1887

Abstract

Crystals were purified from, four serotypes of the insect pathogen . Crystals from these serotypes were similar in amino acid and -terminal analyses, but differed in their toxicity to two species of and in their immunological properties. Toxic polypeptides were obtained following trypsin digestion of solutions of the crystals. In two strains (serotypes 3 and 9) this fraction contained only one polypeptide. Similar results were obtained when dissolved crystals were digested with other proteolytic enzymes or with gut contents from . The trypsin-resistant polypeptide was further purified by gel and ion-exchange chromatography and had a molecular weight of about 70000, estimated by gel chromatography and gel electrophoresis. No evidence was obtained for a toxin of lower molecular weight. This purified toxin accounted for most, if not all, of the toxic activity originally present in the crystal solution and was active by injection and ingestion. The purified toxic fraction from serotype 1 appeared to contain two polypeptides, one of which corresponded to that found with serotypes 3 and 9. There were no major differences in the composition of crystals from different serotypes of and it is concluded that the trypsin-resistant polypeptide represents the active insecticidal toxin of the crystal.

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1980-05-01
2024-11-12
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References

  1. Andrews P. 1965; The gel filtration behaviour of proteins related to their molecular weight over a wide range. Biochemical Journal 96:595–606
    [Google Scholar]
  2. De Barjac H., Bonnefoi A. 1968; Classification des souches de Bacillus thuringiensis . Comptes rendus hebdomadaire des séances de l’Académie des sciences 264:1811–1813
    [Google Scholar]
  3. De Barjac H., Thompson J. V. 1970; A new serotype of Bacillus thuringiensis: B. thuringiensis var. thompsonii (serotype 11). Journal of Invertebrate Pathology 15:141–144
    [Google Scholar]
  4. Bond R. P. M., Boyce C. B. C., Rogoff M. H., Shieh T. P. 1971; The thermostable exotoxin of Bacillus thuringiensis . In Microbial Control of Insects and Mites pp. 275–303 Burges H. D., Hussey N. W. Edited by London: Academic Press;
    [Google Scholar]
  5. Cooksey K. E. 1971; The protein crystal toxin of Bacillus thuringiensis. Biochemistry and mode of action. In Microbial Control of Insects and Mites pp. 247–274 Burges H. D., Hussey N. W. Edited by London: Academic Press;
    [Google Scholar]
  6. Delafield F. P., Somerville H. J., Rittenberg S. C. 1968; Immunological homology between crystals and spores of Bacillus thuringiensis . Journal of Bacteriology 96:713–720
    [Google Scholar]
  7. Edman P., Sjoquist J. 1956; Identification and semiquantitative determination of phenyl thio-hydantoins. Acta chemica scandinavica 10:10–11
    [Google Scholar]
  8. Ellis D. A. 1961; A new universal buffer. Nature; London: 1911099
    [Google Scholar]
  9. Freedman M. H., Slobin L. I., Robbins J. B., Sela M. 1966; Soluble antigen-antibody complexes and intermediates in the purification of antibodies in 8 m urea. Archives of Biochemistry and Biophysics 116:82–91
    [Google Scholar]
  10. Galowalia M. M. S., Gibson N. H. E., Wolf J. 1973; The comparative potencies of the crystal-line endotoxin of et varieties of Bacillus thuringiensis to larvae of Pieris brassicae . Journal of Invertebrate Pathology 21:301–308
    [Google Scholar]
  11. Glatron M. F., Lecadet M. M., Dedonder R. 1969; Etude de la dissolution du cristal de Bacillus thuringiensis par ultracentrifugation et identification des extrémités N et C terminales. Comptes rendus hebdomadaire des séances de l’Académie des sciences 269:1338–1341
    [Google Scholar]
  12. Grant G. F., Simon M. 1968; Use of radioactive antibodies for characterizing antigens and applications to the study of flagella synthesis. Journal of Bacteriology 95:81–86
    [Google Scholar]
  13. Herbert B. N., Gould H. J., Chain E. B. 1971; Crystal protein of Bacillus thuringiensis var. tolworth. Sub-unit structure and toxicity of Pieris brassicae . European Journal of Biochemistry 24:366–375
    [Google Scholar]
  14. Krywienczyk J., Angus T. A. 1966; Serological studies of the protein parasporal inclusions of Bacillus thuringiensis var. entomocidus . Journal of Invertebrate Pathology 8:439–441
    [Google Scholar]
  15. Norris J. R. 1969; Macromolecule synthesis during sporulation of Bacillus thuringiensis . In Spores IV pp. 45–58 Campbell L. L. Edited by Bethesda: American Society for Microbiology;
    [Google Scholar]
  16. Pendleton I. R., Morrison R. B. 1967; Analysis of the crystal antigens of Bacillus thuringiensis by gel diffusion. Journal of Applied Bacteriology 29:519–528
    [Google Scholar]
  17. Sanger F. 1949; Fractionation of oxidized insulin. Biochemical Journal 44:126–128
    [Google Scholar]
  18. Shapiro A. L., Viñuela E., Maizel J. V. 1967; Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochemical and Biophysical Research Communications 28:815–820
    [Google Scholar]
  19. Somerville H. J. 1973; Microbial toxins. Annals of the New York Academy of Sciences 217:93–108
    [Google Scholar]
  20. Somerville H. J. 1977; The insecticidal toxin of Bacillus thuringiensis . Proceedings Vatican Academy of Sciences, Study Week14 1976 Natural Products and the Protection of Plants pp. 253–274
    [Google Scholar]
  21. Somerville H. J., Jones M. L. 1972; DNA competition studies within the Bacillus cereus group of bacilli. Journal of General Microbiology 73:257–263
    [Google Scholar]
  22. Somerville H. J., Tanada Y., Omi E. M. 1970; Lethal effect of purified spore and crystalline endotoxin preparations of Bacillus thuringiensis on several lepidopterous insects. Journal of Invertebrate Pathology 16:241–248
    [Google Scholar]
  23. Somerville H. J., James C. R., Ruffell R. N., Norris J. R. 1972; The parasporal inclusion of Bacillus finitimus . Spore Research 1971 pp. 193–200 Barker A. N., Gould G. W., Wolf J. Edited by London: Academic Press;
    [Google Scholar]
  24. Weber K., Osborne M. 1969; The reliability of molecular weight determinations by dodecylsulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
  25. Young I. E., Fitz-James P. C. 1959; Chemical and morphological studies of bacterial spore formation. II. Spore and parasporal formation in Bacillus cereus var. alesti . Journal of Biophysical and Biochemical Cytology 6:483–498
    [Google Scholar]
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