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Cloning and expression of 130-kd mosquito-larvicidal δ-endotoxin gene of Bacillus thuringiensis var. Israelensis in Escherichia coli

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Summary

Five recombinant E. coli clones exhibiting toxicity to Aedes aegypti larvae were obtained from a library of 800 clones containing XbaI DNA fragments of 110 kb plasmid from B. thuringiensis var. israelensis. All the five clones (pMU 14/258/303/388/679) had the same 3.8-kb insert and encoded a major protein of 130 kDa which was highly toxic to A. aegypti larvae. Three clones (pMU 258/303/388) transcribed the 130 kD a gene in the same direction as that of lac Z promoter of pUC12 vector whereas the transcription of the other two (pMU 14/679) was in the opposite direction. A 1.9-kb fragment of the 3.8 kb insert coded for a protein of 65 kDa. Partial DNA sequence of the 3.8 kb insert, corresponding to the 5′-terminal of the 130 kDa gene, revealed a continuous reading frame, a Shine-Dalgarno sequence and a tentative 5′-regulatory region. These results demonstrated that the 3.8 kb insert is a minimal DNA fragment containing a regulatory region plus the coding sequence of the 130 kDa protein that is highly toxic to mosquito larvae.

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Authors and Affiliations

  1. Center for Molecular Genetics-Genetic Engineering and Department of Biochemistry, Faculty of Science, Mahidol University, Rama VI Road, 10400, Bangkok, Thailand

    Chanan Angsuthanasombat, Wipa Chungjatupornchai, Sunee Kertbundit, Plernpis Luxananil, Chatri Settasatian, Prapon Wilairat & Sakol Panyim

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  1. Chanan Angsuthanasombat
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  2. Wipa Chungjatupornchai
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  3. Sunee Kertbundit
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  4. Plernpis Luxananil
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  5. Chatri Settasatian
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  6. Prapon Wilairat
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Communicated by M. Takanami

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Angsuthanasombat, C., Chungjatupornchai, W., Kertbundit, S. et al. Cloning and expression of 130-kd mosquito-larvicidal δ-endotoxin gene of Bacillus thuringiensis var. Israelensis in Escherichia coli . Mol Gen Genet 208, 384–389 (1987). https://doi.org/10.1007/BF00328128

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  • DOI: https://doi.org/10.1007/BF00328128

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