Single-molecule FRET analysis of protein-DNA complexes

Methods Mol Biol. 2009:543:503-21. doi: 10.1007/978-1-60327-015-1_29.

Abstract

We present a single-molecule method for studying protein-DNA interactions based on fluorescence resonance energy transfer (FRET) and alternating-laser excitation (ALEX) of single diffusing molecules. An application of this method to the study of a bacterial transcription initiation complex is presented.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Buffers
  • Chromatography, High Pressure Liquid
  • DNA / isolation & purification
  • DNA / metabolism*
  • DNA-Directed RNA Polymerases / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescence Resonance Energy Transfer / methods*
  • Fluorescent Dyes / metabolism
  • Lasers
  • Proteins / metabolism*
  • Spectrophotometry, Ultraviolet
  • Staining and Labeling
  • Transcription, Genetic

Substances

  • Buffers
  • Fluorescent Dyes
  • Proteins
  • DNA
  • DNA-Directed RNA Polymerases