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Applied and Environmental Microbiology, July 1999, p. 3021-3026, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Production of Cry11A and Cry11Ba Toxins in Bacillus sphaericus Confers Toxicity towards Aedes aegypti and Resistant Culex Populations

Pascale Servant,1,* Marie-Laure Rosso,2 Sylviane Hamon,2 Sandrine Poncet,3 Armelle Delécluse,2 and Georges Rapoport1

Unité de Biochimie Microbienne, URA 1300 du Centre National de la Recherche Scientifique1 and Laboratoire des Bactéries et Champignons Entomopathogènes,2 Institut Pasteur, 75724 Paris, and INRA-CNRS, Laboratoire de Génétique des Microorganismes, 78550 Thivernal-Grignon,3 France

Received 4 February 1999/Accepted 4 May 1999

Cry11A from Bacillus thuringiensis subsp. israelensis and Cry11Ba from Bacillus thuringiensis subsp. jegathesan were introduced, separately and in combination, into the chromosome of Bacillus sphaericus 2297 by in vivo recombination. Two loci on the B. sphaericus chromosome were chosen as target sites for recombination: the binary toxin locus and the gene encoding the 36-kDa protease that may be responsible for the cleavage of the Mtx protein. Disruption of the protease gene did not increase the larvicidal activity of the recombinant strain against Aedes aegypti and Culex pipiens. Synthesis of the Cry11A and Cry11Ba toxins made the recombinant strains toxic to A. aegypti larvae to which the parental strain was not toxic. The strain containing Cry11Ba was more toxic than strains containing the added Cry11A or both Cry11A and Cry11Ba. The production of the two toxins together with the binary toxin did not significantly increase the toxicity of the recombinant strain to susceptible C. pipiens larvae. However, the production of Cry11A and/or Cry11Ba partially overcame the resistance of C. pipiens SPHAE and Culex quinquefasciatus GeoR to B. sphaericus strain 2297.

* Corresponding author. Mailing address: Unité de Biochimie Microbienne, Institut Pasteur, 25, rue du Dr Roux, 75724 Paris Cedex 15, France. Phone: (33) 01 45 68 88 48. Fax: (33) 01 45 68 89 38. E-mail: pservant{at}pasteur.fr.

Applied and Environmental Microbiology, July 1999, p. 3021-3026, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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