Negroni M, Ricchetti M, Nouvel P, Buc H
Unité de Physicochimie des Macromolécules Biologiques (Unité de Recherche Associée), Institut Pasteur, Paris, France.
Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6971-5. doi: 10.1073/pnas.92.15.6971.
Retroviruses are known to mutate at high rates. An important source of genetic variability is recombination taking place during reverse transcription of internal regions of the two genomic RNAs. We have designed an in vitro model system, involving genetic markers carried on two RNA templates, to allow a search for individual recombination events and to score their frequency of occurrence. We show that Moloney murine leukemia virus reverse transcriptase alone promotes homologous recombination efficiently. While RNA concentration has little effect on recombination frequency, there is a clear correlation between the amount of reverse transcriptase used in the assay and the extent of recombination observed. Under conditions mimicking the in vivo situation, a rate compatible with ex vivo estimates has been obtained.
已知逆转录病毒会以高频率发生突变。基因变异性的一个重要来源是在两条基因组RNA内部区域的逆转录过程中发生的重组。我们设计了一个体外模型系统，其中涉及两个RNA模板上携带的遗传标记，以寻找单个重组事件并对其发生频率进行评分。我们表明，单独的莫洛尼鼠白血病病毒逆转录酶能有效地促进同源重组。虽然RNA浓度对重组频率影响不大，但在测定中使用的逆转录酶量与观察到的重组程度之间存在明显的相关性。在模拟体内情况的条件下，获得了与体外估计值相符的速率。
