关键词 :人Era;人Era C端蛋白;大肠杆菌;基因表达

    摘 要:目的 在大肠杆菌中的高表达人Era和人Era C端蛋白. 方法 PCR扩增人era基因(h-era)的全长cDNA和h-era cDNA的C端区域基因.h-era cDNA克隆到(His)6融合表达载体pRSET-C中，构建融合表达质粒并转化大肠杆菌BL21(DE3)，经IPTG诱导表达(His)6-h-Era融合蛋白;h-era cDNA的C端区域基因克隆到非融合表达载体pDH中PL 启动子下游，转化大肠杆菌TAP106，42℃热诱导表达人Era C端蛋白(h-Era-C).SDS-PAGE电泳、凝胶薄层扫描检测蛋白的表达. 结果 表达的(His)6-h-Era融合蛋白产物占全菌总蛋白的80%;人Era C端蛋白占全菌总蛋白的40%. 结论 人Era蛋白和Era C端蛋白在大肠杆菌中获得了高表达.

    High expression of human Era and human Era C┐terminal domain in E.coli

    ZHANG Jun-Jie，WU Yuan-Ming，JI Zong-Ling，CHEN Nan-Chun，CHEN Su-Min

    Department of Biochemistry and Molecular Biology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:human Era;human Era C-terminal domain;Es-cherichia.coli;gene expression

    Abstract:AIM To highly express human Era(h-Era)and h-Era C-terminal domain protein in E.coli.METHODS Hu-man era(h-era)cDNA and h-era cDNA C-terminal domain gene were amplified by PCR by using a plasmid containing h-era cDNA as a template，and ligated with prokaryotic expres-sion vector pRSET-C and pDH respectively.E.coli BL21(DE3)transformed with the recombinant plasmid pRSET-C-h-era was induced with IPTG to express(His)6-h-Era fusion protein.E.coli TAP106transformed with the recombinant plasmid pDH-h-era-C was induced at42℃to express h-Era C-terminal domain protein.The expression products were i-dentified by SDS-PAGE and gel thin-layer chromatography-scanning.RESULTS the expressed(His)6-h-Era fusion protein was about80%of total bacterial protein.The ex-pressed h-Era C-terminal domain protein was about40%of total bacterial protein.CONCLUSION (His)6-h-Era fusion protein and h-Era C-terminal domain protein had been suc-cessfully highly expressed in E.coli. ......