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

Butala, Matej and Busby, Stephen J. W. and Lee, David J (2009) DNA Sampling: a method for probing protein binding at specific loci on bacterial chromosomes. Nucleic Acids Research, 37 (5). ISSN 0305-1048

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

Identification Number/DOI: 10.1093/nar/gkp043

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.

Type of Work:Article
Date:01 April 2009 (Publication)
School/Faculty:Colleges (2008 onwards) > College of Life & Environmental Sciences
Department:School of Biosciences
Subjects:Q Science (General)
R Medicine (General)
Institution:University of Birmingham
Copyright Holders:Oxford University Press
ID Code:249
Refereed:YES
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