Construction of eukaryotic expression plasmid of VEGF_(165) gene and its expression in myocardial cells

AIM:To construct a eukaryotic expression vector of human vascular endothelial growth factor(VEGF 165 ) gene, and to investigate the transfection and expression of pcDNA3.1( )/hVEGF 165 eukaryotic expression plasmid in myocardial cells. METHODS: pcDNA3.1( )/hVEGF 165 eukaryotic expression plasmid was constructed. Primarily cultured rat myocardial cells were transiently transfected with LipofectAMINE2000. RT PCR, ELISA, Western blot and immunohistochemical methods were used to detect the expression of VEGF gene and MTT to detect the biological activity of the conditioned medium after the transfection. RESULTS: There were significant increases of VEGF mRNA and protein in the myocardial cells transfected with pcDNA3.1( )/hVEGF 165 . The conditioned medium after the transfection showed the biological activity to stimulate the proliferation of endothelial cells. CONCLUSION: The pcDNA3.1( )/VEGF 165 , a eukaryotic expression plasmid for hVEGF 165 gene is constructed. High levels of VEGF mRNA and protein expression can be obtained in the myocardial cells transfected with pcDNA3.1( )/hVEGF 165 eukaryotic expression plamid. The expressed protein has the biological activity.