[發(fā)明專利]制備由肝炎病毒因子產(chǎn)生的蛋白質(zhì)的方法

無(wú)效

• 申請(qǐng)?zhí)枺?/td>: 89104064.1
• 申請(qǐng)日: 1989-06-16
• 公開(kāi)（公告）號(hào): CN1048523C
• 公開(kāi)（公告）日: 2000-01-19
• 發(fā)明（設(shè)計(jì)）人: 格雷戈里·Ｒ·雷耶斯;丹尼爾·Ｗ·布雷德利
• 申請(qǐng)（專利權(quán)）人: 基因?qū)嶒?yàn)室技術(shù)有限公司;美國(guó)政府
• 主分類號(hào): C12N15/51
• 分類號(hào): C12N15/51;C12N15/63
• 代理公司: 中國(guó)專利代理(香港)有限公司
• 代理人: 汪洋,曹恒興

1.一種制備由腸道傳播性非甲非乙型肝炎病毒因子產(chǎn)生的蛋白質(zhì)的方法，所述因子的基因組含有一個(gè)與質(zhì)粒pTZ-KF1(ET1.1)中1.33kb?DNA的EcoRI插段同源的區(qū)域，該質(zhì)粒攜于保藏號(hào)為CCTCCNo：M89041的大腸桿菌BB4株中，所述方法包括：

