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拟南芥中用于组蛋白修饰和蛋白质-DNA结合分析的染色质免疫沉淀方案

Chromatin Immunoprecipitation Protocol for Histone Modifications and Protein-DNA Binding Analyses in Arabidopsis.

作者信息

You Wanhui, Pien Stéphane, Grossniklaus Ueli

机构信息

Department of Plant and Microbial Biology & Zurich-Basel Plant Science Center, University of Zurich, Zollikerstrasse 107, 8008, Zurich, Switzerland.

Bayer CropScience AG, Alfred-Nobel-Straße 50, 40789, Monheim am Rhein, Germany.

出版信息

Methods Mol Biol. 2017;1456:1-13. doi: 10.1007/978-1-4899-7708-3_1.

Abstract

Epigenetic control of plant development via histone modifications is involved in different processes ranging from embryonic development, vegetative development, flowering time control, floral organ development, to pollen tube growth. The identification of an increasing number of epigenetically regulated processes was greatly advanced by methods allowing the survey of genome-wide histone modifications and chromatin-protein interactions. However, genome-wide approaches are too broad to access in detail a large number of histone modifications taking place at a single locus. Here, we provide a robust chromatin immunoprecipitation (ChIP) protocol, allowing in vivo analyses of multiple chromatin modifications and binding of histone modifiers in different plant organs and tissues. This method is quantitative and provides a way to study the dynamic state of chromatin during plant development and also in response to different environmental stimuli.

摘要

通过组蛋白修饰对植物发育进行的表观遗传调控涉及从胚胎发育、营养发育、开花时间控制、花器官发育到花粉管生长等不同过程。能够检测全基因组组蛋白修饰和染色质-蛋白质相互作用的方法极大地推动了越来越多表观遗传调控过程的鉴定。然而,全基因组方法过于宽泛,无法详细研究单个位点发生的大量组蛋白修饰。在此,我们提供了一个可靠的染色质免疫沉淀(ChIP)方案,可用于在体内分析不同植物器官和组织中的多种染色质修饰以及组蛋白修饰因子的结合情况。该方法具有定量性,为研究植物发育过程中以及对不同环境刺激响应时染色质的动态状态提供了一种途径。

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