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首页> 外文期刊>Molecular and Cellular Biology >Intramolecular recombination between transfected repeated sequences in mammalian cells is nonconservative.
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Intramolecular recombination between transfected repeated sequences in mammalian cells is nonconservative.

机译：哺乳动物细胞中转染的重复序列之间的分子内重组是非保守的。

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When plasmids carrying a fragmented gene with segments present as direct repeats are introduced into mammalian cells, recombination or gene conversion between the repeated sequences can reconstruct the gene. Intramolecular recombination leads to the deletion of the intervening sequences and the loss of one copy of the repeat. This process is known to be stimulated by double-strand breaks. Two current models for recombination in eucaryotic cells propose that the reaction is initiated by double-strand breaks, but differ in their predictions as to the fate of the intervening sequences. One model suggests that these sequences are always lost, while the other indicates that the reaction will be conservative as a function of the position of the double-strand break. We have constructed a plasmid in which two overlapping portions of the simian virus 40 early region, which contains the origin and T-antigen gene, are present as direct repeats separated by sequences containing a plasmid with a simian virus 40 origin of replication. Recombination across the repeated segments could produce a plasmid with an origin of replication and/or a plasmid with a gene for a functional T-antigen which would drive the replication of both. Introduction of this construction into African green monkey kidney cells, without coinfection, establishes a condition in which the products of the recombination or gene conversion can be interpreted unambiguously. We find that the majority of the reconstruction reactions are nonconservative.
机译：当将带有片段的基因的质粒以直接重复的形式存在的片段引入哺乳动物细胞时，重复序列之间的重组或基因转化可以重建基因。分子内重组导致插入序列的缺失和重复序列的一个拷贝的丢失。已知该过程受双链断裂刺激。目前在真核细胞中重组的两种模型表明，该反应是由双链断裂引发的，但它们对中间序列命运的预测不同。一种模型表明这些序列总是丢失的，而另一种模型则表明该反应将是保守的，取决于双链断裂的位置。我们构建了一个质粒，其中猿猴病毒40早期区域的两个重叠部分（包含起点和T抗原基因）以直接重复的形式存在，并被含有猿猴病毒40复制起点的质粒的序列隔开。跨重复片段的重组可以产生具有复制起点的质粒和/或具有功能性T抗原的基因的质粒，该基因将驱动两者的复制。将该结构引入非洲绿猴肾细胞而不进行共感染，建立了可以明确解释重组或基因转化产物的条件。我们发现大多数重建反应是非保守的。

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