Abstract
In order to investigate the capacity of the N-terminal half of large T antigen to convert primary cells to continuous cell lines conditions were standardized for transforming primary mouse embryo fibroblasts of C57B1/6 origin (B6/MEF) with DNA. Using these conditions the transforming capability of SV40 virions, viral DNA, and a plasmid, pSV3T3-20-GV ( C. E. Clayton, D. Murphey, M. Lovett, and P. W. J. Rigby, Nature (London), 299, 59–61, 1982 ) containing the 5′ half of the SV40 early region, was determined in a quantitative immortalization assay. The plasmid pSV3T3-20-GV transformed B6/MEF at only 1 16 the frequency of a plasmid containing the entire early region. These results suggest that the 3′ half of large T antigen which cannot be produced by this plasmid strongly influences the frequency of immortalization. It is not known whether this influence reflects the presence of a transformation domain in the carboxy terminus of large T antigen or, alternatively, results from an altered conformation or stability of the truncated polypeptide.
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