Key result
Undiluted passages of the Prague Strain of Rous Sarcoma Virus resulted in heterogeneity of genomic RNA, which was decreased by diluted passage or cloning.
Undiluted passages of PR-RSV A in embryo fibroblasts lead to RNA heterogeneity, including incomplete RNAs containing the src sequence.
May confound retroviral model interpretation; leaves open optimal passage protocols for consistent src analyses.
The size of genomic RNA in PR-RSV A passaged in chick embryo fibroblasts (CEF) or quail embryo fibroblasts (QEF) was determined by gel electrophoresis. The results showed that 3 undiluted passages resulted in heterogeneity of RNA. The heterogeneity of the smaller incomplete RNAs in the virus stock was decreased by diluted passage or cloning, but RNA of the b subunit size and a subunit RNA of complete genome size were relatively stable. These heterogeneous RNAs were characterized by hybridization analysis. The RNAs from 4 peaks hybridized with both cDNAtotal and cDNAsrc to appreciable extents, indicating that they were derived from viral RNA and that at least some of them contained the src sequence. This finding and the failure to isolate a td mutant from the undiluted-passaged virus stock or from some subclones that had a and b subunits of RNA indicate that the td virus was only a minor constituent of the incomplete virus population caused by undiluted passages. Some viruses with incomplete RNA in the virus stock could produce foci with the aid of td B77 or RAV-60. The emergence of rd viruses by undiluted passages was indicated.
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Nasu et al. (1979) studied this question. Undiluted passages vs. Diluted passage or cloning was evaluated on Heterogeneity of genomic RNA. Undiluted passages of the Prague Strain of Rous Sarcoma Virus resulted in heterogeneity of genomic RNA, which was decreased by diluted passage or cloning.
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