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Nucleic Acids Res. 2009 Apr;37(5):e37. doi: 10.1093/nar/gkp043. Epub 2009 Jan 30.

DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes.

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  • 1School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.

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.

PMID:
19181705
[PubMed - indexed for MEDLINE]
PMCID:
PMC2655658
Free PMC Article
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