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Nucleic Acids Res. 2006 Aug 7;34(13):e96.

Engineering a high-affinity methyl-CpG-binding protein.

Author information

  • 1The Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK. helle.jorgensen@csc.mrc.ac.uk

Abstract

Core members of the MBD protein family (MeCP2, MBD1, MBD2 and MBD4) share a methyl-CpG-binding domain that has a specific affinity for methylated CpG sites in double-stranded DNA. By multimerizing the MDB domain of Mbd1, we engineered a poly-MBD protein that displays methyl-CpG-specific binding in vitro with a dissociation constant that is >50-fold higher than that of a monomeric MBD. Poly-MBD proteins also localize to methylated foci in cells and can deliver a functional domain to reporter constructs in vivo. We propose that poly-MBD proteins are sensitive reagents for the detection of DNA methylation levels in isolated native DNA and for cytological detection of chromosomal CpG methylation.

PMID:
16893950
PMCID:
PMC1540740
DOI:
10.1093/nar/gkl527
[PubMed - indexed for MEDLINE]
Free PMC Article
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