MOLECULAR GENETIC ANALYSIS OF CHLAMYDIA PATHOGENICITY

描述（改编自申请人的摘要）： 衣原体属是动物和人类的重要病原体。 由衣原体属引起的疾病在包括肺炎人类中， 心内膜炎，多关节炎，失明，以及各种各样的性疾病。 传播性疾病，包括宫颈炎、输卵管炎、盆腔炎 炎症性疾病和女性不孕症;以及非淋菌性疾病 男性尿道炎和急性附睾炎。尽管多年来 努力，衣原体仍然难以遗传分析，由于 它们的细胞内生活方式和复杂的发育周期。 没有人能够将外源DNA引入这种生物体， 实现外源基因表达的稳定遗传。几 已经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是有问题的， 克隆衣原体基因的缺乏或表达不足， 大肠埃希菌我们的长期目标是运用遗传学的力量 来研究衣原体的致病机制这个目标 一项提案是开发用于衣原体分析的遗传工具 生物学和发病机制，并使用这些工具来解决特定的 衣原体致病机理的问题。具体目标是：1）设计 一种有效的、可重复的用于导入、表达和 外源DNA在衣原体中稳定维持; 2）设计遗传工具 用于衣原体的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 衣原体基因置换系统。每一个目标都将 包括开发遗传工具，演示工具， 有效性，并将该工具应用于一个基本问题， 衣原体生物学。成功实现这些目标将对 对衣原体研究的重大影响，使新的工具，遗传 衣原体分析可用。我们对自然的理解 衣原体的发病机制和生物学以及构建 衣原体突变体的疫苗开发将有可能通过这些 新技术

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.