Isolation and properties of Moloney murine leukemia virus mutants: use of a rapid assay for release of virion reverse transcriptase.
A rapid assay for retroviral reverse transcriptase activity released into the culture medium by infected cells was developed. With the assay, 4,000 clonally infected cell lines could be tested in a few hours. We have adapted the assay for use as a screen for the detection of spontaneous viral mutants. Mutants of Moloney murine leukemia virus have been isolated which (i) produce a thermolabile reverse transcriptase, (ii) are temperature sensitive for release of enzyme activity, or (iii) can only productively infect cells already producing gag-related polypeptides. The assay has also been useful for the isolation of nonproducer cells infected with various replication-defective transforming viruses.
D003470:Culture Media; D008970:Molecular Weight; D008979:Moloney murine leukemia virus / Q000201:enzymology / Q000235:genetics*; D009154:Mutation; D012194:RNA-Directed DNA Polymerase / Q000032:analysis* / Q000235:genetics; D013696:Temperature; D010948:Viral Plaque Assay / Q000379:methods
D003470:Culture Media; D012194:RNA-Directed DNA Polymerase