Aim To investigate vascular endothelial growth factor(VEGF) gene expression and bioactivity in rat cardiomyocytes transfected by a recombinant adeno-associated viral vector 2. Methods Recombinant adeno-associated viral vector 2 encoding the human VEGF 165 were constructed and confected in intro to the neonate rat ventricular myocytes.RT-PCR,Western blot and ELISA were used to detect the expression of VEGF gene,and MTT to detect the biological activity of the conditioned medium after transfection. Results The presence of VEGF165 mRNA in transfected cells was demonstrated by RT-PCR,the expression of VEGF165 protein was confirmed by western blot.The level of VEGF165 protein expressed by transfected cells was higher than that by control accroding to an enzyme-linked immunosorbent assay.The conditioned medium after transfection could stimulate the proliferation of endothelial cells. Conclusion Rat cardiomyocytes transfected by Recombinant adeno-associated viral vector 2 containing the human vascular endothelial growth factor 165 cDNA(rAAV-2/hVEGF165) could express VEGF165 with highly biologial activity,which might be useful for gene therapy of myocardial ischemia.