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Sequence-Specific Affinity Chromatography of Bacterial Small Regulatory RNA-Binding Proteins from Bacterial Cells

  • Jonathan Gans
  • Jonathan Osborne
  • Juliet Cheng
  • Louise Djapgne
  • Amanda G. Oglesby-Sherrouse
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1737)

Abstract

Bacterial small RNA molecules (sRNAs) are increasingly recognized as central regulators of bacterial stress responses and pathogenesis. In many cases, RNA-binding proteins are critical for the stability and function of sRNAs. Previous studies have adopted strategies to genetically tag an sRNA of interest, allowing isolation of RNA–protein complexes from cells. Here we present a sequence-specific affinity purification protocol that requires no prior genetic manipulation of bacterial cells, allowing isolation of RNA-binding proteins bound to native RNA molecules.

Keywords

Bacterial small RNAs Hfq RNA-binding proteins Affinity chromatography In vivo crosslinking Western blot 

Notes

Acknowledgments

This work was supported by NIH grant AI123320 and funding from the University of Maryland School of Pharmacy (to A.G.O.-S.).

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Jonathan Gans
    • 1
  • Jonathan Osborne
    • 1
  • Juliet Cheng
    • 1
  • Louise Djapgne
    • 1
  • Amanda G. Oglesby-Sherrouse
    • 1
  1. 1.Department of Pharmaceutical SciencesUniversity of Maryland, School of PharmacyBaltimoreUSA

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