UNIT 21.21 DamIP: Using Mutant DNA Adenine Methyltransferase to Study DNA-Protein Interactions In Vivo
Published Online: 1 APR 2011
Copyright © 2011 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Xiao, R. and Moore, D. D. 2011. DamIP: Using Mutant DNA Adenine Methyltransferase to Study DNA-Protein Interactions In Vivo. Current Protocols in Molecular Biology. 94:21.21:21.21.1–21.21.10.
- Published Online: 1 APR 2011
- Published Print: APR 2011
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 N6-adenine methylation in nearby sites in the genomic DNA. Methylated DNA fragments are enriched by immunopreciptation with an antibody that recognizes N6-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. Curr. Protoc. Mol. Biol. 94:21.21.1-21.21.10. © 2011 by John Wiley & Sons, Inc.
- DNA adenine methyltransferase;
- transcription factor binding sites;
- chromatin immunoprecipitation