Investigation of Viral and Host Chromatin by ChIP-PCR or ChIP-Seq Analysis

Thomas Günther, Juliane M. Theiss, Nicole Fischer, Adam Grundhoff
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引用次数: 10

Abstract

Complex regulation of viral transcription patterns and DNA replication levels is a feature of many DNA viruses. This is especially true for those viruses which establish latent or persistent infections (e.g., herpesviruses, papillomaviruses, polyomaviruses, or adenovirus), as long-term persistence often requires adaptation of gene expression programs and/or replication levels to the cellular milieu. A key factor in the control of such processes is the establishment of a specific chromatin state on promoters or replication origins, which in turn will determine whether or not the underlying DNA is accessible for other factors that mediate downstream processes. Chromatin immunoprecipitation (ChIP) is a powerful technique to investigate viral chromatin, in particular to study binding patterns of modified histones, transcription factors or other DNA-/chromatin-binding proteins that regulate the viral lifecycle. Here, we provide protocols that are suitable for performing ChIP-PCR and ChIP-Seq studies on chromatin of large and small viral genomes.© 2016 by John Wiley & Sons, Inc.

用ChIP-PCR或ChIP-Seq分析研究病毒和宿主染色质
病毒转录模式和DNA复制水平的复杂调控是许多DNA病毒的特征。对于那些建立潜伏性或持续性感染的病毒(如疱疹病毒、乳头瘤病毒、多瘤病毒或腺病毒)尤其如此,因为长期持续感染通常需要适应细胞环境的基因表达程序和/或复制水平。控制这些过程的一个关键因素是在启动子或复制起点上建立特定的染色质状态,这反过来将决定介导下游过程的其他因素是否可以访问潜在的DNA。染色质免疫沉淀(ChIP)是研究病毒染色质的一种强有力的技术,特别是研究修饰组蛋白、转录因子或其他调节病毒生命周期的DNA /染色质结合蛋白的结合模式。在这里,我们提供了适用于对大小病毒基因组的染色质进行ChIP-PCR和ChIP-Seq研究的方案。©2016 by John Wiley &儿子,Inc。
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来源期刊
Current Protocols in Microbiology
Current Protocols in Microbiology Immunology and Microbiology-Parasitology
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期刊介绍: Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.
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