MOLECULAR GENETIC ANALYSIS OF CHLAMYDIA PATHOGENICITY

衣原体致病性的分子遗传学分析

• 批准号: 2728334
• 负责人: Anthony T Maurelli
• 金额: $ 26.1万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2728334
• 关键词: Chlamydiaceae; bacterial DNA; bacterial cytopathogenic effect; bacterial genetics; genetic manipulation; genetic techniques; molecular cloning; molecular genetics; mutant; phenotype; plasmids; technology /technique development; transposon /insertion element

DESCRIPTION (Adapted from the Applicant's Abstract): Bacteria of the genus Chlamydia are significant pathogens of animals and man. The diseases caused by Chlamydia spp. in man include pneumonitis, endocarditis, polyarthritis, blindness, and a wide range of sexually transmitted diseases including cervicitis, salpingitis, pelvic inflammatory disease, and infertility in females; and non-gonococcal urethritis and acute epididymitis in males. Despite many years of effort, the Chlamydia remain intractable to genetic analysis due to their obligate intracellular lifestyle and complex developmental cycle. No one has been able to introduce foreign DNA into this organism and achieve stable inheritance of the expression of the foreign genes. Few attempts at isolation of Chlamydia mutants have been reported. Even cloning of Chlamydia genes by complementation has been problematic due to the absence of, or poor expression of cloned Chlamydia genes in Escherichia coli. Our long term goal is to apply the power of genetics to study the pathogenic mechanisms of Chlamydia. The goal of this proposal is to develop genetic tools for the analysis of Chlamydia biology and pathogenesis and to use these tools to address specific problems of Chlamydia pathogenesis. The Specific aims are to: 1) design an efficient, reproducible method for introduction, expression, and stable maintenance of foreign DNA in Chlamydia; 2) design genetic tools for mutagenesis and selection of mutant phenotypes of Chlamydia ; 3) clone Chlamydia genes by functional complementation; and, 4) develop a system for gene replacement in Chlamydia. Each of these aims will include development of a genetic tool, demonstration of the tools effectiveness, and application of the tool to a fundamental question of Chlamydia biology. Success in achieving these goals will have a significant impact on Chlamydia research by making new tools for genetic analysis of Chlamydia available. Rapid advances in our understanding of Chlamydia pathogenesis and biology as well as the ability to construct Chlamydia mutants for vaccine development will be made possible by these new techniques.

描述（改编自申请人的摘要）：细菌 衣原体属是动物和人类的重要病原体。这 由衣原体属引起的疾病。男性包括肺炎， 心内膜炎、多关节炎、失明和广泛的性病 传染性疾病包括宫颈炎、输卵管炎、盆腔炎 炎症性疾病和女性不孕症；和非淋菌性 男性尿道炎、急性附睾炎。尽管多年来 尽管努力，衣原体仍然难以进行遗传分析，因为 他们的专性细胞内生活方式和复杂的发育周期。 没有人能够将外源 DNA 引入到这种生物体中，并且 实现外源基因表达的稳定遗传。很少 已报道了分离衣原体突变体的尝试。甚至 通过互补克隆衣原体基因一直存在问题，因为 克隆衣原体基因的缺失或表达不良 大肠杆菌。我们的长期目标是运用遗传学的力量 研究衣原体的致病机制。此举的目标 建议开发用于分析衣原体的遗传工具 生物学和发病机制，并使用这些工具来解决特定的问题 衣原体发病机制的问题。具体目标是：1）设计 一种高效、可重复的引入、表达和分析方法 衣原体中外源DNA的稳定维持； 2）设计遗传工具 用于衣原体突变表型的诱变和选择； 3） 通过功能互补克隆衣原体基因；以及，4) 开发一个 衣原体基因替换系统。这些目标中的每一个都将 包括遗传工具的开发、工具的演示 有效性以及该工具在解决基本问题上的应用 衣原体生物学。成功实现这些目标将带来 通过开发新的遗传工具对衣原体研究产生重大影响 可获得衣原体分析。我们的认识迅速进步 衣原体发病机制和生物学以及构建能力 用于疫苗开发的衣原体突变体将通过这些成为可能 新技术。