DNA vaccine approaches have been applied to generate the protective immunity against various pathogens. However, the strength of immune responses induced by DNA vaccine is weak compared with conventional vaccines. The prime-boost vaccination using DNA vaccine and other viral vector has been suggested as one way to circumvent this limitation. In the present study, we used in vivo CTL activity assay to determine $CD^{8+}$ T cell-mediated immunity induced by prime-boost vaccination with a DNA vaccine ($gB_{498-505}$ DNA) and recombinant vaccinia virus ($VVg_{B498-505}$) expressing $gB_{498-505}$ epitope peptide (SSIEFARL) of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB). The most potent in vivo CTL activity was induced in mice received $VVgB_{498-505}$ when both $gB_{498-505};and;VVgB_{498-505}$ were used at priming step and boosted with the alternative vaccine vector expressing whole antigen protein (gBw). Priming with vaccine vector expressing gBw followed by the use of $VVgB_{498-505}$ at boosting step also induced strong in vivo CTL activity. We also examined in vivo CTL activity after immunization of mice with epitope-expressing vaccine vector at both priming and boosting step. Curiously, in vivo CTL activity mediated by $CD^{8+}$ T cells was strongly elicited at memory stage when animals were primed with $VVgB_{498-505}$ and subsequently boosted with $gB_{498-505}$ DNA. Because the use of $VVgB_{498-505}$ at priming followed by boosting with $gB_{498-505}$ DNA induced most optimal immunity, these results suggest that the order of vaccine type should be carefully considered when used vaccine type expressing only epitope for prime-boost vaccination.