Val Leu Ala Leu Leu Ser Asp His Gln Asp Leu Lys Trp Ala Arg Phe85 90 95Pro Lys Ser Asp Gly Thr Gly Thr Ile Tyr Thr Glu Leu Glu Pro Pro100 105 110Cys Arg Phe Val Thr Asp Thr Pro Lys Gly Pro Lys Val Lys Tyr Leu115120125Tyr Phe Ile Lys Gly Leu Asn Asn Leu Asn Arg Gly Met Val Leu Gly130 135 140Ser Leu Ala Ala Thr Val Arg Leu Gln145 150 153<210>10<211>148<212>PRT<213>SARS coronavirus<400>10Gly Leu Asn Asn Leu Asn Arg Gly Met Val Leu Gly Ser Leu Ala Ala1 51015Thr Val Arg Leu Gln Ala Gly Asn Ala Thr Glu Val Pro Ala Asn Ser20 25 30Thr Val Leu Ser Phe Cys Ala Phe Ala Val Asp Pro Ala Lys Ala Tyr35 40 45Lys Asp Tyr Leu Ala Ser Gly Gly Gln Pro Ile Thr Asn Cys Val Lys50 5560Met Leu Cys Thr His Thr Gly Thr Gly Gln Ala Ile Thr Val Thr Pro65 70 75 80Glu Ala Asn Met Asp Gln Glu Ser Phe Gly Gly Ala Ser Cys Cys Leu85 90 95Tyr Cys Arg Cys His Ile Asp His Pro Asn Pro Lys Gly Phe Cys Asp100 105 110Leu Lys Gly Lys Tyr Val Gln Ile Pro Thr Thr Cys Ala Asn Asp Pro115 120 125Val Gly Phe Thr Leu Arg Asn Thr Val Cys Thr Val Cys Gly Met Trp130 135 140Lys Gly Tyr Gly145148<210>11<211>40<212>PRT<213>SARS coronavirus<400>11Arg Asn Thr Val Cys Thr Val Cys Gly Met Trp Lys Gly Tyr Gly Cys1 5 10 15Ser Cys Asp Gln Leu Arg Glu Pro Leu Met Gln Ser Ala Asp Ala Ser20 25 30Thr Phe Leu Asn Gly Phe Ala Val35 40<210>12<211>150<212>PRT<213>SARS coronavirus<400>12Met Val Pro His Ile Ser Arg Gln Arg Leu Thr Lys Tyr Thr Met Ala1 5 1015Asp Leu Val Tyr Ala Leu Arg His Phe Asp Glu Gly Asn Cys Asp Thr20 2530Leu Lys Glu Ile Leu Val Thr Tyr Asn Cys Cys Asp Asp Asp Tyr Phe35 4045Asn Lys Lys Asp Trp Tyr Asp Phe Val Glu Asn Pro Asp Ile Leu Arg5055 60Val Tyr Ala Asn Leu Gly Glu Arg Val Arg Gln Ser Leu Leu Lys Thr65 70 7580Val Gln Phe Cys Asp Ala Met Arg Asp Ala Gly Ile Val Gly Val Leu85 90 95Thr Leu Asp Asn Gln Asp Leu Asn Gly Asn Trp Tyr Asp Phe Gly Asp100 105 110Phe Val Gln Val Ala Pro Gly Cys Gly Val Pro Ile Val Asp Ser Tyr115120 125Tyr Ser Leu Leu Met Pro Ile Leu Thr Leu Thr Arg Ala Leu Ala Ala130 135 140Glu Ser His Met Asp Ala145 150<210>13<211>146<212>PRT<213>SARS coronavirus<400>13Ile Leu Thr Leu Thr Arg Ala Leu Ala Ala Glu Ser His Met Asp Ala1 5 1015Asp Leu Ala Lys Pro Leu Ile Lys Trp Asp Leu Leu Lys Tyr Asp Phe20 25 30Thr Glu Glu Arg Leu Cys Leu Phe Asp Arg Tyr Phe Lys Tyr Trp Asp3540 45Gln Thr Tyr His Pro Asn Cys Ile Asn Cys Leu Asp Asp Arg Cys Ile505560Leu His Cys Ala Asn Phe Asn Val Leu Phe Ser Thr Val Phe Pro Pro65 70 75 80Thr Ser Phe Gly Pro Leu Val Arg Lys Ile Phe Val Asp Gly Val Pro85 90 95Phe Val Val Ser Thr Gly Tyr His Phe Arg Glu Leu Gly Val Val His100 105110Asn Gln Asp Val Asn Leu His Ser Ser Arg Leu Ser Phe Lys Glu Leu115 120 125Leu Val Tyr Ala Ala Asp Pro Ala Met His Ala Ala Ser Gly Asn Leu130 135140Leu Leu145 146<210>14<211>151<212>PRT<213>SARS coronavirus<400>14Ala Ala Asp Pro Ala Met His Ala Ala Ser Gly Asn Leu Leu Leu Asp1 5 10 15Lys Arg Thr Thr Cys Phe Ser Val Ala Ala Leu Thr Asn Asn Val Ala20 25 30Phe Gln Thr Val Lys Pro Gly Asn Phe Asn Lys Asp Phe Tyr Asp Phe35 4045Ala Val Ser Lys Gly Phe Phe Lys Glu Gly Ser Ser Val Glu Leu Lys50 5560His Phe Phe Phe Ala Gln Asp Gly Asn Ala Ala Ile Ser Asp Tyr Asp65 70 7580Tyr Tyr Arg Tyr Asn Leu Pro Thr Met Cys Asp Ile Arg Gln Leu Leu85 90 95Phe Val Val Glu Val Val Asp Lys Tyr Phe Asp Cys Tyr Asp Gly Gly100 105 110Cys Ile Asn Ala Asn Gln Val Ile Val Asn Asn Leu Asp Lys Ser Ala115 120125Gly Phe Pro Phe Asn Lys Trp Gly Lys Ala Arg Leu Tyr Tyt Asp Ser130 135 140Met Ser Tyr Glu Asp Gln Asp145 150 151<210>15<211>148<212>PRT<213>SARS coronavirus<400>15Lys Trp Gly Lys Ala Arg Leu Tyr Tyr Asp Ser Met Ser Tyr Glu Asp1 5 10 15Gln Asp Ala Leu Phe Ala Tyr Thr Lys Arg Asn Val Ile Pro Thr Ile20 25 30Thr Gln Met Asn Leu Lys Tyr Ala Ile Ser Ala Lys Asn Arg Ala Arg35 40 45Thr Val Ala Gly Val Ser Ile Cys Ser Thr Met Thr Asn Arg Gln Phe50 5560His Gln Lys Leu Leu Lys Ser Ile Ala Ala Thr Arg Gly Ala Thr Val65 70 75 80Val Ile Gly Thr Ser Lys Phe Tyr Gly Gly Trp His Asn Met Leu Lys85 90 95Thr Val Tyr Ser Asp Val Glu Thr Pro His Leu Met Gly Trp Asp Tyr100 105 110Pro Lys Cys Asp Arg Ala Met Pro Asn Met Leu Arg Ile Met Ala Ser115 120 125Leu Val Leu Ala Arg Lys His Asn Thr Cys Cys Asn Leu Ser His Arg130 135 140Phe Tyr Arg Leu145148<210>16<211>154<212>PRT<213>SARS coronavirus<400>16His Asn Thr Cys Cys Asn Leu Ser His Arg Phe Tyr Arg Leu Ala Asn1 5 10 15Glu Cys Ala Gln Val Leu Ser Glu Met Val Met Cys Gly Gly Ser Leu20 25 30Tyr Val Lys Pro Gly Gly Thr Ser Ser Gly Asp Ala Thr Thr Ala Tyr35 40 45Ala Asn Ser Val Phe Asn Ile Cys Gln Ala Val Thr Ala Asn Val Asn50 55 60Ala Leu Leu Ser Thr Asp Gly Asn Lys Ile Ala Asp Lys Tyr Val Arg65 70 75 80Asn Leu Gln His Arg Leu Tyr Glu Cys Leu Tyr Arg Asn Arg Asp Val85 9095Asp His Glu Phe Val Asp Glu Phe Tyr Ala Tyr Leu Arg Lys His Phe100 105 110Ser Met Met Ile Leu Ser Asp Asp Ala Val Val Cys Tyr Asn Ser Asn115 120 125Tyr Ala Ala Gln Gly Leu Val Ala Ser Ile Lys Asn Phe Lys Ala Val130 135 140Leu Tyr Tyr Gln Asn Asn Val Phe Met Ser145 150 154<210>17<211>148<212>PRT<213>SARS coronavirus<400>17Val Ala Ser Ile Lys Asn Phe Lys Ala Val Leu Tyr Tyr Gln Asn Asn1 5 10 15Val Phe Met Ser Glu Ala Lys Cys Trp Thr Glu Thr Asp Leu Thr Lys20 25 30Gly Pro His Glu Phe Cys Ser Gln His Thr Met Leu Val Lys Gln Gly35 40 45Asp Asp Tyr Val Tyr Leu Pro Tyr Pro Asp Pro Ser Arg Ile Leu Gly50 55 60Ala Gly Cys Phe Val Asp Asp Ile Val Lys Thr Asp Gly Thr Leu Met65 70 75 80Ile Glu Arg Phe Val Ser Leu Ala Ile Asp Ala Tyr Pro Leu Thr Lys85 90 95His Pro Asn Gln Glu Tyr Ala Asp Val Phe His Leu Tyr Leu Gln Tyr100 105 110Ile Arg Lys Leu His Asp Glu Leu Thr Gly His Met Leu Asp Met Tyr115 120 125Ser Val Met Leu Thr Asn Asp Asn Thr Ser Arg Tyr Trp Glu Pro Glu130 135140Phe Tyr Glu Ala145148<210>18<211>146<212>PRT<213>SARS coronavirus<400>18Ser Phe Ser Ala Ser Thr Ser Ala Phe Ile Asp Thr Ile Lys Ser Leu1 5 1015Asp Tyr Lys Ser Phe Lys Thr Ile Val Glu Ser Cys Gly Asn Tyr Lys20 25 30Val Thr Lys Gly Lys Pro Val Lys Gly Ala Trp Asn Ile Gly Gln Gln35 40 45Arg Ser Val Leu Thr Pro Leu Cys Gly Phe Pro Ser Gln Ala Ala Gly50 55 60Val Ile Arg Ser Ile Phe Ala Arg Thr Leu Asp Ala Ala Asn His Ser65 70 7580Ile Pro Asp Leu Gln Arg Ala Ala Val Thr Ile Leu Asp Gly Ile Ser85 90 95Glu Gln Ser Leu Arg Leu Val Asp Ala Met Val Tyr Thr Ser Asp Leu100 105 110Leu Thr Asn Ser Val Ile Ile Met Ala Tyr Val Thr Gly Gly Leu Val115 120 125Gln Gln Thr Ser Gln Trp Leu Ser Asn Leu Leu Gly Thr Thr Val Glu130 135 140Lys Leu145 146<210>19<211>151<212>PRT<213>SARS coronavirus<400>19Thr Asn Asp Asn Thr Ser Arg Tyr Trp Glu Pro Glu Phe Tyr Glu Ala1 5 10 15Met Tyr Thr Pro His Thr Val Leu Gln Ala Val Gly Ala Cys Val Leu20 25 30Cys Asn Ser Gln Thr Ser Leu Arg Cys Gly Ala Cys Ile Arg Arg Pro35 40 45Phe Leu Cys Cys Lys Cys Cys Tyr Asp His Val Ile Ser Thr Ser His50 55 60Lys Leu Val Leu Ser Val Asn