中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 猪A组轮状病毒Vp7基因与双标记表达载体pW425et的重组及在大肠杆菌中的表达 吉林农业大学学报 2008,30 (1) 88-92
Position: Home > Articles > Expression of Recombinant Vp7 Gene of Porcine Rotavirus A with Double Labeling Expression Vector pW425et in Escherichia coli Journal of Jilin Agricultural University 2008,30 (1) 88-92

猪A组轮状病毒Vp7基因与双标记表达载体pW425et的重组及在大肠杆菌中的表达

作  者：
胡静涛;宁军;肖冲;王春凤
单  位：
吉林农业大学动物科技学院
关键词：
轮状病毒;Vp7基因;表达载体;大肠杆菌;表达
摘  要：
以猪A组轮状病毒mRNA为模板,应用反转录聚合酶链式反应(RT-PCR)技术,扩增了981 bp的Vp7基因,通过T-A克隆技术,将PCR产物克隆至pGEM-T载体中,构建出克隆质粒pGEM-T-Vp7。以SacⅠ和KpnⅠ双酶切pGEM-T-Vp7及双标记表达载体pW425et,并将纯化的Vp7基因亚克隆至双标记表达载体pW425et中,构建出可以在乳酸菌与大肠杆菌之间穿梭表达的原核表达重组质粒pW425et-Vp7。将pW425et-Vp7转化至thyA基因缺陷型的大肠杆菌感受态Escherichia coliX13中,经生长功能弥补筛选阳性克隆和SDS-PAGE分析,可见约40 kD的融合蛋白。Western blot分析表明该蛋白具有与猪轮状病毒多克隆抗体的反应原性。
译  名：
Expression of Recombinant Vp7 Gene of Porcine Rotavirus A with Double Labeling Expression Vector pW425et in Escherichia coli
作  者：
HU Jing-tao,NING Jun,XIAO Chong,WANG Chun-feng (College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China)
关键词：
Rotavirus;Vp7 gene;expression vector;E.coli;expression
摘  要：
Vp7 gene of Porcine Rotavirus A was amplified by the reverse transcription-polymerase chain reaction(RT-PCR),the product of which was an approximately 981bp cDNA segment.The PCR product was cloned into pGEM-T vector,by T-A cloning technique.Cloning plasmid pGEM-T-Vp7 and the double labeling expression vector pW425et were digested by Sac Ⅰ and Kpn Ⅰ double enzymes respectively.The purified Vp7 gene was subcloned into the vector pW425et.Thus,the recombinant pW425et-Vp7 was constructed,which then was transformed into the competence thyA gene-mutant E.coli X13.Treated lysates of bacterium were loaded directly onto SDSPAGE,on which approximately 40 kD fusion protein was observed.The protein was further analyzed by Western blot,which indicated that the protein was reactive with the antibody of Rotavirus A.The results lay foundation for further studies on the expression in the lactobacillus.

相似文章

导出引用
计量
文章访问数: 5
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46