Homologous recombination in mammalian cells mediates formation of a functional gene from two overlapping gene fragments.
Homologous recombination in mammalian cells mediates formation of a functional gene from two overlapping gene fragments.
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哺乳动物细胞中的同源重组介导两个重叠基因片段形成功能基因。
DOI：
10.1073/pnas.80.7.2002
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发表时间：
1983
影响因子：
11.1
通讯作者：
Wahl,GM
中科院分区：
文献类型：
--
作者：
deSaintVincent,BR;Wahl,GM
CAD基因受损的中国仓鼠细胞（细胞系ud - a）需要外源性尿苷才能存活。引入两个含有叙利亚仓鼠CAD基因重叠片段的重组质粒，可分离出尿苷原生营养物。相比之下，引入只含有两个重叠片段之一的质粒后，没有获得尿苷原生营养物。DNA限制性内切分析表明，原营养转化子含有一个功能CAD基因，该基因是由两个克隆重叠区域的重组事件形成的。大多数重组事件涉及同源交换，其中一些明显是相互的。原位杂交分析表明，所捐赠的序列在单个染色体位点整合，在每次转化中是不同的。这些结果表明，在哺乳动物细胞中存在一个重组系统，可以催化同源交换。捐赠序列之间的重组是一种手段，通过该系统可以表征，也用于生产新的基因融合。
Chinese hamster cells with a lesion in the CAD gene (cell line Urd-A) require exogenous uridine to survive. Uridine prototrophs could be isolated after introducing two recombinant plasmids containing overlapping fragments of a cloned Syrian hamster CAD gene. In contrast, no uridine prototrophs were obtained after introducing a plasmid containing only one of the two overlapping fragments. DNA restriction analysis showed that the prototrophic transformants contain a functional CAD gene which was formed by a recombination event in the overlapping region of the two clones. Most of the recombination events involved homologous exchanges, and some of them apparently were reciprocal. In situ hybridization analysis revealed that the donated sequences were integrated at a single chromosomal site which was different in each transformant. These results demonstrate the existence of a recombination system(s) in mammalian cells that can catalyze homologous exchanges. Recombination between donated sequences is a means by which this system can be characterized and also utilized for the production of novel gene fusions.