(a)將編碼所述蛋白質(zhì)的DNA片段插入適當(dāng)?shù)谋磉_(dá)載體中，生成重組載體；

(b)將重組載體導(dǎo)入適當(dāng)?shù)乃拗骷?xì)胞中；

(c)在可有效地在宿主細(xì)胞中產(chǎn)生所述蛋白質(zhì)的條件下培養(yǎng)宿主細(xì)胞；

(d)從宿主細(xì)胞中純化所述蛋白質(zhì)。

2.權(quán)利要求1的方法，其中蛋白質(zhì)由所述1.33kb?DNA?EcoRI插段中的編碼區(qū)編碼。

3.權(quán)利要求1的方法，其中所述基因組含有一個(gè)與具有第一順序：1?????????11????????21????????31????????41????????51*?????????*?????????*?????????*?????????*?????????*CGAATTCCGCCAACTGATGGAAGGCACTAATCTGGCAAGACCTGTCCCTGTTGCAGCTGT61????????71????????81????????91????????101???????111*?????????*?????????*?????????*?????????*?????????*TCTACCACCCTGCCCCGAGCTCGAACAGGGCCTTCTCTACCTGCCCCAGGAGCTCACCAC121???????131???????141???????151???????161???????171*?????????*?????????*?????????*?????????*?????????*CTGTGATAGTGTCGTAACATTTGAATTAACAGACATTGTGCACTGCCGCATGGCCGCCCC181???????191???????201???????211???????221???????231*?????????*?????????*?????????*?????????*?????????*GAGCCAGCGCAAGGCCGTGCTGTCCACACTCGTGGGCCGCTACGGCGTCGCACAAAGCTC241???????251???????261???????271???????281???????291*?????????*?????????*?????????*?????????*?????????*TACAATGCTTCCCACTCTGATGTTCGCGACTCTCTCGCCCGTTTTATCCCGGCCATTGGC301???????311???????321???????331???????341???????351*?????????*?????????*?????????*?????????*??????????*CCCGTACAGGTTACAACTTGTGAATTGTACGAGCTAGTGGAGGCCATGGTCGAGAAGGGC361???????371???????381???????391???????401???????411?*????????*?????????*?????????*?????????*?????????*CAGGATGGCTCCGCCGTCCTTGAGCTTGATCTTTGCAACCGTGACGTGTCCAGGATCACC421???????431???????441???????451???????461???????471*?????????*?????????*?????????*?????????*?????????*TTCTTCCAGAAAGATTGTAACAAGTTCACCACAGGTGAGACCATTGCCCATGGTAAAGTG481???????491???????501???????511???????521???????531*?????????*?????????*?????????*?????????*?????????*GGCCAGGGCATCTCGGCCTGGAGCAAGACCTTCTGCGCCCTCTTTGGCCCTTGGTTCCGC541???????551???????561???????571???????581???????591*?????????*?????????*?????????*?????????*?????????*GCTATTGAGAAGGCTATTCTGGCCCTGCTCCCTCAGGGTGTGTTTTACGGTGATGCCTTT601???????611???????621???????631???????641???????651*?????????*?????????*?????????*?????????*?????????*GATGACACCGTCTTCTCGGCGGCTGTGGCCGCAGCAAAGGCATCCATGGTGTTTGAGAAT661???????671???????681???????691???????701???????711*?????????*?????????*?????????*?????????*?????????*GACTTTTCTGAGTTTGACTCCACCCAGAATAACTTTTCTCTGGGTCTAGAGTGTGCTATT721???????731???????741???????751???????761???????771*?????????*?????????*?????????*?????????*?????????*ATGGAGGAGTGTGGGATGCCGCAGTGGCTCATCCGCCTGTATCACCTTATAAGGTCTGCG781???????791???????801???????811???????821???????831*?????????*?????????*?????????*?????????*?????????*TGGATCTTGCAGGCCCCGAAGGAGTCTCTGCGAGGGTTTTGGAAGAAACACTCCGGTGAG841???????851???????861???????871???????881???????891*?????????*?????????*?????????*?????????*?????????*CCCGGCACTCTTCTATGGAATACTGTCTGGAATATGGCCGTTATTACCCACTGTTATGAC901???????911???????921???????931???????941???????951*?????????*?????????*?????????*?????????*?????????*TTCCGCGATTTTCAGGTGGCTGCCTTTAAAGGTGATGATTCGATAGTGCTTTGCAGTGAG961???????971???????981???????991???????1001??????1011?*????????*?????????*?????????*?????????