Citation
- Authors: Jorgensen, H. F., Adie, K., Chaubert, P., Bird, A. P.
- Year: 2006
- Journal: Nucleic Acids Res 34 e96
- Applications: in vitro / DNA / jetPEI
- Cell type: MEF
Description: Murine embryonic fibroblast cells
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.