Papillomaviruses are a diverse group of pathogens that infect the skin and mucosal tissues of humans and various animal species. The viral genome is a circular, double‐stranded DNA molecule ∼8‐kb in length. The non‐enveloped papillomavirus capsid is composed of a virally encoded major coat protein, L1, and a minor coat protein, L2. L1 and L2 co‐assemble when expressed in mammalian cells, and can promiscuously encapsidate essentially any <8‐kb plasmid present in the cell nucleus. In the last several years, there has been rapid development of techniques for intracellular production of papillomavirus‐based gene transfer vectors (also known as pseudoviruses). This unit outlines the production and propagative amplification of papillomaviral vectors. The system represents a highly tractable method for converting pre‐existing mammalian expression plasmids into infectious virus stocks. The resulting vectors have utility for in vitro, as well as in vivo gene delivery applications. Curr. Protoc. Cell Biol. 37:26.1.1‐26.1.19. © 2007 by John Wiley & Sons, Inc.