EXITING STATIONARY PHASE IN E COLI

退出大肠杆菌中的固定相

• 批准号: 6386646
• 负责人: DEBORAH A. SIEGELE
• 金额: $ 15.74万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6386646
• 关键词: Escherichia coli; bacterial genetics; bacterial proteins; gene expression; gene mutation; mass spectrometry; microorganism growth; polymerase chain reaction; protein sequence

DESCRIPTION: Bacterial infections remain a leading cause of illness and death. To control bacterial pathogens it is important to understand not only host-pathogen interactions, but also how the pathogen's life cycle is affected by the environment. Bacteria can survive a variety of environmental stresses and still rapidly resume growth when conditions become favorable. This ability is an important factor in disease transmission and can also affect the initial stages of infection. The long-term goal of this project is to understand the mechanisms that regulate gene expression as the enteric bacterium E coli recovers from starvation stress. Mechanisms we identify are likely to be important in the infection cycle of pathogenic E. coli and other pathogenic bacteria. A group of nine E. coli proteins are transiently expressed during the initial period of outgrowth from stationary phase. The limited time when these proteins are made suggests that they are controlled by factors involved in regulating important aspects of the outgrowth response. The immediate goal of this proposal is to identify the genes encoding outgrowth-specific proteins. Once identified, these genes will be used as reporters to study the regulation of the outgrowth response. The first specific aim is to identify genes encoding outgrowth-specific proteins by obtaining amino acid sequence information from the outgrowth-specific proteins already described. State-of-the-art mass spectrometry methods will be used to analyze gel-purified proteins. The sequence information obtained will be used to identify the genes in the completed E. coli genome sequence. The second aim is to identify transcripts made specifically during outgrowth using PCR based differential display. The sequence of the differentially expressed cDNAs will be compared to the E. coli genome sequence to identify the genes. Once outgrowth-specific genes are identified, their function. and the mechanisms regulating their expression will be studied. Mutations will be constructed and the phenotypes of mutants characterized. Standard molecular methods, such as Northern blots and fusions to reporter genes, will be used to characterize how expression is the outgrowth-specific genes is controlled.

描述：细菌感染仍然是疾病的主要原因， 死亡。要控制细菌病原体，重要的是要了解 不仅是宿主-病原体的相互作用，而且还有病原体的生命周期是如何 受环境影响。细菌可以存活于各种不同的 环境压力，并在以下条件下仍快速恢复增长 变得有利。这种能力是疾病的一个重要因素。 传播，也会影响感染的初期阶段。这个 这个项目的长期目标是了解调节 肠道细菌大肠杆菌从饥饿中恢复时的基因表达 压力。我们确定的机制很可能在感染中起重要作用 致病性大肠杆菌和其他致病细菌的循环。九个人组成的小组 大肠埃希氏菌的蛋白在感染的最初阶段瞬时表达 从固定相生长出来的。这些蛋白质存在的有限时间 Made表明，它们是由参与调控的因素控制的 成长反应的重要方面。这样做的直接目标是 该计划的目的是确定编码生长特异蛋白的基因。 一旦确定，这些基因将被用作研究 对生长反应的调节。 第一个特定的目标是识别编码特定于生长的基因。 蛋白质通过获取氨基酸序列信息 已经描述过的生长特异蛋白。最先进的弥撒 光谱分析方法将用于分析凝胶纯化的蛋白质。这个 获得的序列信息将被用来识别 已完成的大肠杆菌基因组序列。第二个目标是确定 利用基于聚合酶链式反应的差异技术在生长发育过程中获得特定的转录本 展示。差异表达的cDNA的序列将是 与大肠杆菌基因组序列进行比对，以鉴定基因。 一旦确定了生长发育的特定基因，它们的功能就会改变。以及 调控它们表达的机制将被研究。突变将是 构建并鉴定了突变体的表型。标准分子 将使用Northern杂交和融合报告基因等方法 为了表征生长特异型基因的表达方式， 控制住了。