- DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes.
DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes.
Nucleic acids research (2009-02-03)
Matej Butala, Stephen J W Busby, David J Lee
PMID19181705
ABSTRACT
We describe a protocol, DNA sampling, for the rapid isolation of specific segments of DNA, together with bound proteins, from Escherichia coli K-12. The DNA to be sampled is generated as a discrete fragment within cells by the yeast I-SceI meganuclease, and is purified using FLAG-tagged LacI repressor and beads carrying anti-FLAG antibody. We illustrate the method by investigating the proteins bound to the colicin K gene regulatory region, either before or after induction of the colicin K gene promoter.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)