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© Research
Publication : Proceedings of the National Academy of Sciences of the United States of America

Homologous recombination promoted by reverse transcriptase during copying of two distinct RNA templates

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Proceedings of the National Academy of Sciences of the United States of America - 18 Jul 1995

Negroni M, Ricchetti M, Nouvel P, Buc H

Link to Pubmed [PMID] – 7542781

Proc. Natl. Acad. Sci. U.S.A. 1995 Jul;92(15):6971-5

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.