Open AccessTemperature-dependent template switching duringin vitrocDNA synthesis by the AMV-reverse transcriptase
Author(s) -
Mohamed Ouhammouch,
Edward N. Brody
Publication year - 1992
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/20.20.5443
Subject(s) - reverse transcriptase , biology , complementary dna , rna directed dna polymerase , primer (cosmetics) , murine leukemia virus , microbiology and biotechnology , in vitro , virology , primer extension , chimera (genetics) , dna , virus , rna , genetics , gene , base sequence , chemistry , organic chemistry
Reverse transcriptase template switching has been invoked to explain several aspects of retroviral 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 degrees C to 50 degrees 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 degrees C).