Counting proteins bound to a single DNA molecule

被引：11
|
作者：
Graham, John S. [1 ]
Johnson, Reid C. [2 ]
Marko, John F. [1 ,3 ]
机构：
[1] Northwestern Univ, Dept Mol Biosci, Evanston, IL 60208 USA
[2] Univ Calif Los Angeles, David Geffen Sch Med, Dept Biol Chem, Los Angeles, CA 90095 USA
[3] Northwestern Univ, Dept Phys & Astron, Evanston, IL 60208 USA
来源：
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2011年 / 415卷 / 01期
基金：
美国国家科学基金会;
关键词：
DNA; Fluorescence; Protein counting; Single molecule; ESCHERICHIA-COLI; HU;
D O I：
10.1016/j.bbrc.2011.10.029
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：
Chromosomes contain DNA covered with proteins performing functions such as architectural organization and transcriptional regulation. The ability to count the number of proteins bound to various regions of the genome is essential for understanding both architectural and regulatory functions. We present a straightforward method of counting gfp-conjugated proteins bound to an individual duplex DNA molecule by calibrating to a commercially available fluorescence standard using wide-field fluorescence microscopy. We demonstrate our method using the E. coli nucleoid-associated protein Fis. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码：131 / 134
页数：4
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