1887

Abstract

Summary: We have previously shown that subsp. serotype 3, produces two extracellular inhibitors of the immune system of Saturniid pupae (designated inhibitors A and B; Edlund ., 1976 ). Starting from the culture supernatant of a new mutant of with a decreased extracellular proteolytic activity, we have now purified immune inhibitor A (InA). The procedure described consists of three steps: ultrafiltration, precipitation with ammonium sulphate and chromatography on hydroxylapatite. Purified InA gave a single band on polyacrylamide gel electrophoresis using either a gel concentration of 7·5 % (w/v) and reducing and denaturing conditions or a gradient gel and native conditions. In both cases the apparent molecular weight was 78000. A certain amount of proteolytic activity was always co-purified with InA but the two activities could be dissociated by heat or EDTA treatment. Antiserum against purified InA gave only one sharp precipitation band on immunodiffusion against InA with or without EDTA. InA inhibited the killing of by immune haemolymph but did not affect the killing of InA was toxic for Drosophila when injected into the abdomen of adult male flies.

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/content/journal/micro/10.1099/00221287-114-1-45
1979-09-01
2024-12-08
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References

  1. Bertani G. 1951; Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. Journal of Bacteriology 62:293–300
    [Google Scholar]
  2. Boman H. G., Nilsson-Faye I., Paul K., Rasmuson T. 1974; Insect immunity. I. Characteristics of an inducible cell-free antibacterial reaction in hemolymph of Samia cynthia pupae. Infection and Immunity 10:136–145
    [Google Scholar]
  3. Brown M. R. W., Scott Foster J. H. 1970; A simple diagnostic milk medium for Pseudomonas aeruginosa. Journal of Clinical Pathology 23:172–177
    [Google Scholar]
  4. Bulla L. A., Rhodes R. A., St Julian G. 1975; Bacteria as insect pathogens. Annual Review of Microbiology 29:163–190
    [Google Scholar]
  5. Delafield F. P., Somerville H. J., Rittenberg S. C. 1968; Immunological homology between crystal and spore protein of Bacillus thuringiensis. Journal of Bacteriology 96:713–720
    [Google Scholar]
  6. Edlund T., Sidén I., Boman H. G. 1976; Evidence for two immune inhibitors from Bacillus thuringiensis interfering with the humoral defense system of Saturniid pupae. Infection and Immunity 14:934–941
    [Google Scholar]
  7. Faust R. M. 1975; Toxins of Bacillus thuringiensis: mode of action. In Biological Regulation of Vectors pp. 31–48 Briggs J. D. Edited by Washington, D. C.: U. S. Department of Health, Education and Welfare;
    [Google Scholar]
  8. Faye I., Pye A., Rasmuson T., Boman H. G., Boman I. A. 1975; Insect immunity. II. Simultaneous induction of antibacterial activity and and selective synthesis of some hemolymph proteins in diapausing pupae of Hyalophora cecropia and Samia cynthia. Infection and Immunity 12:1426–1438
    [Google Scholar]
  9. Kunitz M. 1946; Crystalline soybean trypsin inhibitor. II. General properties. Journal of General Physiology 30:291–310
    [Google Scholar]
  10. Li E., Yousten A. A. 1975; Metalloprotease from Bacillus thuringiensis. Applied Microbiology 30:354–361
    [Google Scholar]
  11. Maizel J. V. 1969; Acrylamide gel electrophoresis of proteins and nucleic acids. In Fundamental Techniques in Virology pp. 334–362 Habel K., Salzman N. P. Edited by New York: Academic Press;
    [Google Scholar]
  12. Pendelton I. R., Bernheimer A. W., Grushoff P. 1973; Purification and partial characterization of hemolysins from Bacillus thuringiensis. Journal of Invertebrate Pathology 21:131–135
    [Google Scholar]
  13. Rasmuson T., Boman H. G. 1977; The assay and the specificity problem in insect immunity. In Developmental Immunobiology pp. 83–90 Solomon J. B., Horton J. D. Edited by Amsterdam: Elsevier/North Holland;
    [Google Scholar]
  14. Somerville H. J. 1971; Formation of the para- sporal inclusion of Bacillus thuringiensis. European Journal of Biochemistry 18:226–237
    [Google Scholar]
  15. Somerville H. J., Pockett H. V. 1975; An insect toxin from spores of Bacillus thuringiensis and Bacillus cereus. Journal of General Microbiology 87:359–369
    [Google Scholar]
  16. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
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