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

M Butala; Stephen Busby; David Lee

doi:10.1093/nar/gkp043

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

M Butala, Stephen Busby, David Lee

Biosciences

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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.

Original language	English
Journal	Nucleic Acids Research
DOIs	https://doi.org/10.1093/nar/gkp043
Publication status	Published - 30 Jan 2009

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http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=19181705

Molecular Mechanism of Triggering of a Bacterial Transcription Activator
Busby, S. (Principal Investigator)
Biotechnology & Biological Sciences Research Council
1/12/04 → 14/09/08
Project: Research Councils

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title = "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.",

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

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Molecular Mechanism of Triggering of a Bacterial Transcription Activator

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