聚合酶链式反应期间的DNA重组。
DNA recombination during PCR.
作者信息
Meyerhans A, Vartanian J P, Wain-Hobson S
机构信息
Laboratoire de Biologie et Immunologie Moléculaires des Rétrovirus, Institut Pasteur, Paris, France.
出版信息
Nucleic Acids Res. 1990 Apr 11;18(7):1687-91. doi: 10.1093/nar/18.7.1687.
Abstract
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.
摘要
对两个不同的HIV1 tat基因序列进行PCR共扩增会导致重组DNA分子的形成。此类重组体的频率高达所有扩增分子的5.4%，通过将Taq DNA聚合酶延伸时间增加6倍，可使其降低2.7倍。交叉位点基本上定位在三个离散区域，表明存在特定的Taq DNA聚合酶停顿或终止位点。在研究诸如RNA病毒、多基因家族或重复序列等异质遗传物质时，PCR介导的重组可能是一个问题。这一现象可被利用来从相关序列创建嵌合分子。
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