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

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.