{"title":"TGEV HB06株核衣壳蛋白基因的克隆与表达。","authors":"Jinghui Fan, Yuzhu Zuo, Yuelan Zhao, Tanqing Li, Xiaobo Zhang","doi":"10.1007/s11703-007-0060-5","DOIUrl":null,"url":null,"abstract":"<p><p>The nucleocapsid protein gene of transmissible gastroenteritis virus, 1 149 bp in length, was amplified by RT-PCR from isolated strain HB06 and cloned into pMD18-T. Sequence comparison with other transmissible gastroenteritis virus (TGEV) strains selected from the Gene Bank revealed that the homology of <i>N</i> gene complete sequence shares more than 97% in nucleotide. <i>N</i> gene was cloned into <i>Bam</i>HI and <i>Eco</i>RI multiple cloning sites of the prokaryotic expression vector pET 20 b, and named pETN. After being induced by isopropyl-<i>β</i>-D-thiogalactopyranoside (IPTG), the recombinant nucleocapsid protein was expressed. The result of SDS-PAGE and Western-blot showed that the recombinant nucleocapsid protein was 47 kDa and had strong positive reactions with TGEV-specific antibody.</p>","PeriodicalId":58797,"journal":{"name":"中国农学前沿","volume":"1 3","pages":"357-360"},"PeriodicalIF":0.0000,"publicationDate":"2007-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11703-007-0060-5","citationCount":"4","resultStr":"{\"title\":\"Cloning and expression of nucleocapsid protein gene of TGEV HB06 strain.\",\"authors\":\"Jinghui Fan, Yuzhu Zuo, Yuelan Zhao, Tanqing Li, Xiaobo Zhang\",\"doi\":\"10.1007/s11703-007-0060-5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The nucleocapsid protein gene of transmissible gastroenteritis virus, 1 149 bp in length, was amplified by RT-PCR from isolated strain HB06 and cloned into pMD18-T. Sequence comparison with other transmissible gastroenteritis virus (TGEV) strains selected from the Gene Bank revealed that the homology of <i>N</i> gene complete sequence shares more than 97% in nucleotide. <i>N</i> gene was cloned into <i>Bam</i>HI and <i>Eco</i>RI multiple cloning sites of the prokaryotic expression vector pET 20 b, and named pETN. After being induced by isopropyl-<i>β</i>-D-thiogalactopyranoside (IPTG), the recombinant nucleocapsid protein was expressed. The result of SDS-PAGE and Western-blot showed that the recombinant nucleocapsid protein was 47 kDa and had strong positive reactions with TGEV-specific antibody.</p>\",\"PeriodicalId\":58797,\"journal\":{\"name\":\"中国农学前沿\",\"volume\":\"1 3\",\"pages\":\"357-360\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2007-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1007/s11703-007-0060-5\",\"citationCount\":\"4\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中国农学前沿\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1007/s11703-007-0060-5\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中国农学前沿","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/s11703-007-0060-5","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 4
摘要
从分离株HB06中扩增出传染性胃肠炎病毒核衣壳蛋白基因，全长1 149 bp，并将其克隆到pMD18-T中。与从基因库中选择的其他传染性胃肠炎病毒(TGEV)株进行序列比较，发现N基因全序列同源性大于97%。将N基因克隆到原核表达载体pET 20b的BamHI和EcoRI多个克隆位点，命名为pETN。经异丙基-β- d -硫代半乳糖苷(IPTG)诱导后，表达重组核衣壳蛋白。SDS-PAGE和Western-blot结果显示重组核衣壳蛋白为47 kDa，与tgev特异性抗体有强烈的阳性反应。
Cloning and expression of nucleocapsid protein gene of TGEV HB06 strain.
The nucleocapsid protein gene of transmissible gastroenteritis virus, 1 149 bp in length, was amplified by RT-PCR from isolated strain HB06 and cloned into pMD18-T. Sequence comparison with other transmissible gastroenteritis virus (TGEV) strains selected from the Gene Bank revealed that the homology of N gene complete sequence shares more than 97% in nucleotide. N gene was cloned into BamHI and EcoRI multiple cloning sites of the prokaryotic expression vector pET 20 b, and named pETN. After being induced by isopropyl-β-D-thiogalactopyranoside (IPTG), the recombinant nucleocapsid protein was expressed. The result of SDS-PAGE and Western-blot showed that the recombinant nucleocapsid protein was 47 kDa and had strong positive reactions with TGEV-specific antibody.
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