Engineering a high-affinity methyl-CpG-binding protein

被引：66

作者：

Jorgensen, Helle F.

Adie, Karen

Chaubert, Pascal

Bird, Adrian P.

机构：

[1] Univ Edinburgh, Wellcome Trust Ctr Cell Biol, Edinburgh EH9 3JR, Midlothian, Scotland

[2] CHU Vaudois, Inst Pathol, CH-1011 Lausanne, Switzerland

来源：

NUCLEIC ACIDS RESEARCH | 2006年 / 34卷 / 13期

基金：

英国惠康基金;

关键词：

D O I：

10.1093/nar/gkl527

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

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页数：7

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