Isolation and characterization of transposon-derived fimbria-deficient mutants of Porphyromonas gingivalis

牙龈卟啉单胞菌转座子源性菌毛缺陷突变体的分离和表征

• 批准号: 09470398
• 负责人: YOSHIMURA Fuminobu
• 金额: $ 3.26万
• 依托单位: Aichi-Gakuin University School of Dentistry
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470398/
• 关键词: Porphyromonas gingivalis; Formation of fimbriae; Fimbria-deficient mutants; Transposon; Two-component signal transduction system; Anaerobe; Periodontal disease; Gram-negative rod; 線毛; 付着因子

Transposon-derived, fimbria-deficient mutants of Porphyromonas gingivalis have been isolated and characterized to identify a new chromosomal locus essential for fimbriation. The fimA gene coding for fimbrilin, a major subunit protein of fimbriae, was located on the largest NotI fragment (780kb). At least 8 independent, fimbria-deficient mutants were found to have insertions on a 300kb NotI fragment. Almost all the mutants had insertions in a limited area, judged by genomic Southern analysis. Therefore we cloned the Tn4351 containing fragment of the chromosome from AG30-4, one of the 7 mutants in Escherichia coli. A cloned P. gingivalis DNA fragment adjacent to Tn4351 was sequenced. The nucleotide sequence obtained revealed that a two component signal transduction system was interrupted by the transposon insertion, and two genes, fimS, a sensor histidine kinase gene and fimR, a response regulator gene were found as a tandem arrangement in this region. Among the 7 mutants 6 mutants had insertions in fimS and only one in fimR. This two-component system is the first one found in this organism and is involved in fimbriation.

转座子衍生的牙龈卟啉单胞菌纤维缺陷突变体已经被分离出来并鉴定出一个新的染色体位点。编码纤维蛋白的fimA基因位于最大的NotI片段（780kb）上。至少有8个独立的菌毛缺陷突变体被发现在300kb NotI片段上有插入。根据基因组南方分析，几乎所有的突变体在有限的区域内都有插入。因此，我们从大肠杆菌7个突变体之一的AG30-4中克隆了含有该染色体片段的Tn4351。对邻近Tn4351的克隆牙龈假单胞菌DNA片段进行了测序。获得的核苷酸序列表明，该转座子插入中断了一个双组分信号转导系统，在该区域发现了两个基因，即组氨酸激酶传感器基因fimS和反应调节基因fimR串联排列。7个突变体中有6个突变体在fimS中插入，只有1个突变体在fimR中插入。这种双组分系统是在这种生物体中发现的第一个系统，它与颤振有关。

项目成果

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Hongo,H., et al.：“牙龈卟啉单胞菌外膜蛋白基因 pamA 及其基因产物的表征”Microbiol.Immunol.. 43. 937-946 (1999)

Hayashi,J. et al.: "Advance in Microbiology : Application of transposon to Porphyromonas gingivalis"Japanese J.Bacteriol. 53, in Japanese. 509-518 (1998)

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Hayashi, J. et al: "Identification of a two-component signal transduction system involved in fimbriation of Porphyromonas gingivalis"Microbiol. Immunol.. 44(). (2000)

Hayashi, J. 等人：“鉴定参与牙龈卟啉单胞菌菌毛形成的双组分信号转导系统”微生物学。

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