【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.更多还原

【Abstract】 The coding sequence of truncate gp160 was amplified by polymerase chain reaction(PCR) and cloned into yeast expression vector YEpFLAG-1 to construct plasmid of YEp-gp160Δ12.The plasmids were transformed into yeast Bj3505,and the recombinant strains were identified by cultivating in SC media with-out 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 gp160Δ12(rgp160Δ12) has good bioactivity.更多还原