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J. Biol. Chem., Vol. 279, Issue 15, 14595-14601, April 9, 2004

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A Novel Secreted Protein Toxin from the Insect Pathogenic Bacterium Xenorhabdus nematophila^*

Susan E. Brown, Anh T. Cao, Eric R. Hines, Raymond J. Akhurst, and Peter D. East

From the CSIRO Entomology, GPO Box 1700, Acton, Australian Capital Territory 2601, Australia

The bacterium Xenorhabdus nematophila is an insect pathogen that produces several proteins that enable it to kill insects. Screening of a cosmid library constructed from X. nematophila strain A24 identified a gene that encoded a novel protein that was toxic to insects. The 42-kDa protein encoded by the toxin gene was expressed and purified from a recombinant system, and was shown to kill the larvae of insects such as Galleria mellonella and Helicoverpa armigera when injected at doses of around 30–40 ng/g larvae. Sequencing and bioinformatic analysis suggested that the toxin was a novel protein, and that it was likely to be part of a genomic island involved in pathogenicity. When the native bacteria were grown under laboratory conditions, a soluble form of the 42-kDa toxin was secreted only by bacteria in the phase II state. Preliminary histological analysis of larvae injected with recombinant protein suggested that the toxin primarily acted on the midgut of the insect. Finally, some of the common strategies used by the bacterial pathogens of insects, animals, and plants are discussed.

Received for publication, September 5, 2003 , and in revised form, December 19, 2003.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 61-2-6246-4053; Fax: 61-2-6246-4173; E-mail: sue.brown{at}csiro.au.

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