The minimum amount of homology required for homologous recombination in mammalian cells.
The minimum amount of homology required for homologous recombination in mammalian cells.
复制标题
哺乳动物细胞中同源重组所需的最小同源量。
DOI：
10.1128/mcb.4.11.2253-2258.1984
复制
发表时间：
1984
影响因子：
5.3
通讯作者：
Subramani,S
中科院分区：
文献类型：
--
作者：
Rubnitz,J;Subramani,S
尽管DNA序列同源性被认为是原核生物和真核生物中同源重组事件的先决条件，但尚未对哺乳动物细胞中同源重组所需的最低同源性进行系统研究。我们使用体外构建的猴病毒40-pBR 322杂合质粒作为底物，定量培养猴细胞中的分子内同源重组。通过同源重组切除野生型猿猴病毒40 DNA，通过病毒空斑试验进行评分。使用一系列含有0至243个碱基对的同源性的质粒，我们已经表明，重组频率随着同源性的降低而降低，当同源性从214个碱基对降低到163个碱基对时，重组频率下降最快。然而，也观察到低重组频率，只有14个碱基对的同源性。
Although DNA sequence homology is believed to be a prerequisite for homologous recombination events in procaryotes and eucaryotes, no systematic study has been done on the minimum amount of homology required for homologous recombination in mammalian cells. We have used simian virus 40-pBR322 hybrid plasmids constructed in vitro as substrates to quantitate intramolecular homologous recombination in cultured monkey cells. Excision of wild-type simian virus 40 DNA by homologous recombination was scored by the viral plaque assay. Using a series of plasmids containing 0 to 243 base pairs of homology, we have shown that the recombination frequency decreases as the homology is reduced, with the sharpest drop in recombination frequency occurring when the homology was reduced from 214 to 163 base pairs. However, low recombination frequencies were also observed with as little as 14 base pairs of homology.