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

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.

SUBMITTER: Butala M

PROVIDER: S-EPMC2655658 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature

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

Butala Matej M Busby Stephen J W SJ Lee David J DJ

Nucleic acids research 20090130 5

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 p ...[more]

PMID: 19181705

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