Induction and Purification of C. difficile Phage Tail-Like Particles

Methods Mol Biol. 2016;1476:167-75. doi: 10.1007/978-1-4939-6361-4_12.

Abstract

Due to the inherent limitations of conventional antibiotics for the treatment of C. difficile infection (CDI), there is a growing interest in the development of alternative treatment strategies. Both bacteriophages and R-type bacteriocins, also known as phage tail-like particles (PTLPs), show promise as potential antibacterial alternatives for treating CDI. Similar to bacteriophages, but lacking a viral capsid and genome, PTLPs remain capable of killing target bacteria. Here we describe our experience in the induction and purification of C. difficile PTLPs. These methods have been optimized to allow production of concentrated, non-contractile, and non-aggregated samples for both sensitivity testing and structural electron microscopy studies.

Keywords: Clostridium difficile; Induction; Microscopy; Phage tail-like particles; Purification.

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Bacteriocins / genetics
  • Bacteriocins / metabolism
  • Bacteriophages / drug effects*
  • Bacteriophages / growth & development
  • Bacteriophages / pathogenicity
  • Bacteriophages / ultrastructure
  • Centrifugation, Density Gradient
  • Cesium / chemistry
  • Chlorides / chemistry
  • Clostridioides difficile / virology*
  • Magnesium Sulfate / pharmacology
  • Microscopy, Electron, Transmission
  • Norfloxacin / pharmacology*
  • Polyethylene Glycols / pharmacology
  • Protein Binding
  • Receptors, Virus / genetics
  • Receptors, Virus / metabolism
  • Virion / drug effects*
  • Virion / growth & development
  • Virion / pathogenicity
  • Virion / ultrastructure

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Bacteriocins
  • Chlorides
  • Receptors, Virus
  • Cesium
  • Polyethylene Glycols
  • Magnesium Sulfate
  • cesium chloride
  • Norfloxacin
  • polyethylene glycol 8000