*?????????*TATCGTCAGAGTCCAGGAGCTGCTGTCCTGATCGCCGGCTGTGGCTTGAAGTTGAAGGTA1021??????1031??????1041??????1051??????1061??????1071*?????????*?????????*?????????*?????????*?????????*GATTTCCGCCCGATCGGTTTGTATGCAGGTGTTGTGGTGGCCCCCGGCCTTGGCGCGCTC1081??????1091??????1101??????1111??????1121??????1131*?????????*?????????*?????????*?????????*?????????*CCTGATGTTGTGCGCTTCGCCGGCCGGCTTACCGAGAAGAATTGGGGCCCTGGCCCTGAG1141??????1151??????1161??????1171??????1181??????1191*?????????*?????????*?????????*?????????*??????????*CGGGCGGAGCAGCTCCGCCTCGCTGTTAGTGATTTCCTCCGCAAGCTCACGAATGTAGCT1201??????1211??????1221??????1231??????1241??????1251*?????????*?????????*?????????*?????????*?????????*CAGATGTGTGTGGATGTTGTTTCCCGTGTTTATGGGGTTTCCCCTGGACTCGTTCATAAC1261??????1271??????1281??????1291??????1301??????1311*?????????*?????????*?????????*?????????*?????????*CTGATTGGCATGCTACAGGCTGTTGCTGATGGCAAGGCACATTTCACTGAGTCAGTAAAA1321??????1331??????1341*?????????*?????????*CCAGTGCTCGACCGGAATTCGAGC或第二順序1?????????11????????21?????????31???????41????????51*?????????*?????????*??????????*????????*?????????*GCTCGAATTCCGGTCGAGCACTGGTTTTACTGACTCAGTGAAATGTGCCTTGCCATCAGC61????????71????????81?????????91???????101???????111*?????????*?????????*??????????*????????*?????????*AACAGCCTGTAGCATGCCAATCAGGTTATGAACGAGTCCAGGGGAAACCCCATAAACACG121???????131???????141????????151??????161???????171*?????????*?????????*??????????*????????*?????????*GGAAACAACATCCACACACATCTGAGCTACATTCGTGAGCTTGCGGAGGAAATCACTAAC181???????191???????201????????211??????221???????231*?????????*?????????*??????????*????????*?????????*AGCGAGGCGGAGCTGCTCCGCCCGCTCAGGGCCAGGGCCCCAATTCTTCTCGGTAAGCCG241???????251???????261????????271??????281???????291*?????????*?????????*??????????*????????*?????????*GCCGGCGAAGCGCACAACATCAGGGAGCGCGCCAAGGCCGGGGGCCACCACAACACCTGC301???????311???????321????????331??????341???????351*?????????*?????????*??????????*????????*?????????*ATACAAACCGATCGGGCGGAAATCTACCTTCAACTTCAAGCCACAGCCGGCGATCAGGAC361???????371???????381????????391??????401???????411*?????????*?????????*??????????*????????*?????????*AGCAGCTCCTGGACTCTGACGATACTCACTGCAAAGCACTATCGAATCATCACCTTTAAA421???????431???????441???????451???????461???????471*?????????*?????????*?????????*?????????*?????????*GGCAGCCACCTGAAAATCGCGGAAGTCATAACAGTGGGTAATAACGGCCATATTCCAGAC481???????491???????501???????511???????521???????531*?????????*?????????*?????????*?????????*?????????*AGTATTCCATAGAAGAGTGCCGGGCTCACCGGAGTGTTTCTTCCAAAACCCTCGCAGAGA541???????551???????561???????571???????581???????591*?????????*?????????*?????????*?????????*?????????*CTCCTTCGGGGCCTGCAAGATCCACGCAGACCTTATAAGGTGATACAGGCGGATGAGCCA601???????611???????621???????631???????641???????651*?????????*?????????*?????????*?????????*?????????*CTGCGGCATCCCACACTCCTCCATAATAGCACACTCTAGACCCAGAGAAAAGTTATTCTG661???????671???????681???????691???????701???????711*?????????*?????????*?????????*?????????*?????????*GGTGGAGTCAAACTCAGAAAAGTCATTCTCAAACACCATGGATGCCTTTGCTGCGGCCAC721???????731???????741???????751???????761???????771*?????????