Peptide and naked DNA vaccines, aimed at generating strong cytotoxic T lymphocyte (CTL) responses capable of mediating tumor regression, have been considered of the most rapidly evolving technologies for tumor vaccination. Results from clinical trials of these strategies are encouraging, but unfortunately, most of those trials have been proved as partly successful. Given the distinct dynamics of antigen presentation for peptide and gene forms of antigens, we explored a novel concept that collocation of the antigen with the encoding gene in the same vaccine could effectively elicit anti-tumor immunity, and developed a novel peptide-DNA dual vaccine (PDDV), which combines the benefits of peptide and DNA vaccines and could induce tumor-specific CTL response. Furthermore, PDDV effectively protected mice against fatal P815 tumor challenge and cured tumor-bearing DBA/2 mice, suggesting PDDV as a potential formulation of tumor vaccine.