Transposon mutagenesis for Chlamydia trachomatis

项目概要 沙眼衣原体对美国和美国的公共卫生产生巨大影响 全世界。尽管存在这种影响，但仍存在很少的毒力因子 实验定义和评估。这主要是由于最近的发展 遗传工具和能力。转座子是一种尚未开发出来的工具 诱变。这是一种非常强大的方法来产生单基因缺失和 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 himar 转座子系统在生成 衣原体转座子插入突变株。该提案旨在扩大 并生成确定的沙眼衣原体转座子文库（具体目标 1）。 由于衣原体的专性细胞内性质，破坏性转座子插入 支持这些基因产物对于体外（组织培养）生长不是必需的。 鉴于高度进化和浓缩的遗传库，我们假设一些 这些非必需的体外基因对于体内（脊椎动物）很重要 感染。为了检验这一假设，我们正在评估以下物质的相对细菌负荷： 小鼠模型中的突变株。通过这些努力，预计将通过实验 发现沙眼衣原体编码的新毒力因子。这些发现 可能会导致衣原体或其他微生物的新治疗或预防策略 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.