Unit

UNIT 21.21 DamIP: Using Mutant DNA Adenine Methyltransferase to Study DNA-Protein Interactions In Vivo

  1. Rui Xiao,
  2. David D. Moore

Published Online: 1 APR 2011

DOI: 10.1002/0471142727.mb2121s94

Current Protocols in Molecular Biology

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.

Author Information

  1. Baylor College of Medicine, Houston, Texas

Publication History

  1. Published Online: 1 APR 2011
  2. Published Print: APR 2011

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 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.

Keywords:

  • DNA adenine methyltransferase;
  • transcription factor binding sites;
  • DamIP;
  • immunoprecipitation;
  • chromatin immunoprecipitation