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

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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Jørgensen HF,Adie K,Chaubert P,Bird AP

doi

10.1093/nar/gkl527

subject

Has Abstract

pub_date

2006-08-07 00:00:00

pages

e96

issue

13

eissn

0305-1048

issn

1362-4962

pii

34/13/e96

journal_volume

34

pub_type

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