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首页> 外文期刊>Molecular and Cellular Biology >Somatic cells efficiently join unrelated DNA segments end-to-end.
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Somatic cells efficiently join unrelated DNA segments end-to-end.

机译：体细胞有效地端对端连接无关的DNA片段。

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摘要

Molecular substrates for probing nonhomologous recombination in somatic cells were constructed by inserting pBR322 sequences at selected sites on the simian virus 40 (SV40) genome. The chimeric products are too large to be packaged into an SV40 capsid. Therefore, production of viable progeny requires that most of the pBR322 sequences be deleted without altering any SV40 sequences that are essential for lytic infection. As judged by plaque assay, these recombination events occur at readily detectable frequencies after transfection into CV1 monkey kidney cells. Depending on the site of pBR322 insertion, the infectivities of the full-length circular or linear chimeras ranged from 0.02 to 2% of the infectivity of linear wild-type SV40 DNA. Nucleotide sequence analysis of several recombinant progeny revealed three distinct classes of recombination junction and indicated that the causative recombination events were minimally dependent on sequence homology. Potential mechanisms involving recombination at internal sites or at ends were distinguished by measuring the infectivity of chimeric molecules from which various lengths of pBR322 had been removed. These data support end-to-end joining as the primary mechanism by which DNA segments recombine nonhomologously in somatic cells. This end joining appears to be very efficient, since SV40 genomes with complementary single-stranded tails or with short non-complementary pBR322 tails were comparably infectious. Overall, this study indicates that mammalian somatic cells are quite efficient at the willy-nilly end-to-end joining of unrelated DNA segments.
机译：通过在猿猴病毒40（SV40）基因组的选定位点插入pBR322序列，构建了探测体细胞中非同源重组的分子底物。嵌合产品太大，无法包装到SV40衣壳中。因此，有活力的后代的产生需要删除大多数pBR322序列，而不改变任何溶菌感染必不可少的SV40序列。通过噬菌斑测定判断，这些重组事件在转染到CV1猴肾细胞后以易于检测的频率发生。取决于pBR322插入的位置，全长圆形或线性嵌合体的感染性为线性野生型SV40 DNA的感染性的0.02％至2％。几个重组子代的核苷酸序列分析揭示了三类不同的重组连接，并表明致病性重组事件对序列同源性的依赖性最小。通过测量已除去各种长度的pBR322的嵌合分子的感染力，可以区分涉及内部位点或末端重组的潜在机制。这些数据支持端到端连接，这是DNA片段在体细胞中非同源重组的主要机制。这种末端连接似乎非常有效，因为具有互补单链尾部或短非互补pBR322尾部的SV40基因组具有相当的感染力。总体而言，这项研究表明，哺乳动物的体细胞在不相关的DNA片段的无意识的端到端连接中非常有效。

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