Abstract
In order to study N-acyl homoserine lactone (AHL)-based quorum sensing in vivo, we present a protocol using an Escherichia coli strain equipped with a luxR-based monitor system, which in the presence of exogenous AHL molecules expresses a green fluorescent protein (GFP). Lungs from mice challenged intratracheally with alginate beads containing both a Pseudomonas aeruginosa strain together with the E. coli monitor strain can be investigated at different time points postinfection. Epifluorescent or confocal scanning laser microscopy (CSLM) is used to detect the GFP-expressing E. coli monitor strain in the lung tissues, indicating production and excretion of AHLs in vivo by the infecting P. aeruginosa.
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Acknowledgement
We acknowledge the contributions made by Drs. Morten Hentzer, Hong Wu, Thomas Bovbjerg Rasmussen, Jens Bo Andersen, and Allan Bech Christensen with regard to the initial developments of the model systems.
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Hultqvist, L.D., Alhede, M., Jakobsen, T.H., Givskov, M., Bjarnsholt, T. (2018). Imaging N-Acyl Homoserine Lactone Quorum Sensing In Vivo. In: Leoni, L., Rampioni, G. (eds) Quorum Sensing. Methods in Molecular Biology, vol 1673. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7309-5_16
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DOI: https://doi.org/10.1007/978-1-4939-7309-5_16
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