STAR Protocols (Dec 2024)

Protocol for analyzing the function of the type VI secretion system of the oral symbiont Aggregatibacter aphrophilus in targeting pathobionts

  • Jan Oscarsson,
  • Kai Bao,
  • Akiko Shiratsuchi,
  • Jonas Grossmann,
  • Witold Wolski,
  • Kyaw Min Aung,
  • Mark Lindholm,
  • Anders Johansson,
  • Ferdousi Rahman Mowsumi,
  • Sun Nyunt Wai,
  • Georgios N. Belibasakis,
  • Nagihan Bostanci

Journal volume & issue
Vol. 5, no. 4
p. 103415

Abstract

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Summary: Here, we present a protocol for evaluating type VI secretion system (T6SS)-dependent fitness of the oral symbiont A. aphrophilus using biofilm competition assays and metaproteomics. We describe steps for designing T6SS-specific mutants. We then detail procedures for using them in competition assays with the pathobiont A. actinomycetemcomitans and in biofilm models, analyzing metaproteomes to assess the impact of the T6SS on multiple pathobionts. The biofilm model is designed to mimic the oral plaque ecosystem and includes seven species.For complete details on the use and execution of this protocol, please refer to Oscarsson et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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