HOMOLOGOUS RECOMBINATION PROMOTED BY REVERSE-TRANSCRIPTASE DURING COPYING OF 2 DISTINCT RNA TEMPLATES
HOMOLOGOUS RECOMBINATION PROMOTED BY REVERSE-TRANSCRIPTASE DURING COPYING OF 2 DISTINCT RNA TEMPLATES
复制标题
DOI：
10.1073/pnas.92.15.6971
复制
发表时间：
1995-07-18
影响因子：
11.1
通讯作者：
BUC, H
中科院分区：
文献类型：
--
作者：
NEGRONI, M;RICCHETTI, M;BUC, H
众所周知，逆转录病毒的变异率很高。遗传变异的一个重要来源是在两个基因组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，包括携带在两个RNA模板上的遗传标记，以允许搜索单个重组事件并对其发生频率进行评分。结果表明，单用Moloney小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，但在检测中使用的逆转录酶的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，获得了与离体估计相容的速率。
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.