Construction of eukaryotic expression vector of VEGF_(165) gene and its expression in rat cardiomyocytes

Objective To construct an eukaryotic expression vector of human vascular endothelial growth factor (VEGF 165) gene, and to investigate the transfection and expression of pCR3.hVEGF 165 eukaryotic expression vector in rat myocardial cells.Methods pCR3.hVEGF 165 eukaryotic expression vector was constructed. Primarily cultured rat cardiomyocytes were transfected with liposome/plasmid pCR3.hVEGF 165. RT-PCR, Southern blot, immunohistochemical method and ELISA were used to detect the expression and secretion of VEGFgene.Result There were significant increases in VEGFmRNA and protein in the myocardial cells transfected with pCR3.hVEGF 165.Conclusions The pCR3.hVEGF 165, an eukaryotic expression plasmid for hVEGF 165 gene,was constructed. High levels of VEGFmRNA and protein expression could be obtained in the myocardial cells transfected with pCR3.hVEGF 165 eukaryotic expression plasmid.