PERIPLASMIC FLAGELLA OF TREPONEMA DENTICOLA

齿螺旋体周质鞭毛

• 批准号: 2128679
• 负责人: JOHN Dawson RUBY
• 金额: $ 9.04万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2128679
• 关键词: Treponema; Treponema infection; anaerobiosis; chromosome walking; cilium /flagellum motility; dental plaque; flagellum; gel electrophoresis; gingival sulcus; laboratory mouse; microorganism classification; microorganism genetics; oral bacteria; periodontium disorder; protein structure; southern blotting; virulence

Treponema denticola is a member of the oral flora with its primary habitat being the gingival sulcus/periodontal pocket. This spirochete's potential for causing periodontal destruction along with being associated with anaerobic infections of the oral cavity is well documented. Movement of T. denticola through crevicular fluid and possibly periodontal tissue may not only be necessary for the survival of the organism, but also contribute to its virulence. A fundamental understanding of motility would be helpful in defining T. denticola's role as a pathogen in oral disease. Spirochete swimming is mediated by the periplasmic flagella (PFs) which reside between the protoplasmic cylinder and outer membrane sheath. It was recently shown that the PFs of T. denticola are antigenically similar to those of the well studied but genetically different spirochete T. phagedenis. The goals of the present investigation are to analyze the PFs of T. denticola in detail, and compare these results to those of other spirochetes. In this way, a better understanding of spirochete motility will be obtained. We propose the following: (1) Isolate the PFs of denticola; (2) characterize the major structural proteins comprising the PFs and their arrangement on the PFs; (3) clone and sequence the PF genes of T. denticola, and compare these results to those of T. phagedenis and other spirochetes. PFs will be isolated using established procedures. The separation and analysis of PF proteins will be done using sodium dodecyl sulfate- polyacrylamide gel electrophoresis, two dimensional gel electrophoresis, and Western blotting using both polyclonal and monoclonal antibody. The distribution of these proteins on the PFs will be determined by immunocytochemical methods using either monoclonal antibodies conjugated to protein A-colloidal gold, or undecagold-Fab probes. The PF genes of T. denticola will be cloned into Escherichia coli using PF specific monoclonal and polyclonal antisera and lambda gtll. Restriction enzyme maps along with Southern blotting and chromosome walking of cloned gene fragments will be used to determine the number of PF genes. The PF gene will be sequenced by the dideoxy chain termination method. Nucleotide and deduced amino acid sequences will be compared to data-bases, with a special emphasis on the PF genes from other spirochetes. Regions of homology will define conserved sequences among the spirochetes. These regions are likely to be necessary for PF structure and motility.

treponema denticola是其主要栖息地的口腔菌群的成员 是牙龈沟/牙周口袋。 这个螺旋体的潜力 造成牙周破坏以及与 口腔的厌氧感染已充分记录。 T的运动。 通过缝隙液和牙周组织的牙齿可能不会 仅对于生物的生存是必要的，但也有助于 它的毒力。 对运动的基本理解将有所帮助 定义牙霉菌在口腔疾病中的病原体作用。 螺旋体 游泳是由驻留在 原生质缸和外膜鞘。 最近显示了 牙霉菌的PFS在抗原上与井的PF相似 研究但在遗传上不同的螺旋体T. phagedenis。 目标 本研究是为了详细分析牙霉菌的PFS， 并将这些结果与其他螺旋体的结果进行比较。 这样， 将获得对螺旋体运动的更好理解。 我们建议 以下内容：（1）隔离牙本质的PF； （2）表征 主要结构蛋白包括PFS及其在 PFS; （3）克隆并序列的牙霉菌的PF基因，并比较这些 结果是thephagedenis和其他螺旋体的结果。 PF将使用既定程序隔离。 分离和 PF蛋白的分析将使用十二烷基硫酸钠进行。 聚丙烯酰胺凝胶电泳，二维凝胶电泳， 以及使用多克隆和单克隆抗体的蛋白质印迹。 这 这些蛋白质在PFS上的分布将由 使用一种共轭的单克隆抗体的免疫细胞化学方法 蛋白质A-胶体黄金或未结合的探针。 T的PF基因。 牙齿将使用PF特异性单克隆克隆到大肠杆菌中 以及多克隆抗血清和lambda gtll。 限制酶图 随着南部印迹和克隆基因片段的染色体步行将 用于确定PF基因的数量。 PF基因将进行测序 通过Dideoxy链终止方法。 核苷酸和推导的氨基酸 将比较序列与数据库，并特别强调PF 来自其他螺旋体的基因。 同源性区域将定义保守 螺旋体之间的序列。 这些区域可能是必要的 用于PF结构和运动。

项目成果

Effect of temperature and viscosity on the motility of the spirochete Treponema denticola.

温度和粘度对螺旋体齿垢密螺旋体运动的影响。

• DOI: 10.1111/j.1574-6968.1998.tb13325.x
• 发表时间: 1998
• 期刊: FEMS microbiology letters
• 影响因子: 2.1
• 作者: Ruby,JD;Charon,NW
• 通讯作者: Charon,NW