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(改编自申请人摘要)：细菌 衣原体属是动物和人的重要病原体。这个 衣原体引起的疾病。在人类中包括肺炎， 心内膜炎、多发性关节炎、失明和广泛的性行为 子宫颈炎、输卵管炎、盆腔炎等传染病 女性的炎症性疾病和不孕不育；以及非淋球菌性 男性的尿道炎和急性附睾炎。尽管多年来 努力，衣原体仍然很难进行基因分析，因为 他们专属的细胞内生活方式和复杂的发育周期。 还没有人能够将外来DNA导入到这个有机体中 实现外源基因表达的稳定遗传。一些 据报道，有人试图分离衣原体突变株。连 通过互补克隆衣原体基因一直是一个问题，因为 克隆衣原体基因的缺失或表达不佳 大肠埃希菌。我们的长期目标是应用遗传学的力量 目的：探讨衣原体的致病机制。这样做的目的是 建议开发用于衣原体分析的基因工具 生物学和发病机制，并使用这些工具来解决特定的 衣原体致病机制的几个问题。具体目标是：1)设计 一种高效、可重现的介绍、表达和 外源DNA在衣原体中的稳定维持；2)设计基因工具 用于衣原体突变表型的诱变和筛选；3) 通过功能互补克隆衣原体基因；以及4)开发一种 衣原体基因替换系统。这些目标中的每一个将 包括基因工具的开发、工具的演示 有效性，以及该工具在以下基本问题上的应用 衣原体生物学。成功地实现这些目标将有一个 制造新的基因工具对衣原体研究产生重大影响 可获得衣原体分析。我们对……的理解有了迅速的进步 衣原体的致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株将通过这些 新技术。