DNA recombination during PCR.
PCR co-amplification of two distinct HIV1 tat gene sequences lead to the formation of recombinant DNA molecules. The frequency of such recombinants, up to 5.4% of all amplified molecules, could be decreased 2.7 fold by a 6 fold increase in Taq DNA polymerase elongation time. Crossover sites mapped essentially to three discrete regions suggesting specific Taq DNA polymerase pause or termination sites. PCR mediated recombination may be a problem when studying heterogeneous genetic material such as RNA viruses, multigene families, or repetitive sequences. This phenomenon can be exploited to create chimeric molecules from related sequences.
Laboratoire de Biologie et Immunologie Moléculaires des Rétrovirus, Institut Pasteur, Paris, France.
A
A
Meyerhans
J P
JP
Vartanian
S
S
Wain-Hobson
D000595:Amino Acid Sequence; D001483:Base Sequence; D002678:Chimera; D003001:Cloning, Molecular / Q000379:methods; D004274:DNA, Recombinant / Q000378:metabolism*; D005784:Gene Amplification; D015696:Gene Products, tat / Q000235:genetics; D005814:Genes, Viral; D015756:Genes, tat; D015497:HIV-1 / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D054322:tat Gene Products, Human Immunodeficiency Virus
D004274:DNA, Recombinant; D015696:Gene Products, tat; D015345:Oligonucleotide Probes; D054322:tat Gene Products, Human Immunodeficiency Virus