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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

Pien Stéphane, Grossniklaus Ueli

机构信息

Institute of Plant Biology & Zürich-Basel Plant Science Center, University of Zürich, Zürich, Switzerland.

出版信息

Methods Mol Biol. 2010;631:209-20. doi: 10.1007/978-1-60761-646-7_15.

Abstract

Epigenetic gene regulation via histone modifications controls different processes ranging from embryonic development, vegetative development, floral induction, floral organ development, to pollen tube growth. The identification of an increasing number of epigenetically regulated processes was greatly advanced by genome-wide histone modification and chromatin-protein interaction surveys. However, genome-wide approaches are too global 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

通过组蛋白修饰进行的表观遗传基因调控控制着从胚胎发育、营养生长、花诱导、花器官发育到花粉管生长等不同过程。全基因组组蛋白修饰和染色质-蛋白质相互作用研究极大地推动了对越来越多表观遗传调控过程的识别。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。在这里,我们提供了一个可靠的染色质免疫沉淀(ChIP)方案,允许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。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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