Comparative Genomics of Francisella

弗朗西斯菌的比较基因组学

• 批准号: 7083568
• 负责人: GEORGE M WEINSTOCK
• 金额: $ 29.3万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7083568
• 关键词: Francisella tularensis; bacterial genetics; biotechnology; bioterrorism /chemical warfare; expression cloning; functional /structural genomics; gene deletion mutation; genetic library; genetic strain; immunologic assay /test; microarray technology; nucleic acid sequence; proteomics; single nucleotide polymorphism; virulence

The focus of this project is Francisella tularensis, the causative agent of tularemia and a Category A Bioterrorism Select Agent. There is an urgent need for data and reagents that immediately can be directed toward vaccine and therapeutic agent discovery as well as immunological diagnostic assay development for F. tularensis, and other Category A organisms. Our overall goal is to use genomic and proteomic approaches to identify genes involved in F. tularensis pathogenesis and antigens important for tularemia immunity. In this proposal, we will merge novel sequencing and oligosynthesis/array technologies (developed by our subcontract partners) with traditional Sanger sequencing strategies to create a cost effective method to identify genetic differences between important Francisella strains with different virulence and lifestyle phenotypes. As a first step toward this goal, we will determine the genomic sequences of two type A and one type B Francisella strain. These sequences will be compared to three already completed Francisella genome sequences to identify the insertions, deletions, and single-nucleotide polymorphisms that may lead to differing Francisella phenotypes. Genome sequencing will be coupled to proteome expression using a novel shotgun library/expression plasmid to both host the clones for Francisella sequencing and to express peptides from each Francisella proteome. Peptides from the three proteomes will be screened with immunoreactive sera to identify differences in their antigen profiles. The comparative genetic analysis of virulent and avirulent Francisella strains in conjunction with an antigen screen against peptides from the proteomes of these bacteria will identify putative virulence factors and antigens for future study as vaccine and immunodiagnostic reagents.

该项目的重点是土拉热弗朗西斯菌，土拉菌病的病原体和A类生物恐怖主义选择剂。目前迫切需要的数据和试剂，立即可以直接对疫苗和治疗剂的发现，以及免疫诊断检测的发展，为F。土拉菌和其他A类生物。我们的总体目标是利用基因组学和蛋白质组学的方法来确定参与F。土拉菌病发病机制和对土拉菌免疫重要的抗原。在这项提案中，我们将把新的测序和寡核苷酸合成/阵列技术（由我们的合作伙伴开发）与传统的桑格测序策略相结合，以创造一种具有成本效益的方法， 鉴定具有不同毒力和生活方式表型的重要弗朗西斯菌菌株之间的遗传差异。作为实现这一目标的第一步，我们将确定两个A型和一个B型弗朗西斯菌株的基因组序列。将这些序列与三个已经完成的弗朗西斯菌基因组序列进行比较，以确定可能导致不同弗朗西斯菌表型的插入、缺失和单核苷酸多态性。 将使用新型鸟枪文库/表达质粒将基因组测序与蛋白质组表达偶联，以宿主用于弗朗西斯菌测序的克隆并表达来自每个弗朗西斯菌蛋白质组的肽。将用免疫反应性血清筛选来自三种蛋白质组的肽，以鉴定其抗原谱的差异。对强毒和无毒弗朗西斯菌菌株进行比较遗传分析，并结合针对这些细菌蛋白质组中的肽进行抗原筛选，将鉴定推定的毒力 因子和抗原作为疫苗和免疫诊断试剂的未来研究。