1990 Fiscal Year Final Research Report Summary
Site-Directed Mutagenesis of HlyA of Vibrio Cholerae at Common Domain Found in Vibroio Vulnificus Cytolysin.
| Project/Area Number |
01570236
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | Research Institute for Microbial Diseases, Osaka University |
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Principal Investigator |
YAMAMOTO Koichiro Biken, Osaka Univ., Research Instructor, 微生物病研究所, 助手 (30158274)
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| Co-Investigator(Kenkyū-buntansha) |
YOH Myonsun Biken, Osaka Univ., Research Instructor, 微生物病研究所, 助手 (70093482)
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| Project Period (FY) |
1989 – 1990
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| Keywords | mutation / Amino Acid Sequence / hemolysin / Pathogenic Factor / cloning / domain / vibrio / Nucleotide Sequence |
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| Research Abstract |
Site-directed mutagenesis of hlyA of Vibrio cholerae at common domain found in Vibrio vulnificus cytolysin was carried out.
1. Oligonucleotide primers were synthesized as one of the amino acids of the domain in substituted to another amino acid.
The mutations were designed to substitute to another amino acid of which
(A) hydropathy and
(B) charge was greatly changed and
(C) the third nucleotide of the codons was changed
where it gives amino acid substitution.
2. Site-directed mutations were carried out using these oligonucleotide primers and the mutation was confirmed by DNA sequencing and ten mutants were obtained.
3. The mutant precursors were over-produced in E. Coli. The mutant cytolysins were extracted and partially purified. The quantities of the cytolysin antigens and the protein contents were determined.
4. The specific activities of the mutant cytolysins were 1/10 - 1/100 of the native cytolysin. This result suggest that the common domain found in V. Cholerae and V. Vulnificus play an important role(s) in cytolytic activities of the cytolysins.
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Research Products
(32 results)
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[Publications] Yamamoto, K., Ichinose, Y., Shinagawa, H., Makino, K., Nakata, A., Iwanaga, M., Honda, T. and Miwatani, T.: "Two-step processing for activation of the cytolysin/hemolysin of Vibrio cholerae O1 biotype E1 Tor : Nucleotide sequence of the structural gene (hlyA) and characterization of the processed products." Infection and Immunity. 58. 4106-4116 (1990)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Honda, T., Ni, Y., Yoh, M., Nishibuchi, M., Miwatani, T., Ishibashi, M., Kinoshita, Y., Honda, S., Goto, I., Minematsu, I., Shimoirisa, K., Ikeda, N., and Asano, N.: "Characterization of a TDH-related hemolysin produced by Kanagawa phenomenon-negative Vibrio parahaemolyticus which caused an outbreak of traveller's diarrhea." Jpn. J. Med. Sci. Biol.,.
Description
「研究成果報告書概要(欧文)」より
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[Publications] Yoh, M., Honda, T., Miwatani, T., and Nishibuchi, M.: "Characterization of four thermostable direct hemolysins encoded by four representative tdh genes of Vibrio parahaemolyticus." Advances in Research on Cholera and Related Diarrheas, 7, (Eds., R. B. Sack and Y. Zinnaka). KTK Scientific Publishers, Tokyo. 277-286 (1990)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Naka, A., Lertpocasombat, K., Yamamoto, K., Honda, T., and Miwatani, T.,: "Characterization of Vibrio cholerae non-O1 hemagglulutinin/protease." Jpn. Med. Sci. Biol.
Description
「研究成果報告書概要(欧文)」より
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[Publications] Honda, T., Ni, Y., Iida, T., Yoshida, H., Yoh, M., Yamamoto, K., Miwatani, T., Okamoto, T., Goshima, K., Takakura, H., Tsunasawa, S. and Sakiyama, F.: "Characterization of TDH-related hemolysins (Vp-TRH) produced by Kanagawa phenomenon-negative clinical isolates of Vibrio parahaemolyticus" Advances in Research on Cholera and Related Diarrheas.
Description
「研究成果報告書概要(欧文)」より
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