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1992 Fiscal Year Final Research Report Summary

Roles of the alpha-,theta-,and k-toxins in histotoxicity of Clostridium perfringens

Research Project

Project/Area Number 03670217
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionKagawa Medical School

Principal Investigator

OKABE Akinobu  Kagawa Medical School Professor, 医学部, 教授 (20093677)

Co-Investigator(Kenkyū-buntansha) KATAYAMA Sei-ichi  Kagawa Medical School Assistant Professor, 医学部, 助手 (70169473)
MINAMI Junzaburo  Kagawa Medical School Lecturer, 医学部, 講師 (40157566)
Project Period (FY) 1991 – 1992
KeywordsClostridium / C.perfringens / Bacterial toxin / Histotoxicity / Toxin genes / Collagenase
Research Abstract

Genes encoding for alpha-and theta-toxins of Clostridium perfringens(type A NCTC 8237)were separately cloned into pHY300PLK and expressed in Bacillus subtilis ISW 1214(spoo^).Levels of both the alpha- and theta-toxins produced by each strain were almost the same as those by C.perfringens. Viability of these two strains within a macrophage was examined using macrophage cell line SV-BP-1.The theta-toxin producing strain increased in number within the macrophage,while the alpha-toxin producing strain decreased similarly to a control.The result indicates that the theta-toxin functions as an antiphagocytic determinant but the alpha-toxin does not.
Virulence of the two strains was examined by determining lethal dose against a mouse. When the alpha-toxin producing strain was injected interperitoneally,it was more virulent than the theta-toxin producing strain. On the other hand,the former was less virulent than the latter when injected intravenously. A plasmid carrying the two toxin genes was also constructed.B subtilis carrying this plasmid produced the theta-toxin in large amounts but the alpha-toxin in a very small amount. However,the strain producing the two toxins exhibited mouse lethality to the same extent as the two alpha- and theta-producing strains used in combination, suggesting that a synergistic effect of the two toxins may be enhanced if they are produced by a single cell into microenvironment surrounding a bacterial cell.
A 120-KDa C.perfringens collagenase(k-toxin)was found and purified. Its encoding gene was cloned and sequenced. The nucleotide sequence shows that the toxin belongs to Zn metalloproteinase and is homologous to other bacterial proteinase and collagenase.

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Toyonaga,T.er al.: "Role of the upstream region containing an intrinsic DNA curvature in the negative regulation of the phospholipase C gene of Clostridium perfringens." Microbiology and Immunology. 36. 603-613 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Katayama,S.et al.: "Comparison of the alpha-toxin genes of Clostridium perfringens type A and C strains:Evidence for extragenic regulation of transcription." Infection and Immunity. 61. 457-463 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsushita,O.et al.: "Purification and characterization of a Clostridium perfrigens 120-KDa collagenase and nucleotide sequence of the corresponding gene." Journal of Bacteriology.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tatsuo Toyonaga et al.: "Role of the upstream region containing an intrinsic DNA curvature in the negative regulation of the phospholipase C gene of Clostridium perfringens." Microbiology and Immunology. 36. 603-613 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sei-ichi Katayama et al.: "Comparison of the alpha-toxin genes of Clostridium perfringens type A and C strains:evidence for extragenic regulation of transcription." Infection and Immunity. 61. 457-463 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Osamu Matsushita et al.: "Purification and characterization of a Clostridium perfringens 120-KDa collagenase and nucleotide sequence of the corresponding gene." Journal of Bacteriology.

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1994-03-24  

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