HSV-1 amplicon vectors have been used as platforms for the generation of genetic vaccines against both DNA and RNA viruses. Mice vaccinated with such vectors encoding structural proteins from both foot-and-mouth disease virus and rotavirus were partially protected from challenge with wild-type virus (D'Antuono et al. Vaccine 28: 7363-7372, 2010; Laimbacher et al. Mol Ther 20: 1810-1820, 2012), indicating that HSV-1 amplicon vectors are attractive tools for the development of complex and safe genetic vaccines. This chapter describes the use of HSV-1 amplicon vectors that encode individual or multiple viral structural proteins from a polycistronic transgene cassette in mammalian cells. More precisely, amplicon vectors that encode multiple structural viral proteins support the in situ production of viruslike particles (VLPs) in vector-infected cells. The expression of the viral genes is confirmed by Western blot and immune fluorescence analysis, and the generation of VLPs in vector-infected cells is demonstrated by electron microscopy.