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)


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.


Dimethyl Sulfate Saran Wrap Vortex Briefly Footprinting Assay Chemical Nuclease 
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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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