Molecular Genetic Analysis of Chlamydia Pathogenicity

衣原体属细菌是动物和人类的重要病原体。 衣原体属物种在人类中包括肺炎、心内膜炎、多关节炎、失明以及各种各样疾病， 性传播疾病，包括宫颈炎、输卵管炎、盆腔炎和不孕症， 女性;男性非淋菌性尿道炎和急性附睾炎。衣原体也与 作为多种慢性疾病如冠心病的辅助因子。尽管经过多年努力， 衣原体由于其专性的细胞内生活方式和复杂的 发展周期没有人能够将外源DNA引入这种生物并实现稳定 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。该提案的目的包括： 用于分析衣原体发病机制的遗传工具和假设驱动的目的，以解决特定的 衣原体生物学的问题。具体目标是：1）开发一种介绍，表达， 衣原体中外源DNA的稳定维持和衣原体中基因置换系统; 2） 表征肽聚糖合成mChlamydia的途径;和，3）鉴定用于 衣原体生长的基本成分的摄取，特别是甲基供体的来源， 甲基转移酶我们将采用遗传学，生物化学和细胞生物学策略来实现每个目标。成功 实现第一个目标将对衣原体研究产生重大影响， 衣原体的基因分析。衣原体致病机制的快速进展 和生物学以及构建用于疫苗开发的衣原体突变体的能力 这些新技术可能。aim 2的成功将最终解决衣原体异常，而aim 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 一些长期存在的问题衣原体生物学和扩展我们的知识的细胞内的生活方式 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..