Pro Tyr Val Cys Asn Ala Pro Gly Cys6570 75 80Asp Val Thr Asp Val Thr Gln Leu Tyr Leu Gly Gly Met Ser Tyr Tyr85 9095Cys Lys Ser His Lys Pro Pro Ile Ser Phe Pro Leu Cys Ala Asn Gly100 105 110Gln Val Phe Gly Leu Tyr Lys Asn Thr Cys Val Gly Ser Asp Asn Val115120125Thr Asp Phe Asn Ala Ile Ala Thr Cys Asp Trp Thr Asn Ala Gly Asp130 135 140Tyr Ile Leu Ala Asn Thr Cys145 150 151<210>20<211>148<212>PRT<213>SARS coronavirus<400>20Ala Thr Cys Asp Trp Thr Asn Ala Gly Asp Tyr Ile Leu Ala Asn Thr1 510 15Cys Thr Glu Arg Leu Lys Leu Phe Ala Ala Glu Thr Leu Lys Ala Thr20 25 30Glu Glu Thr Phe Lys Leu Ser Tyr Gly Ile Ala Thr Val Arg Glu Val35 40 45Leu Ser Asp Arg Glu Leu His Leu Ser Trp Glu Val Gly Lys Pro Arg50 55 60Pro Pro Leu Asn Arg Asn Tyr Val Phe Thr Gly Tyr Arg Val Thr Lys65 70 7580Asn Ser Lys Val Gln Ile Gly Glu Tyr Thr Phe Glu Lys Gly Asp Tyr85 90 95Gly Asp Ala Val Val Tyr ArgGly Thr Thr Thr Tyr Lys Leu Asn Val100 105 110Gly Asp Tyr Phe Val Leu Thr Ser His Thr Val Met Pro Leu Ser Ala115120 125Pro Thr Leu Val Pro Gln Glu His Tyr Val Arg Ile Thr Gly Leu Tyr130 135 140Pro Thr Leu Asn145 148<210>21<211>147<212>PRT<213>SARS coronavirus<400>21Glu His Tyr Val Arg Ile Thr Gly Leu Tyr Pro Thr Leu Asn Ile Ser1 5 10 15Asp Glu Phe Ser Ser Asn Val Ala Asn Tyr Gln Lys Val Gly Met Gln20 25 30Lys Tyr Ser Thr Leu Gln Gly Pro Pro Gly Thr Gly Lys Ser His Phe35 40 45Ala Ile Gly Leu Ala Leu Tyr Tyr Pro Ser Ala Arg Ile Val Tyr Thr50 55 60Ala Cys Ser His Ala Ala Val Asp Ala Leu Cys Glu Lys Ala Leu Lys65 70 75 80Tyr Leu Pro Ile Asp Lys Cys Ser Arg Ile Ile Pro Ala Arg Ala Arg85 90 95Val Glu Cys Phe Asp Lys Phe Lys Val Asn Ser Thr Leu Glu Gln Tyr100 105 110Val Phe Cys Thr Val Asn Ala Leu Pro Glu Thr Thr Ala Asp Ile Val115 120 125Val Phe Asp Glu Ile Ser Met Ala Thr Asn Tyr Asp Leu Ser Val Val130 135 140Asn Ala Arg145 147<210>22<211>149<212>PRT<213>SARS coronavirus<400>22Glu Ile Ser Met Ala Thr Asn Tyr Asp Leu Ser Val Val Asn Ala Arg1 5 1015Leu Arg Ala Lys His Tyr Val Tyr Ile Gly Asp Pro Ala Gln Leu Pro2025 30Ala Pro Arg Thr Leu Leu Thr Lys Gly Thr Leu Glu Pro Glu Tyr Phe35 4045Asn Ser Val Cys Arg Leu Met Lys Thr Ile Gly Pro Asp Met Phe Leu50 55 60Gly Thr Cys Arg Arg Cys Pro Ala Glu Ile Val Asp Thr Val Ser Ala65 70 7580Leu Val Tyr Asp Asn Lys Leu Lys Ala His Lys Asp Lys Ser Ala Gln85 90 95Cys Phe Lys Met Phe Tyr Lys Gly Val Ile Thr His Asp Val Ser Ser100105 110Ala Ile Asn Arg Pro Gln Ile Gly Val Val Arg Glu Phe Leu Thr Arg115 120125Asn Pro Ala Trp Arg Lys Ala Val Phe Ile Ser Pro Tyr Asn Ser Gln130 135 140Asn Ala Val Ala Ser145149<210>23<211>150<212>PRT<213>SARS coronavirus<400>23Ala Val Phe Ile Ser Pro Tyr Asn Ser Gln Asn Ala Val Ala Ser Lys1510 15Ile Leu Gly Leu Pro Thr Gln Thr Val Asp Ser Ser Gln Gly Ser Glu20 25 30Tyr Asp Tyr Val Ile Phe Thr Gln Thr Thr Glu Thr Ala His Ser Cys35 40 45Asn Val Asn Arg Phe Asn Val Ala Ile Thr Arg Ala Lys Ile Gly Ile50 55 60Leu Cys Ile Met Ser Asp Arg Asp Leu Tyr Asp Lys Leu Gln Phe Thr65 70 75 80Ser Leu Glu Ile Pro Arg Arg Asn Val Ala Thr Leu Gln Ala Glu Asn85 90 95Val Thr Gly Leu Phe Lys Asp Cys Ser Lys Ile Ile Thr Gly Leu His100 105 110Pro Thr Gln Ala Pro Thr His Leu Ser Val Asp Ile Lys Phe Lys Thr115 120125Glu Gly Leu Cys Val Asp Ile Pro Gly Ile Pro Lys Asp Met Thr Tyr130 135 140Arg Arg Leu Ile Ser Met145150<210>24<211>158<212>PRT<213>SARS coronavirus<400>24Pro Gly Ile Pro Lys Asp Met Thr Tyr Arg Arg Leu Ile Ser Met Met1 5 10 15Gly Phe Lys Met Asn Tyr Gln Val Asn Gly Tyr Pro Asn Met Phe Ile20 25 30Thr Arg Glu Glu Ala Ile Arg His Val Arg Ala Trp Ile Gly Phe Asp35 40 45Val Glu Gly Cys His Ala Thr Arg Asp Ala Val Gly Thr Asn Leu Pro50 55 60Leu Gln Leu Gly Phe Ser Thr Gly Val Asn Leu Val Ala Val Pro Thr65 7075 80Gly Tyr Val Asp Thr Glu Asn Asn Thr Glu Phe Thr Arg Val Asn Ala85 90 95Lys Pro Pro Pro Gly Asp Gln Phe Lys His Leu Ile Pro Leu Met Tyr100 105110Lys Gly Leu Pro Trp Asn Val Val Arg Ile Lys Ile Val Gln Met Leu115 120 125Ser Asp Thr Leu Lys Gly Leu Ser Asp Arg Val Val Phe Val Leu Trp130 135 140Ala His Gly Phe Glu Leu Thr Ser Met Lys Tyr Phe Val Lys145150 155 158<210>25<211>146<212>PRT<213>SARS coronavirus<400>25Leu Ser Asp Arg Val Val Phe Val Leu Trp Ala His Gly Phe Glu Leu1 5 10 15Thr Ser Met Lys Tyr Phe Val Lys Ile Gly Pro Glu Arg Thr Cys Cys2025 30Leu Cys Asp Lys Arg Ala Thr Cys Phe Ser Thr Ser Ser Asp Thr Tyr35 40 45Ala Cys Trp Asn His Ser Val Gly Phe Asp Tyr Val Tyr Asn Pro Phe50 55 60Met Ile Asp Val Gln Gln Trp Gly Phe Thr Gly Asn Leu Gln Ser Asn65 70 75 80His Asp Gln His Cys Gln Val His Gly Asn Ala His Val Ala Ser Cys85 9095Asp Ala Ile Met Thr Arg Cys Leu Ala Val His Glu Cys Phe Val Lys100105 110Arg Val Asp Trp Ser Val Glu Tyr Pro Ile Ile Gly Asp Glu Leu Arg115 120125Val Asn Ser Ala Cys Arg Lys Val Gln His Met Val Val Lys Ser Ala130 135140Leu Leu145 146<210>26<211>146<212>PRT<213>SARS coronavirus<400>26Arg Lys Val Gln His Met Val Val Lys Ser Ala Leu Leu Ala Asp Lys1 5 1015Phe Pro Val Leu His Asp Ile Gly Asn Pro Lys Ala Ile Lys Cys Val20 25 30Pro Gln Ala Glu Val Glu Trp Lys Phe Tyr Asp Ala Gln Pro Cys Ser35 40 45Asp Lys Ala Tyr Lys Ile Glu Glu Leu Phe Tyr Ser Tyr Ala Thr His50 55 60His Asp Lys Phe Thr Asp Gly Val Cys Leu Phe Trp Asn Cys Asn Val65 70 75 80Asp Arg Tyr Pro Ala Asn Ala Ile Val Cys Arg Phe Asp Thr Arg Val85 9095Leu Ser Asn Leu Asn Leu Pro Gly Cys Asp Gly Gly Ser Leu Tyr Val100 105110Asn Lys His Ala Phe His Thr Pro Ala Phe Asp Lys Ser Ala Phe Thr115 120125Asn Leu Lys Gln Leu Pro Phe Phe Tyr Tyr Ser Asp Ser Pro Cys Glu130 135 140Ser His145 146<210>27<211>151<212>PRT<2l3>SARS coronavirus<400>27Pro Phe Phe Tyr Tyr Ser Asp Ser Pro Cys Glu Ser His Gly Lys Gln1 5 10 15Val Val Ser Asp Ile Asp Tyr Val Pro Leu Lys Ser Ala Thr Cys Ile20 25 30Thr Arg Cys Asn Leu Gly Gly Ala Val Cys Arg His His Ala Asn Glu35 40 45Tyr Arg Gln Tyr Leu Asp Ala Tyr Asn Met Met Ile Ser Ala Gly Phe50 55 60Ser Leu Trp Ile Tyr Lys Gln Phe Asp Thr Tyr Asn Leu Trp Asn Thr65 70 75 80Phe Thr Arg Leu Gln Ser Leu Glu Asn Val Ala Tyr Asn Val Val Asn85 9095Lys Gly His Phe Asp Gly His Ala Gly Glu Ala Pro Val Ser Ile Ile100 105 110Asn Asn Ala Val Tyr Thr Lys Val Asp Gly Ile Asp Val Glu Ile Phe115120 125Glu Asn Lys Thr Thr Leu Pro Val Asn Val Ala Phe Glu Leu Trp Ala130 135 140Lys Arg Asn Ile Lys Pro Val145 150 151<210>28<211>148<212>PRT<213>SARS coronavirus<400>28Pro Val Asn Val Ala Phe Glu Leu Trp Ala Lys Arg Asn Ile Lys Pro1 5 10 15Val Pro Glu Ile Lys Ile Leu Asn Asn Leu Gly Val Asp Ile Ala Ala20 25 30Asn Thr Val Ile Trp Asp Tyr Lys Arg Glu Ala Pro Ala His Val Ser35 40 45Thr Ile Gly Val Cys Thr Met Thr Asp Ile Ala Lys Lys Pro Thr Glu50 55 60Ser Ala Cys Ser Ser Leu Thr Val Leu Phe Asp Gly Arg Val Glu Gly65 70 75 80Gln Val Asp Leu Phe Arg Asn Ala Arg Asn Gly Val Leu Ile Thr Glu85 90 95Gly Ser Val Lys Gly Leu Thr Pro Ser Lys Gly Pro Ala Gln