P4: SPATIOTEMPORAL EXPRESSION OF BURKHOLDERIA PSEUDOMALLEI GENES

P4: 伯克霍尔德氏菌基因的时空表达

• 批准号: 8360756
• 负责人: Tung T Hoang
• 金额: $ 21.2万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360756
• 关键词: Actins; American; Bacteria; Bioterrorism; Burkholderia pseudomallei; Categories; Cells; Cytoplasm; Drug Delivery Systems; Emerging Communicable Diseases; Endocytic Vesicle; Environment; Essential Genes; Funding; Gene Expression; Gene Fusion; Genes; Grant; Infection; Infectious Diseases Research; Intervention; Melioidosis; Membrane; Military Personnel; Modeling; Molecular; National Center for Research Resources; Pathogenesis; Prevention; Principal Investigator; Process; Reporter Genes; Reporting; Research; Research Infrastructure; Resources; Source; Staging; Time; United States National Institutes of Health; cost; macrophage; novel; spatiotemporal; vaccine candidate

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Burkholderia pseudomallei is a category B potential bioterrorism agent, which causes melioidosis, a global emerging infectious disease. Thousands of cases of melioidosis have occurred throughout the tropics, and several cases have been reported in American travelers (civilian and military). B. pseudomallei infects host-cells through various stages: i) entry into host cell endocytic vesicles, ii) vesicular escape into the cytoplasm, iii) cytoplasmic replication, iv) utilize host actin for cytoplasmic mobility to v) catapult themselves into neighboring host-cells to spread the infection through membrane protrusions. We hypothesized that various genes are expressed to facilitate the transit of this bacterium through these different stages of cellular infection. Through gene-expression analyses, the long-term objective is to understand the molecular mechanism of pathogenesis of B. pseudomallei with respect to the intracellular environment of an infected cell. Two aims will identify essential genes and genes expressed on a spatiotemporal scale in a eukaryotic macrophage model. Aim 1 will identify genes expressed on a temporal scale during the various cellular infection processes. The expression of these genes will further be confirmed by reporter-gene-fusion and mutational analyses and video time-lapse recording. Aim 2 will employ a novel high-throughput negative-selection approach to identify genes essential for the infection process, which will be confirmed by real-time PCR, reporter-gene-fusion, and time-lapse recording of the infection process. We expect that achieving the objective will yield critical intervention strategies, novel drug targets, and vaccine candidates for the treatment and prevention of melioidosis.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 鼻疽伯克霍尔德菌是一种 B 类潜在生物恐怖主义制剂，可导致类鼻疽，一种全球新发传染病。整个热带地区已发生数千例类鼻疽病例，美国旅行者（平民和军人）中报告了几例病例。 B. 假鼻疽通过不同阶段感染宿主细胞：i) 进入宿主细胞内吞囊泡，ii) 囊泡逃逸到细胞质中，iii) 细胞质复制，iv) 利用宿主肌动蛋白进行细胞质移动，v) 将自身弹射到邻近宿主中细胞通过膜突起传播感染。我们假设表达了多种基因以促进这种细菌通过细胞感染的不同阶段。通过基因表达分析，长期目标是了解类鼻疽伯克氏菌与受感染细胞的细胞内环境相关的发病机制的分子机制。两个目标将确定真核巨噬细胞模型中的必需基因和时空尺度表达的基因。目标 1 将鉴定在各种细胞感染过程中在时间尺度上表达的基因。这些基因的表达将通过报告基因融合、突变分析和视频延时记录进一步证实。目标2将采用一种新颖的高通量阴性选择方法来鉴定感染过程所必需的基因，并将通过实时PCR、报告基因融合和感染过程的延时记录来证实。我们预计，实现这一目标将产生治疗和预防类鼻疽的关键干预策略、新药物靶点和候选疫苗。