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.
Meyerhans
A
A
Laboratoire de Biologie et Immunologie Moléculaires des Rétrovirus, Institut Pasteur, Paris, France.
Vartanian
J P
JP
Wain-Hobson
S
S
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