Identification of Novel DNA-Binding proteins Using DNA-Affinity Chromatography/Pull Down

Brandon L. Jutras, Ashutosh Verma, Brian Stevenson

Research output: Contribution to journalArticlepeer-review

71 Scopus citations

Abstract

This units presents methods through which one may isolate and identify novel bacterial DNA-binding proteins. Briefly, the DNA sequence of interest is affixed to beads, and then incubated with bacterial cytoplasmic extract. Washes with buffers containing nonspecific DNA and low-salt concentrations will remove non-adhering and low-specificity DNAbinding proteins, while subsequent washes with higher salt concentrations will elute more specific DNA-binding proteins. Eluted proteins may then be identified by standard proteomic techniques.

Original languageEnglish
Article number1F.1
JournalCurrent Protocols in Microbiology
Issue numberSUPPL.24
DOIs
StatePublished - Feb 2012

Keywords

  • DNA-binding protein
  • Gene regulation
  • Prokaryote

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Virology

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