Expression of mSDF-1?/GM-CSF fusion protein and its protective effect on radiotherapy-induced hematopoiesis damages / 中国肿瘤生物治疗杂志

ABSTRACT

Objective: To express mSDF-1?/GM-CS fusion protein in the Pichia pastoris expression system and investigate its in vivo promoting effects on hematopoiesis function and immune function.Methods: The mSDF-1?/GM-CS fusion gene was chemically synthesized and was cloned into pichia expression vector;the vector was then transfected into Pichia pastoris expression system.Then the expressed products were detected by SDS-PAGE,Western blotting and purified by ion exchange columns.Rat irradiation model was established by 60Co ? ray and were subcutaneously injected with the fusion protein.The hematopoietic and chemotactic activities of the fusion protein were investigated in vivo.Results: The expression vector pPIC9K-SDF1-L-GM-CSF was successfully constructed,and mSDF-1?/GM-CS fusion protein was successfully expressed by Pichia pastoris strain GSl15.The molecular weight of the protein was about 32000 D,with a concentration of 78 ng/ml.The numbers of bone marrow mononuclear cells and GM-CFU were significantly increased in rats after subcutaneous injection of the fusion protein(P