Virulence Genes in Mycobacterium tuberculosis

结核分枝杆菌的毒力基因

• 批准号: 6585813
• 负责人: Mark A Yoder
• 金额: $ 5.39万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6585813
• 关键词: Mycobacterium tuberculosis; bacteria infection mechanism; bacterial genetics; functional /structural genomics; gene expression profiling; in situ hybridization; laboratory mouse; laboratory rabbit; microarray technology; microorganism culture; mutant; postdoctoral investigator; respiratory infections; virulence

DESCRIPTION (provided by applicant): In order to identify mycobacterial genes important for survival within the host, a rabbit model of pulmonary tuberculosis will be developed. Rabbits will be infected by transthoracic injection and intrabronchial instillation of mycobacteria. Bacillary multiplication within cavities will allow for isolation of sufficient quantities of RNA for analysis using a Mycobacterium tuberculosis oligonucleotide microarray. Comparison of gene expression patterns between mycobacteria growing in culture and those growing in vivo will provide information on genes important in mycobacterial survival within the host. A second approach will be used concurrently to determine virulence genes. A Mariner-derived transposon that has been shown to insert randomly into the M. tuberculosis genome will be used to generate libraries of mycobacterial mutants. The technique of transposon site hybridization will be used to compare pools of mutants before and after animal infection. Microarray analysis will be used to determine deletion mutants which are absent from the output pools. These mutants potentially have deletions in virulence genes, and these genes, in addition to those identified by in vivo gene expression profiling, will be further characterized individually.

描述（由申请人提供）：为了鉴定对宿主存活重要的分枝杆菌基因，将开发兔肺结核模型。家兔经胸注射和支气管内滴注支杆菌感染。在空腔内的细菌增殖将允许分离足够数量的RNA，以便使用结核分枝杆菌寡核苷酸微阵列进行分析。比较在培养物中生长的分枝杆菌和在体内生长的分枝杆菌的基因表达模式，将为分枝杆菌在宿主体内存活提供重要的基因信息。第二种方法将同时用于确定毒力基因。一个水手衍生的转座子已被证明可以随机插入结核分枝杆菌基因组，将用于生成分枝杆菌突变体文库。转座子位点杂交技术将用于比较动物感染前后的突变体池。微阵列分析将用于确定输出池中缺失的缺失突变体。这些突变体可能在毒力基因中有缺失，这些基因，除了那些通过体内基因表达谱鉴定的基因外，将进一步单独表征。