PMID:21472695: Difference between revisions

Revision as of 19:12, 1 September 2013

Citation	No author listed (2011) DamIP: using mutant DNA adenine methyltransferase to study DNA-protein interactions in vivo. Curr Protoc Mol Biol Chapter 21:Unit21.21
Abstract	DamIP is a new method for studying DNA-protein interaction in vivo. A mutant form of DNA adenine methyltransferase (DamK9A) from E. coli is fused to the protein of interest and expressed. The fusion protein will bind to target binding sites and introduce N(6)-adenine methylation in nearby sites in the genomic DNA. Methylated DNA fragments are enriched by immunopreciptation with an antibody that recognizes N(6)-methyladenine, and can then be used for further analysis, e.g., real-time PCR, microarray, or high-throughput sequencing. This method is simple and does not require protein-DNA crosslinking or a specific antibody to the protein of interest. This unit describes the application of this method for identification of DNA binding sites in vivo.
Links	PubMed PMC3085922 Online version:10.1002/0471142727.mb2121s94
Keywords	Artificial Gene Fusion; Chromatin Immunoprecipitation; DNA/metabolism; DNA-Binding Proteins/metabolism; Escherichia coli/enzymology; Gene Expression; Mutant Proteins/genetics; Mutant Proteins/metabolism; Protein Binding; Site-Specific DNA-Methyltransferase (Adenine-Specific)/genetics; Site-Specific DNA-Methyltransferase (Adenine-Specific)/metabolism
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