Identification of in vivo DNA targets of chromatin proteins using tethered dam methyltransferase.

摘要: We have developed a novel technique, named DamID, for the identification of DNA loci that interact in vivo with specific nuclear proteins eukaryotes. By tethering Escherichia coli adenine methyltransferase (Dam) to chromatin protein, Dam can be targeted native binding sites this resulting local methylation. Sites methylation subsequently mapped using methylation-specific restriction enzymes or antibodies. demonstrate successful application DamID both Drosophila cell cultures and whole flies. When is tethered DNA-binding domain GAL4, limited region few kilobases surrounding GAL4 sequence. Using we identified number expected unexpected target heterochromatin protein 1. has potential genome-wide mapping targets various