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Nucleic Acids Research, 1979, Vol. 7, No. 4 1091-1108
© 1979

Articles

Sub-genomic RNA in Moloney leukemia virus grown in Iymphoid-derived cell lines consists primarily of homologous viral RNA

J.K. Ball, G.A. Dekaban, S.M. Loosmore, S.K. Chan and J.A. McCarter

Cancer Research Laboratory, University of Western Ontario London, Ontario, Canada

Received March 22, 1979. Moloney murine leukemia virus (MoMuLV) grown in 2 lymphoid-derived murine cell lines (JLS-V9 and TB) contained 2 size classes of RNA subunits: 2.8 ±0.4x106 (n=30)and 1.6±0.1x106 (n=15)daltons. Detectable levels of low molecular weight viral RNA (LMW vRNA) were not present in MoMuLV grown in mouse embryo fibroblasts, nor rat cells, nor was it produced by them when they were infected with MoMuLV containing both species of vRNA. DNA probes complementary to both subunits were synthesized separately using purified reverse transcriptase (R.T.) and calf thymus DNA as primer. Polyadenylated LMW vRNA was selected and, by molecular hybridization was found to be completely homologous with the large RNA subunit. This was confirmed using cDNA probes representing defined regions of the genome The LMW vRNA therefore contains multiple subsets of the viral RNA, which could correspond to multiple deletion mutants possibly generated by an efficient RNA splicing mechanism. In addition, low levels of non-homologous virus-like RNA were detected in MoMuLV grown in TB (2%) and NIH/3T3 cells (0.4%).


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