Abstract
The cytA gene encoding the 28-kDa polypeptide of Bacillus thuringiensis subsp. israelensis crystals was disrupted in the 72-MDa resident plasmid by in vivo recombination, thus indicating that homologous recombination occurs in B. thuringiensis. The absence of the 28-kDa protein in B. thuringiensis did not affect the crystallization of the other toxic components of the parasporal body (68-, 125-, and 135-kDa polypeptides). The absence of the 28-kDa protein abolished the hemolytic activity of B. thuringiensis subsp. israelensis crystals. However, the mosquitocidal activity of the 28-kDa protein-free crystals did not differ significantly from that of the wild-type crystals when tested on Aedes aegypti and Culex pipiens larvae. The 28-kDa protein contributed slightly to the toxicity to Anopheles stephensi larvae. This indicates that the 28-kDa protein is not essential for mosquitocidal activity, at least against the three species tested.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adams L. F., Visick J. E., Whiteley H. R. A 20-kilodalton protein is required for efficient production of the Bacillus thuringiensis subsp. israelensis 27-kilodalton crystal protein in Escherichia coli. J Bacteriol. 1989 Jan;171(1):521–530. doi: 10.1128/jb.171.1.521-530.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Albertini A. M., Galizzi A. Amplification of a chromosomal region in Bacillus subtilis. J Bacteriol. 1985 Jun;162(3):1203–1211. doi: 10.1128/jb.162.3.1203-1211.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alouf J. E., Viette M., Corvazier R., Raynaud M. Préparation et propriétés de sérums de chevaux antistreptolysine O. Ann Inst Pasteur (Paris) 1965 Apr;108(4):476–500. [PubMed] [Google Scholar]
- Armstrong J. L., Rohrmann G. F., Beaudreau G. S. Delta endotoxin of Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1985 Jan;161(1):39–46. doi: 10.1128/jb.161.1.39-46.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bourgouin C., Delécluse A., Ribier J., Klier A., Rapoport G. A Bacillus thuringiensis subsp. israelensis gene encoding a 125-kilodalton larvicidal polypeptide is associated with inverted repeat sequences. J Bacteriol. 1988 Aug;170(8):3575–3583. doi: 10.1128/jb.170.8.3575-3583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bourgouin C., Klier A., Rapoport G. Characterization of the genes encoding the haemolytic toxin and the mosquitocidal delta-endotoxin of Bacillus thuringiensis israelensis. Mol Gen Genet. 1986 Dec;205(3):390–397. doi: 10.1007/BF00338072. [DOI] [PubMed] [Google Scholar]
- Charles J. F., Nicolas L., Sebald M., de Barjac H. Clostridium bifermentans serovar malaysia: sporulation, biogenesis of inclusion bodies and larvicidal effect on mosquito. Res Microbiol. 1990 Jul-Aug;141(6):721–733. doi: 10.1016/0923-2508(90)90066-y. [DOI] [PubMed] [Google Scholar]
- Chilcott C. N., Ellar D. J. Comparative toxicity of Bacillus thuringiensis var. israelensis crystal proteins in vivo and in vitro. J Gen Microbiol. 1988 Sep;134(9):2551–2558. doi: 10.1099/00221287-134-9-2551. [DOI] [PubMed] [Google Scholar]
- Delécluse A., Bourgouin C., Klier A., Rapoport G. Specificity of action on mosquito larvae of Bacillus thuringiensis israelensis toxins encoded by two different genes. Mol Gen Genet. 1988 Sep;214(1):42–47. doi: 10.1007/BF00340177. [DOI] [PubMed] [Google Scholar]
- Ferrari F. A., Nguyen A., Lang D., Hoch J. A. Construction and properties of an integrable plasmid for Bacillus subtilis. J Bacteriol. 1983 Jun;154(3):1513–1515. doi: 10.1128/jb.154.3.1513-1515.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- González J. M., Jr, Carlton B. C. A large transmissible plasmid is required for crystal toxin production in Bacillus thuringiensis variety israelensis. Plasmid. 1984 Jan;11(1):28–38. doi: 10.1016/0147-619x(84)90004-0. [DOI] [PubMed] [Google Scholar]
- Heierson A., Landén R., Lövgren A., Dalhammar G., Boman H. G. Transformation of vegetative cells of Bacillus thuringiensis by plasmid DNA. J Bacteriol. 1987 Mar;169(3):1147–1152. doi: 10.1128/jb.169.3.1147-1152.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Held G. A., Huang Y. S., Kawanishi C. Y. Effect of removal of the cytolytic factor of Bacillus thuringiensis subsp. israelensis on mosquito toxicity. Biochem Biophys Res Commun. 1986 Dec 30;141(3):937–941. doi: 10.1016/s0006-291x(86)80133-4. [DOI] [PubMed] [Google Scholar]
