Isolation and characterization of T antigen-negative revertants from a line of transformed rat cells containing one copy of the SV40 genome. |
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Authors: | B Steinberg R Pollack W Topp M Botchan |
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Affiliation: | 1. SUNY at Stonybrook Stonybrook, New York 11794 USA;2. Cold Spring Harbor Laboratory Cold Spring Harbor, New York 11724 USA |
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Abstract: | Negative selection with FUdR produced revertants from the transformed rat line 14B, which contains one insertion of the SV40 viral genome (Botchan, Topp and Sambrook, 1976). 14B contains nuclear T antigen, grows to a high density, grows in low serum and is anchorage-independent. The revertants fall into three classes with regard to viral DNA sequences: the SV40 DNA is retained; the SV40 DNA is retained but has undergone a deletion; and the SV40 DNA is lost, generating a cured cell. This heterogeneity is not a result of long-term passage. The revertants arise with a frequency of one in 8.4 X 10(5) cells after as few as 12 passages. All three classes of revertants are T antigen-negative, density-sensitive, more serum sensitive than 14B and anchorage-dependent. These data argue for a direct role of the functioning viral genome in the maintenance of the transformed state, and that with 14B, the phenotypes of transformation are not virus gene dosage-dependent. |
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