Identification of Salmonella Genes Involved in Persistence in the Murine Intestin

沙门氏菌在小鼠小肠中持续存在的基因鉴定

• 批准号: 7914838
• 负责人: HELENE L ANDREWS-POLYMENIS
• 金额: $ 42.27万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7914838
• 关键词: Animal Experiments; Animal Feed; Animal Model; Animals; Antibiotics; Biological; Biological Assay; Candidate Disease Gene; Cecum; Cessation of life; Cistrons; Clinical; Collection; Complement; Complex; Data; Development; Disadvantaged; Disease; Dose; Epithelium; Essential Genes; Food Supply; Gene Deletion; Genes; Genetic; Genetic Screening; Genome; Goals; Growth; Human; In Vitro; Individual; Infection; Intestines; Lead; Libraries; Livestock; Measures; Methods; Modeling; Molecular; Molecular Genetics; Monitor; Mus; Oligonucleotide Microarrays; Organism; Pathway interactions; Phenotype; Population; Process; Proteins; Public Health; Relative (related person); Resistance; Salmonella; Salmonella enterica; Salmonella infections; Salmonella typhimurium; Screening procedure; Series; Serotyping; Study models; Systemic infection; Testing; Therapeutic Agents; Transcript; United States; base; deletion library; fitness; foodborne; foodborne illness; gene function; genetic manipulation; genome sequencing; high throughput screening; in vivo; mutant; new technology; novel; novel therapeutics; oral infection; pathogen; promoter; recombinase; tool; tool development; transmission process; vector

We will use the power of complete genome sequences and forward genetics to identify novel genes that allow a bacterial pathogen to colonize and persist in the mammalian intestine. Non-typhoidal Salmonella are food borne bacterial pathogens that cause hundreds of millions of cases of diarrheal disease and hundreds of thousands of deaths worldwide each year. Salmonella enterica serotype Typhimurium is an excellent model for study of intestinal fitness because of the availability of complete genome sequence information, the ease of genetic manipulation, and the availability of numerous animal models. We have generated a collection of 1150 targeted deletion strains (approximately 25% of the genome), developed a novel oligonucleotide microarray, and begun forward genetic screening this mutant collection both in vitro and in vivo in murine and chick models. We wish to complete the development of these tools and progress to more comprehensive studies of a subset of genes involved in intestinal persistence and fecal shedding, processes which are critical for transmission of this organism. We will approach this goal by: 1. Completion of our deletion library in all non-essential genes in Salmonella Typhimurium, 2. Screen our deletion collection in murine models to identify new genes involved in intestinal colonization and persistence, 3. Confirm each mutant as defective for intestinal persistence, 4. Characterize mutants that are defective for intestinal persistence.

我们将利用全基因组序列和前向遗传学的力量 识别允许细菌病原体定植并持续存在的新基因 哺乳动物的肠道。非伤寒沙门氏菌是食源性细菌 导致数亿例腹泻疾病的病原体和 全球每年有数十万人死亡。肠沙门氏菌 血清型鼠伤寒沙门氏菌是研究肠道健康的极佳模型 由于可获得完整的基因组序列信息，因此 以及众多动物模型的可获得性。我们 产生了1150个目标缺失菌株(约25% 基因组)，开发了一种新的寡核苷酸微阵列，并开始 在小鼠体内和体外对该突变体进行正向遗传筛选 还有小鸡模特。我们希望完成这些工具的开发，并 对参与基因子集的更全面研究的进展 肠道持久性和粪便排出，这两个过程对于 这种有机体的传播。我们将通过以下方式实现这一目标：1.完成 我们的鼠伤寒沙门氏菌非必需基因缺失文库， 2.在小鼠模型中筛选我们的缺失集合以确定新的基因 参与肠道定植和持久性，3.确认每个突变体 对肠道持久性有缺陷，4.描述符合以下条件的突变 有肠道顽固性缺陷。