Counting proteins bound to a single DNA molecule

John S. Graham; Reid C. Johnson; John F. Marko

doi:10.1016/j.bbrc.2011.10.029

Counting proteins bound to a single DNA molecule

John S. Graham^*, Reid C. Johnson, John F. Marko

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

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.

Original language	English (US)
Pages (from-to)	131-134
Number of pages	4
Journal	Biochemical and Biophysical Research Communications
Volume	415
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2011.10.029
State	Published - Nov 11 2011

Funding

Work at NU was supported by NSF Grants DMR-0715099, MCB-1022117, and by NIH Grant 1U54CA143869-01 [NU-PS-OC]. Work at UCLA was supported by NIH Grant GM038509. The authors thank Dr. Botao Xiao for generously providing the equilibrium data for figure 4.

Keywords

DNA
Fluorescence
Protein counting
Single molecule

ASJC Scopus subject areas

Molecular Biology
Biophysics
Biochemistry
Cell Biology

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10.1016/j.bbrc.2011.10.029

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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.",

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AU - Marko, John F.

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AB - 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.

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Counting proteins bound to a single DNA molecule

Abstract

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Keywords

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