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DNA-Footprinting using Crude Embryonic Extracts from Xenopus laevis

  • Rob Orford
  • Darren Mernagh
  • Matt Guille
Part of the Methods in Molecular Biology™ book series (MIMB, volume 127)

Abstract

The interactions of trans-acting factors with cis-acting DNA elements have been a mainstay of molecular biology. With the emergence of new techniques, it is becoming easier to investigate how specific genes are regulated in differentiating cells. In 1978, Galas and Schmitz developed a modified employment of the enzyme deoxyribonuclease I (DNase I) that made it possible to map the DNA-binding site of a putative transcription factor (1). With the advent of Maxam and Gilbert sequencing, it also became apparent that chemical nucleases could be used to investigate the nature of protein-DNA interactions (2). Although these techniques are applicable to other types of DNA-binding proteins, we will specifically consider transcription factors.

Keywords

Dimethyl Sulfate Saran Wrap Vortex Briefly Footprinting Assay Chemical Nuclease 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  • Rob Orford
    • 1
  • Darren Mernagh
    • 1
  • Matt Guille
    • 1
  1. 1.Biophysics Laboratories, Institute of Biomolecular and Biomedical SciencesUniversity of PortsmouthPortsmouthUK

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