Spin-column isolation of DNA-protein interactions from complex protein mixtures for AFM imaging

Peter R. Hoyt, Mitchel J. Doktycz, Robert J. Warmack, David P. Allison

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Applications of atomic force microscopy (AFM) to investigate structural-functional interactions between DNA and proteins, at the molecular level, should prove valuable for gaining a better understanding of gene expression. Specific genomic DNA-protein interactions occur within a sea of intracellular proteins. Successful AFM imaging requires isolating the specific DNA-protein complex free of background protein contamination. Using spin-column chromatography, we report the successful isolation and AFM imaging of transcription factor DNA complexes from DNA molecules incubated with crude cell lysates. This method should be applicable for the isolation and imaging of other specific DNA-protein complexes pertinent to functional genomic research.

Original languageEnglish
Pages (from-to)139-143
Number of pages5
JournalUltramicroscopy
Volume86
Issue number1-2
DOIs
StatePublished - 2001

Keywords

  • Atomic force microscopy
  • DNA
  • Protein binding
  • Transcription factors

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