*?????????*?????????*?????????*?????????*AGCCGCCGAGAAGACGGTGTCATCAAAGGCATCACCGTAAAACACACCCTGAGGGAGCAG781???????791???????801???????811???????821???????831*?????????*?????????*?????????*?????????*?????????*GGCCAGAATAGCCTTCTCAATAGCGCGGAACCAAGGGCCAAAGAGGGCGCAGAAGGTCTT841???????851???????861???????871???????881???????891*?????????*?????????*?????????*?????????*?????????*GCTCCAGGCCGAGATGCCCTGGCCCACTTTACCATGGGCAATGGTCTCACCTGTGGTGAA901???????911???????921???????931???????941???????951*?????????*?????????*?????????*?????????*?????????*CTTGTTACAATCTTTCTGGAAGAAGGTGATCCTGGACACGTCACGGTTGCAAAGATCAAG961???????971???????981???????991???????1001??????1011*?????????*?????????*?????????*?????????*?????????*CTCAAGGACGGCGGAGCCATCCTGGCCCTTCTCGACCATGGCCTCCACTAGCTCGTACAA1021??????1031??????1041??????1051??????1061??????1071*?????????*?????????*?????????*?????????*?????????*TTCACAAGTTGTAACCTGTACGGGGCCAATGGCCGGGATAAAACGGGCGAGAGAGTCGCG1081??????1091??????1101??????1111??????1121??????1131*?????????*?????????*?????????*?????????*?????????*AACATCAGAGTGGGAAGCATTGTAGAGCTTTGTGCGACGCCGTAGCGGCCCACGAGTGTG1141??????1151??????1161??????1171??????1181??????1191*?????????*?????????*?????????*?????????*?????????*GACAGCACGGCCTTGCGCTGGCTCGGGGCGGCCATGCGGCAGTGCACAATGTCTGTTAAT1201??????1211??????1221??????1231??????1241??????1251*?????????*?????????*?????????*?????????*?????????*TCAAATGTTACGACACTATCACAGGTGGTGAGCTCCTGGGGCAGGTAGAGAAGGCCCTGT1261??????1271??????1281??????1291??????1301??????1311*?????????*?????????*?????????*?????????*?????????*TCGAGCTCGGGGCAGGGTGGTAGAACAGCTGCAACAGGGACAGGTCTTGCCAGATTAGTG1321??????1331??????1341*?????????*?????????*CCTTCCATCAGTTGGCGGAATTCG.或第三順序CGGTTGTTCAGTACCAGTTTACTGCAGGTGTGCCTGGATCCGGCAAGTCCCGCTCTATCACCCAAGCCGATGTGGACGTTGTCGTGGTCCCGACGCGTGA?????100GTTGCGTAATGCCTGGCGCCGTCGCGGCTTTGCTGCTTTTACCCCGCATACTGCCGCCAGAGTCACCCAGGGGCGCCGGGTTGTCATTGATGAGGCTCCA?????200TCCCTCCCCCCTCACCTGCTGCTGCTCCACATGCAGCGGGCCGCCACCGTCCACCTTCTTGGCGCCCCGAACCAGATCCCAGCCATCGACTTTGAGCACG?????300CTGGGCTCGTCCCCGCCATCAGGCCCGACTTAGCCCCACCTCCTGGTGGCATGTTACCCATCGCTGCCTGCGGATGTATGCGAGCTCATCCGTGGTGCAT?????400ACCCCATGATCCAGACCACTAGCCGGGTTCTCCGTTCGTTGTTCTGGGGTGAGCCTGCCGTCGGGCAGAAACTAGTGTTCACCCAGGCGGCCAAGCCCGC?????500CAACCCCGGCTCAGTGACGGTCCACGAGGCGCAGGGCGCTACCTACACGGAGACCACTATTATTGCCACAGCAGATGCCCGGGGCCTTATTCAGTCGTCT?????600CGGGCTCATGCCATTGTTGCTCTGACGCGCCACACTGAGAAGTGCGTCATCATTGACGCACCAGGCCTGCTTCGCGAGGTGGGCATCTCCGATGCAATCG?????700TTAATAACTTTTTCCTCGCTGGTGGCGAAATTGGTCACCAGCGCCCATCAGTTATTCCCCGTGGCAACCCTGACGCCAATGTTGACACCCTGGCTGCCTT?????800CCCGCCGTCTTGCCAGATTAGTGCCTTCCATCAGTTGGCTGAGGAGCTTGGCCACAGACCTGTCCCTGTTGCAGCTGTTCTACCACCCTGCCCCGAGCTC?????900GAACAGGGCCTTCTCTACCTGCCCCAGGAGCTCACACCACCTGTGATAGTGTCGTAACATTTGAATTAACAGACATTGTGCACTGCCGCATGGCCGCCCC????1000GAGCCAGCGCAAGGCCGTGCTGTCCACACTCGTGGGCCGCTACGGCGTCGCACAAAGCTCTACAATGCTTCCCACTCTGATGTTCGCGACTCTCTCATCC????1100CCGGATTGGCCCGGATTGGCCCCGTACAGGTTACAACTTGTGAATTGTACGAGCTAGTGGAGGCCATGGTCGAGAAGGGCCAGGATCTGCCGCCGTCCTT????1200GAGCTTGATCTTTGCAACCGTGACGTGTCCAGGATCACCTTCTTCCAGAAAGATTGTAACAAGTTCACCACAGGTGAGACCATTGCCCATGGTAAAGTGG????1300GCCAGGGCATCTCGGCCTGGAGCAAGACCTTCTGCGCCCTCTTTGGCCCTTGGTTCCGCGCTATTGAGAAGGCTATTCTGGCCCTGCTCCCTCAGGGTGT????1400GTTTTACGGTGATGCCTTTGATGACACCGTCTTCTCGGCGGCTGTCGAGGCCAAAGGCATCCATGGTGTTTGAGAATGACTTTTCTGAGTTTGACTCCAC????1500CCAGAATAACTTTTCTCTGGGTCTAGAGTGTGCTATTATGGAGGAGTGTCGGGATGCCGCAGTGGCTCATCCGCCTGTATCACCTTATAAGGTCTGCGTG????1600GATCTTGCAGGCCCCGAAGGAGTCTCTGCGAGGGTTTTGGAAGAAACACTCCGGTGAGCCCGGCACTCTTCTATGGAATACTGTCTGGAATATGGCCGTT????1700ATTACCCACTGTTATGACTTCCGCGATTTTCAGGTGGCTGCCTTTAAAGGTGATGATTCGATAGTGCTTTGCAGTGAGTATCGTCAGAGTCCAGGAGCTG????1800CTGTCCTGATCGCCGGCTGTGGCTTGAAGTTGAAGGTAGATTTCCGCCCGATCGGTTTGTATGCAGGTGTTGTGGTGGCCCCCGGCCTTGGCGCGCTCCC????1900TGATGTTGTGCCGTTCGCCGGCCGGCTTACCGAGAAGAATTGGCCCTGGCCTGAGCGGCGAAGACACGTGCGCTCGCTGTTGTTAGTGATTTCCTCCGCA????2000AGCTCACGAATGTAGCTCAGAGTGTGTGGATGTTGTTTCCCGTGTTTATGGGGTTTCCCCTGGACTCGTTCATAACCTGATTGGCATGCTACAGGCTGTT????2100GCTGATGGCAAGGCACATTTCACTGAGTCAGTAAAACCAGTGCTCGACTTGACAAATTCAATCTTGTGTCGGGTGGAATGAATAACATGTCTTTTGCTGC????2200GCCCATGGGTTCGCGACCATGCCCCTCGGCCTATTTTGTTGCTGCTCCTCATGTTTTTGCCTATGCTGCCCGCGCCACCGCCCCGTCAGCCGTCTGGCCG???2300CCGTCGTGGGCGGCGCAGCGGCGGTTCCGGCGGTGGTTTCTGGGGTGACCGGGTTGATTCTCAGCCCTTCGCAATCCCCTATATTCATCCAACCAACCCC???2400TTCGCCCCCGATGTCACCGCTGCGGCCGGGGCTGGACCTCGTGTTCGCCAACCCGCCCGACCACTCGGCTCCGCTTGGCGTGACCAGGCCCAGCGCCCCG???2500CCGTTGCCTCACGTCGTAGACCGACCACAGCTGGGGCCGCGCCGCTAACCGCGGTCGCTCCGGCCCCG???????????????????????????????????2568或第四順序GCGGCCGCTCGGCAAAAGAGGCCTAGGGCGCATGGTGGCGAACCCATGGGCGCACGAAACCACATGTTATTCATTCAGACCGGTGCATAATTGAGTGTGT????100AAGGTCAAGTATAGGCTTAACAGACTCTGTAAAATGCGCCTTACCATCACCAATAGTCTGGAGCATGCCTATCAGGTTATGAACCAGACCCGGGGAAACC????200CCGTAAACTCTAGACACCACCTCAACACAAATCTGGGCCACATTCGTTAACCTACGGAGGAAATCCTGCACGGCGAGGCGGACGTGCTCTGCCCGCTCCG????300GATCAGGCCCCCAGTTCTTCTCCGAAAGCCGTCCGGCGAATCGAACGACATCGGGTAGGGCCCCGAGCCCGGGGCGACGACAACCCCGGCATACAGCCCA????400ATCGGCCGGAAGTCAGCCTTCAACTTCGGGCCACAGCCTGCTATAAGCGAACCGGCGCCTGGGCTCTGGCGGTATTCACTACAGAAAGGAACCGACCGAG????500TCGTCGCCCTTGAAGGCGGCAACCTGGAGGTCCCGGAACTCATAGCAATGGGCAATGATTGCCATGTTCCACACCGTATTCCAGAGCAACGTGCCCGGCT????600CACCAGAATGCTTCTTCCAGAACCCTCTCAAAGACTCTTTTGGGGCCTGCAGGGATCCACGCCGACCGGACGGCATGGTACAACCTGACAAGCCACTGGG????700GCATACCACACTCTTCCATAATGGCGCACTCAAGACCTAGGGAAAAGTTATTCTGAGTCGAGTCAAACTCAGAAAAATCATTTTCAAACACCATGGCATG????800GCTGGCGCCAGCCACGGCAGCAGAGAATACTGAGTCGTCATAAGCATCCCCGTAGAACACAGCTTGTGGTAAAAGGGATAGAATAGCCTTCTCAATCGCA????900CGGAACCAGGGGCCAAACAGGGCACAAAAGGTCTTACTCCAGGCGGAGATACCCTGACCGACTTTGCCATGCGCAATTGTCTCGCCGGTCGTGAACTTGT???1000TACAATCCTTCTGGAAAAAGGTTATGCGGGAGACATCTCGGCTGCACAAATCCAACTCGAGGACGGCTGAACCGTCTTGGCCCTTCTCCACCATCGCCTC???1100TACAAGCTCAAAGAGTTCACAGGTGGTGGCAGTAACCCGCCGAGAGTGGGAATAAAGCGCGCAAGGGAGGCGCGGACATCGGTGTGACCCGCATCATAAA???1200GCCTTGTGCGTCTGCCATACCGGCCTACCAGCATGACAAAACAGCTTTCCTTTGGCTAGGGGCCGCCATGGGCCTGACAACTGTCAGTTAGCTCAAATGT???1300CACA?????????????????????????????????????????????????1304的雙鏈DNA同源的區(qū)域。

4.權(quán)利要求1的方法，其中所述蛋白質(zhì)具有對(duì)存在于腸道傳播性非甲非乙型肝炎病毒因子感染的個(gè)體中的抗體的免疫活性。