Tuberculosis caused by infection with Mycobacterium tuberculosis or M. bovis remains one of the most important infectious diseases of man and animals, and continues to inflict a huge cost in both health and financial terms. The current vaccine, BCG demonstrates variable efficacy and so a more robust vaccine strategy to either replace or supplement BCG is required. We have utilised a DNA prime-BCG boost strategy in a murine M. bovis challenge model using a cocktail of 3 DNA vaccines (encoding Hsp65, Hsp70 and Apa) followed by BCG. Controls were inoculated with vector DNA only, coding DNA only, BCG only or vector DNA followed by BCG boost. Analysis of immune responses by ELISpot prior to challenge, revealed that the coding DNA/BCG prime boost resulted in an increased frequency of antigen-specific IFNgamma producing cells compared to the other regimes. When spleen cell cytokine production to BCG antigens was analysed, significantly more IFNgamma and IL-12 was seen in those groups primed with DNA (coding or vector) prior to BCG than those receiving BCG alone. Analysis of bacterial counts revealed that DNA priming followed by BCG boost further improved the protective immunity induced by BCG alone. Surprisingly, inoculation with vector DNA was as efficacious as the coding DNA in enhancing BCG protection. Taken together these results indicate that whilst the coding DNA vaccines induce antigen specific responses, treatment with the vector DNA is sufficient for the increase in protective immunity over that induced by BCG, suggesting that the vector DNA may be acting as a non-specific adjuvant for BCG immunization.