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Curr Protoc Microbiol. 2011 Nov;Chapter 1:Unit1C.1. doi: 10.1002/9780471729259.mc01c01s23.

Methods for analysis of bacterial autoinducer-2 production.

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1
Department of Plant and Microbial Biology, University of California, Berkeley, California, USA.

Abstract

Quorum sensing is a cell-cell signaling process that many bacteria use to regulate gene expression as a function of the density of the population. This phenomenon involves the production, release, and response to small chemical molecules termed autoinducers. Most autoinducers are species-specific; however, one autoinducer called autoinducer-2 (AI-2) is produced and detected by many species of bacteria and thus can foster inter-species communication. This unit describes two assays to detect and quantify AI-2 from biological samples. The first uses a bacterial reporter strain, which produces bioluminescence in response to AI-2. The second is an in vitro assay based on a modified version of an AI-2 receptor fused to a cyan fluorescent protein and a yellow fluorescent protein. Binding of AI-2 to this fusion protein induces a dose-dependent decrease in fluorescence resonance energy transfer (FRET), enabling quantification of the AI-2 concentration in the samples.

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