HIV-1 gp160截短蛋白在酿酒酵母中的表达  

Expression of Truncate HIV-1 Envelope Glycoprotein gp160 in Saccharomyces cerevisiae

作　　者：杨坤宇[1,2] 李少伟[2] 张毅[1] 李庶甘[1] 何芳萍[2] 林庆山[2] 周正[3] 张军[2] 夏宁邵[2] 

机构地区：[1]厦门出入境检验检疫局,中国福建厦门361012 [2]厦门大学国家传染病诊断试剂与疫苗工程技术研究中心,中国福建厦门361005 [3]武汉大学,中国湖北武汉430072

出　　处：《生命科学研究》2011年第4期298-302,共5页Life Science Research

基　　金：国家高技术研究发展计划重点项目(863计划)(2006AA020905);国家自然科学基金资助项目(3050092);教育部新世纪优秀人才培养计划项目(NCET-0500567)

摘　　要：利用PCR技术从pNL-43上扩增出截短的编码gp160蛋白的基因片断,克隆到酿酒酵母表达载体YEpFLAG-1上构建表达质粒YEp-gp160Δ12,电转化到酿酒酵母中,用缺色氨酸的SC培养基筛选出阳性克隆,重组子经YP培养基诱导后进行全菌蛋白的SDS-PAGE和Western Blotting分析,筛选出高表达菌株.纯化后的重组gp160Δ12(rgp160Δ12)蛋白经ELISA鉴定显示具有良好的生物活性.The coding sequence of truncate gpl60 was amplified by polymerase chain reaction （PCR） and cloned into yeast expression vector YEpFLAG-1 to construct plasmid of YEp-gpl60A12. The plasmids were transformed into yeast Bj3505, and the recombinant strains were identified by cultivating in SC media without tryptophan. After induction in YP media, the whole proteins of recombinant cells were analyzed by SDS- PAGE and Western blotting, and the high-yielding cells were selected. The ELISA performed with a panel of human sera suggested that the purified recombinant gpl60A12 （rgpl60A12） has good bioactivity.

关 键 词：人免疫缺陷病毒Ⅰ型(HIV-1) gp160 酿酒酵母 表达 

分 类 号：Q819[生物学—生物工程]