Transposon mutagenesis for Chlamydia trachomatis

沙眼衣原体转座子诱变

• 批准号: 9165577
• 负责人: P Scott Hefty
• 金额: $ 22.23万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-06 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9165577
• 关键词: Animal Model; Applied Genetics; Bacteria; Bacterial Sexually Transmitted Diseases; Biology; Blindness; Cell Culture Techniques; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Cis-Acting Sequence; Code; Communicable Diseases; Coupled; DNA Insertion Elements; Data; Data Reporting; Defect; Development; Disease; Environment; Evaluation; Future; Gene Deletion; Genes; Genetic; Genetic Screening; Genome; Growth; Human; Immune response; In Vitro; Infection; Intercistronic Region; Lead; Libraries; Mammals; Mus; Mutagenesis; Nature; Pathogenesis; Phenotype; Physiological; Play; Prevention; Prevention strategy; Process; Public Health; Role; Specificity; Staging; System; Techniques; Testing; Transposase; United States; Untranslated RNA; Virulence Factors; design; gene product; genetic approach; genetic manipulation; in vivo; microbial; mouse model; mutant; novel; pathogen; reproductive tract; tissue culture; tool; treatment strategy

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.

项目摘要 沙眼衣原体对美国的公共卫生有着巨大的影响， 国际吧尽管有这种影响，但缺乏毒力因子， 实验定义和评估。这在很大程度上是由于最近的发展， 基因工具和能力。转座子是一种尚未开发的工具 诱变这是一种非常强大的方法来产生单基因缺失， 评价对特定表型的影响。作为我们的初步数据报告， 证明了himar转座子系统在产生 衣原体的转座子插入突变株。该提案旨在扩大 并产生确定的沙眼衣原体转座子文库（特异性目的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。 考虑到高度进化和浓缩的基因库，我们假设一些 这些非必需的体外基因中的一个对于体内（脊椎动物哺乳动物） 感染为了验证这一假设，我们正在评估 在小鼠模型中的突变株。通过这些努力，有望在实验上 发现沙眼衣原体编码的新毒力因子。这些发现 可能导致新的治疗或预防衣原体或其他微生物感染的策略， 传染病