The host restriction imposed on polyoma virus by embryonal carcinoma cells can be circumvented by mutations in the enhancer region of the viral genome. In addition, expression of the early viral genes can be induced by differentiating cells transfected with purified viral DNA. Although no host-range mutants of SV40 have been isolated, expression of T antigen from episomal genomes can be induced by differentiating the embryonal carcinoma cells transfected with microgram quantities of DNA. Further, transient expression of T antigen can be observed in the embryonal carcinoma cells following transfection with large amounts of viral DNA. In addition, replication of the A2 strain of polyoma in F9 cells is enhancer dependent but the SV40 enhancer can functionally substitute for the polyoma enhancer. The F9 host-range mutant TT340 contains five tandem repeats of the region surrounding the origin of replication, and it requires T antigen for replication. The parental strain (Toronto) of the mutant is able to replicate at low levels in a T antigen-dependent manner in F9 cells. This strain also has an unselected host range for PCC4 cells and the mutation in TT340 required for growth on F9 cells does not alter this inherent host range.