PLASMID BIOLOGY OF ACTINOBACILIUS ACTINOMYCETEMCOMITANS

伴放线杆菌的质粒生物学

• 批准号: 2430145
• 负责人: DONALD J LEBLANC
• 金额: $ 15.41万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2430145
• 关键词: Actinobacillus actinomycetemcomitans; Escherichia coli; Pseudomonas aeruginosa; Streptococcus lactis; bacterial DNA; bacterial genetics; electroporation; molecular cloning; plasmids; recombinant DNA; replicon; tissue /cell culture; transfection /expression vector

DESCRIPTION (Adapted from the Applicant's Abstract): Actinobacillus actinomycetemcomitans (Aa) has been implicated in the pathogenesis of juvenile and adult periodontitis. This microorganism also is capable of causing more serious human infections such as endocarditis and soft tissue abscesses, and the likely source of these infections is Aa derived from oral sources. The ability of Aa to cause these disease states clearly depends on its ability to colonize and/or invade oral, cardiac, and soft tissues, and to elaborate various products that contribute to its survival in the host, or to cause damage to these tissues. The identification of Aa virulence traits, and tests for their actual roles in pathogenesis, will depend on the availability of modern genetic and molecular tools, and suitable animal models. With regard to the former, genetic and molecular analyses of Aa and its virulence have been lacking, due in large measure to the paucity of genetic transfer systems for this microorganism, and the total absence of appropriate host-vector systems for the application for recombinant DNA technology, including the delivery of manipulated genetic information for mutagenesis and studies of gene regulation. The applicant has described the isolation of plasmid DNA from two strains of Aa, and the construction of E. coli/Aa shuttle plasmids from one of the Aa replicons (pVT736-1), and the use of one of the shuttles to establish an electroporation-based transformation system for this species. More recently, the applicant has examined the properties of pVT736-1 in greater detail, and obtained preliminary data on the replication in Aa of three additional plasmids, two of Gram-negative and one of Gram- positive bacterial origin. The goal of this proposal is to assess the potential of all four plasmids as molecular and genetic tools for the analysis of Aa. Two specific aims are: 1) to evaluate the properties of each plasmid relative to its potential as an E. coli/Aa shuttle vector; and 2) to construct a vector with properties that enhance recombinational events between Aa genomic DNA and plasmid DNA.

描述（改编自申请人的摘要）：actinobacillus 放线症患者（AA）与 少年和成人牙周炎。这种微生物也能够 引起更严重的人类感染，例如心内膜炎和软感染 组织脓肿，这些感染的可能来源是AA 源自口服来源。 AA引起这些疾病的能力 各州显然取决于其殖民和/或入侵口头的能力， 心脏和软组织，并详细说明各种产品 促进其在宿主中的生存，或造成这些损害 组织。识别AA毒力性状，并测试其 在发病机理中的实际作用将取决于现代的可用性 遗传和分子工具以及合适的动物模型。关于 前者的AA及其毒力的遗传和分子分析具有 缺乏，很大程度上是由于遗传转移的匮乏 这种微生物的系统，总缺乏适当的 重组DNA技术应用的宿主矢量系统， 包括传递操纵的遗传信息 基因调节的诱变和研究。申请人已经描述了 从两个AA菌株中分离质粒DNA，然后 来自AA复制品之一的大肠杆菌/AA班车质粒 （pvt736-1），并使用其中一辆班车来建立一个 该物种的基于电穿孔的转换系统。 更多的 最近，申请人检查了pvt736-1在 更大的详细信息，并获得了有关AA复制的初步数据 在另外三个质粒中，革兰氏阴性剂和革兰氏 阳性细菌起源。该提议的目的是评估 所有四个质粒的潜力作为分子和遗传工具 AA分析。两个具体目的是：1）评估 每个质粒相对于其作为大肠杆菌/AA班车载体的潜力； 2）构建具有增强属性的向量 AA基因组DNA和质粒DNA之间的重组事件。