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Nucleic Acids Research, 1992, Vol. 20, No. 20 5443-5450
© 1992

MOLECULAR BIOLOGY

Temperature-dependent template switching during in vitro cDNA synthesis by the AMV-reverse transcriptase

Mohamed Ouhammouch and Edward N. Brody*

CNRS Centre de Génétiqu Moléculaire, Laboratoir Propre Associé è I'Université Pierre et Marie Curie 91198 Gif-sur-Yvette, France

* To whom correspondence should be addressed at: State University of New York at Buffalo, Department of Biological Sciences, Cooke Hall, Buffalo, NY 14260, USA

Received June 17, 1992. Revised September 24, 1992. Accepted September 24, 1992.

Reverse transcriptase template switching has been invoked to explain several aspects of retrovlral replication and recombination, and has been reported in vitro for the Moloney murine leukemia virus (M-MuLV) reverse transcriptase. During in vitro cDNA synthesis, the avian myeloblastosis virus (AMV) reverse transcriptase can switch from one template to another in a homology-dependent and temperature-dependent manner. Chimeric cDNA molecules are generated within 30 min at high incubation temperatures, with an increasing efficiency from 42°C to 50°C. Such products are detectable only after much longer incubation times when primer extension reactions are carried out at lower temperatures (90 min at 37°C).


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