The immune efficacy of DNA vaccines containing three plasmids encoding gB, gC, and gD glycoproteins (Mix DNA) of Pseudorabies virus (PRV) or the plasmid for gC only (gC DNA), killed virus (KV) vaccine or combination of gC DNA, Mix DNA and KV vaccines was evaluated in mice using primeboost strategy. The mice vaccinated twice with Mix DNA, and once with KV generated higher levels of gCspecific and virus neutralization (VN) antibodies and a stronger cellular immune response than the mice vaccinated three times with the Mix DNA vaccine only. The highest level of VN antibodies were detected in mice vaccinated twice with KV vaccines alone or with combination of DNA and KV vaccines. The challenge of vaccinated mice with the lethal dose of PRV showed that the complete protection against PRV was achieved in the group of mice immunized with the DNA and KV vaccines combined. The results suggested that DNA priming followed by KV vaccine boosting could enhance the antibody response and cellular immunity against PRV infection in mice.