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

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

Current protocols in microbiology (2016-02-09)
Thomas Günther, Juliane M Theiss, Nicole Fischer, Adam Grundhoff
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-trimethyl-Histone H3 (Lys4) Antibody, clone MC315, rabbit monoclonal, culture supernatant, clone MC315, Upstate®
Sigma-Aldrich
Normal Rabbit IgG, Normal Rabbit IgG Polyclonal Antibody control validated for use in Immunoprecipitation & Western Blotting.
Sigma-Aldrich
Normal Mouse IgG, Normal Mouse IgG Polyclonal Antibody control validated for use in Immunoprecipitation & Western Blotting.