Transposon mutagenesis for Chlamydia trachomatis

项目摘要 沙眼衣原体对美国和美国的公共健康有巨大的影响 全世界。尽管有这种影响，但仍然缺乏具有 通过实验定义和评估。这在很大程度上是由于最近的发展 遗传工具和能力。一个尚未开发的工具是转座子 诱变。这是一种产生单基因缺失的非常有效的方法 评估对特定表型的影响。作为我们的初步数据报告，我们有 证明了HIMAR转座子系统在产生 衣原体转座子插入突变株。这项提议旨在扩大 并产生定义的沙眼衣原体转座子文库(特定目标1)。 由于衣原体的专性胞内性质，破坏性的转座子插入 支持这些基因产物不是体外(组织培养)生长所必需的。 鉴于高度进化和浓缩的基因谱系，我们假设一些 在这些非必需的体外基因中，对体内(脊椎动物)很重要 感染。为了验证这一假设，我们正在评估 小鼠模型中的突变株。通过这些努力，它有望在实验中 发现沙眼衣原体编码的新毒力因子。这些发现 可能导致针对衣原体或其他微生物的新的治疗或预防策略 传染病。

Project Summary Chlamydia trachomatis has an immense impact on public health in the US and worldwide. Despite this impact, there is a paucity of virulence factors that have experimentally defined and evaluated. This is largely due to the very recent development of genetic tools and capabilities. One tool that has not been developed yet is transposon mutagenesis. This is a very powerful approach to generate single gene deletions and evaluate influence on specific phenotype. As our preliminary data report, we have demonstrated that the himar transposon system is functional and effective in generating transposon insertion mutant strains of Chlamydia. This proposal is designed to expand and generate a defined transposon library of Chlamydia trachomatis (Specific Aim 1). Due to the obligate intracellular nature of Chlamydia, disruptive transposon insertions support that these gene products are not essential for in vitro (tissue culture) growth. Given the highly evolved and condensed genetic repertoire, we hypothesize that some of these unessential in vitro genes are important for in vivo (vertebrate mammal) infection. To test this hypothesis, we are evaluating the relative bacterial burden of mutant strains in a mouse model. Through these efforts it is expect to experimentally discover novel virulence factors encoded by Chlamydia trachomatis. These discoveries may lead to new treatment or prevention strategies for Chlamydia or other microbial infectious diseases.