Transposon mutagenesis for Chlamydia trachomatis

沙眼衣原体转座子诱变

基本信息

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
项目类别：
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

项目摘要

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和全世界。尽管有这种影响，但毒力因素很少实验定义和评估。这主要是由于最近的发展遗传工具和能力。尚未开发的一种工具是转座子诱变。这是一种非常有力的方法来产生单基因删除，并且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转座系统在生成中有效有效衣原体的转座子插入突变株。该建议旨在扩大并产生一个定义的沙眼衣原体的转座库（特定目标1）。由于衣原体的细胞内性质，破坏性转座子插入支持这些基因产物对于体外（组织培养）生长不是必不可少的。鉴于高度发展和凝结的遗传库，我们假设有些这些不必要的体外基因对体内（脊椎动物哺乳动物）很重要感染。为了检验这一假设，我们正在评估小鼠模型中的突变菌株。通过这些努力，可以实验发现由沙眼衣原体编码的新型毒力因子。这些发现可能导致针对衣原体或其他微生物的新治疗或预防策略传染病。