核酸-蛋白质互作生物化学研究方法00581.pdfVIP

HEREDITAS (Beijing) 2009 3 , 31(3): 325―336 ISSN 0253-9772 技术与方法 DOI: 10.3724/SP.J.1005.2009.00325 核酸－蛋白质互作的生物化学研究方法 张金璧, 潘增祥, 林飞, 马雪山, 刘红林 , 210095 研究核酸－蛋白质的互作是揭示生命活动机理的基础, 文章简要综述了用于研究核酸－蛋白质互作的 各种生物化学方法。从体内、体外两个研究角度, 针对核酸、蛋白以及复合物 3 个研究水平, 概述了硝化纤维 膜过滤实验、足迹法、EMSA 、Southwestern 杂交等经典分析方法的原理、发展和运用。还着重介绍了最近在 表观遗传学领域中广泛运用的 nChIP 、xChIP 等基本染色质免疫沉淀(ChIP)技术及其衍生出的 ChIP-on-chip 等 方法。 核酸; 蛋白质; 互作; 足迹; 染色质免疫沉淀 Biochemical methods for the analysis of DNA-protein interactions ZHANG Jin-Bi, PAN Zeng-Xiang, LIN Fei, MA Xue-Shan, LIU Hong-Lin College of animal science and technology, Nanjing Agricultural University, Nanjing 210095, China Abstract: Investigation of DNA-protein interactions is fundamental to understand the mechanism underlying a variety of life processes. In this article, various types of biochemical methods in DNA-protein interaction study in vivo and in vitro at the level of DNA, protein, and the complex, respectively were briefly reviewed. Traditional assays including Nitrocellulose filter-binding assay, Footprinting, EMSA, and Southwestern blotting were summarized. In addition, chromatin immunopre- cipitation techniques including nChIP, xChIP, and ChIP-on-chip, which were widely used in epigenetics, were particularly introduced. Keywords: nucleic acid; protein; interaction; footprinting; chromatin immunoprecipitation (ChIP) , , , , DNase Exo [1] , , / , DNA , ,