Ala Ser100 105 110Val Asn Gly Val Thr Leu Ile Gly Glu Ser Val Lys Thr Gln Phe Asn115 120 125Tyr Phe Lys Lys Val Asp Gly Ile Ile Gln Gln Leu Pro Glu Thr Tyr130 135 140Phe Thr Gln Ser145148<210>29<211>144<212>PRT<213>SARS coronavirus<400>29Ser Gly Cys Leu Ile Phe Trp Ala Leu Leu Leu Lys Asn Ser Gly Leu1 5 10 15Ser Leu Asn Gly Leu Arg Arg Asn Leu Val Gln Glu Leu Asn Phe Ser20 25 30Arg Met Leu Gly Arg Phe Ser Asn Phe Ser Leu Gln Val Phe Leu Thr35 40 45Ser Ser Arg Val Lys Tyr Arg Leu Leu Gln Ile Thr Ser Arg Ile Val50 5560Asn Ala Ser Leu Met Leu Leu Thr Arg His Ser Lys Cys Ala Leu Ile65 70 75 80Lys Ser Leu Ser Leu Ala Gln Ser Cys Asp His Ser Thr Val Lys Ser859095Ser Ser Leu Lys Ala Arg Asp Phe Thr Val Ser Val Tys Val Ala Arg100 105 110Ser Ser Cys Asn Tyr Ser Cys Leu Leu Arg His Gln Lys Lys Pro Phe115 120125Leu Lys Val Ile His Met Thr Gln Tyr Leu Pro Leu Arg Arg Leu Phe144<210>30<211>150<212>PRT<213>SARS coronavirus<400>30Asp Gly Ile Ile Gln Gln Leu Pro Glu Thr Tyr Phe Thr Gln Ser Arg1 5 1015Asp Leu Glu Asp Phe Lys Pro Arg Ser Gln Met Glu Thr Asp Phe Leu20 2530Glu Leu Ala Met Asp Glu Phe Ile Gln Arg Tyr Lys Leu Glu Gly Tyr35 40 45Ala Phe Glu His Ile Val Tyr Gly Asp Phe Ser His Gly Gln Leu Gly50 55 60Gly Leu His Leu Met Ile Gly Leu Ala Lys Arg Ser Gln Asp Ser Pro65 70 75 80Leu Lys Leu Glu Asp Phe Ile Pro Met Asp Ser Thr Val Lys Asn Tyr85 90 95Phe Ile Thr Asp Ala Gln Thr Gly Ser Ser Lys Cys Val Cys Ser Val100 105 110Ile Asp Leu Leu Leu Asp Asp Phe Val Glu Ile Ile Lys Ser Gln Asp115 120 125Leu Ser Val Ile Ser Lys Val Val Lys Val Thr Ile Asp Tyr Ala Glu130 135 140Ile Ser Phe Met Leu Trp145150<210>31<211>146<212>PRT<213>SARS coronavirus<400>31Val Val Lys Val Thr Ile Asp Tyr Ala Glu Ile Ser Phe Met Leu Trp1 5 10 15Cys Lys Asp Gly His Val Glu Thr Phe Tyr Pro Lys Leu Gln Ala Ser20 2530Arg Ala Trp Gln Pro Gly Val Ala Met Pro Asn Leu Tyr Lys Met Gln35 40 45Arg Met Leu Leu Glu Lys Cys Asp Leu Gln Asn Tyr Gly Glu Asn Ala50 5560Val Ile Pro Lys Gly Ile Met Met Asn Val Ala Lys Tyr Thr Gln Leu6570 75 80Cys Gln Tyr Leu Asn Thr Leu Thr Leu Ala Val Pro Tyr Asn Met Arg85 9095Val Ile His Phe Gly Ala Gly Ser Asp Lys Gly Val Ala Pro Gly Thr100105 110Ala Val Leu Arg Gln Trp Leu Pro Thr Gly Thr Leu Leu Val Asp Ser115120 125Asp Leu Asn Asp Phe Val Ser Asp Ala Tyr Ser Thr Leu Ile Gly Asp130 135140Cys Ala145 146<210>32<211>151<212>PRT<213>SARS coronavirus<400>32Asp Leu Asn Asp Phe Val Ser Asp Ala Tyr Ser Thr Leu Ile Gly Asp1 5 10 15Cys Ala Thr Val His Thr Ala Asn Lys Trp Asp Leu Ile Ile Ser Asp20 25 30Met Tyr Asp Pro Arg Thr Lys His Val Thr Lys Glu Asn Asp Ser Lys35 40 45Glu Gly Phe Phe Thr Tyr Leu Cys Gly Phe Ile Lys Gln Lys Leu Ala50 55 60Leu Gly Gly Ser Ile Ala Val Lys Ile Thr Glu His Ser Trp Asn Ala65 7075 80Asp Leu Tyr Lys Leu Met Gly His Phe Ser Trp Trp Thr Ala Phe Val85 90 95Thr Asn Val Asn Ala Ser Ser Ser Glu Ala Phe Leu Ile Gly Ala Asn100 105 110Tyr Leu Gly Lys Pro Lys Glu Gln Ile Asp Gly Tyr Thr Met His Ala115120125Asn Tyr Ile Phe Trp Arg Asn Thr Asn Pro Ile Gln Leu Ser Ser Tyr130 135 140Ser Leu Phe Asp Met Ser Lys145 150 151<210>33<21l>149<212>PRT<2l3>SARS coronavirus<400>33Ile Gln Leu Ser Ser Tyr Ser Leu Phe Asp Met Ser Lys Phe Pro Leu1 5 10 15Lys Leu Arg Gly Thr Ala Val Met Ser Leu Lys Glu Asn Gln Ile Asn20 2530Asp Met Ile Tyr Ser Leu Leu Glu Lys Gly Arg Leu Ile Ile Arg Glu35 40 45Asn Asn Arg Val Val Val Ser Ser Asp Ile Leu Val Asn Asn Thr Asn50 55 60Met Phe Ile Phe Leu Leu Phe Leu Thr Leu Thr Ser Gly Ser Asp Leu65 70 75 80Asp Arg Cys Thr Thr Phe Asp Asp Val Gln Ala Pro Asn Tyr Thr Gln85 9095His Thr Ser Ser Met Arg Gly Val Tyr Tyr Pro Asp Glu Ile Phe Arg100 105 110Ser Asp Thr Leu Tyr Leu Thr Gln Asp Leu Phe Leu Pro Phe Tyr Ser115 120 125Asn Val Thr Gly Phe His Thr Ile Asn His Thr Phe Gly Asn Pro Val130 135 140Ile Pro Phe Lys Asp145149<210>34<211>152<212>PRT<213>SARS coronavirus<400>34Asn Val Thr Gly Phe His Thr Ile Asn His Thr Phe Gly Asn Pro Val1 5 10 15Ile Pro Phe Lys Asp Gly Ile Tyr Phe Ala Ala Thr Glu Lys Ser Asn20 25 30Val Val Arg Gly Trp Val Phe Gly Ser Thr Met Asn Asn Lys Ser Gln35 40 45Ser Val Ile Ile Ile Asn Asn Ser Thr Asn Val Val Ile Arg Ala Cys50 55 60Asn Phe Glu Leu Cys Asp Asn Pro Phe Phe Ala Val Ser Lys Pro Met65 70 7580Gly Thr Gln Thr His Thr Met Ile Phe Asp Asn Ala Phe Asn Cys Thr85 9095Phe Glu Tyr Ile Ser Asp Ala Phe Ser Leu Asp Val Ser Glu Lys Ser100 105 110Gly Asn Phe Lys His Leu Arg Glu Phe Val Phe Lys Asn Lys Asp Gly115 120 125Phe Leu Tyr Val Tyr Lys Gly Tyr Gln Pro Ile Asp Val Val Arg Asp130 135 140Leu Pro Ser Gly Phe Asn Thr Leu145 150152<210>35<211>152<212>PRT<213>SARS coronavirus<400>35Tyr Gln Pro Ile Asp Val Val Arg Asp Leu Pro Ser Gly Phe Asn Thr1 510 15Leu Lys Pro Ile Phe Lys Leu Pro Leu Gly Ile Asn Ile Thr Asn Phe2025 30Arg Ala Ile Leu Thr Ala Phe Ser Pro Ala Gln Asp Ile Trp Gly Thr35 40 45Ser Ala Ala Ala Tyr Phe Val Gly Tyr Leu Lys Pro Thr Thr Phe Met50 55 60Leu Lys Tyr Asp Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ser65 70 7580Gln Asn Pro Leu Ala Glu Leu Lys Cys Ser Val Lys Ser Phe Glu Ile85 90 95Asp Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Val Pro Ser Gly100 105 110Asp Val Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu115120 125Val Phe Asn Ala Thr Lys Phe Pro Ser Val Tyr Ala Trp Glu Arg Lys130 135 140Lys Ile Ser Asn Cys Val Ala Asp145 150152<210>36<211>153<212>PRT<213>SARS coronavirus<400>36Lys Phe Pro Ser Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys1 5 10 15Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe20 25 30Lys Cys Tyr Gly Val Ser A1a Thr Lys Leu Asn Asp Leu Cys Phe Ser35 40 45Asn Val Tyr Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln50 55 60Ile Ala Pro Gly Gln Thr Gly ValIle Ala Asp Tyr Asn Tyr Lys Leu65 70 75 80Pro Asp Asp Phe Met Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile85 90 95Asp Ala Thr Ser Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg100 105 110His Gly Lys Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe115 120 125Ser Pro Asp Gly Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp130 135 140Pro Leu Asn Asp Tyr Gly Phe Tyr Thr145 150 153<210>37<211>146<212>PRT<213>SARS coronavirus<400>37Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn Asp Tyr Gly Phe Tyr Thr1 5 10 15Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe20 25 30Glu Leu Leu Asn Ala Pro Ala Thr Val Cys Gly Pro Lys Leu Ser Thr35 40 45Asp Leu Ile Lys Asn Gln Cys Val Asn Phe Asn Phe Asn Gly Leu Thr50 5560Gly Thr Gly Val Leu Thr Pro Ser Ser Lys Arg Phe Gln Pro Phe Gln65 70 75 80Gln Phe Gly Arg Asp Val Ser Asp Phe Thr Asp Ser Val Arg Asp Pro85 90 95Lys Thr Ser Glu Ile Leu Asp Ile Ser Pro Cys Ala Phe Gly Gly Val100 105110Ser Val Ile Thr Pro Gly Thr Asn Ala Ser Ser Glu Val Ala Val Leu115 120125Tyr Gln Asp Val Asn Cys Thr Asp Val Ser Thr Ala Ile