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Discovery of Bacillus thuringiensis Virulence Genes Using Signature-Tagged Mutagenesis in an Insect Model of Septicaemia

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Abstract

Transposon Tn917 was used to identify Bacillus thuringiensis genes required for virulence and survival in a Manduca sexta (tobacco hornworm) septicaemia model. Uniquely tagged transposons, n = 72, were constructed and used to generate 1152 insertion mutants. Sixteen pools of 72 mutants were screened in the infection model, and 12 virulence-attenuated mutants were unable to survive the infection. Analysis of the mutated DNA sequences implicated an arsR family transcriptional regulator, a histone-like DNA-binding protein, a transposon, and several sequences of unknown function in B. thuringiensis pathogenesis.

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Acknowledgments

We thank Daniel Guttmann for providing protocols, David Holden for supplying pID408, Didier Lereclus for supplying pRN5101 and pDG783, and Jorge Ibarra for providing Bt MEX312. This project was funded by the BBSRC (Biotechnology and Biological Sciences Research Council).

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Authors and Affiliations

  1. Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, England

    James R. Steggles, Jinhong Wang & David J. Ellar

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  1. James R. Steggles
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  2. Jinhong Wang
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  3. David J. Ellar
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Correspondence to Jinhong Wang.

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Steggles, J.R., Wang, J. & Ellar, D.J. Discovery of Bacillus thuringiensis Virulence Genes Using Signature-Tagged Mutagenesis in an Insect Model of Septicaemia. Curr Microbiol 53, 303–310 (2006). https://doi.org/10.1007/s00284-006-0037-2

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  • DOI: https://doi.org/10.1007/s00284-006-0037-2

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