Gene Expression and Manipulation of Coxiella Burnetii

伯内氏柯克斯体的基因表达和操作

基本信息

批准号：
7641034
负责人：
Michael F Minnick
金额：
$ 70.15万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7641034
关键词：
Acute Address Aerosols Alveolar Macrophages Antigens Attenuated Attenuated Vaccines Bacteria Candidate Disease Gene Cells Clinical Cloning Complement Coxiella Coxiella burnetii Cytolysis Data Development Dust Endocarditis Environment Future Gene Expression Gene Proteins General Practitioners Generations Genes Hepatitis Human In Vitro Individual Infection Insect Bites Invasive Knock-out Lead Life Cycle Stages Mammals Milk Mind Modeling Molecular Molecular Biology Molecular Profiling Morphogenesis Mus Mutagenesis Natural History Nature Open Reading Frames Organism Parasites Pathogenesis Phagolysosome Phase Pneumonia Polyvalent Vaccine Population Proteins Q Fever Range Relative (related person)Reporting Research Role Route Shuttle Vectors Site Site-Directed Mutagenesis Spontaneous abortion Staging System Technology Testing Ticks Time Vaccine Design Vaccine Production Variant Virulence Virulence Factors base cell type design extracellular genetic manipulation improved in vivo infancy mouse model mutant pathogen research study tool trait transmission process

项目摘要

Introduction- Coxiella burnetii is the etiologic agent of Q fever, a potential bioweapon and select agent, and one of the most infectious pathogens known. However, there are few reports concerning this obligate intracellular agent's molecular pathogenesis or developmental cycle, and lack of a system for site-directed genetic manipulation has severely hampered research progress. To address this dearth of information and to improve the current state of molecular biology technology, we propose to: 1) examine the expression profiles of small cell variants (SCVs) and large cell variants (LCVs) of the developmental cycle to identify genes and proteins that allow Coxiella to survive for extended periods of time in the environment (SCV stage) and to subsequently grow in the inhospitable confines of the host cell phagolysosome (LCV stage); 2) analyze and compare expression profiles of in vitro versus in vivo-cultivated Coxiella and phase I versus phase II organisms to identify potential virulence determinants, and 3) develop a system of site-directed mutagenesis for this organism to allow for the routine genetic manipulation and future development of an attenuated vaccine strain. These data are expected to provide valuable information on the life cycle of Coxiella and the respective genes and proteins involved, and should lead to strategies for disrupting cellular development to control Q fever. Finally, a system for routine genetic manipulation of Coxiella is an essential molecular biology tool for examining suspected virulence genes and performing molecular Koch's postulates in vivo. Project Interactions- Project 3 (Harmsen) will interact with Project 1 (this study) as it relates to in vivocultivated Coxiella from the mouse model. Projects 4 (Pascual) and 2 (Jutila) will benefit from Project 1 data on stage-specific gene products; SCV-specific genes / antigens are hypothesized to be involved in transmission / early infection and LCV-specific genes / antigens are involved in intracellular replication.

简介 - 伯纳特柯克斯体是 Q 热的病原体，是一种潜在的生物武器和选择剂，并且已知最具传染性的病原体之一。然而，关于这一义务的报道却很少。细胞内因子的分子发病机制或发育周期，以及缺乏定点系统基因操纵严重阻碍了研究进展。为了解决信息缺乏的问题并为了改善分子生物学技术的现状，我们建议：1）检查表达谱发育周期的小细胞变体 (SCV) 和大细胞变体 (LCV) 来识别基因和使柯克斯体能够在环境中长时间存活（SCV 阶段）并随后在宿主细胞吞噬溶酶体的恶劣环境中生长（LCV 阶段）； 2）分析和比较体外与体内培养的柯克斯体以及 I 期与 II 期的表达谱生物体以确定潜在的毒力决定因素，3) 开发定点诱变系统让这种生物体能够进行常规基因操作和未来发展减毒疫苗株。这些数据有望提供有关柯克斯体生命周期和涉及各自的基因和蛋白质，并且应该导致破坏细胞发育的策略控制Q热。最后，柯克斯体的常规遗传操作系统是一个重要的分子用于检查可疑毒力基因并在体内执行分子科赫假设的生物学工具。项目互动 - 项目 3（Harmsen）将与项目 1（本研究）互动，因为它涉及体内培养来自小鼠模型的柯克斯体。项目 4 (Pascual) 和 2 (Jutila) 将受益于项目 1 的数据关于特定阶段的基因产物；假设 SCV 特异性基因/抗原参与传播/早期感染和LCV特异性基因/抗原参与细胞内复制。