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Cloning and Characterization of Two Novel Genes, cry24B and s1orf2, from a Mosquitocidal Strain of Bacillus thuringiensis serovar sotto

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Abstract

Two new crystal protein genes, cry24B and s1orf2, were cloned from a mosquitocidal Bacillus thuringiensis serovar sotto strain. The cry24B and s1orf2 genes encoded a 76-kDa and 62-kDa protein, respectively. The Cry24B protein retained five conserved regions commonly found in the existing Cry proteins. The amino acid sequence of the S1ORF2 had a high homology to that of the ORF2 protein of B. thuringiensis serovar jegathesan. Southern hybridization experiments with a cry24B gene-specific probe revealed that these genes are located on two large plasmids of > 100 kb. When the two genes, cry24B and s1orf2, were expressed in an acrystalliferous B. thuringiensis host, the proteins were synthesized and accumulated as inclusions. These inclusions exhibited no larvicidal activities against three mosquito species: Aedes aegypti, Anopheles stephensi, and Culex pipiens molestus. Likewise, the inclusions contained no cytocidal activity against HeLa cells.

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Acknowledgments

We thank Dr. Daniel Zeigler and Dr. Neil Crickmore, Bacillus thuringiensis Pesticidal Crystal Protein Nomenclature Committee, for invaluable advice on the designation of the protein class. We also thank Dr. Minoru Maeda for support in in vitro cytotoxicity tests.

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Authors and Affiliations

  1. Graduate School of Agriculture, Kyushu University, Fukuoka, 812-8581, Japan

    Akira Ohgushi, Akiko Uemori & Michio Ohba

  2. Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, Kurume, 839-0861, Japan

    Hiroyuki Saitoh & Naoya Wasano

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  1. Akira Ohgushi
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  2. Hiroyuki Saitoh
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  3. Naoya Wasano
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  4. Akiko Uemori
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  5. Michio Ohba
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Correspondence to Michio Ohba.

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Ohgushi, A., Saitoh, H., Wasano, N. et al. Cloning and Characterization of Two Novel Genes, cry24B and s1orf2, from a Mosquitocidal Strain of Bacillus thuringiensis serovar sotto. Curr Microbiol 51, 131–136 (2005). https://doi.org/10.1007/s00284-005-7529-3

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  • DOI: https://doi.org/10.1007/s00284-005-7529-3

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