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

By Matej Butala, Stephen J. W. Busby and David J Lee

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

Topics: Q Science (General), R Medicine (General)
Publisher: Oxford University Press
Year: 2009
OAI identifier: oai:eprints.bham.ac.uk:249

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