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

被引：21

作者：

Butala, Matej ^{[1
,2
]}

Busby, Stephen J. W. ^{[1
]}

Lee, David J. ^{[1
]}

机构：

[1] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England

[2] Univ Ljubljana, Dept Biol, Biotechn Fac, Ljubljana, Slovenia

来源：

NUCLEIC ACIDS RESEARCH | 2009年 / 37卷 / 05期

基金：

英国惠康基金; 英国生物技术与生命科学研究理事会;

关键词：

ESCHERICHIA-COLI K-12; MOBILITY SHIFT ASSAY; RNA-POLYMERASE; TRANSCRIPTION FACTORS; MASS-SPECTROMETRY; IDENTIFICATION; GENOME; GENE; PURIFICATION; REPRESSOR;

D O I：

10.1093/nar/gkp043

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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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页数：6

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