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Signal Mimics Derived from a Metagenomic Analysis of the Gypsy Moth Gut Microbiota ^,

Department of Plant Pathology, University of Wisconsin—Madison, Madison, Wisconsin 53706,¹ Division of Pharmaceutical Sciences, University of Wisconsin—Madison, Madison, Wisconsin 53705,² Department of Chemistry,³ Department of Entomology, University of Wisconsin—Madison, Madison, Wisconsin 53706⁴

Received 9 November 2006/ Accepted 2 April 2007

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 midguts 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 and a biosensor reside in the same cell. The biosensor consists of a quorum-sensing promoter, which requires an acylhomoserine lactone or other small molecule ligand for activation, driving the 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 the production of a quorum-sensing inducing compound. The signal from this clone induces the activities 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.

Published ahead of print on 13 April 2007.

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