We constructed simian virus 40 (SV40)/polyomavirus recombinants by replacing in SV40 the T antigen coding region with polyoma early region sequences, either cDNAs encoding small, middle or large T antigen or the wild-type sequence coding all three proteins. The recombinants maintained the SV40 late region and origin of replication and were propagated in COS cells yielding recombinant virus preparations with titers of 10(6)-10(7) infectious particles per milliliter. These viruses were characterized in productive infections of COS cells by analyzing early and late mRNA levels and by following synthesis of polyoma early proteins. In the absence of viral DNA replication, i.e. in infected monkey or mouse cells, expression of the polyoma T antigens was weak. Further experiments indicated that this was mostly due to high genomic instability during amplification, to lower levels of cDNA transcripts as compared to spliced mRNA, and possibly also to lower infectivity of the recombinant virions. It remains to be determined, whether these handicaps are unique to SV40/polyoma recombinants or whether SV40 is in general inadequate as a viral vector.