DNA vaccines have frequently been associated with poor efficacy in large animals. In the present study, one administration of an inactivated marker vaccine to cattle considerably boosted both humoral and cellular arms of the immune response primed with Bovine herpesvirus-1 (BoHV-1) DNA vaccines encoding glycoprotein D (gD) or gC+gD. Calves vaccinated according to the DNA prime-inactivated boost also showed significantly enhanced virological protection as compared to controls. The 4-logarithms reduction of virus shedding observed in primed-boosted animals was comparable to the one previously reported in calves immunized twice with marker vaccines. Intradermal immunization of cattle with DNA vaccines promoted a Th2-biased immune response but also primed a cellular component that was further boosted by the inactivated vaccine. Individual IgG2 titers of vaccinated calves were significantly correlated to IFN-gamma production. The immunization protocol described in the present study demonstrates the complementarity between DNA and conventional marker vaccines.