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Appl. Environ. Microbiol. doi:10.1128/AEM.02617-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Signal mimics derived from a metagenomic analysis of gypsy moth gut microbiota

Department of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin 53706, Division of Pharmaceutical Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53705, and Department of Chemistry; and Department of Entomology, University of Wisconsin-Madison, Madison, Wisconsin 53706

^* To whom correspondence should be addressed. Email: joh{at}plantpath.wisc.edu.

	Abstract

Bacterial signaling is an important part of community life, but little is known about the signal transduction pathways of the as-yet-uncultured members of microbial communities. To address this gap, we aimed to identify genes directing the synthesis of signals in uncultured bacteria associated with the midgut of gypsy moth larvae. We constructed a metagenomic library consisting of DNA extracted directly from the midgut microbiota and analyzed it using an intracellular screen, designated METREX, which detects inducers of quorum sensing. In this screen, the metagenomic DNA resides in the same cell as a biosensor. The biosensor consists of a quorum sensing promoter, which requires an acylhomoserine lactone or other small molecule ligand for activation, driving expression of the reporter gene, gfp. We identified an active metagenomic clone encoding a monooxygenase homologue that mediates a pathway of indole oxidation that leads to production of a quorum sensing inducing compound. The signal from this clone induces activity of LuxR from Vibrio fischeri and CviR from Chromobacterium violaceum. This study is the first to identify a new structural class of quorum sensing inducer from uncultured bacteria.

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