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

Matej Butala | Zendy; Stephen Busby | Zendy; David J. Lee | Zendy

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

Author(s) -

Matej Butala,

Stephen Busby,

David J. Lee

Publication year - 2009

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkp043

Subject(s) - biology , lac repressor , colicin , dna , repressor , gene , genetics , circular bacterial chromosome , escherichia coli , microbiology and biotechnology , dna binding protein , lac operon , gene expression , transcription factor

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.

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