Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 415, Issue 1, Pages 131-134Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2011.10.029
Keywords
DNA; Fluorescence; Protein counting; Single molecule
Categories
Funding
- NSF [DMR-0715099, MCB-1022117]
- NIH [1U54CA143869-01 [NU-PS-OC], GM038509]
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [1022117] Funding Source: National Science Foundation
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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.
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