His Ala Asp130 135 140Gln Leu145 146<210>38<211>147<212>PRT<213>SARS coronavirus<400>38Cys Thr Asp Val Ser Thr Ala Ile His Ala Asp Gln Leu Thr Pro Ala1 510 15Trp Arg Ile Tyr Ser Thr Gly Asn Asn Val Phe Gln Thr Gln Ala Gly20 25 30Cys Leu Ile Gly Ala Glu His Val Asp Thr Ser Tyr Glu Cys Asp Ile35 40 45Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr His Thr Val Ser Leu Leu50 55 60Arg Ser Thr Ser Gln Lys Ser Ile Val Ala Tyr Thr Met Ser Leu Gly65 70 7580Ala Asp Ser Ser Ile Ala Tyr Ser Asn Asn Thr Ile Ala Ile Pro Thr85 90 95Asn Phe Ser Ile Ser Ile Thr Thr Glu Val Met Pro Val Ser Met Ala100 105 110Lys Thr Ser Val Asp Cys Asn Met Tyr Ile Cys Gly Asp Ser Thr Glu115 120 125Cys Ala Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn130 135 140Arg Ala Leu145 147<210>39<211>147<212>PRT<213>SARS coronavirus<400>39Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Ser Gly1 5 10 15Ile Ala Ala Glu Gln Asp Arg Asn Thr Arg Glu Val Phe Ala Gln Val2025 30Lys Gln Met Tyr Lys Thr Pro Thr Leu Lys Tyr Phe Gly Gly Phe Asn35 40 45Phe Ser Gln Ile Leu Pro Asp Pro Leu Lys Pro Thr Lys Arg Ser Phe50 55 60Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe65 70 75 80Met Lys Gln Tyr Gly Glu Cys Leu Gly Asp Ile Asn Ala Arg Asp Leu85 90 95Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu100 105 110Thr Asp Asp Met Ile Ala Ala Tyr Thr Ala Ala Leu Val Ser Gly Thr115 120 125Ala Thr Ala Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro130 135 140Phe Ala Met145 147<210>40<211>146<212>PRT<213>SARS coronavirus<400>40Asp Ser His Asp Thr Val Leu Thr Ser Glu Glu Val Val Leu Lys Asn1 5 10 15Gly Glu Leu Glu Ala Leu Glu Thr Pro Val Asp Ser Phe Thr Asn Gly20 25 30Ala Ile Val Gly Thr Pro Val Cys Val Asn Gly Leu Met Leu Leu Glu35 40 45Ile Lys Asp Lys Glu Gln Tyr Cys Ala Leu Ser Pro Gly Leu Leu Ala505560Thr Asn Asn Val Phe Arg Leu Lys Gly Gly Ala Pro Ile Lys Gly Val65 70 7580Thr Phe Gly Glu Asp Thr Val Trp Glu Val Gln Gly Tyr Lys Asn Val85 90 95Arg Ile Thr Phe Glu Leu Asp Glu Arg Val Asp Lys Val Leu Asn Glu100 105 110Lys Cys Ser Val Tyr Thr Val Glu Ser Gly Thr Glu Val Thr Glu Phe115 120 125Ala Cys Val Val Ala Glu Ala Val Val Lys Thr Leu Gln Pro Val Ser130 135 140Asp Leu145 146<210>41<211>149<212>PRT<213>SARS coronavirus<400>41Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln1 5 10 15Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr20 25 30Glu Asn Gln Lys Gln Ile Ala Asn Gln Phe Asn Lys Ala Ile Ser Gln35 40 45Ile Gln Glu Ser Leu Thr Thr Thr Ser Thr Ala Leu Gly Lys Leu Gln50 5560Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln65 70 75 80Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu8590 95Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile100 105 110Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile115 120 125Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met130 135 140Ser Glu Cys Val Leu145 149<210>42<211>150<212>PRT<213>SARS coronavirus<400>42Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu1 5 10 15Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met202530Ser Phe Pro Gln Ala Ala Pro His Gly Val Val Phe Leu His Val Thr35 4045Tyr Val Pro Ser Gln Glu Arg Asn Phe Thr Thr Ala Pro Ala Ile Cys50 55 60His Glu Gly Lys Ala Tyr Phe Pro Arg Glu Gly Val Phe Val Phe Asn65 70 75 80Gly Thr Ser Trp Phe Ile Thr Gln Arg Asn Phe Phe Ser Pro Gln Ile85 9095Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val ValIle100 105 110Gly Ile Ile Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp115120 125Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro130 135 140Asp Val Asp Leu Gly Asp145150<210>43<211>120<212>PRT<213>SARS coronavirus<400>43Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile1 510 15Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg20 25 30Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln35 40 45Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Val Trp50 55 60Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Leu65 70 75 80Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Ala Cys Ser85 9095Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu100105 110Lys Gly Val Lys Leu His Tyr Thr115 120<210>44<211>15l<212>PRT<213>SARS coronavirus<400>44Thr Leu Gly Ser Ile Thr Ala Gln Pro Val Lys Ile Asp Asn Ala Ser1 51015Pro Ala Ser Thr Val His Ala Thr Ala Thr Ile Pro Leu Gln Ala Ser20 25 30Leu Pro Phe Gly Trp Leu Val Ile Gly Val Ala Phe Leu Ala Val Phe35 40 45Gln Ser Ala Thr Lys Ile Ile Ala Leu Asn Lys Arg Trp Gln Leu Ala50 55 60Leu Tyr Lys Gly Phe Gln Phe Ile Cys Asn Leu Leu Leu Leu Phe Val65 70 7580Thr Ile Tyr Ser His Leu Leu Leu Val Ala Ala Gly Met Glu Ala Gln85 90 95Phe Leu Tyr Leu Tyr Ala Leu Ile Tyr Phe Leu Gln Cys Ile Asn Ala100 105 110Cys Arg Ile Ile Met Arg Cys Trp Leu Cys Trp Lys Cys Lys Ser Lys115120 125Asn Pro Leu Leu Tyr Asp Ala Asn Tyr Phe Val Cys Trp His Thr His130 135 140Asn Tyr Asp Tyr Cys Ile Pro145 150 151<210>45<211>133<212>PRT<213>SARS coronavirus<400>45Asp Ala Asn Tyr Phe Val Cys Trp His Thr His Asn Tyr Asp Tyr Cys1 5 10 15Ile Pro Tyr Asn Ser Val Thr Asp Thr Ile Val Val Thr Glu Gly Asp20 25 30Gly Ile Ser Thr Pro Lys Leu Lys Glu Asp Tyr Gln Ile Gly Gly Tyr35 4045Ser Glu Asp Arg His Ser Gly Val Lys Asp Tyr Val Val Val His Gly50 55 60Tyr Phe Thr Glu Val Tyr Tyr Gln Leu Glu Ser Thr Gln Ile Thr Thr65 70 75 80Asp Thr Gly Ile Glu Asn Ala Thr Phe Phe Ile Phe Asn Lys Leu Val85 90 95Lys Asp Pro Pro Asn Val Gln Ile His Thr Ile Asp Gly Ser Ser Gly100 105 110Val Ala Asn Pro Ala Met Asp Pro Ile Tyr Asp Glu Pro Thr Thr Thr115 120125Thr Ser Val Pro Leu130 133<210>46<211>83<212>PRT<213>SARS coronavirus<400>46Gln Glu Ser Glu Tyr Glu Leu Met Tyr Ser Phe Val Ser Glu Glu Thr1 5 10 15Glv Thr Leu Ile Val Asn Ser Val Leu Leu Phe Leu Ala Phe Val Val20 25 30Phe Leu Leu Val Thr Leu Ala Ile Leu Thr Ala Leu Arg Leu Cys Ala35 40 45Tyr Cys Cys Asn Ile Val Asn Val Ser Leu Val Lys Pro Thr Val Tyr50 55 60Val Tyr Ser Arg Val Lys Asn Leu Asn Ser Ser Glu Gly Val Pro Asp65 70 7580Leu Leu Val83<210>47<211>150<212>PRT<213>SARS coronavirus<400>47Leu Leu Gln Phe Ala Tyr Ser Asn Arg Asn Arg Phe Leu Tyr Ile Ile1 5 10 15Lys Leu Val Phe Leu Trp Leu Leu Trp Pro Val Thr Leu Ala Cys Phe20 25 30Val Leu Ala Ala Val Tyr Arg Ile Asn Trp Val Thr Gly Gly Ile Ala354045Ile Ala Met Ala Cys Ile Val Gly Leu Met Trp Leu Ser Tyr Phe Val50 55 60Ala Ser Phe Arg Leu Phe Ala Arg Thr Arg Ser Met Trp Ser Phe Asn65 70 75 80Pro Glu Thr Asn Ile Leu Leu Asn Val Pro Leu Arg Gly Thr Ile Val85 9095Thr Arg Pro Leu Met Glu Ser Glu Leu Val Ile Gly Ala ValIle Ile100105 110Arg Gly His Leu Arg Met Ala Gly His Ser Leu Gly Arg Cys Asp Ile115 120125Lys Asp Leu Pro Lys Glu Ile Thr Val Ala Thr Ser Arg Thr Leu Ser130 135 140Tyr Tyr Lys Leu Gly Ala145 150<210>48<211>55<212>PRT<213>SARS coronavirus<400>48Ile Thr Val Ala Thr Ser Arg Thr Leu Ser