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Specificity of action on mosquito larvae of Bacillus thuringiensis israelensis toxins encoded by two different genes

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Summary

A 135 kDa protein gene and two open reading frames (ORF1 and ORF2) have been cloned from a large plasmid of Bacillus thuringiensis israelensis (Bourgouin et al. 1986). The Escherichia coli recombinant clones containing these genes were highly toxic to larvae of Aedes aegypti, Anopheles stephensi and Culex pipiens. From subcloning experiments it was deduced that the 135 kDa polypeptide alone was responsible for the toxic activity on both A. aegypti and An. stephensi larvae. In contrast, the presence of two polypeptides, the 135 kDa protein and the ORF1 product was required for toxicity to C. pipiens larvae. The minimal toxic fragment of the 135 kDa polypeptide has been delineated. The results indicate that a polypeptide of about 65 kDa, corresponding to an amino-terminal part of the 135 kDa protein is sufficient for toxicity. Sequence comparisons indicate that the ORF1 product may correspond to an N-terminal part of a rearranged 130 kDa protein.

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References

  • Angsuthanasombat C, Chungjatupornchai W, Kertbundit S, Luxananil P, Settasatian C, Wilairat P, Panyim S (1987) 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

    Google Scholar 

  • Bourgouin C, Klier A, Rapoport G (1986) Characterization of the genes encoding the haemolytic toxin and the mosquitocidal delta-endotoxin of Bacillus thuringiensis israelensis. Mol Gen Genet 205:390–397

    Google Scholar 

  • Chestukhina GG, Zalunin IA, Kostina LI, Bormatova ME, Klepikova FS, Khodova OM, Stepanov VM (1985) Structural features of crystal-forming proteins produced by Bacillus thuringiensis subspecies israelensis. FEBS Lett 190:345–348

    Google Scholar 

  • Dulmage HT (1981) Insecticidal activity of isolates of Bacillus thuringiensis and their potential for pest control. In: Burges HD (ed) Microbial control of pests and plant diseases. Academic Press, London, p 193–223

    Google Scholar 

  • Goldberg LH, Margalit J (1977) A bacterial spore demonstrating rapid larvicidal activity against Anopheles sergentii, Uranotaenia unguiculata, Culex univitatus, Aedes aegypti and Culex pipiens. Mosq News 37:355–358

    Google Scholar 

  • Herrnstadt C, Gilroy TE, Sobieski DA, Bennett BD, Gaertner FH (1987) Nucleotide sequence and deduced amino acid sequence of a coleopteran-active delta-endotoxin gene from Bacillus thuringiensis subsp. san diego. Gene 57:37–46

    Google Scholar 

  • Höfte H, de Greve H, Seurinck J, Jansens S, Mahillon J, Ampe C, Vandekerckhove J, Vanderbruggen H, Van Montagu M, Zabeau M, Vaeck M (1986) Structural and functional analysis of a cloned delta endotoxin of Bacillus thuringiensis berliner 1715. Eur J Biochem 161:273–280

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    Google Scholar 

  • Hurley JM, Lee SG, Andrews Jr RE, Klowden MJ, Bullar Jr LA (1985) Separation of the cytolytic and mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis. Biochem Biophys Res Commun 126:961–965

    Google Scholar 

  • Ibarra JE, Federici BA (1986) Parasporal bodies of Bacillus thuringiensis subsp. morrisoni (PG-14) and Bacillus thuringiensis subsp. israelensis are similar in protein composition and toxicity. FEMS Microbiol Lett 34:79–84

    Google Scholar 

  • Lederberg EM, Cohen SN (1974) Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol 119:1072–1074

    Google Scholar 

  • Lee SG, Eckblad W, Bulla Jr LA (1985) Diversity of protein inclusion bodies and identification of mosquitocidal protein in Bacillus thuringiensis subsp. israelensis. Biochem Biophys Res Commun 126:953–960

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Pfannenstiel MA, Couche GA, Ross EJ, Nickerson KW (1986) Immunological relationships among proteins making up the Bacillus thuringiensis subsp. israelensis crystalline toxin. Appl Environ Microbiol 52:644–649

    Google Scholar 

  • Schnepf HE, Whiteley HR (1985) Delineation of a toxin-encoding segment of a Bacillus thuringiensis crystal protein gene. J Biol Chem 260:6273–6280

    Google Scholar 

  • Sekar V, Carlton BC (1985) Molecular cloning of the delta-endotoxin gene of Bacillus thuringiensis var. israelensis. Gene 33:151–158

    Google Scholar 

  • Sekar V, Carlton BC (1986) Molecular cloning of the delta-endotoxin gene of Bacillus thuringiensis var. israelensis. European Patent Application EP 0 195 285 A2

  • Thomas WE, Ellar DJ (1983) Bacillus thuringiensis var. israelensis crystal δ-endotoxin: effects on insect and mammalian cells in vitro and in vivo. J Cell Sci 60:181–197

    Google Scholar 

  • Thorne L, Garduno F, Thompson T, Decker D, Zounes M, Wild M, Walfield AM, Pollock TJ (1986) Structural similarity between the Lepidoptera- and Diptera- specific insecticidal endotoxin genes of Bacillus thuringiensis subsp. “kurstaki” and “israelensis”. J Bacteriol 166:801–811

    Google Scholar 

  • Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268

    Google Scholar 

  • Visser B, Van Workum M, Dullemans A, Waalwijk C (1986) The mosquitocidal activity of Bacillus thuringiensis var. israelensis is associated with Mr 230,000 and 130,000 crystal proteins. FEMS Microbiol Lett 30:211–214

    Google Scholar 

  • Ward ES, Ellar DJ (1987) Nucleotide sequence of a Bacillus thuringiensis var. israelensis gene encoding a 130 kDa delta-endotoxin. Nucleic Acids Res 15:7195

    Google Scholar 

  • Wu D, Chang FN (1985) Synergism in mosquitocidal activity of 26 and 65 kDa proteins from Bacillus thuringiensis subsp. israelensis crystal. FEBS Lett 190:232–236

    Google Scholar 

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Communicated by W. Goebel

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Delécluse, A., Bourgouin, C., Klier, A. et al. Specificity of action on mosquito larvae of Bacillus thuringiensis israelensis toxins encoded by two different genes. Mol Gen Genet 214, 42–47 (1988). https://doi.org/10.1007/BF00340177

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  • DOI: https://doi.org/10.1007/BF00340177

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