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Controlling the progression of pneumonic plague

控制肺鼠疫的进展

基本信息

批准号：
8234195
负责人：
WILLIAM E GOLDMAN
金额：
$ 22.39万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-03-01 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8234195
关键词：
Aerosols Animal Model Animals Bacteria Bacterial Genes Biological Models Bioterrorism Cessation of life Collaborations Collection Development Disease Disease Progression Gene Expression Gene Expression Profile Genes Genetic Genome Genomics Goals Histopathology Hour Hybridization Array Infection Inflammation Inflammatory Response Insertional Mutagenesis Instruction Kinetics Lead Libraries Lung Lung diseases Methods Microarray Analysis Modeling Monitor Mus Mutagenesis Oligonucleotides Organism Output Paper Pathology Peptide Hydrolases Phase Plague Plasminogen Pneumonia Pneumonic Plague Publishing Research Reverse Transcriptase Polymerase Chain Reaction Role Route Spleen Staging Symptoms Syndrome Testing Time Virginia Virulence Yersinia pestis antimicrobial base biodefense comparative gene discovery in vivo mouse model mutant novel therapeutics response transposon site hybridization

项目摘要

Our overall goal for this research plan is to use a mouse model system for pneumonic plague to discover and evaluate Y. pestis genes critical for the development and progression of disease. We will pinpoint these candidates using two methods: transcriptional profiling to reveal genes that are differentially regulated in the various stages of pneumonic plague; and forward genetics approaches to screen/select for Y. pestis genes that are indispensable for development of pulmonary disease. Specific Aim 1. Comparative transcriptional responses by Y. pestis during the stages of pneumonic plague. We previously developed a whole genome microarray to characterize the bacterial transcriptome during pneumonic plague, but this analysis was technically limited to a late stage of infection. Therefore, we will use quantitative RT-PCR to examine a subset of Y. peso's genes throughout the entire time course of disease. This subset of approximately 250 genes is based on genes that show evidence of differential expression during infection, as well as genes that were not sufficiently explored by microarray technology. Specific Aim 2. Forward genetics to identify bacterial genes important in the development of pneumonic plague. Transposon site hybridization (TraSH) is a gene discovery strategy using negative selection to dentify bacterial genes that are essential during infection. The microarrays we have constructed will allow us to take advantage of a a TraSH-based approach using array hybridizations to identify Y. pestis genes triplicated in various stages of the pulmonary infection. Specific Aim 3. Analyzing the importance and role of candidate virulence-associated genes. The genes selected in the first two Aims will be targeted for further analysis by creating defined mutant strains of Y. pestis. Mutant and control strains will be tested for virulence in the murine model of pneumonic plague, monitoring bacterial proliferation in the lung, dissemination to the spleen, and histopathology to evaluate differences in the manifestation or kinetics of disease. The characterization of mutant strains will be extended to a microarray analysis of host transcriptional responses during infection, done in collaboration with Dr. Virginia Miller.

本研究计划的总体目标是利用小鼠模型系统对肺鼠疫进行发现