Investigation and characterization for the periodontopathic bacterial genes specifically expressing in vivo

体内特异性表达的牙周病细菌基因的研究和表征

• 批准号: 13671901
• 负责人: KOKEGUCHI Susumu
• 金额: $ 1.92万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671901/
• 关键词: periodontopathic bacteria; Porphyromonas gingivalis; Actinobacillus actinomycetemcomitans; in vivo-expressing; bacterial genes; Actimobacillus actinomycetemcomitans; 遺伝子解析

Pathogenic bacteria could express specific genes and its proteins under the specific growth condition or especially in vivo, which might be related to bacterial growth and pathogenicity. In this study, we aimed to investigate and characterize such specific genes and proteins of periodontopathic bacteria, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans expressing in vivo.The major results of this study are as follows:1)P. gingivalis expresses specific 53-kDa outer membrane protein(Ag53) which evokes strong humoral immune responses in evokes strong humoral immune responses in many patients with periodontal disease. Three sequences, RAAIRAS, YYLQ and MSPARR, were identified as major B-cell epitopes in Ag53, which might influence individual differences in antibody titer against Ag53.2)A. actinomycetemcomitans express fimbriae in vivo that plays an important role in the ability of this bacterium to colonize teeth and other surfaces. The flp-1 gene encodes the major fimbriae protein component of A. actinomycetemcomitans. We compared flp-1 DNA sequences from 43 A. actinomycetemcomitans clinical isolates. DNA and predicted protein sequences were highly divergent in the C-terminus of the flp-1 protein that has been shown to be important for the bundling and adhesive properties of the fimbriae, suggesting that these diverse strains carry pathogenic potential.3) Two genes encoding ferritin-like protein were identified in the upstream region of actX from A. actinomycetemcomitans which controls its growth and virulence. Two ferritin genes constituted a bicistronic operon and the accumulation of its mRNA was upregulated under aerobic conditions, suggested that the operon may have important roles in the adaptation of the bacteria to oxidative environmental changes.

病原菌在特定的生长条件下，尤其是在体内，可以表达特定的基因及其蛋白，这可能与细菌的生长和致病性有关。本研究的主要结果如下：1）牙龈卟啉单胞菌（Porphyromonas gingivalis）表达特异性的53-kDa外膜蛋白（Ag 53），该蛋白在牙周病患者中引起强烈的体液免疫应答。RAAIRAS、YYLQ和MSPARR序列是Ag 53的主要B细胞表位，可能影响抗Ag53.2）A抗体效价的个体差异。伴放线菌在体内表达菌毛，该菌毛在该细菌定殖牙齿和其它表面的能力中起重要作用。flp-1基因编码A.伴放线菌我们比较了43株A.伴放线菌临床分离株。flp-1蛋白C端的DNA序列与预测的蛋白序列高度不同，该蛋白对菌毛的成束和粘附特性具有重要作用，表明这些不同的菌株具有致病潜力。放线菌共生体，控制其生长和毒力。两个铁蛋白基因构成了一个双顺反子操纵子，其mRNA的积累在有氧条件下被上调，表明该操纵子可能在细菌适应氧化环境变化中起重要作用。

项目成果

Fujimoto C, Maeda H, Kokeguchi S, et al.: "Application of denaturing gradient gel electrophoresis(DGGE) of the analysis of microbial communities of subgingival plaque"J Periodont Res. (in press). (2003)

Fujimoto C，Maeda H，Kokeguchi S，等：“应用变性梯度凝胶电泳（DGGE）分析龈下菌斑微生物群落”J periodont Res。

Oyaizu K, Ohyama H, Nishimura F, Kurihara H, Matsushita S, Maeda H, Kokeguchi S, Hongyo H, Takashiba S, Murayama Y: "Identification and characterization of B-cell epitopes of a 53-kDa outer membrane protein from Porphyromonas gingivalis"Oral Microbiol & I

Oyaizu K、Ohyama H、Nishimura F、Kurihara H、Matsushita S、Maeda H、Kokeguchi S、Hongyo H、Takashiba S、Murayama Y：“牙龈卟啉单胞菌 53 kDa 外膜蛋白 B 细胞表位的鉴定和表征

Kaplan JB, Kokeguchi S, Murayama Y, et al.: "Sequence diversity in the major fimbrial gene(flp-1) of Actinobacillus actinomycetemcomitans"Oral Microbiol & Immunol. 17. 354-359 (2002)

Kaplan JB、Kokeguchi S、Murayama Y 等人：“Actinobacillus actinomycetemcomitans 的主要菌毛基因 (flp-1) 的序列多样性”口腔微生物

Fujimoto C, Maeda H, Kokeguchi S, Nishimura F, Takashiba S, Fukui K, Murayama Y: "Application of denaturing gradient gel electrophoresis(DGGE) of the analysis of microbial communities of subgingival plaque"J Periodont Res. (in press). (2003)

Fujimoto C、Maeda H、Kokeguchi S、Nishimura F、Takashiba S、Fukui K、Murayama Y：“变性梯度凝胶电泳（DGGE）在龈下菌斑微生物群落分析中的应用”J periodont Res。

Hirosue M, Kokeguchi S, Maeda H, et al.: "Characterization of two genes encoding ferritin-like protein in Actinobacillus actinomycetemcomitans"Microbiol Immunol. 45. 721-727 (2001)

Hirosue M、Kokeguchi S、Maeda H 等人：“Actinobacillus actinomycetemcomitans 中编码铁蛋白样蛋白的两个基因的特征”Microbiol Immunol。