Expression and identification of recombinant fusion protein of thrombopoietin mimetic peptide and growth hormone in Pichia pastoris / 第三军医大学学报

ABSTRACT

Objective To fulfill high-efficient expression of rTMP-GH,a recombinant fusion protein of thrombopoietin mimetic peptide and human growth hormone,in Pichia pastoris.Methods cDNA fragment coding for rTMP-GH was amplified by PCR,and subsequently cloned into the expression vector pPIC9K.The linearlized plasmid pPIC9K-rTMP-GH was transformed into Pichia pastoris GS115 cells and screened in MD/MM plates under pressure of G418.The recombinant fusion protein rTMP-GH was identified by Western blotting,and its biological activity was analyzed by its ability on colony formation of megakaryoblasts.Results The pPIC9K-rTMP-GH expression plasmid was successfully constructed.GS115 cells with high expression of rTMP-GH was screened out,and the rough purified rTMP-GH showed promotive effect on megakaryoblast colony formation.Conclusion High-efficient expression of rTMP-GH in Pichia pastoris is achieved.