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将独特和重复的DNA片段插入仓鼠细胞的aprt基因座。

Insertion of unique and repetitive DNA fragments into the aprt locus of hamster cells.

作者信息

Nalbantoglu J, Miles C, Meuth M

机构信息

Imperial Cancer Research Fund, Clare Hall Laboratories, Hertfordshire, U.K.

出版信息

J Mol Biol. 1988 Apr 5;200(3):449-59. doi: 10.1016/0022-2836(88)90535-9.

Abstract

Two insertion mutations occurring at the aprt locus of Chinese hamster ovary cells were analyzed at the nucleotide level by cloning and sequencing the mutant genes. The insertions are similar with respect to events at the target site in that both are accompanied by small deletions. The nature of the DNA introduced, on the other hand, is very different, a unique fragment in a spontaneously occurring mutant and a highly dispersed, repetitive fragment in a gamma radiation-induced strain. The inserts are small (285 and 58 base-pairs) and have none of the structural features or sequences related to putative mobile sequences in mammalian cells. The mechanism of transposition was further examined by cloning the unique donor fragment of the spontaneous mutant. These analyses revealed that the insert in the mutant gene was a precise duplicate of the donor DNA fragment.

摘要

通过对突变基因进行克隆和测序,在核苷酸水平上分析了中国仓鼠卵巢细胞aprt基因座发生的两个插入突变。这两个插入突变在靶位点事件方面相似,因为两者都伴有小的缺失。另一方面,所引入的DNA的性质非常不同,自发突变体中的是一个独特片段,而γ辐射诱导菌株中的是一个高度分散的重复片段。插入片段很小(285和58个碱基对),没有与哺乳动物细胞中假定的移动序列相关的结构特征或序列。通过克隆自发突变体的独特供体片段,进一步研究了转座机制。这些分析表明,突变基因中的插入片段是供体DNA片段的精确复制品。

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