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仓鼠细胞APRT基因座处γ辐射诱导的缺失和插入的DNA序列分析。

DNA sequence analysis of gamma radiation-induced deletions and insertions at the APRT locus of hamster cells.

作者信息

Miles C, Sargent G, Phear G, Meuth M

机构信息

Imperial Cancer Research Fund, Clare Hall Laboratories, Hertfordshire, England.

出版信息

Mol Carcinog. 1990;3(4):233-42. doi: 10.1002/mc.2940030411.

Abstract

Gamma radiation-induced gene rearrangements at the Chinese hamster ovary cell locus coding for the purine salvage enzyme adenine phosphoribosyl transferase (APRT) consist of both simple deletions and more complex alterations that are presumably the result of multiple strand breaks. To characterize these mutations at the DNA sequence level, fragments altered by deletion and insertion mutations were obtained by cloning in lambda phage vectors or by using the polymerase chain reaction. The radiation-induced deletions characterized here eliminate 3-4 kb and have at least one breakpoint in an AT-rich region or near short direct or inverted repeats. Insertions involve small fragments (102 and 456 bp) of repetitive DNA that appear to be related to B2 (short interspersed repetitive) and long interspersed repeat families. The novel fragments bear little resemblance to each other or to sequences at the integration sites, and their introduction is accompanied by a small target site deletion.

摘要

γ辐射诱导中国仓鼠卵巢细胞中编码嘌呤补救酶腺嘌呤磷酸核糖转移酶(APRT)的基因发生重排,包括简单缺失和更复杂的改变,这些改变可能是多链断裂的结果。为了在DNA序列水平上表征这些突变,通过克隆到λ噬菌体载体中或使用聚合酶链反应获得了因缺失和插入突变而改变的片段。这里表征的辐射诱导缺失消除了3 - 4 kb,并且在富含AT的区域或短直接重复或反向重复序列附近至少有一个断点。插入涉及重复DNA的小片段(102和456 bp),这些片段似乎与B2(短散在重复序列)和长散在重复序列家族有关。这些新片段彼此之间以及与整合位点的序列几乎没有相似之处,并且它们的引入伴随着一个小的靶位点缺失。

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