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Methods for Measuring the Production of Quorum Sensing Signal Molecules

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Antibiotic Resistance Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1736))

Abstract

One relevant aspect for understanding the bottlenecks that modulate the spread of resistance among bacterial pathogens consists in the effect that the acquisition of resistance may have on the microbial physiology . Whereas studies on the effect of acquiring resistance of bacterial growth are frequently performed, more detailed analyses aiming to understand in depth the cross talk between resistance and virulence, including bacterial communication are less frequent. The bacterial quorum sensing system, is an important intraspecific and interspecific communication system highly relevant for many physiological processes, including virulence and bacterial/host interactions. Some works have shown that the acquisition of antibiotic resistance may impair the quorum sensing response. In addition, some antibiotics as antimicrobial peptides can affect the production and accumulation of the quorum sensing signal molecules. Given the relevance that this system has in the bacterial behavior in the human host, it is important to study the effect that the acquisition of antibiotic resistance may have on the production of quorum sensing signals. In this chapter we present a set of methods for measuring quorum sensing signals based on the use of biosensor strains, either coupled to Thin Layer Chromatography or for performing automated luminometry/spectrophotometry assays. We use Pseudomonas aeruginosa as bacterial model because it has a complex quorum system than encloses different signals. Namely, P. aeruginosa quorum sensing system consists in three different interconnected regulatory networks, each one presenting a specific autoinducer molecule: the las system, which signal is N-(3-oxo-dodecanoyl)-l-homoserine lactone, the rhl system, which signal is N-butanoyl-homoserine lactone and the pqs system, which signals are 2-heptyl-3-hydroxy-4(1H)-quinolone together with its immediate precursor 2-heptyl-4-hydroxy-quinoline.

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Acknowledgments

The work in our laboratory is supported by grants BIO2011-25255 and BIO2014-54507-R from the Spanish Ministry of Economy and Competitiveness, S2010/BMD2414 (PROMPT) from CAM, Spanish Network for Research on Infectious Diseases (REIPI RD12/0015) from the Instituto de Salud Carlos III, and HEALTH-F3-2011-282004 (EVOTAR) from the European Union. MAR has been the recipient of an FPI fellowship. Special thanks are given to Miguel Cámara, Paul Williams, and Robert Hancock for providing control strains and QSSMs.

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Authors and Affiliations

  1. Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Madrid, Spain

    Manuel Alcalde-Rico & José Luis Martínez

Authors
  1. Manuel Alcalde-Rico
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  2. José Luis Martínez
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Correspondence to José Luis Martínez .

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Editors and Affiliations

  1. School of Medicine, University of St. Andrews, North Haugh, United Kingdom

    Stephen H. Gillespie

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Alcalde-Rico, M., Martínez, J.L. (2018). Methods for Measuring the Production of Quorum Sensing Signal Molecules. In: Gillespie, S. (eds) Antibiotic Resistance Protocols. Methods in Molecular Biology, vol 1736. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7638-6_1

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  • DOI: https://doi.org/10.1007/978-1-4939-7638-6_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7636-2

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