Tyr Tyr Lys Leu Gly Ala1 510 15Ser Gln Arg Val Gly Thr Asp Ser Gly Phe Ala Ala Tyr Asn Arg Tyr202530Arg Ile Gly Asn Tyr Lys Leu Asn Thr Asp His Ala Gly Ser Asn Asp35 40 45Asn Ile Ala Leu Leu Val Gln50 55<210>49<211>117<212>PRT<213>SARS coronavirus<400>49Phe Leu Thr Leu Ile Val Phe Thr Ser Cys Glu Leu Tyr His Tyr Gln1 5 10 15Glu Cys Val Arg Gly Thr Thr Val Leu Leu Lys Glu Pro Cys Pro Ser20 25 30Gly Thr Tyr Glu Gly Asn Ser Pro Phe His Pro Leu Ala Asp Asn Lys35 40 45Phe Ala Leu Thr Cys Thr Ser Thr His Phe Ala Phe Ala Cys Ala Asp50 55 60Gly Thr Arg His Thr Tyr Gln Leu Arg Ala Arg Ser Val Ser Pro Lys65 70 75 80Leu Phe Ile Arg Gln Glu Glu Val Gln Gln Glu Leu Tyr Ser Pro Leu85 90 95Phe Leu Ile Val Ala Ala Leu Val Phe Leu Ile Leu Cys Phe Thr Ile100 105 110Lys Arg Lys Thr Glu115 117<210>50<211>55<212>PRT<213>SARS coronavirus<400>50Ala Ile Pro Cys Phe Asn Asn Ala Tyr Tyr Ile Leu Val Phe Thr Arg1 5 1015Asn Pro Gly Ser Arg Arg Thr Leu Tyr Gln Ser Leu Asn Glu His Glu20 25 30Thr Ser His Cys Phe Asp Leu Tyr Phe Ser Met Gln Leu His Met His35 4045Cys Ser Thr Ala Leu Cys Ile50 55<210>51<211>147<212>PRT<213>SARS coronavirus<400>51Ala Val Val Lys Thr Leu Gln Pro Val Ser Asp Leu Leu Thr Asn Met1 5 1015Gly Ile Asp Leu Asp Glu Trp Ser Val Ala Thr Phe Tyr Leu Phe Asp20 25 30Asp Ala Gly Glu Glu Asn Phe Ser Ser Arg Met Tyr Cys Ser Phe Tyr35 40 45Pro Pro Asp Glu Glu Glu Glu Asp Asp Ala Glu Cys Glu Glu Glu Glu50 55 60Ile Asp Glu Thr Cys Glu His Glu Tyr Gly Thr Glu Asp Asp Tyr Gln65 70 75 80Gly Leu Pro Leu Glu Phe Gly Ala Ser Ala Glu Thr Val Arg Val Glu85 9095Glu Glu Glu Glu Glu Asp Trp Leu Asp Asp Thr Thr Glu Gln Ser Glu100 105 110Ile Glu Pro Glu Pro Glu Pro Thr Pro Glu Glu Pro Val Asn Gln Phe115 120 125Thr Gly Tyr Leu Lys Leu Thr Asp Asn Val Ala Ile Lys Cys Val Asp130 135 140Ile Val Lys145 147<210>52<211>146<212>PRT<213>SARS coronavirus<400>52Asn Glu Gln Ile Lys Met Ser Asp Asn Gly Pro Gln Ser Asn Gln Arg1 5 10 15Ser Ala Pro Arg Ile Thr Phe Gly Gly Pro Thr Asp Ser Thr Asp Asn20 25 30Asn Gln Asn Gly Gly Arg Asn Gly Ala Arg Pro Lys Gln Arg Arg Pro35 40 45Gln Gly Leu Pro Asn Asn Thr Ala Ser Trp Phe Thr Ala Leu Thr Gln50 55 60His Gly Lys Glu Glu Leu Arg Phe Pro Arg Gly Gln Gly Val Pro Ile65 70 75 80Asn Thr Asn Ser Gly Pro Asp Asp Gln Ile Gly Tyr Tyr Arg Arg Ala85 90 95Thr Arg Arg Val Arg Gly Gly Asp Gly Lys Met Lys Glu Leu Ser Pro100 105 110Arg Trp Tyr Phe Tyr Tyr Leu Gly Thr Gly Pro Glu Ala Ser Leu Pro115 120 125Tyr Gly Ala Asn Lys Glu Gly Ile Val Trp Val Ala Thr Glu Gly Ala130 135 140Leu Asn145 146<210>53<211>147<212>PRT<213>SARS coronavirus<400>53Asn Lys Glu Gly Ile Val Trp Val Ala Thr Glu Gly Ala Leu Asn Thr1 5 1015Pro Lys Asp His Ile Gly Thr Arg Asn Pro Asn Asn Asn Ala Ala Thr20 25 30Val Leu Gln Leu Pro Gln Gly Thr Thr Leu Pro Lys Gly Phe Tyr Ala35 40 45Glu Gly Ser Arg Gly Gly Ser Gln Ala Ser Ser Arg Ser Ser Ser Arg50 5560Ser Arg Gly Asn Ser Arg Asn Ser Thr Pro Gly Ser Ser Arg Gly Asn65 70 75 80Ser Pro Ala Arg Met Ala Ser Gly Gly Gly Glu Thr Ala Leu Ala Leu85 90 95Leu Leu Leu Asp Arg Leu Asn Gln Leu Glu Ser Lys Val Ser Gly Lys100105 110Gly Gln Gln Gln Gln Gly Gln Thr Val Thr Lys Lys Ser Ala Ala Glu115 120 125Ala Ser Lys Lys Pro Arg Gln Lys Arg Thr Ala Thr Lys Gln Tyr Asn130 135 140Val Thr Gln145 147<210>54<211>146<212>PRT<213>SARS coronavirus<400>54Arg Gln Lys Arg Thr Ala Thr Lys Gln Tyr Asn Val Thr Gln Ala Phe1 5 10 15Gly Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp Gln Asp20 2530Leu Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile Ala Gln35 40 45Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile Gly Met50 55 60Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr His Gly Ala Ile Lys65 70 7580Leu Asp Asp Lys Asp Pro Gln Phe Lys Asp Asn Val Ile Leu Leu Asn85 90 95Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys100 105 110Asp Lys Lys Lys Lys Thr Asp Glu Ala Gln Pro Leu Pro Gln Arg Gln115 120 125Lys Lys Gln Pro Thr Val Thr Leu Leu Pro Ala Ala Asp Met Asp Asp130 135 140Phe Ser145 146<210>55<211>51<212>PRT<213>SARS coronavirus<400>55Asp Lys Lys Lys Lys Thr Asp Glu Ala Gln Pro Leu Pro Gln Arg Gln1 5 10 15Lys Lys Gln Pro Thr Val Thr Leu Leu Pro Ala Ala Asp Met Asp Asp20 25 30Phe Ser Arg Gln Leu Gln Asn Ser Met Ser Gly Ala Ser Ala Asp Ser35 40 45Thr Gln Ala50 51<210>56<211>51<212>PRT<213>SARS coronavirus<400>56Asp Lys Lys Lys Lys Thr Asp Glu Ala Gln Pro Leu Pro Gln Arg Gln1 5 10 15Lys Lys Gln Pro Thr Val Thr Leu Leu Pro Ala Ala Asp Met Asp Asp20 25 30Phe Ser Arg Gln Leu Gln Asn Ser Met Ser Gly Ala Ser Ala Asp Ser3540 45Thr Gln Ala50 51<210>57<211>150<212>PRT<213>SARS coronavirus<400>57Tyr Leu Lys Leu Thr Asp Asn Val Ala Ile Lys Cys Val Asp Ile Val1 510 15Lys Glu Ala Gln Ser Ala Asn Pro Met Val Ile Val Asn Ala Ala Asn20 2530Ile His Leu Lys His Gly Gly Gly Val Ala Gly Ala Leu Asn Lys Ala35 40 45Thr Asn Gly Ala Met Gln Lys Glu Ser Asp Asp Tyr Ile Lys Leu Asn50 55 60Gly Pro Leu Thr Val Gly Gly Ser Cys Leu Leu Ser Gly His Asn Leu65 70 75 80Ala Lys Lys Cys Leu His Val Val Gly Pro Asn Leu Asn Ala Gly Glu85 90 95Asp Ile Gln Leu Leu Lys Ala Ala Tyr Glu Asn Phe Asn Ser Gln Asp100 105 110Ile Leu Leu Ala Pro Leu Leu Ser Ala Gly Ile Phe Gly Ala Lys Pro115120 125Leu Gln Ser Leu Gln Val Cys Val Gln Thr Val Arg Thr Gln Val Tyr130 135 140Ile Ala Val Asn Asp Lys145 150<210>58<211>155<212>PRT<213>SARS coronavirus<400>58Leu Gln Ser Leu Gln Val Cys Val Gln Thr Val Arg Thr Gln Val Tyr1 5 10 15Ile Ala Val Asn Asp Lys Ala Leu Tyr Glu Gln Val Val Met Asp Tyr20 25 30Leu Asp Asn Leu Lys Pro Arg Val Glu Ala Pro Lys Gln Glu Glu Pro35 40 45Pro Asn Thr Glu Asp Ser Lys Thr Glu Glu Lys Ser Val Val Gln Lys50 55 60Pro Val Asp Val Lys Pro Lys Ile Lys Ala Cys Ile Asp Glu Val Thr65 7075 80Thr Thr Leu Glu Glu Thr Lys Phe Leu Thr Asn Lys Leu Leu Leu Phe859095Ala Asp Ile Asn Gly Lys Leu Tyr His Asp Ser Gln Asn Met Leu Arg100105 110Gly Glu Asp Met Ser Phe Leu Glu Lys Asp Ala Pro Tyr Met Val Gly115 120 125Asp Val Ile Thr Ser Gly Asp Ile Thr Cys Val Val Ile Pro Ser Lys130 135 140Lys Ala Gly Gly Thr Thr Glu Met Leu Ser Arg145 150 155<210>59<211>149<212>PRT<213>SARS coronavirus<400>59Leu Ser Thr Asn His Gly His Lys Val Val Glu Leu Val Ala Glu Met1 5 10 15Asp Gly Ile Gln Tyr Gly Arg Ser Gly Ile Thr Leu Gly Val Leu Val20 25 30Pro His Val Gly Glu Thr Pro Ile Ala Tyr Arg Asn Val Leu Leu Arg35 40 45Lys Asn Gly Asn Lys Gly Ala Gly Gly His Ser Tyr Gly Ile Asp Leu50 55 60Lys Ser Tyr Asp Leu Gly Asp Glu Leu Gly Thr Asp Pro Ile Glu Asp65 70 75 80Tyr Glu Gln Asn Trp Asn Thr Lys His Gly Ser Gly Ala Leu Arg Glu85 9095Leu Thr Arg Glu Leu Asn Gly Gly Ala Val Thr Arg Tyr Val Asp Asn100 105 110Asn Phe Cys Gly Pro Asp Gly Tyr Pro Leu Asp Cys Ile Lys Asp Phe115 120 125Leu Ala Arg Ala Gly Lys Ser Met Cys Thr Leu Ser Glu Gln Leu Asp130 135 140Tyr Ile Glu Ser Lys145149<210>60<211>148<212>PRT<213>SARS