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原核生物DNA甲基转移酶在现代生物学中作为实验和分析工具的应用。

The use of prokaryotic DNA methyltransferases as experimental and analytical tools in modern biology.

作者信息

Buryanov Yaroslav, Shevchuk Taras

机构信息

Shemyakin-Ovchinnikov Institute of Bioorganik Chemistry, Pushchino Branch, Russian Academy of Sciences, 142290 Pushchino, Moscow oblast, Russia.

出版信息

Anal Biochem. 2005 Mar 1;338(1):1-11. doi: 10.1016/j.ab.2004.02.048.

Abstract

Prokaryotic DNA methyltransferases (MTases) are used as experimental and research tools in molecular biology and molecular genetics due to their ability to recognize and transfer methyl groups to target bases in specific DNA sequences. As a practical tool, prokaryotic DNA MTases can be used in recombinant DNA technology for in vitro alteration and enhancing of cleavage specificity of restriction endonucleases. The ability of prokaryotic DNA MTases to methylate cytosine residues in specific sequences, which are also methylated in eukaryotic DNA, makes it possible to use them as analytical reagent for determination of the site-specific level of methylation in eukaryotic DNA. In vivo DNA methylation by prokaryotic DNA MTases is used in different techniques for probing chromatin structure and protein-DNA interactions. Additional prospects are opened by development of the methods of DNA methylation targeted to predetermined DNA sequences by fusion of DNA MTases to DNA binding proteins. This review will discuss the application of prokaryotic DNA MTases of Type II in the methods and approaches mentioned above.

摘要

原核生物DNA甲基转移酶(MTases)因其能够识别甲基基团并将其转移至特定DNA序列中的靶碱基,而被用作分子生物学和分子遗传学的实验及研究工具。作为一种实用工具,原核生物DNA MTases可用于重组DNA技术,以在体外改变和增强限制性内切核酸酶的切割特异性。原核生物DNA MTases能够甲基化特定序列中的胞嘧啶残基,而这些序列在真核生物DNA中也会被甲基化,这使得它们能够用作分析试剂来测定真核生物DNA中位点特异性甲基化水平。原核生物DNA MTases在体内的DNA甲基化被用于多种技术,以探究染色质结构和蛋白质-DNA相互作用。通过将DNA MTases与DNA结合蛋白融合,开发针对预定DNA序列的DNA甲基化方法,开辟了更多前景。本综述将讨论II型原核生物DNA MTases在上述方法和途径中的应用。

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