Homologous recombination promoted by reverse transcriptase during copying of two distinct RNA templates.
Retroviruses are known to mutate at high rates. An important source of genetic variability is recombination taking place during reverse transcription of internal regions of the two genomic RNAs. We have designed an in vitro model system, involving genetic markers carried on two RNA templates, to allow a search for individual recombination events and to score their frequency of occurrence. We show that Moloney murine leukemia virus reverse transcriptase alone promotes homologous recombination efficiently. While RNA concentration has little effect on recombination frequency, there is a clear correlation between the amount of reverse transcriptase used in the assay and the extent of recombination observed. Under conditions mimicking the in vivo situation, a rate compatible with ex vivo estimates has been obtained.
Negroni
M
M
Unité de Physicochimie des Macromolécules Biologiques (Unité de Recherche Associée), Institut Pasteur, Paris, France.
Ricchetti
M
M
Nouvel
P
P
Buc
H
H
D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D008957:Models, Genetic; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000201:enzymology; D012313:RNA / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011995:Recombination, Genetic; D014158:Transcription, Genetic
D017931:DNA Primers; D012313:RNA; D012194:RNA-Directed DNA Polymerase