- Hurley J. M., Lee S. G., Andrews R. E., Jr, Klowden M. J., Bulla L. A., Jr Separation of the cytolytic and mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis. Biochem Biophys Res Commun. 1985 Jan 31;126(2):961–965. doi: 10.1016/0006-291x(85)90279-7. [DOI] [PubMed] [Google Scholar]
- Höfte H., Whiteley H. R. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev. 1989 Jun;53(2):242–255. doi: 10.1128/mr.53.2.242-255.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ibarra J. E., Federici B. A. Isolation of a relatively nontoxic 65-kilodalton protein inclusion from the parasporal body of Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1986 Feb;165(2):527–533. doi: 10.1128/jb.165.2.527-533.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Insell J. P., Fitz-James P. C. Composition and Toxicity of the Inclusion of Bacillus thuringiensis subsp. israelensis. Appl Environ Microbiol. 1985 Jul;50(1):56–62. doi: 10.1128/aem.50.1.56-62.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jannière L., Niaudet B., Pierre E., Ehrlich S. D. Stable gene amplification in the chromosome of Bacillus subtilis. Gene. 1985;40(1):47–55. doi: 10.1016/0378-1119(85)90023-x. [DOI] [PubMed] [Google Scholar]
- Lecadet M. M., Blondel M. O., Ribier J. Generalized transduction in Bacillus thuringiensis var. berliner 1715 using bacteriophage CP-54Ber. J Gen Microbiol. 1980 Nov;121(1):203–212. doi: 10.1099/00221287-121-1-203. [DOI] [PubMed] [Google Scholar]
- Lereclus D., Arantès O., Chaufaux J., Lecadet M. Transformation and expression of a cloned delta-endotoxin gene in Bacillus thuringiensis. FEMS Microbiol Lett. 1989 Jul 15;51(1):211–217. doi: 10.1016/0378-1097(89)90511-9. [DOI] [PubMed] [Google Scholar]
- McLean K. M., Whiteley H. R. Expression in Escherichia coli of a cloned crystal protein gene of Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1987 Mar;169(3):1017–1023. doi: 10.1128/jb.169.3.1017-1023.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nishiura J. T. Fractionation of two mosquitocidal activities from alkali-solubilized extracts of Bacillus thuringiensis subspecies israelensis spores and parasporal inclusions. J Invertebr Pathol. 1988 Jan;51(1):15–22. doi: 10.1016/0022-2011(88)90083-3. [DOI] [PubMed] [Google Scholar]
- Pfannenstiel M. A., Couche G. A., Ross E. J., Nickerson K. W. Immunological relationships among proteins making up the Bacillus thuringiensis subsp. israelensis crystalline toxin. Appl Environ Microbiol. 1986 Oct;52(4):644–649. doi: 10.1128/aem.52.4.644-649.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Thomas W. E., Ellar D. J. Bacillus thuringiensis var israelensis crystal delta-endotoxin: effects on insect and mammalian cells in vitro and in vivo. J Cell Sci. 1983 Mar;60:181–197. doi: 10.1242/jcs.60.1.181. [DOI] [PubMed] [Google Scholar]
- Trieu-Cuot P., Poyart-Salmeron C., Carlier C., Courvalin P. Nucleotide sequence of the erythromycin resistance gene of the conjugative transposon Tn1545. Nucleic Acids Res. 1990 Jun 25;18(12):3660–3660. doi: 10.1093/nar/18.12.3660. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ward E. S., Ellar D. J. Bacillus thuringiensis var. israelensis delta-endotoxin. Nucleotide sequence and characterization of the transcripts in Bacillus thuringiensis and Escherichia coli. J Mol Biol. 1986 Sep 5;191(1):1–11. doi: 10.1016/0022-2836(86)90417-1. [DOI] [PubMed] [Google Scholar]
- Ward E. S., Ellar D. J. Cloning and expression of two homologous genes of Bacillus thuringiensis subsp. israelensis which encode 130-kilodalton mosquitocidal proteins. J Bacteriol. 1988 Feb;170(2):727–735. doi: 10.1128/jb.170.2.727-735.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ward E. S., Ellar D. J., Todd J. A. Cloning and expression in Escherichia coli of the insecticidal delta-endotoxin gene of Bacillus thuringiensis var. israelensis. FEBS Lett. 1984 Oct 1;175(2):377–382. doi: 10.1016/0014-5793(84)80772-3. [DOI] [PubMed] [Google Scholar]
- Ward E. S., Ridley A. R., Ellar D. J., Todd J. A. Bacillus thuringiensis var. israelensis delta-endotoxin. Cloning and expression of the toxin in sporogenic and asporogenic strains of Bacillus subtilis. J Mol Biol. 1986 Sep 5;191(1):13–22. doi: 10.1016/0022-2836(86)90418-3. [DOI] [PubMed] [Google Scholar]
- Young M. Gene amplification in Bacillus subtilis. J Gen Microbiol. 1984 Jul;130(7):1613–1621. doi: 10.1099/00221287-130-7-1613. [DOI] [PubMed] [Google Scholar]