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 c DNA和 h-era c DNA的 C端区域基因 . h- era c DNA克隆到 (His) 6融合表达载体 p RSET- C中 ,构建融合表达质粒并转化大肠杆菌BL 2 1(DE3) ,经 IPTG诱导表达 (His) 6 - h- Era融合蛋白 ;h- erac DNA的 C端区域基因克隆到非融合表达载体 p DH中 PL 启动子下游 ,转化大肠杆菌 TAP10 6 ,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 .
AIM To highly express human Era (h Era) and h Era C terminal domain protein in E.coli . METHODS Human 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 expression 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 TAP106 transformed with the recombinant plasmid pDH h era C was induced at 42℃ to express h Era C terminal domain protein. The expression products were identified by SDS PAGE and gel thin layer chromatography scanning. RESULTS the expressed (His)6 h Era fusion protein was about 80% of total bacterial protein. The expressed h Era C terminal domain protein was about 40% of total bacterial protein. CONCLUSION (His)6 h Era fusion protein and h Era C terminal domain protein had been successfully highly expressed in E.coli .
出处
《第四军医大学学报》
北大核心
2001年第8期759-762,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金!资助项目 (39870 380 )
全军医药卫生科研基金!资助项目(98M10 8)
关键词
人ERA
人Era端蛋白
大肠杆菌
基因表达
human Era
human Era C terminal domain
Escherichia.coli
gene expression