coronavirus<400>60Val Ile Pro Ser Lys Lys Ala Gly Gly Thr Thr Glu Met Leu Ser Arg15 10 15Ala Leu Lys Lys Val Pro Val Asp Glu Tyr Ile Thr Thr Tyr Pro Gly2025 30Gln Gly Cys Ala Gly Tyr Thr Leu Glu Glu Ala Lys Thr Ala Leu Lys35 4045Lys Cys Lys Ser Ala Phe Tyr Val Leu Pro Ser Glu Ala Pro Asn Ala50 55 60Lys Glu Glu Ile Leu Gly Thr Val Ser Trp Asn Leu Arg Glu Met Leu65 70 7580Ala His Ala Glu Glu Thr Arg Lys Leu Met Pro Ile Cys Met Asp Val85 90 95Arg Ala Ile Met Ala Thr Ile Gln Arg Lys Tyr Lys Gly Ile Lys Ile100 105 110Gln Glu Gly Ile Val Asp Tyr Gly Val Arg Phe Phe Phe Tyr Thr Ser115 120 125Lys Glu Pro Val Ala Ser Ile Ile Thr Lys Leu Asn Ser Leu Asn Glu130 135 140Pro Leu Val Thr145 148<210>61<211>147<212>PRT<213>SARS coronavirus<400>61Thr Lys Leu Asn Ser Leu Asn Glu Pro Leu Val Thr Met Pro Ile Gly1 510 15Tyr Val Thr His Gly Phe Asn Leu Glu Glu Ala Ala Arg Cys Met Arg2025 30Ser Leu Lys Ala Pro Ala Val Val Ser Val Ser Ser Pro Asp Ala Val35 40 45Thr Thr Tyr Asn Gly Tyr Leu Thr Ser Ser Ser Lys Thr Ser Glu Glu50 55 60His Phe Val Glu Thr Val Ser Leu Ala Gly Ser Tyr Arg Asp Trp Ser65 70 75 80Tyr Ser Gly Gln Arg Thr Glu Leu Gly Val Glu Phe Leu Lys Arg Gly85 90 95Asp Lys Ile Val Tyr His Thr Leu Glu Ser Pro Val Glu Phe His Leu100 105 110Asp Gly Glu Val Leu Ser Leu Asp Lys Leu Lys Ser Leu Leu Ser Leu115120 125Arg Glu Val Lys Thr Ile Lys Val Phe Thr Thr Val Asp Asn Thr Asn130 135 140Leu His Thr145 147<210>62<211>147<212>PRT<213>SARS coronavirus<400>62Thr Lys Leu Asn Ser Leu Asn Glu Pro Leu Val Thr Met Pro Ile Gly1 5 10 15Tyr Val Thr His Gly Phe Asn Leu Glu Glu Ala Ala Arg Cys Met Arg20 2530Ser Leu Lys Ala Pro Ala Val Val Ser Val Ser Ser Pro Asp Ala Val35 40 45Thr Thr Tyr Asn Gly Tyr Leu Thr Ser Ser Ser Lys Thr Ser Glu Glu50 55 60His Phe Val Glu Thr Val Ser Leu Ala Gly Ser Tyr Arg Asp Trp Ser65 70 75 80Tyr Ser Gly Gln Arg Thr Glu Leu Gly Val Glu Phe Leu Lys Arg Gly85 90 95Asp Lys Ile Val Tyr His Thr Leu Glu Ser Pro Val Glu Phe His Leu100 105 110Asp Gly Glu Val Leu Ser Leu Asp Lys Leu Lys Ser Leu Leu Ser Leu115120 125Arg Glu Val Lys Thr Ile Lys Val Phe Thr Thr Val Asp Asn Thr Asn130 135140Leu His Thr145 147<210>63<211>149<212>PRT<213>SARS coronavirus<400>63Tyr Arg Ala Arg Ala Gly Asp Ala Ala Asn Phe Cys Ala Leu Ile Leu1 5 10 15Ala Tyr Ser Asn Lys Thr Val Gly Glu Leu Gly Asp Val Arg Glu Thr20 25 30Met Thr His Leu Leu Gln His Ala Asn Leu Glu Ser Ala Lys Arg Val35 40 45Leu Asn Val Val Cys Lys His Cys Gly Gln Lys Thr Thr Thr Leu Thr5055 60Gly Val Glu Ala Val Met Tyr Met Gly Thr Leu Ser Tyr Asp Asn Leu65 70 75 80Lys Thr Gly Val Ser Ile Pro Cys Val Cys Gly Arg Asp Ala Thr Gln85 90 95Tyr Leu Val Gln Gln Glu Ser Ser Phe Val Met Met Ser Ala Pro Pro100 105 110Ala Glu Tyr Lys Leu Gln Gln Gly Thr Phe Leu Cys Ala Asn Glu Tyr115 120 125Thr Gly Asn Tyr Gln Cys Gly His Tyr Thr His Ile Thr Ala Lys Glu130 135 140Thr Leu Tyr Arg Ile145149<210>64<211>148<212>PRT<213>SARS coronavirus<400>64Cys Gly His Tyr Thr His Ile Thr Ala Lys Glu Thr Leu Tyr Arg Ile1 51015Asp Gly Ala His Leu Thr Lys Met Ser Glu Tyr Lys Gly Pro Val Thr20 25 30Asp Val Phe Tyr Lys Glu Thr Ser Tyr Thr Thr Thr Ile Lys Pro Val35 40 45Ser Tyr Lys Leu Asp Gly Val Thr Tyr Thr Glu Ile Glu Pro Lys Leu5055 60Asp Gly Tyr Tyr Lys Lys Asp Asn Ala Tyr Tyr Thr Glu Gln Pro Ile65 70 7580Asp Leu Val Pro Thr Gln Pro Leu Pro Asn Ala Ser Phe Asp Asn Phe85 90 95Lys Leu Thr Cys Ser Asn Thr Lys Phe Ala Asp Asp Leu Asn Gln Met100105 110Thr Gly Phe Thr Lys Pro Ala Ser Arg Glu Leu Ser Val Thr Phe Phe115 120 125Pro Asp Leu Asn Gly Asp Val Val Ala Ile Asp Tyr Arg His Tyr Ser130 135 140Ala Ser Phe Lys145 148<210>65<211>151<212>PRT<213>SARS coronavirus<400>65Val Val Ala Ile Asp Tyr Arg His Tyr Ser Ala Ser Phe Lys Lys Gly1 510 15Ala Lys Leu Leu His Lys Pro Ile Val Trp His Ile Asn Gln Ala Thr20 25 30Thr Lys Thr Thr Phe Lys Pro Asn Thr Trp Cys Leu Arg Cys Leu Trp35 40 45Ser Thr Lys Pro Val Asp Thr Ser Asn Ser Phe Glu Val Leu Ala Val505560Glu Asp Thr Gln Gly Met Asp Asn Leu Ala Cys Glu Ser Gln Gln Pro65 70 75 80Thr Ser Glu Glu Val Val Glu Asn Pro Thr Ile Gln Lys Glu Val Ile85 90 95Glu Cys Asp Val Lys Thr Thr Glu Val Val Gly Asn Val Ile Leu Lys100 105 110Pro Ser Asp Glu Gly Val Lys Val Thr Gln Glu Leu Gly His Glu Asp115120 125Leu Met Ala Ala Tyt Val Glu Asn Thr Ser Ile Thr Ile Lys Lys Pro130 135 140Asn Glu Leu Ser Leu Ala Leu145 150 151<210>66<211>146<212>PRT<213>SARS coronavirus<400>66Asn Thr Ser Ile Thr Ile Lys Lys Pro Asn Glu Leu Ser Leu Ala Leu1 5 10 15Gly Leu Lys Thr Ile Ala Thr His Gly Ile Ala Ala Ile Asn Ser Val20 25 30Pro Trp Ser Lys Ile Leu Ala Tyr Val Lys Pro Phe Leu Gly Gln Ala35 40 45Ala Ile Thr Thr Ser Asn Cys Ala Lys Arg Leu Ala Gln Arg Val Phe50 5560Asn Asn Tyr Met Pro Tyr Val Phe Thr Leu Leu Phe Gln Leu Cys Thr65 70 7580Phe Thr Lys Ser Thr Asn Ser Arg Ile Arg Ala Ser Leu Pro Thr Thr85 90 95Ile Ala Lys Asn Ser Val Lys Ser Val Ala Lys Leu Cys Leu Asp Ala100 105 110Gly Ile Asn Tyr Val Lys Ser Pro Lys Phe Ser Lys Leu Phe Thr Ile115120 125Ala Met Trp Leu Leu Leu Leu Ser Ile Cys Leu Gly Ser Leu Ile Cys130135 140Val Thr145 146<210>67<211>151<212>PRT<213>SARS coronavirus<400>67Leu Leu Leu Leu Ser Ile Cys Leu Gly Ser Leu Ile Cys Val Thr Ala1 5 1015Ala Phe Gly Val Leu Leu Ser Asn Phe Gly Ala Pro Ser Tyr Cys Asn20 25 30Gly Val Arg Glu Leu Tyr Leu Asn Ser Ser Asn Val Thr Thr Met Asp35 40 45Phe Cys Glu Gly Ser Phe Pro Cys Ser Ile Cys Leu Ser Gly Leu Asp50 55 60Ser Leu Asp Ser Tyr Pro Ala Leu Glu Thr Ile Gln Val Thr Ile Ser65 70 75 80Ser Tyr Lys Leu Asp Leu Thr Ile Leu Gly Leu Ala Ala Glu Trp Val85 90 95Leu Ala Tyr Met Leu Phe Thr Lys Phe Phe Tyr Leu Leu Gly Leu Ser100105110Ala Ile Met Gln Val Phe Phe Gly Tyr Phe Ala Ser His Phe Ile Ser115 120125Asn Ser Trp Leu Met Trp Phe Ile Ile Ser Ile Val Gln Met Ala Pro130 135 140Val Ser Ala Met Val Arg Met145 150 151<210>68<211>150<212>PRT<213>SARS coronavirus<400>68Leu Ser His Phe Val Asn Leu Asp Asn Leu Arg Ala Asn Asn Thr Lys1 5 10 15Gly Ser Leu Pro Ile Asn Val Ile Val Phe Asp Gly Lys Ser Lys Cys20 25 30Asp Glu Ser Ala Ser Lys Ser Ala Ser Val Tyr Tyr Ser Gln Leu Met35 40 45Cys Gln Pro Ile Leu Leu Leu Asp Gln Ala Leu Val Ser Asp Val Gly50 55 60Asp Ser Thr Glu Val Ser Val Lys Met Phe Asp Ala Tyr Val Asp Thr65 70 75 80Phe Ser Ala Thr Phe Ser Val Pro Met Glu Lys Leu Lys Ala Leu Val85 90 95Ala Thr Ala His Ser Glu Leu Ala Lys Gly Val Ala Leu Asp Gly Val100 105 110Leu Ser Thr Phe Val Ser Ala Ala Arg Gln Gly Val Val Asp Thr Asp115120 125Val Asp Thr Lys Asp ValIle Glu Cys Leu Lys Leu Ser His His Ser130 135 140Asp Leu Glu Val Thr Gly145 150<210>69<211>146<212>PRT<213>SARS coronavirus<400>69Cys Leu Lys Leu Ser His His Ser Asp Leu Glu Val Thr Gly Asp Ser1 5 10 15Cys Asn Asn Phe Met Leu Thr Tyr Asn Lys Val Glu Asn Met Thr Pro2025 30Arg Asp Leu Gly Ala Cys Ile Asp Cys