Counting proteins bound to a single DNA molecule

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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 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 languageEnglish (US)
Pages (from-to)131-134
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume415
Issue number1
DOIs
StatePublished - Nov 11 2011

Keywords

  • DNA
  • Fluorescence
  • Protein counting
  • Single molecule

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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