HTPP-MDC原核表达载体的构建及在大肠杆菌中的表达  

Construction of HTPP-MDC's Prokaryotic Expression Vector and Its Expression in Escherichia Coli

作　　者：卢雪梅[1,2] 黄演婷[1,2] 金小宝[1,2] 汪洁[1,2] 朱家勇[1,2] 

机构地区：[1]广东药学院药用生物活性物质研究所,广东广州510006 [2]广东省生物活性药物研究重点实验室,广东广州510006

出　　处：《化学与生物工程》2012年第5期16-18,45,共4页Chemistry & Bioengineering

基　　金：国家自然科学基金资助项目(30671832)

摘　　要：通过双酶切、连接、转化等方法将肝靶向穿膜肽-家蝇天蚕素(HTPP-MDC)融合基因克隆至原核表达载体pET32a(+)上,构建重组表达质粒HTPP-MDC/pET32a。重组质粒转化E.coli BL21(DE3)后,以IPTG诱导,SDS-PAGE分析表明重组融合蛋白得到了可溶性表达,Western blot杂交证实了表达蛋白的抗原活性。为HTPP-MDC的生物学活性研究及产业化开发奠定了基础。The fusion gene HTPP-MDC was cloned into prokaryotic expression vector pET32a（＋） to con- struct prokaryotic expression recombinant plasmid HTPP-MDC/pET32a. The recombinant plasmids were trans- formed into E. coli BL21 （DE3） for protein expression. Analysis of fusion protein expressed in E. coli after being induced by IPTG was conducted with SDS-PAGE and Western blot. SDS-PAGE analysis showed that the re- combinant fusion protein HTPP-MDC-Trx was solubly expressed successfully, and Western blot analysis con- firmed its antigenic activity on mouse-His monoclonal antibody. These results may provide foundation for fur- ther study of its biological activity and applications.

关 键 词：肝靶向穿膜肽 家蝇天蚕素 融合基因 原核表达 

分 类 号：Q786[生物学—分子生物学]