Butala Matej, Busby Stephen J W, Lee David J
School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.
Nucleic Acids Res. 2009 Apr;37(5):e37. doi: 10.1093/nar/gkp043. Epub 2009 Jan 30.
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.
我们描述了一种名为DNA采样的方案,用于从大肠杆菌K-12中快速分离特定的DNA片段以及与之结合的蛋白质。待采样的DNA通过酵母I-SceI巨核酸酶在细胞内生成离散片段,并使用带有FLAG标签的LacI阻遏物和携带抗FLAG抗体的磁珠进行纯化。我们通过研究在大肠杆菌K基因启动子诱导之前或之后与大肠杆菌K基因调控区域结合的蛋白质来说明该方法。