| 公開(公告)號(hào) | CN1560066A |
| 公開(公告)日 | 2005.01.05 |
| 申請(qǐng)(專利)號(hào) | CN200410010714.7 |
| 申請(qǐng)日期 | 2004.03.03 |
| 專利名稱 | 抗隱孢子蟲重組免疫毒素蛋白及其制備工藝和醫(yī)用用途 |
| 主分類號(hào) | C07H21/04 |
| 分類號(hào) | C07H21/04;C12N15/11;C12N15/09;C12N15/63;A61K38/16;A61P33/02 |
| 分案原申請(qǐng)?zhí)? | |
| 優(yōu)先權(quán) | |
| 申請(qǐng)(專利權(quán))人 | 中國人民解放軍軍需大學(xué) |
| 發(fā)明(設(shè)計(jì))人 | 張西臣;李建華;蔡輝 |
| 地址 | 130062吉林省長春市西安大路175號(hào) |
| 頒證日 | |
| 國際申請(qǐng) | |
| 進(jìn)入國家日期 | |
| 專利代理機(jī)構(gòu) | 吉林長春新紀(jì)元專利代理有限責(zé)任公司 |
| 代理人 | 陳宏偉 |
| 國省代碼 | 吉林;22 |
| 主權(quán)項(xiàng) | 抗隱孢子蟲重組免疫毒素蛋白的基因序列如下：CAGGTGAAGCTGCAGGAGTCAGGACCTGGCCTGGTGGCACCCTCA45GlnValLysLeuGlnGluSerGlyProGlyLeuValAlaProSer151015CAGAGCCTGTCCATCACATGCACTGTCTCTGGGTTCTCATTATCC90GlnSerLeuSerIleThrCysThrValSerGlyPheSerLeuSer16202530AGATATAGTGTACACTGGGTTCGCCAGCCTCCAGGAAAGGGTCTG135ArgTyrSerValHisTrpValArgGlnProProGlyLysGlyLeu31354045GAGTGGCTGGGAATGATATGGGGTGGTGGAAGCACAGACTATAAT180GluTrpLeuGlyMetIleTrpGlyGlyGlySerThrAspTyrAsn46505560TCAGCTCTCAAATCCAGACTGAGCATCAGCAAGGACAACTCCAAG225SerAlaLeuLysSerArgLeuSerIleSerLysAspAsnSerLys61657075AGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACA270SerGlnValPheLeuLysMetAsnSerLeuGlnThrAspAspThr76808590GCCATGTACTACTGTGCCAGAAACCGGTATGGTAATGCTATGGAC315AlaMetTyrTyrCysAlaArgAsnArgTyrGlyAsnAlaMetAsp9195100105TACTGGGGCCAAGGCACCACGGTCACCGTCTCCTCAGGTGGAGGC360TyrTrpGlyGlnGlyThrThrValThrValSerSerGlyGlyGly106110115120GGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCGAG405GlySerGlyGlyGlyGlySerGlyGlyGlyGlySerAspIleGlu121125130135CTCACTCAGTCTCCAGCCTCCCTATCTGTATCTGTGGGAGAAACT450LeuThrGlnSerProAlaSerLeuSerValSerValGlyGluThr136140145150GTCACCATCACATGTCGAGCAAGTGAGAATATTTACAGTAATTTA495ValThrIleThrCysArgAlaSerGlnAsnIleTyrSerAsnLeu151155160165GCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAGCTCCTGGTC540AlaTrpTyrGlnGlnLysGlnGlyLysSerProGlnLeuLeuVal166170175180TATGCTGCAACAAACTTAGCAGATGGTGTGCCATCAAGGTTCAGT585TyrAlaAlaThrAsnLeuAlaAspGlyValProSerArgPheSer181185190195GGCAGTGGATCAGGCACACAGTATTCCCTCAAGATCAACAGCCTG630GlySerGlySerGlyThrGlnTyrSerLeuLysIleAsnSerLeu196200205210CAGTCTGAAGATTTTGGGAGTTATTACTGTCAACATTTTTGGGGT675GlnSerGluAspPheGlySerTyrTyrCysGlnHisPheTrpGly211215220225ACTCCGTACACGTTCGGAGGGGGGAGCAAGCTGGAGCTGAAACGG720ThrProTyrThrPheGlyGlyGlySerLysLeuGluLeuLysArg226230235240CATATGGCCGAAGAGGGCGGCAGCCTGGCCGCGCTGACCGCGCAC765HisMetAlaGluGluGlyGlySerLeuAlaAlaLeuThrAlaHis241245250255CAGGCTTGCCACCTGCCGCTGGAGACTTTCACCCGTCATCGCCAG810GlnAlaCysHisLeuProLeuGluThrPheThrArgHisArgGln256260265270CCGCGCGGCTGGGAACAACTGGAGCAGTGCGGCTATCCGGTGCAG855ProArgGlyTrpGluGlnLeuGluGlnCysGlyThrProValGln271275280285CGGCTGGTCGCCCTCTACCTGGCGGCGCGGCTGTCGTGGAACCAG900ArgLeuValAlaLeuTyrLeuAlaAlaArgLeuSerTrpAsnGln286290295300GTCGACCAGGTGATCCGCAACGCCCTGGCCAGCCCCGGCAGCGGC945ValAspGlnValIleArgAsnAlaLeuAlaSerProGlySerGly301305310315GGCGACCTGGGCGAAGCGATCCGCGAGCAGCCGGAGCAGGCCCTG990GlyAspLeuGlyGluAlaIleArgGluGlnProGluGlnAlaLeu316320325330GCCCTGACCCTGGCCGCCGCCGAGAGCGAGCGCTTCGTCCGGCAG1035AlaLeuThrLeuAlaAlaAlaGluSerGluArgPheValArgGln331335340345GGCACCGGCAACGACGAGGCCGGCGCGGCCAACGCCGACGTGGTG1080GlyThrGlyAsnAspGluAlaGlyAlaAlaAsnAlaAspValVal346350355360AGCCTGACCTGCCCGGTCGCCGCCGGTGAATGCGCGGGCCCGGCG1125SerLeuThrCysProValAlaAlaGlyGluCysAlaGlyProAla361365370375GACAGCGGCGACGCCCTGCTGGAGCGCAACTATCCCACTGGCGCG1170AspSerGlyAspAlaLeuLeuGluArgAsnTyrProThrGlyAla376380385390GAGTTCCTCGGCGACGGCGGCGACGTCAGCTTCAGCACCCGCGGC1215GluPheLeuGlyAspGlyGlyAspValSerPheSerThrArgGly391395400405ACGCAGAACTGGACGGTGGAGCGGCTGCTCCAGGCGCACCGCCAA1260ThrGluAsnTrpThrValGluArgLeuLeuGlnAlaHisArgGln406410415420CTGGAGGAGCGCGGCTATGTGTTCGTCGGCTACCACGGCACCTTC1305LeuGluGluArgGlyTyrValPheValGlyTyrHisGlyThrPhe42l425430435CTCGAAGCGGCGCAAAGCATCGTCTTCGGCGGGGTGCGCGCGCGC1350LeuGluAlaAlaGlnSerIleValPheGlyGlyValArgAlaArg436440445450AGCCAGGACCTCGACGCGATCTGGCGCGGTTTCTATATCGCCGGC1395SerGlnAspLeuAspAlaIleTrpArgGlyPheTyrIleAlaGly451455460465GATCCGGCGCTGGCCTACGGCTACGCCCAGGACCAGGAACCCGAC1440AspProAlaLeuAlaTyrGlyTyrAlaGlnAspGlnGluProAsp466470475480GCACGCGGCCGGATCCGCAACGGTGCCCTGCTGCGGGTCTATGTG1485AlaArgGlyArgIleArgAsnGlyAlaLeuLeuArgValTyrVal481485490495CCGCGCTCGAGCCTGCCGGGCTTCTACCGCACCAGCCTGACCCTG1530ProArgSerSerLeuProGlyPheTyrArgThrSerLeuThrLeu496500505510GCCGCGCCGGAGGCGGCGGGCGAGGTCGAACGGCTGATCGGCCAT1575AlaAlaProGluAlaAlaGlyGluValGluArgLeuIleGlyHis511515520525CCGCTGCCGCTGCGCCTGGACGCCATCACCGGCCCCGAGGAGGAA1620ProleuProLeuArgleuAspAlaIleThrGlyProGluGluGlu526530535540GGCGGGCGCCTGGAGACCATTCTCGGCTGGCCGCTGGCCGAGCGC1665GlyGlyArgLeuGluThrIleLeuGlyTrpProLeuAlaGluArg541545550555ACCGTGGTGATTCCCTCGGCGATCCCCACCGACCCGCGCAACGTC1710ThrValValIleProSerAlaIleProThrAspProArg |
| 摘要 | 本發(fā)明公開一種抗隱孢子蟲重組免疫毒素蛋白及其制備工藝和醫(yī)用用途，是一種預(yù)防和治療隱孢子蟲病的藥物，利用單克隆抗體技術(shù)和基因重組技術(shù)，從分泌抗微小隱孢子蟲子孢子表膜抗體的雜交瘤細(xì)胞株中克隆其可變區(qū)基因，再連接成單鏈抗體基因，然后將其插入原核表達(dá)載體的相應(yīng)位點(diǎn)上，構(gòu)建了重組原核表達(dá)質(zhì)粒，再分別采用酶切和PCR方法將去除受體結(jié)合區(qū)的細(xì)菌外毒素插入重組原核質(zhì)粒中單鏈抗體基因的下游位點(diǎn)，構(gòu)建重組表達(dá)質(zhì)粒。將上述構(gòu)建的重組表達(dá)質(zhì)粒分別轉(zhuǎn)化受體菌，經(jīng)IPTG誘導(dǎo)，SDS-PAGE分析表達(dá)產(chǎn)物，后對(duì)表達(dá)產(chǎn)物包涵體進(jìn)行純化和復(fù)性，并對(duì)其進(jìn)行體外抗蟲活性測(cè)定和模型動(dòng)物隱孢子蟲病治療。 |
| 國際公布 |
