Background: Toxoplasma gondii is an obligate intracellular protozoan parasite that can infect large scale of warm-blooded animals including humans, livestock and marine mammals.The heavy incidence and severe or lethal damage caused by T.gondii infection clearly indicates the need for the development of an effective vaccine.The aim of this study was to construct a DNA vaccine expressing several epitopes of T.gondii and evaluate the immune protective efficacy against acute T.gondii infection in mice.Methods: In the present study, several T-and B-cell epitopes from SAG1, GRA2, GRA7 and ROP16 of T.gondii were predicted by bioinformatics analysis, and a multi-epitopes DNA vaccine was constructed.After intramuscular immunization in BALB/c mice with a genetic adjuvant PEGFP-RANTES, we evaluated the systemic responses including survival time, cytokines and special antibodies produced in BALB/c mice.Results: Mice immunized with the multi-epitopes DNA vaccine showed significantly increased serum IgG2a as well as total IgG titers and production of gamma interferon (IFN-γ).Compared with mice immunized with the empty plasmid, mice accepted multi-epitopes DNA vaccine produced a stronger immune response and showed an obviously prolonged survival time.Moreover, mice immunized with the vaccine plus RANTES represent more effective immune responses than any other groups.Conclusions: Our data suggested that the multi-epitopes DNA vaccine was effective in stimulating mice protective immune responses against toxoplasmosis, and that RANTES served as a good genetic adjuvant.