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DNA采样：一种探测细菌染色体上特定位点蛋白质结合情况的方法。

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

作者信息

Butala Matej, Busby Stephen J W, Lee David J

机构信息

School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.

出版信息

Nucleic Acids Res. 2009 Apr;37(5):e37. doi: 10.1093/nar/gkp043. Epub 2009 Jan 30.

DOI:10.1093/nar/gkp043
PMID:19181705
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2655658/
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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/959b76446697/gkp043f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/9510e9d9b54f/gkp043f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/b99aa0906604/gkp043f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/0d0911fb6d58/gkp043f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/959b76446697/gkp043f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/9510e9d9b54f/gkp043f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/b99aa0906604/gkp043f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/0d0911fb6d58/gkp043f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78f/2655658/959b76446697/gkp043f4.jpg

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