Cloning of the Gene Encoding a Major Immunodominant Surface Protein of a Peridontopathogen

1991 Fiscal Year Final Research Report Summary

• Project/Area Number: 02670821
• Research Category: Grant-in-Aid for General Scientific Research (C)
• Allocation Type: Single-year Grants
• Research Field: Functional basic dentistry
• Research Institution: Aichi-Gakuin University
• Principal Investigator: YOSHIMURA Fuminobu Aichi-Gakuin Univ., Dept. Microbiology, Professor, 歯学部, 教授 (50001962)
• Co-Investigator(Kenkyū-buntansha): KATO Hiroshi Hokkaido Univ., Dept. of Periodont., Professor, 歯学部, 教授 (60001020) ; IKEDA Takeshi Aichi-Gakuin Univ., Dept. of Microbiology, Ass. Prof., 歯学部, 講師 ; OZEKI Masami Aichi-Gakuin Univ., Dept. of Microbiology, Ass. Prof., 歯学部, 講師 (80090124) ; OSANO Etsuo Aichi-Gakuin Univ., Dept. of Microbiology, Ass. Prof., 歯学部, 講師 (80110998) ; HIBI Eiko Aichi-Gakuin Univ., Dept. of Microbiology, Ass. Prof., 歯学部, 講師 (50097606)
• Project Period (FY): 1990 – 1991
• Keywords: Periodontopathogen / Surface protein / Cloning of a gene / Porphyromonas gingivalis / Immnodominat protein / 75K蛋白質

Research Abstract

A major immunodominant surface protein (the 75K protein) of Porphyromonas gingivalis 381 has been purified and its amino-terminal amino acid sequence has been determined. Using oligonucleotide probes corresponding to the sequence, we identified a recombinant plasmid clone carrying a single 4.2-kilobase pair Bam HI fragment from pUC19 libraries of P. gingivalis genomic DNA. The Ram HI fragment transferred to the bacteriophage T7 RNA polymerase/promoter expression vector system produced a slightly larger 77-kilodalton protein, a precursor form, immunoreactive to the antibody against the 75K protein, suggesting that the cloned DNA fragment probably carried an entire gene for the 75K protein. Genomic Southern analysis revealed a single copy of the 75K protein gene per genome among all P. gingivalis strains tested, and that no homologous genes are present in other black-pigmented Bacteroides species. These observations suggest that the 75K protein gene may be useful as a specific DNA probe to classify or to detect this organism.

Research Products

• [Publications] Watanabe,K.,T.Takasawa,F.Yoshimura,M.Ozeki,M.Kawanami and H.Kato: "Molecular Cloning and Expression of a Major Surface Protein (the 75 Kilodalton Protein) of Porphyromonas(Bacteroides)gingivalis in Esherichia coli" FEMS Microbiology Letters. (1992)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Watanabe, K., T. Takasawa, F. Yoshimura, M. Ozeki, M. Kawanami and H. Kato: "Molecular Cloning and Expression of the Gene Encoding a Major Surface Protein of Porphyromonas gingivalis in Escherichia coli." FEMS Microbiology Letters. (1992)
  • Description: 「研究成果報告書概要(欧文)」より