核酸-蛋白质互作的生物化学研究方法

Biochemical methods for the analysis of DNA-protein interactions

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。

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 immunoprecipitation techniques including nChIP, xChIP, and ChiP-on-chip, which were widely used in epigenetics, were particularly introduced.