Asn Ala Arg His Ile Asn Ala35 40 45Gln Val Ala Lys Ser His Asn Val Ser Leu Ile Trp Asn Val Lys Asp50 5560Tyr Met Ser Leu Ser Glu Gln Leu Arg Lys Gln Ile Arg Ser Ala Ala65 70 75 80Lys Lys Asn Asn Ile Pro Phe Arg Leu Thr Cys Ala Thr Thr Arg Gln85 90 95Val Val Asn Val Ile Thr Thr Lys Ile Ser Leu Lys Gly Gly Lys Ile100 105 110Val Ser Thr Cys Phe Lys Leu Met Leu Lys Ala Thr Leu Leu Cys Val115 120125Leu Ala Ala Leu Val Cys Tyr Ile Val Met Pro Val His Thr Leu Ser130 135 140Ile His145 146<210>70<211>150<212>PRT<213>SARS coronavirus<400>70Ser Met Cys Thr Leu Ser Glu Gln Leu Asp Tyr Ile Glu Ser Lys Arg1 510 15Gly Val Tyr Cys Cys Arg Asp His Glu His Glu Ile Ala Trp Phe Thr202530Glu Arg Ser Asp Lys Ser Tyr Glu His Gln Thr Pro Phe Glu Ile Lys35 40 45Ser Ala Lys Lys Phe Asp Thr Phe Lys Gly Glu Cys Pro Lys Phe Val50 55 60Phe Pro Leu Asn Ser Lys Val Lys Val Ile Gln Pro Arg Val Glu Lys65 70 75 80Lys Lys Thr Glu Gly Phe Met Gly Arg Ile Arg Ser Val Tyr Pro Val8590 95Ala Ser Pro Gln Glu Cys Asn Asn Met His Leu Ser Thr Leu Met Lys100 105110Cys Asn His Cys Asp Glu Val Ser Trp Gln Thr Cys Asp Phe Leu Lys115 120 125Ala Thr Cys Glu His Cys Gly Thr Glu Asn Leu Val Ile Glu Gly Pro130 135 140Thr Thr Cys Gly Tyr Leu145 150<210>71<211>150<212>PRT<213>SARS coronavirus<400>71Leu Leu Leu Ser Ile Cys Leu Gly Ser Leu Ile Cys Val Thr Ala Ala1 5 10 15Phe Gly Val Leu Leu Ser Asn Phe Gly Ala Pro Ser Tyr Cys Asn Gly2025 30Val Arg Glu Leu Tyr Leu Asn Ser Ser Asn Val Thr Thr Met Asp Phe35 40 45Cys Glu Gly Ser Phe Pro Cys Ser Ile Cys Leu Ser Gly Leu Asp Ser50 55 60Leu Asp Ser Tyr Pro Ala Leu Glu Thr Ile Gln Val Thr Ile Ser Ser65 70 75 80Tyr Lys Leu Asp Leu Thr Ile Leu Gly Leu Ala Ala Glu Trp Val Leu85 90 95Ala Tyr Met Leu Phe Thr Lys Phe Phe Tyr Leu Leu Gly Leu Ser Ala100105 110Ile Met Gln Val Phe Phe Gly Tyr Phe Ala Ser His Phe Ile Ser Asn115 120 125Ser Trp Leu Met Trp Phe Ile Ile Ser Ile Val Gln Met Ala Pro Val130 135140Ser Ala Met Val Arg Met145 150<210>72<211>149<212>PRT<213>SARS coronavirus<400>72Ser Thr Cys Phe Lys Leu Met Leu Lys Ala Thr Leu Leu Cys Val Leu1 5 10 15Ala Ala Leu Val Cys Tyr Ile Val Met Pro Val His Thr Leu Ser Ile20 25 30His Asp Gly Tyr Thr Asn Glu Ile Ile Gly Tyr Lys Ala Ile Gln Asp3540 45Gly Val Thr Arg Asp Ile Ile Ser Thr Asp Asp Cys Phe Ala Asn Lys50 55 60His Ala Gly Phe Asp Ala Trp Phe Ser Gln Arg Gly Gly Ser Tyr Lys65 70 75 80Asn Asp Lys Ser Cys Pro Val Val Ala Ala Ile Ile Thr Arg Glu Ile85 90 95Gly Phe Ile Val Pro Gly Leu Pro Gly Thr Val Leu Arg Ala Ile Asn100105 110Gly Asp Phe Leu His Phe Leu Pro Arg Val Phe Ser Ala Val Gly Asn115120 125Ile Cys Tyr Thr Pro Ser Lys Leu Ile Glu Tyr Ser Asp Phe Ala Thr130 135 140Ser Ala Cys Val Leu145 149<210>73<211>151<212>PRT<213>SARS coronavirus<400>73Thr Pro Ser Lys Leu Ile Glu Tyr Ser Asp Phe Ala Thr Ser Ala Cys1 5 10 15Val Leu Ala Ala Glu Cys Thr Ile Phe Lys Asp Ala Met Gly Lys Pro20 25 30Val Pro Tyr Cys Tyr Asp Thr Asn Leu Leu Glu Gly Ser Ile Ser Tyr35 40 45Ser Glu Leu Arg Pro Asp Thr Arg Tyr Val Leu Met Asp Gly Ser Ile50 55 60Ile Gln Phe Pro Asn Thr Tyr Leu Glu Gly Ser Val Arg Val Val Thr65 70 75 80Thr Phe Asp Ala Glu Tyr Cys Arg His Gly Thr Cys Glu Arg Ser Glu85 90 95Val Gly Ile Cys Leu Ser Thr Ser Gly Arg Trp Val Leu Asn Asn Glu100105 110His Tyr Arg Ala Leu Ser Gly Val Phe Cys Gly Val Asp Ala Met Asn115 120125Leu Ile Ala Asn Ile Phe Thr Pro Leu Val Gln Pro Val Gly Ala Leu130 135 140Asp Val Ser Ala Ser Val Val145 150 151<210>74<211>154<212>PRT<213>SARS coronavirus<400>74Pro Leu Val Gln Pro Val Gly Ala Leu Asp Val Ser Ala Ser Val Val1 5 10 15Ala Gly Gly Ile Ile Ala Ile Leu Val Thr Cys Ala Ala Tyr Tyr Phe20 25 30Met Lys Phe Arg Arg Val Phe Gly Glu Tyr Asn His Val Val Ala Ala3540 45Asn Ala Leu Leu Phe Leu Met Ser Phe Thr Ile Leu Cys Leu Val Pro50 55 60Ala Tyr Ser Phe Leu Pro Gly Val Tyr Ser Val Phe Tyr Leu Tyr Leu65 70 75 80Thr Phe Tyr Phe Thr Asn Asp Val Ser Phe Leu Ala His Leu Gln Trp85 9095Phe Ala Met Phe Ser Pro Ile Val Pro Phe Trp Ile Thr Ala Ile Tyr100 105 110Val Phe Cys Ile Ser Leu Lys His Cys His Trp Phe Phe Asn Asn Tyr115 120 125Leu Arg Lys Arg Val Met Phe Asn Gly Val Thr Phe Ser Thr Phe Glu130 135 140Glu Ala Ala Leu Cys Thr Phe Leu Leu Asn145 150 154<210>75<211>147<212>PRT<213>SARS coronavirus<400>75Phe Asn Gly Val Thr Phe Ser Thr Phe Glu Glu Ala Ala Leu Cys Thr1 5 10 15Phe Leu Leu Asn Lys Glu Met Tyr Leu Lys Leu Arg Ser Glu Thr Leu20 2530Leu Pro Leu Thr Gln Tyr Asn Arg Tyr Leu Ala Leu Tyr Asn Lys Tyr35 40 45Lys Tyr Phe Ser Gly Ala Leu Asp Thr Thr Ser Tyr Arg Glu Ala Ala50 55 60Cys Cys His Leu Ala Lys Ala Leu Asn Asp Phe Ser Asn Ser Gly Ala65 70 75 80Asp Val Leu Tyr Gln Pro Pro Gln Thr Ser Ile Thr Ser Ala Val Leu85 90 95Gln Ser Gly Phe Arg Lys Met Ala Phe Pro Ser Gly Lys Val Glu Gly100 105 110Cys Met Val Gln Val Thr Cys Gly Thr Thr Thr Leu Asn Gly Leu Trp115 120 125Leu Asp Asp Thr Val Tyr Cys Pro Arg His Val Ile Cys Thr Ala Glu130 135 140Asp Met Leu145 147<210>76<211>22<212>PRT<213>SARS coronavirus<400>76Lys Lys Met Ala Asp Asn Gly Thr Ile Thr Val Glu Glu Leu Lys Gln1 510 15Leu Leu Glu Gln Trp Asn20 22<210>77<211>24<212>PRT<213>SARS coronavirus<400>77Lys Lys Leu Asn Val Pro Leu Arg Gly Thr Ile Val Thr Arg Pro Leu1 51015Met Glu Ser Glu Leu Val Ile Gly20 24<210>78<211>20<212>PRT<213>SARS coronavirus<400>78Lys Lys Arg Met Ala Gly His Ser Leu Gly Arg Cys Asp Ile Lys Asp1 5 10 15Leu Pro Lys Glu20<210>79<211>18<212>PRT<213>SARS coronavirus<400>79Lys Lys Asn Thr Asp His Ala Gly Ser Asn Asp Asn Ile Ala Leu Leu1 5 1015Val Gln18<210>80<211>27<212>PRT<213>SARS coronavirus<400>80Lys Lys Lys Asp Gly Ile Tyr Phe Ala Ala Thr Glu Lys Ser Asn Val1 510 15Val Arg Gly Trp Val Phe Gly Ser Thr Met Asn20 27<210>81<211>24<212>PRT<213>SARS coronavirus<400>81Lys Lys Trp Val Phe Gly Ser Thr Met Asn Asn Lys Ser Gln Ser Val1 510 15Ile Ile Ile Asn Asn Ser Thr Asn2024<210>82<211>24<212>PRT<213>SARS coronavirus<400>82Lys Lys Asp Cys Ser Gln Asn Pro Leu Ala Glu Leu Lys Cys Ser Val1 5 1015Lys Ser Phe Glu Ile Asp Lys Gly2024<210>83<211>24<212>PRT<213>SARS coronavirus<400>83Lys Lys Gln Thr Ser Asn Phe Arg Val Val Pro Ser Gly Asp Val Val1 5 1015Arg Phe Pro Asn Ile Thr Asn Leu20 24<210>84<211>23<212>PRT<213>SARS coronavirus<400>84Lys Lys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser Val1 5 1015Tyr Ala Trp Glu Arg Lys Lys20 23<210>85<211>24<212>PRT<213>SARS coronavirus<400>85Lys Lys Gln Asp Val Asn Cys Thr Asp Val Ser Thr Ala Ile His Ala1 5 10 15Asp Gln Leu Thr Pro Ala Trp Arg20 24<210>86<211>24<212>PRT<213>SARS coronavirus<400>86Lys Lys Thr Gly Asn Asn Val Phe Gln Thr Gln Ala Gly Cys Leu Ile1 510 15Gly Ala Glu His Val Asp Thr Ser20 24<210>87<211>24<212>PRT<213>SARS coronavirus<400>87Lys Lys Lys Gly Ala Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu1 5 10 15Asp Asp Ser Glu Pro Val Leu