A novel DNA vaccine pFUW-Ag85B-Rv3425 based on lentiviral vector pFUW was constructed to improve the transfection efficiency in vivo. The DNA injection combined with electroporation in C57BL/6 mice three times with two-week interval was conducted. After the last immunization, the mice were sacrificed. The spleen lymphocytes were isolated and analyzed by flow cytometry. The expression levels of IgG, IgG1, IgG2a, and interleukin 2 (IL-2) in serum were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that DNA injection in combination with electroporation could induce stronger humoral immunity, more Th1-type immune response and significantly enhance CD8⁺ T cell and CD27⁺ B cell proliferation. It is indicated that DNA vaccine in combination with electroporation can be taken as a new therapy strategy against tuberculosis.