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三螺旋导向补骨脂素加合物在SV40穿梭载体中诱导低频重组。

Triple helix directed psoralen adducts induce a low frequency of recombination in an SV40 shuttle vector.

作者信息

Sandor Z, Bredberg A

机构信息

Department of Medical Microbiology, University of Lund, General Hospital, Malmö, Sweden.

出版信息

Biochim Biophys Acta. 1995 Sep 19;1263(3):235-40. doi: 10.1016/0167-4781(95)00109-t.

Abstract

Triple helix forming oligonucleotide directed psoralen adducts in a mammalian shuttle vector have been reported to be repaired efficiently in human cells. In this study we examined the role of intermolecular homologous recombination in triple helix targeted psoralen adduct repair. A simian virus 40 (SV40) shuttle vector carrying a mutated supF gene was treated with a triplex forming oligonucleotide psoralen conjugate and cotransfected into human cells with a second plasmid bearing the wild type supF gene. Recombinants with a reactivated marker gene were detected by an X-gal assay in indicator bacteria. We could observe a low frequency of psoralen adduct induced recombination indicating that recombination does not play a major role in triplex directed psoralen adduct repair. The implications for targeted mutagenesis by triple helix forming oligonucleotides are discussed.

摘要

据报道,在哺乳动物穿梭载体中形成三链螺旋的寡核苷酸导向补骨脂素加合物在人类细胞中能被有效修复。在本研究中,我们检测了分子间同源重组在三链螺旋靶向补骨脂素加合物修复中的作用。用一个携带突变型supF基因的猿猴病毒40(SV40)穿梭载体与一个三链形成寡核苷酸补骨脂素共轭物进行处理,并与携带野生型supF基因的第二个质粒共转染到人类细胞中。通过在指示菌中进行X-gal检测来检测标记基因重新激活的重组体。我们可以观察到补骨脂素加合物诱导重组的频率较低,这表明重组在三链导向补骨脂素加合物修复中并不起主要作用。本文讨论了三链螺旋形成寡核苷酸在靶向诱变方面的意义。

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