Lys2024<210>88<211>25<212>PRT<213>SARS coronavirus<400>88Lys Lys Met Ser Asp Asn Gly Pro Gln Ser Asn Gln Arg Ser Ala Pro1 5 1015Arg Ile Thr Phe Gly Gly Pro Thr Asp20 25<210>89<211>24<212>PRT<213>SARS coronavirus<400>89Lys Lys Pro Thr Asp Ser Thr Asp Asn Asn Gln Asn Gly Gly Arg Asn1 510 15Gly Ala Arg Pro Lys Gln Arg Arg20 24<210>90<211>24<212>PRT<213>SARS coronavirus<400>90Lys Lys Ser Arg Gly Gly Ser Gln Ala Ser Ser Arg Ser Ser Ser Arg1 5 10 15Ser Arg Gly Asn Ser Arg Asn Ser20 24<210>91<211>24<212>PRT<213>SARS coronavirus<400>91Lys Lys Arg Asn Ser Thr Pro Gly Ser Ser Arg Gly Asn Ser Pro Ala1 5 10 15Arg Met Ala Ser Gly Gly Gly Glu20 24<210>92<211>19<212>PRT<213>SARS coronavirus<400>92Lys Lys Lys Val Ser Gly Lys Gly Gln Gln Gln Gln Gly Gln Thr Val1 5 10 15Thr Lys Lys19<210>93<211>22<212>PRT<213>SARS coronavirus<400>93Lys Lys Lys Asp Lys Lys Lys Lys Thr Asp Glu Ala Gln Pro Leu Pro1 5 1015Gln Arg Gln Lys Lys Gln2022<210>94<211>24<212>PRT<213>SARS coronavirus<400>94Lys Lys Met Asp Asp Phe Ser Arg Gln Leu Gln Asn Ser Met Ser Gly15 10 15Ala Ser Ala Asp Ser Thr Gln Ala20 24<210>95<211>25<212>PRT<213>SARS coronavirus<400>95Lys Lys Tyr Phe Val Ala Ser Phe Arg Leu Phe Ala Arg Thr Arg Ser1 5 10 15Met Try Ser Phe Asn Pro Glu Thr Asn20 25<210>96<211>23<212>PRT<213>SARS coronavirus<400>96Lys Lys Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr1 5 1015Glu Asn Gln Lys Gln Ile Ala20 23<210>97<211>20<212>PRT<213>SARS coronavirus<400>97Lys Lys Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser1 5 10 15Pro Asp Val Asp20<210>98<211>24<212>PRT<213>SARS coronavirus<400>98Lys Lys Asp Gly Lys Met Lys Glu Leu Ser Pro Arg Trp Tyr Phe Tyr1 5 10 15Tyr Leu Gly Thr Gly Pro Glu Ala2024<210>99<211>24<212>PRT<213>SARS coronavirus<400>99Lys Lys Lys Pro Arg Gln Lys Arg Thr Ala Thr Lys Gln Tyr Asn Val1 5 10 15Thr Gln Ala Phe Gly Arg Arg Gly20 24<210>100<211>27<212>PRT<213>SARS coronavirus<400>100Lys Lys Glu Gln Trp Asn Leu Val Ile Gly Phe Leu Phe Leu Ala Trp1 5 10 15Ile Met Leu Leu Gln Phe Ala Tyr Ser Asn Arg20 25 27<210>101<211>24<212>PRT<213>SARS coronavirus<400>101Lys Lys Ala Arg Thr Arg Ser Met Trp Ser Phe Asn Pro Glu Thr Asn1 5 10 15Ile Leu Leu Asn Val Pro Leu Arg2024<210>102<211>23<212>PRT<213>SARS coronavirus<400>102Lys Lys Leu Val Ile Gly Ala Val Ile Ile Arg Gly His Leu Arg Met1 510 15Ala Gly His Ser Leu Gly Arg2023<210>103<211>27<212>PRT<213>SARS coronavirus<400>103Lys Lys Lys Asp Leu Pro Lys Glu Ile Thr Val Ala Thr Ser Arg Thr1 5 10 15Leu Ser Tyr Tyr Lys Leu Gly Ala Ser Gln Arg20 25 27<210>104<211>25<212>PRT<213>SARS coronavirus<400>104Lys Lys Asp Ser Gly Phe Ala Ala Tyr Asn Arg Tyr Arg Ile Gly Asn1 5 10 15Tyr Lys Leu Asn Thr Asp His Ala Gly20 25<210>105<211>25<212>PRT<213>SARS coronavirus<400>105Lys Lys Thr Gly Phe His Thr Ile Asn His Thr Phe Gly Asn Pro Val1 5 10 15Ile Pro Phe Lys Asp Gly Ile Tyr Phe20 25<210>106<211>27<212>PRT<213>SARS coronavirus<400>106Lys Lys Asn Asn Ser Thr Asn Val Val Ile Arg Ala Cys Asn Phe Glu1 5 10 15Leu Cys Asp Asn Pro Phe Phe Ala Val Ser Lys20 25 27<210>107<211>24<212>PRT<213>SARS coronavirus<400>107Lys Lys Thr Thr Phe Met Leu Lys Tyr Asp Glu Asn Gly Thr Ile Thr1 5 10 15Asp Ala Val Asp Cys Ser Gln Asn20 24<210>108<211>24<212>PRT<213>SARS coronavirus<400>108Lys Lys Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser1 510 15Thr Phe Phe Ser Thr Phe Lys Cys20 24<210>109<211>25<212>PRT<213>SARS coronavirus<400>109Lys Lys Ser Val Ile Thr Pro Gly Thr Asn Ala Ser Ser Glu Val Ala1 510 15Val Leu Tyr Gln Asp Val Asn Cys Thr2025<210>110<211>25<212>PRT<213>SARS coronavirus<400>110Lys Lys Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala1 5 1015Ser Tyr His Thr Val Ser Leu Leu Arg2025<210>111<211>24<212>PRT<213>SARS coronavirus<400>111Lys Lys Met Val Thr Ile Leu Leu Cys Cys Met Thr Ser Cys Cys Ser15 10 15Cys Leu Lys Gly Ala Cys Ser Cys20 24<210>112<211>24<212>PRT<213>SARS coronavirus<400>112Lys Lys Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu1 5 10 15Lys Gly Val Lys Leu His Tyr Thr20 24<210>113<211>24<212>PRT<213>SARS coronavirus<400>113Lys Lys Gly Ala Arg Pro Lys Gln Arg Arg Pro Gln Gly Leu Pro Asn1 5 10 15Asn Thr Ala Ser Trp Phe Thr Ala20 24<210>114<211>24<212>PRT<213>SARS coronavirus<400>114Lys Lys Gln Leu Pro Gln Gly Thr Thr Leu Pro Lys Gly Phe Tyr Ala1 5 10 15Glu Gly Ser Arg Gly Gly Ser Gln20 24<210>115<211>27<212>PRT<213>SARS coronavirus<400>115Lys Lys Gly Gly Glu Thr Ala Leu Ala Leu leu leu Leu Asp Arg Leu1 5 10 15Asn Gln Leu Glu Ser Lys Val Ser Gly Lys Gly20 25 27<210>116<211>26<212>PRT<213>SARS coronavirus<400>116Lys Lys Gln Thr Val Thr Lys Lys Ser Ala Ala Glu Ala Ser Lys Lys1 5 10 15Pro Arg Gln Lys Arg Thr Ala Thr Lys Gln2026<210>117<211>24<212>PRT<213>SARS coronavirus<400>117Lys Lys Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu1 5 10 15Pro Lys Lys Asp Lys Lys Lys Lys20 24<210>118<211>25<212>PRT<213>SARS coronavirus<400>118Lys Lys Gln Arg Gln Lys Lys Gln Pro Thr Val Thr Leu Leu Pro Ala1 5 10 15Ala Asp Met Asp Asp Phe Ser Arg Gln20 2權利要求
1.一種非典型性肺炎病毒特異蛋白質(zhì)，其特征在于它包括非典型性肺炎病毒的蛋白質(zhì)和經(jīng)修飾的重組非典型性肺炎病毒蛋白質(zhì)，或其具有結合人免疫球蛋白之免疫活性的片段或衍生物，具有通式Seq.ID No.1-Seq.ID No.118的氨基酸序列的蛋白質(zhì)。
2.根據(jù)權利要求1所述的蛋白質(zhì)相對應的抗體，其特征在于所述說的蛋白質(zhì)相對應的抗是單克隆抗體，或是多克隆抗體。
3.一種非典型性肺炎病毒特異蛋白質(zhì)臨床檢測方法，其特征在于其具體步驟如下1)制備如權利要求1所述的蛋白質(zhì)；通過本領域技術人員周知的PCR擴增技術和重組DNA技術，具體如下A、設計相應的PCR引物，同時在所用PCR引物中添加編碼相應數(shù)目氨基酸殘基對應的堿基，分別從臨床非典型性肺炎病毒標準株中擴增編碼上述蛋白質(zhì)的DNA片段；B、將所得PCR產(chǎn)物導入合適的表達載體，構建重組DNA；C、用所得重組DNA轉化合適的宿主細胞，在適于目標蛋白質(zhì)表達的條件下培養(yǎng)如此轉化的宿主細胞；D、回收并純化目標蛋白質(zhì)；2)用純化的如權利要求1所述的蛋白質(zhì)，或如權利要求2所述的抗體包被一種固相支持物；所說的固相支持物包括本領域周知的有機和無機多聚物，為葡聚糖、天然或經(jīng)修飾的纖維素、聚乙烯、聚苯乙烯；或用膠體金的方法包被到試紙上；3)用比色法、分光光度法或熒光法測定的酶標記如權利要求1所述的蛋白質(zhì)或權利要求2所述的抗體，或權利要求2所述的抗體的第二抗體(IgG、IgM、IgA)；標記酶包括氧化還原酶類，為催化酶、過氧化物酶、辣根過氧化物酶、葡萄糖氧化酶、β-半乳糖苷酶；堿性磷酸酶；乙酰膽堿酯酶；使用膠體金，或放射性同位素作為標記；4)將待檢測個體的生物學樣本及由步驟3)標記的蛋白質(zhì)加入經(jīng)包被的固相支持物上，在適合形成抗原抗體免疫結合復合物的條件下孵育一段時間；5)適度清洗去除任何未結合的非典型性肺炎病毒抗體；6)定量檢測孵育混合物中抗原抗體免疫復合物的存在。4、一種非典型性肺炎病毒特異蛋白質(zhì)臨床檢測的試劑盒，其特征在于包括1)包被有純化的如權利要求1所述的蛋白質(zhì)的固相支持物；相應的經(jīng)標記的如權利要求1所述的蛋白質(zhì)和使用該試劑盒的說明；2)包被有純化的如權利要求2所述的抗體的固相支持物；相應的經(jīng)標記的如權利要求2所述的抗體和使用該試劑盒的說明；3)包被有純化的如權利要求1所述的蛋白質(zhì)的固相支持物；如上文所述標記的第二抗體，該第二抗體可以是抗IgG抗體和/或抗IgM抗體和使用該試劑盒的說明；5、根據(jù)權利要求4所述的一種非典型性肺炎病毒特異蛋白質(zhì)臨床檢測的試劑盒，其特征在于所說的快速檢測非典型性肺炎病毒的試紙，是由噴涂過如權利要求1所述的蛋白或權利要求2所述的抗體，及噴涂有膠體金的試紙基質(zhì)粘合而成；在上述檢測試劑盒中，經(jīng)標記的非典型性肺炎病毒蛋白質(zhì)是液體的形式包裝在容器中，或便于重構的凍干物的形式置于的包裝中。在后一種情況中，檢測試劑盒中還優(yōu)選含有用于重構經(jīng)標記的非典型性肺炎病毒蛋白質(zhì)的緩沖液；在上述檢測試劑盒中，還含有提供判斷檢測結果的相應的范圍。
全文摘要
本發(fā)明公開了一種非典型性肺炎病毒特異蛋白質(zhì)和臨床檢測的方法及試劑盒。它提供了非典型性肺炎病毒特異蛋白質(zhì)，或其具有結合人免疫球蛋白之免疫活性的片段或衍生物；并利用該通式蛋白質(zhì)免疫檢測人非典型性肺炎病毒的方法及試劑盒。本發(fā)明解決了臨床對非典型性肺炎病毒感染者的早期快速診斷問題，具有較高的準確性，為臨床檢測本病提供了快速、準確的診斷方法和試劑盒。
文檔編號G01N33/68GK1483737SQ0311665
公開日2004年3月24日 申請日期2003年4月26日 優(yōu)先權日2003年4月26日
發(fā)明者汪建, 方健秋, 文潔, 牟峰, 陳唯軍, 劉斯奇, 胡詠武, 殷劍寧, 王俊, 楊煥明, 汪 建 申請人:杭州華大基因研發(fā)中心