转染到哺乳动物细胞中的DNA分子间同源重组的拓扑学要求。
Topological requirements for homologous recombination among DNA molecules transfected into mammalian cells.
作者信息
Wake C T, Vernaleone F, Wilson J H
出版信息
Mol Cell Biol. 1985 Aug;5(8):2080-9. doi: 10.1128/mcb.5.8.2080-2089.1985.
Abstract
Cultured animal cells rearrange foreign DNA very efficiently by homologous recombination. The individual steps that constitute the mechanism(s) of homologous recombination in transfected DNA are as yet undefined. In this study, we examined the topological requirements by using the genome of simian virus 40 (SV40) as a probe. By assaying homologous recombination between defective SV40 genomes after transfection into CV1 monkey cells, we showed that linear molecules are preferred substrates for homologous exchanges, exchanges are distributed around the SV40 genome, and the frequency of exchange is not diminished significantly by the presence of short stretches of non-SV40 DNA at the ends. These observations are considered in relation to current models of homologous recombination in mammalian cells, and a new model is proposed. The function of somatic cell recombination is discussed.
摘要
培养的动物细胞通过同源重组非常有效地重排外源DNA。构成转染DNA中同源重组机制的各个步骤尚未明确。在本研究中，我们以猴病毒40（SV40）基因组为探针研究了拓扑学要求。通过检测缺陷型SV40基因组转染CV1猴细胞后的同源重组，我们发现线性分子是同源交换的首选底物，交换分布在SV40基因组周围，并且末端存在短片段非SV40 DNA时交换频率不会显著降低。结合当前哺乳动物细胞同源重组模型对这些观察结果进行了探讨，并提出了一个新模型。还讨论了体细胞重组的功能。
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