反转染色质免疫沉淀（R-ChIP）。

Reverse Chromatin Immunoprecipitation (R-ChIP).

机构信息

State Key Laboratory of Desert and Oasis Ecology, Key Laboratory of Ecological Safety and Sustainable Development in Arid Lands, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, Urumqi, China.

Xinjiang Key Laboratory of Conservation and Utilization of Plant Gene Resources, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, Urumqi, China.

出版信息

Methods Mol Biol. 2024;2846:123-132. doi: 10.1007/978-1-0716-4071-5_8.

DOI:10.1007/978-1-0716-4071-5_8

PMID:39141233

Abstract

DNA-protein interactions play fundamental roles in diverse biological functions. The gene-centered method is used to identify the upstream regulators of defined genes. In this study, we developed a novel method for capturing the proteins that bind to certain chromatin fragments or DNA sequences, which is called reverse chromatin immunoprecipitation (R-ChIP). This technology uses a set of specific DNA probes labeled with biotin to isolate chromatin or DNA fragments, and the DNA-associated proteins are then analyzed using mass spectrometry. This method can capture DNA-associated proteins with sufficient quantity and purity for identification.

摘要

DNA-蛋白质相互作用在各种生物功能中发挥着基础性作用。基因中心法被用于鉴定特定基因的上游调控因子。在本研究中，我们开发了一种新的方法来捕获与特定染色质片段或 DNA 序列结合的蛋白质，称为反向染色质免疫沉淀（R-ChIP）。该技术使用一组带有生物素标记的特定 DNA 探针来分离染色质或 DNA 片段，然后使用质谱分析法分析与 DNA 相关的蛋白质。该方法可以捕获足够数量和纯度的与 DNA 相关的蛋白质，以进行鉴定。