HTPP-MDC原核表达载体的构建及在大肠杆菌中的表达

摘要

The fusion gene HTPP-MDC was cloned into prokaryotic expression vector pET32a( + ) to construct prokaryotic expression recombinant plasmid HTPP-MDC/pET32a.The recombinant plasmids were transformed into E.coli BL2KDE3) 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 recombinant fusion protein HTPP-MDC-Trx was solubly expressed successfully,and Western blot analysis confirmed its antigenic activity on mouse-His monoclonal antibody.These results may provide foundation for further study of its biological activity and applications.%通过双酶切、连接、转化等方法将肝靶向穿膜肽(一)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,SDSPAGE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.