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

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 repress...

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Detalles Bibliográficos
Autores principales: Butala, Matej, Busby, Stephen J. W., Lee, David J.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2009
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2655658/
https://www.ncbi.nlm.nih.gov/pubmed/19181705
http://dx.doi.org/10.1093/nar/gkp043
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author Butala, Matej
Busby, Stephen J. W.
Lee, David J.
author_facet Butala, Matej
Busby, Stephen J. W.
Lee, David J.
author_sort Butala, Matej
collection PubMed
description 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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spelling pubmed-26556582009-04-01 DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes Butala, Matej Busby, Stephen J. W. Lee, David J. Nucleic Acids Res Methods Online 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. Oxford University Press 2009-04 2009-01-30 /pmc/articles/PMC2655658/ /pubmed/19181705 http://dx.doi.org/10.1093/nar/gkp043 Text en © 2009 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Butala, Matej
Busby, Stephen J. W.
Lee, David J.
DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes
title DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes
title_full DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes
title_fullStr DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes
title_full_unstemmed DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes
title_short DNA sampling: a method for probing protein binding at specific loci on bacterial chromosomes
title_sort dna sampling: a method for probing protein binding at specific loci on bacterial chromosomes
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2655658/
https://www.ncbi.nlm.nih.gov/pubmed/19181705
http://dx.doi.org/10.1093/nar/gkp043
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AT busbystephenjw dnasamplingamethodforprobingproteinbindingatspecificlocionbacterialchromosomes
AT leedavidj dnasamplingamethodforprobingproteinbindingatspecificlocionbacterialchromosomes

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