RNA 超诱变反应中正向重复序列和反向重复序列的命运。
Fate of direct and inverted repeats in the RNA hypermutagenesis reaction.
作者信息
Pezo V, Martinez M A, Wain-Hobson S
机构信息
Unité de Rétrovirologie Moléculaire, Institute Pasteur, Paris, France.
出版信息
Nucleic Acids Res. 1996 Jan 15;24(2):253-6. doi: 10.1093/nar/24.2.253.
Abstract
RNA hypermutagenesis results from cDNA synthesis in the presence of highly biased dNTP precursor concentrations and preferentially exploits human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. Such reaction conditions slow down DNA synthesis, which might be conducive to strand transfer and deletion. This has been investigated. A 6 bp inverted repeat nested between 10 bp repeats was efficiently deleted at dCTP concentrations typically used. Inter- or intramolecular strand transfer between 10 bp repeated sequences separated by runs of templated G residues occurred, but at lower concentrations. If RNA hypermutagenesis of a sequence containing direct and inverted repeats is unavoidable, avian myeloblastosis virus (AMV) reverse transcriptase could be used, as strand transfer occurs with much diminished dCTP substrate dependence.
摘要
RNA高度诱变是在dNTP前体浓度高度偏向的情况下进行cDNA合成导致的，并且优先利用1型人类免疫缺陷病毒（HIV-1）逆转录酶。这样的反应条件会减慢DNA合成，这可能有利于链转移和缺失。对此进行了研究。在通常使用的dCTP浓度下，嵌套在10 bp重复序列之间的6 bp反向重复序列被有效删除。在由模板化G残基序列分隔的10 bp重复序列之间发生了分子间或分子内链转移，但发生在较低浓度下。如果含有正向和反向重复序列的序列发生RNA高度诱变不可避免，那么可以使用禽成髓细胞瘤病毒（AMV）逆转录酶，因为链转移发生时对dCTP底物的依赖性大大降低。
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