Plasmid DNA vaccination is a very powerful and easy method for the induction of strong humoral and cell-mediated immune responses in mice. The technique has also been successfully applied for the definition of immunodominant, human T-cell epitopes using HLA-transgenic mice. By virtue of its strong capacity to induce CD4(+)-mediated Th1 and CD8(+)-mediated cytotoxic T-lymphocyte responses, this vaccine approach is particularly attractive for the prophylaxis of intracellular pathogens, such as Mycobacterium tuberculosis (TB) and other pathogenic mycobacteria. In small rodents, the potential of mycobacterial DNA vaccines is well established. In humans, DNA vaccines are clearly less immunogenic and, so far, TB-specific DNA vaccines have not been assessed in humans. However, a number of studies in cattle and sheep have demonstrated the potential of mycobacterial DNA vaccines in larger animals. Also, immunization protocols combining the potent priming capacity of plasmid DNA with subsequent boosting with recombinant protein, recombinant pox-viruses or with Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccine are particularly promising for future applications. The potential of mycobacterial DNA vaccines for immunotherapy and post-exposure prophylaxis is still not clear.