This study aims to evaluate the effect of electroporation on the expression of DNA vaccine in vivo and its induced adaptive immune responses. The gene expression in vivo following DNA delivery via electroporation was determined by assessing reporter gene products of firefly luciferase by using in vivo imaging device, which was further dissected as the tissue distribution of gene products, the peak level and the lasting time of expression when compared to the traditional immunization through muscular injection. The data showed that EP did not alter the tissue distribution, but increased the peak level and lowered the plateau dosage of DNA vaccine. Further experiments with DNA vaccine pDRVI1-HIV expressing an ENV from HIV-1 CRF07BC demonstrated that electroporation was able to enhance cellular and humoral immune responses which were gauged by the level of interferon-γ secreting enzyme-linked immunospot (ELISPOT) responses and HIV-1 specific binding antibody titers, respectively. Altogether, electroporation in vivo technology is likely to be a useful tool in the administration of DNA vaccines.