MOLECULAR BIOLOGICAL STUDIES ON ADHERENT FACTORS OF A PERIODONTOPATHOGEN

牙周病原粘附因子的分子生物学研究

• 批准号: 05454494
• 负责人: YOSHIMURA Fuminobu
• 金额: $ 4.29万
• 依托单位: AICHI-GAKUIN UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454494/
• 关键词: PERIODONTOPATHOGEN; Porphyromonas gingivalis; ADHERENT FACTOR; HEMAGGLUTININ; FIMBRIAE; GENE PRODUCT; FIMBRIAL GENES; PROTEASE; Porphyromonas qinqivalis; 欠損変異体; 線毛関連遺伝子; 遺伝子の塩基配列

Porphyromonas gingivalis is a Gram-negative, oral anaerobe and has been considered to be a periodontopathogen. It has several adherent factors such as fimbriae, hemagglutinin, and an outer membrane protein as possible virulent factors. Region downstream from the fimA gene encoding fimbrilin, a major subunit of fimbriae, was seqenced, and four open reading frames (ORFs) were found in this region. ORF2 and ORF3 encoding 50- and 80-kDa protein, respectively, were found to be involved in fimbriae as minor components. A mutant possessing mutation in ORF3 was created by insertional mutagenesis using a suicide vector. This mutant produced fimbrilin in cell, but no fimbriae on the surface of the cell. Hemagglutinin possessing trypsin-like protease activity (HA/P) has also been studied. Gene encoding this unique substance was cloned and sequenced almost completely. The DNA sequence revealed that HA/P is expressed from a large ORF and the hemagglutinin and protease domains seem to be located in the N-terminal region. They seem to share one common active site. Gene encoding a major outer membrane protein (75-kDa) was also cloned and sequenced.

牙龈卟啉单胞菌是一种革兰氏阴性的口腔厌氧菌，被认为是一种牙周病病原体。它有几个粘附因子，如菌毛、血凝素和外膜蛋白可能是致毒因子。对fimA基因编码纤原蛋白的下游区域进行了测序，在该区域发现了4个开放阅读框（orf）。ORF2和ORF3分别编码50- kda和80-kDa蛋白，作为次要组分参与菌毛。利用自杀载体进行插入诱变，获得了ORF3突变体。该突变体在细胞内产生纤原蛋白，但在细胞表面没有纤原。具有胰蛋白酶样蛋白酶活性（HA/P）的血凝素也被研究过。编码这种独特物质的基因被克隆并几乎完全测序。DNA序列显示HA/P是从一个大的ORF中表达的，血凝素和蛋白酶结构域似乎位于n端区域。它们似乎共享一个共同的活性位点。此外，还克隆了一个主要外膜蛋白（75-kDa）的编码基因，并对其进行了测序。

项目成果

Yoshimura, F.: Molecular Biology of P.gingivalis Fimbriae. In H.N.Shah, D.Mayrand, and R.J.Genco (ed.) Biology of the Species Porphyromonas gingivalis. CRC press, Boca Raton, 321-338 (1993)

Yoshimura, F.：牙龈卟啉单胞菌菌毛的分子生物学。

Hamada,H.et al: "Constnuction and characterization of a fimA mulant of Porphyromonas gingivalis" Infection and Immunity. 62. 1696-1704 (1994)

Hamada,H.et al：“牙龈卟啉单胞菌 fimA 突变体的构建和表征”感染和免疫。

Hamada, N., K.Watanabe, C.Sasakawa, M.Yoshikawa, F.Yoshimura, and T.Umemoto: "Construction and Characterization of a fimA Mutant of porphyromonas gingivalis" Infect.Immun.62. 1696-1704 (1994)

Hamada, N.、K.Watanabe、C.Sasakawa、M.Yoshikawa、F.Yoshimura 和 T.Umemoto：“牙龈卟啉单胞菌 fimA 突变体的构建和表征”Infect.Immun.62。

Hoover,C,H.& F.Yoshimura: "Transposon-induced pigment-deficient murants of Podphyromonas gihgivalis" FEMS Microbiology Letters. 124. 43-48 (1994)

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