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[通过在由修饰转录本形成的R环中进行烷基化对T7噬菌体DNA早期区域进行定向修饰]

[Directed modification of the early region of T7 bacteriophage DNA by alkylation in the R-loop formed by a modified transcript].

作者信息

Dianov G L, Mazin A V, Zaĭtsev B N, Vavilin V I, Salganik R I

出版信息

Mol Biol (Mosk). 1980 Mar-Apr;14(2):261-4.

PMID:7383025
Abstract

For addressed modification of T7 phage DNA a polyalkylating derivative of T7 phage early transcript was used. The derivative was obtained by attaching polyfunctional alkylating agent N,N,N,'-tri(beta-chloroethyl)-N'-(p-formylphenyl) propylene diamine-1,3 to the transcript, their molecules being covalently bound by alkylating 4--5% of the RNA adenine and guanine residues. The polyalkylating RNA was used to form R-loops in the complementary site of T7 DNA to alkylate selectively the T7 phage early DNA. Alkylation of DNA by the modified transcript in the region of the R-loop led to the covalent binding of the transcript to the complementary site of DNA. The correct localization of this R-loop was confirmed by electron microscopy. The application of the results for the addressed mutagenesis and inactivation of selected genes is discussed.

摘要

为了对T7噬菌体DNA进行定向修饰,使用了T7噬菌体早期转录本的一种多烷基化衍生物。该衍生物是通过将多官能烷基化剂N,N,N'-三(β-氯乙基)-N'-(对甲酰基苯基)丙二胺-1,3连接到转录本上而获得的,它们的分子通过烷基化4-5%的RNA腺嘌呤和鸟嘌呤残基而共价结合。多烷基化RNA用于在T7 DNA的互补位点形成R环,以选择性地烷基化T7噬菌体早期DNA。在R环区域,经修饰的转录本对DNA的烷基化导致转录本与DNA的互补位点共价结合。通过电子显微镜证实了该R环的正确定位。文中还讨论了将这些结果应用于定向诱变和所选基因失活的情况。

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