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1993 Fiscal Year Final Research Report Summary

Role of of Pro region of El Tor cytolysin

Research Project

Project/Area Number 04670246
Research Category

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

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionOsaka University

Principal Investigator

YAMAMOTO Koichiro  Osaka Univ. RIMD, Associate Prof., 微生物病研究所, 助教授 (30158274)

Co-Investigator(Kenkyū-buntansha) YOH Myonsun  Osaka Univ. RIMD, Res. Associate, 微生物病研究所, 助手 (70093482)
HONDA Takeshi  Osaka Univ. RIMD, Prof., 微生物病研究所, 教授 (60029808)
Project Period (FY) 1992 – 1993
KeywordsVibrio cholerae / hemolysin / gene / DNA sequence / Mutation / Protein toxin / homologous reconbination / Secretion
Research Abstract

Purified protoxin purified from E.coli harboring hlyA.The protoxin was processed to mature hemolysin by HA/protease of V.cholerae and trypsin, resulting increase in hemolytic activity. The region of hlyA coding for Pro region of was deleted and the deleted gene (hlyA-DELTA pro) cloned to a suicide vector plasmid. The plasmid was introduced to the parent strain and the native gene was replaced with hlyA-DELTA pro (N86-hlyA-DELTA pro). The jmutant strain decreased in hemolytic activity. The mutant strain did not produce mature hemolysin in the supernatant. Western blot showed that destruction of protoxin in the periplasm. Protoxin and mature toxin was denatured with guanidine-HC1 and reconstituted by decreasin the concentration of the solution. Protoxin recovered 100% hemolytic activity but mature toxin did not recover the activity. These data suggest that Pro region may play role in protective effect against proteases and renaturation of hemolsin.

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Miyagi,K.et al.: "Cholera diagnosed ub clinical laboratory by DNA hybridization" Lancet. 339. 989 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto,K.et al.: "Enzyme-labeled oligonucleotide probes for detection for genes for thermostable direct hemolysin(TDH)…." Can.J.Microbiol.38. 410-416 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoh,M.et al.: "Development of an enzyme-labeled oligonucleotide probe for the cholera toxin gene." J.Clin.Microbiol.31. 1312-1314 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shinagawa,H.et al.: "Cloning of the genes that control formation of the fimbrial colonization factor antigen III(CFA/III)…." Microbiol.Immunol.37. 689-694 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Iida,T.et al.: "Cholera isolates in relation to the “eight pandemic"." Lancet. 342. 926 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Xu,M.et al.: "Demonstration and characterization of simultaneous production of a thermostable direct heameolysin…." Infect.Immun.62. 166-171 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本耕一郎,本田武司: "ビブリオ属の病原遺伝子(分担)「微生物学の新しい展開1993」" 菜根出版(東京), 16 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyagi, K., Matsumoto, Y., Hayashi, Y., Takarada, Y., Shibata, S., Yoh, M., Yamamoto, K.and Honda, T.: "Cholera diagnosed in clinical laboratory by DNA hybridization" Lancet. 339. 988-989 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Iida, Yamamoto, K., Honda, T.and Miwatani, T.: "Structure jand function of Vibrio parahaemolyticus hemolysins : Analysis jof chimeric toxins between TDH and TRH" Jpn. J.Med. Sci. Biol. 44. 271-272 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto, K., Honda, T., Miwatani, T., S.Tamatsukuri, and S.Shibata: "Enzyme-labeled oligonucleotide probes fordetection of genes for thermo-stable direct hemolysin (TDH) and TDH-related hemolysin (TRH) of Vibrio parahaemolyticus" Can. J.Microbiol. 37(1). 83-86 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Naka, A., Yamamoto, K., Miwatani, T.and Honda, T.: "Characterization of two forms of hemaggluinin/protease produced by Vibrio cholerae non-01" FEMS Microbiol. Lett. 98. 197-200 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoh, M., Miyagi, K., Matsumoto, Y., Hayashi, K., Takarada, Y., Yamamoto, K.and Honda, T.: "Development of an enzyme-labeled oligonucleotide probe for the cholera toxin gene" J.Clin. Microbiol. 31. 1312-1314 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shinagawa, H., Taniguchi, T., Yamaguchi, O., Yamamoto, K.and Honda, T.: "Cloning jof the genes that control formation of the fimbrial colonization factor antigen III (CFA/III) from an enterotoxigenic Escherichia coli" Microbiol. Immunol. 37. 689-694 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Iida, T., Shrestha, J., Yamamoto, K., Honda, T.and Albert, J.M.: "Cholera isolates in relation to the "eighth pandemic"" Lancet. 342. 926 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Xu, M., Iida, T., Yamamoto, K., Takarada, Y., Miwatani, T.and Honda, T.: "Demonstration and characterization of simultaneous production of a tehrmostalbe direct hemolysin (TDH/I) and a TDH-related hemolysin (TRHx) by a clinically isolated Bivrio parahaemolyticus strain" TH3766.

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

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Published: 1995-03-27  

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