Objective：To construct Mycobacterium tuberculosis ESAT6 （pE） antigen DNA vaccine and ubiquitin-ESAT6 fusion gene （pUE） DNA vaccine. Methyls： The constructed DNA vaccines were intramuscularly inoculated into female BALB/c mice separately. The serum antibodies （including IgG, IgG1, and IgG2a ）, Cytokines （IFN-γ and IL-4）, and cytotoxic T lymphocytes response were detected in the immunized mice. Results： Mice in pE group had a higher serum level of IgG （P〈 0.01） and a lower value of IgG2a/IgG1 （[2.28±0.40] vs [3.87±0.60] ,P〈0.05） than mice in pUE group. Besides, mice in pUE group secreted more IFN-γ than those in pE group （P〈0.01）, but secreted less IL-4 （P〈0.01）. Furthermore, pUE enhanced the activity of CTL. The results showed that pUE DNA vaccine induced weaker humoral immune response, but stronger cellular immune responses compared to pE DNA vaccine. Conclusion.. The pUE DNA vaccine constructed in this study sheds new lights on the prophylactic and therapy of tuberculosis.[