LRP16融合蛋白载体的构建及其原核表达  

Construction of human LRP16 fusion protein vector and its expression in E.coli

作　　者：孟元光[1] 韩为东[2] 黄柯[1] 李琦[2] 伍志强[2] 宋磊[1] 

机构地区：[1]解放军总医院妇产科,北京100853 [2]解放军总医院基础所分子生物室

出　　处：《中国肿瘤临床与康复》2006年第3期212-214,共3页Chinese Journal of Clinical Oncology and Rehabilitation

基　　金：国家自然科学基金(编号30471813);北京市自然科学基金(编号7052061)资助项目

摘　　要：目的观察和鉴定LRP16在大肠杆菌中的表达,并分离、纯化其蛋白产物。方法应用DNA重组技术,用RT-PCR方法从正常人血液中扩增出LRP16基因全长及其抗原表位区,构建重组表达质粒pRSET-C-LRP16,IPTG诱导表达并纯化融合蛋白采用SDS-PAGE凝胶电泳进行鉴定。结果成功构建LRP16基因融合蛋白载体,SDS-PAGE显示pRSET-C-LRP16表达于原核细胞,蛋白经电泳分离,切胶后获得纯化的LRP16蛋白。结论制备的高纯度的LRP16蛋白,可在大肠杆菌中稳定表达。Objective To observe and indentify the expression of human LRP16 gene in E. coli, and isolate and purify its protein product. Methods With DNA recombinant technique ,the recombinant expres- sive plasmide pRSET-C-LRP16 was constructed and introduced into E. coli. pRSET-C-LRP16 fusion protein was expressed at the induction of IPTG. After pRSET-C-LRP16 fusion protein was purified by sodium dedecyl sulfate polyacrylamide gel electrophoresis （ SDS-PAGE ）. Results Recombinant plasmid pRSET-C-LRP16 was successfully constructed. The results of SDS-PAGE showed pRSET-C-LRP16 fusion protein was expressed in E. coli cell,and was produced at a level of 50% of the total cellar protein. Conclusions LRP16 protein could be expressed in E. coli system stably.

关 键 词：LRP16基因 融合蛋白 

分 类 号：R346[医药卫生—基础医学]