Coexpression Of Human Endostatin And PredhPK-5 In E.coli By Two Incompatible Plasmids

Objective: To contructre combinant plasmid pET28a/hES and coexpress human Endostatin and predhPK-5 in E. coli.Methods: mRNA from human liver tissue was exracted, and the Endostatin gene was amplified by RT-PCR. Clone it into pET-28a (＋), then cotransformed into E. coliBL21(DE3) with pGEX-1λT/predhPK-5, and coexpessed.Result: The two incompatible plasmids can be coexisted under the pressure of two antibiotics(ampicillin and kanamycin). After induction with IPTG, both human Endostation and predhPK-5 gene were coexpressed.Conclution: The cloning and expessing of human Endostatin lay the foundation of luther study. The two incompatible plasmids pET28a/hES and pGEX-1λT/predhPK-5 coexpressed recombinant protein proteins in E. coli. A new method for coexpression of proteins in E. coli containing two incompatible plasmids was proved to be feasible.