Cloning,heterologous expression,and localization of a novel crystal protein gene fromBacillus thuringiensis serovarjaponensis strain Buibui toxic to scarabaeid insects |
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| Authors: | Sato Ryoichi Takeuchi Katsuyoshi Ogiwara Katsutoshi Minami Masayosi Kaji Yasuko Suzuki Nobukazu Hori Hidetaka Asano Shoji Ohba Michio Iwahana Hidenori |
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| Institution: | (1) Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 183 Fuchu, Tokyo, Japan;(2) Tsukuba Laboratories for Research and Development, Kubota Corp., Kyugasaki, Ibaraki, Japan;(3) Institute of Biological Control, Faculty of Agriculture, Kyushu University, Fukuoka, Japan |
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| Abstract: | RecombinantEscherichia coli strains harboring pAG1, pAG2, pKBB100, and pKBB101 were cloned by using antiserum constructed against 130-kDa crystal protein antigen ofBacillus thuringiensis serovarjaponensis strain Buibui. DNAs in the recombinant strains hybridized to the 26-base oligonucleotide probe corresponding to N-terminal amino acids of the 130-kDa crystal protein of strain Buibui. Cultures of the recombinant strains were toxic to larvae of the cupreous chafer,anomala cuprea. Furthermore, the production of the 130-kDa polypeptide was demonstrated in the cells harboring pAG1 and pAG2 by immunoblot analysis with antiserum against the 130-kDa crystal protein. Southern hybridization analysis showed that the 130-kDa crystal protein gene is located on the chromosomal DNA of strain Buibui. On the other hand, DNA probes derived fromcryIA(a) andcryIIIA genes did not hybridize to the DNA of strain Buibui. |
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