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<p>Immunization of mice with plasmids containing genes of <i>Trypanosoma cruzi</i> induces protective immunity in the murine model of Chagas disease. A cDNA clone that codes for an amastigote-specific surface protein (<i>Tc</i>SSP4) was used as a candidate to develop a DNA vaccine. Mice were immunized with the recombinant protein <i>rTcSSP4 </i>and with cDNA for <i>Tc</i>SSP4<i>,</i> and challenged with bloodstream trypomastigotes. Immunization with <i>rTcSSP4</i> protein makes mice more susceptible to trypomastigote infection, with high mortality rates, whereas mice immunized with a eukaryotic expression plasmid containing the <i>TcSSP4</i> cDNA were able to control the acute phase of infection. Heart tissue of gene-vaccinated animals did not show myocarditis and tissue damage at 365 days following infection, as compared with control animals. INF-&#947; was detected in sera of DNA vaccinated mice shortly after immunization, suggesting the development of a Th1 response. The <i>Tc</i>SSP4 gene is a promising candidate for the development of an anti-<i>T. cruzi </i>DNA vaccine.</p>