Construction of Eukaryotic Expression Plasmid of hIGF-1 Gene and its Expression in Myoblast Cells / 中国运动医学杂志

ABSTRACT

Objective To construct a eukaryotic expression vector of huma n insuli n-like growth factor-1 (hIGF-1) gene,and to investigate the transfection and ex pression of pcDNA3.1(-)/hIGF-1 eukaryotic expression plasmid in myoblast cells. Methods pcDNA3.1(-)/hIGF-1 eukaryotic expression plasmid was co nstructed. Myobl ast C2C12 cells were transiently transfected with lipofectamine. RT-PCR a nd immunohistochemical methods were used to detect the expression of hIGF-1 gene . MTT test was used to detect the biological activity of the conditioned medium after the transfection. Results There were significant increas es of hIGF-1 mRN A and protein in the myoblast cells transfected with pcDNA3.1(-)/hIGF-1. The con ditioned medium after the transfection showed the biological activity which coul d stimula te the proliferation of fibroblast cells. Conclusion The pcDNA 3.1(-)/hIGF-1,a eukaryotic expression plasmid for hIGF-1 gene is constructed. High levels of hI GF-1 mRNA and protein expression can be obtained in the myoblast cells transfect ed with pcDNA3.1(-)/hIGF-1 eukaryotic expression plasmid. The expressed protein has the biological activity.