Molecular Genetic Analysis of Chlamydia Pathogenicity

衣原体属细菌是动物和人的重要病原体。由以下原因引起的疾病 衣原体(Chlamydia spp.)在人类中包括肺炎、心内膜炎、多发性关节炎、失明和多种疾病 性传播疾病，包括子宫颈炎、输卵管炎、盆腔炎和不孕症 女性；男性为非淋菌性尿道炎和急性附睾炎。衣原体感染也受到了牵连 作为多种慢性病的辅因子，如冠心病。尽管多年来的努力， 衣原体由于其固有的细胞内生活方式和复杂性，仍然难以进行遗传分析 发育周期。还没有人能够将外来DNA导入到这个有机体中并获得稳定的 外源基因的遗传和表达。我们的长期目标是将遗传学的力量应用于 研究衣原体的致病机制。这项提议的目的包括发展 分析衣原体发病机制的遗传工具和假说驱动的目的是解决特定的 衣原体生物学问题。其具体目的是：1)开发一种引入、表达、 以及衣原体中外源DNA的稳定维持和衣原体基因替换系统；2) 鉴定肽聚糖合成衣原体的途径；以及，3)鉴定 衣原体生长必需成分的摄取，特别是甲基供体的来源 甲基转移酶。我们将使用遗传、生化和细胞生物学策略来实现每个目标。在以下方面取得成功 实现第一个目标将对衣原体研究产生重大影响，因为它将为 可获得衣原体的遗传分析。我们对衣原体致病机制认识的快速进展 生物学和构建衣原体突变株的能力将用于疫苗开发 通过这些新技术。目标2的成功将最终解决衣原体异常，而目标3 将试图解决衣原体生物学的另一个明显的悖论。因此，这两个目标将解决 衣原体生物学的一些长期存在的问题和扩展我们对细胞内生活方式的认识 这种重要的病原体..

Bacteria of the genus Chlamydia are significant pathogens of animals and man. The diseases caused by Chlamydia spp. in man include pneumonia, 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. Chlamydia has also been implicated as a cofactor in a variety of chronic diseases such as coronary heart disease. 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 and expression of the foreign genes. Our long-term goal is to apply the power of genetics to study the pathogenic mechanisms of Chlamydia. The aims of this proposal include the development of genetic tools for the analysis of Chlamydia pathogenesis and hypothesis-driven aims to address specific questions of Chlamydia biology. The specific aims are to: 1) develop a method for introduction, expression, and stable maintenance of foreign DNA in Chlamydia and a system for gene replacement in Chlamydia; 2) characterize the pathway for peptidoglycan synthesis mChlamydia; and, 3) identify the transport system for uptake of essential constituents for Chlamydia growth, specifically the source of methyl donors for methyltransferases. We will employ genetic, biochemical and cell biology strategies to each aim. Success in achieving the first aim 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. Success in aim 2 will finally resolve the chlamydia anomaly while aim 3 will attempt to resolve another apparent paradox of Chlamydia biology. Thus, these two aims will resolve some long-standing questions of Chlamydia biology and extend our knowledge of the intracellular lifestyle of this important pathogen..