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Biochem Biophys Res Commun. 2011 Nov 11;415(1):131-4. doi: 10.1016/j.bbrc.2011.10.029. Epub 2011 Oct 12.

Counting proteins bound to a single DNA molecule.

Author information

1
Northwestern University, Department of Molecular Biosciences, 2205 Tech Drive, Hogan 2-100, Evanston, IL 60208-3500, United States. john-graham@northwestern.edu

Abstract

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.

PMID:
22020072
PMCID:
PMC3215857
DOI:
10.1016/j.bbrc.2011.10.029
[Indexed for MEDLINE]
Free PMC Article

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