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，重要的是不了解 仅宿主 - 病原体相互作用，也只有病原体的生命周期 受环境的影响。 细菌可以生存多种 环境压力，并在条件下仍然迅速恢复增长 变得有利。 这种能力是疾病的重要因素 传播，也可能影响感染的初始阶段。 这 该项目的长期目标是了解规范的机制 基因表达作为肠细菌e从饥饿中恢复 压力。 我们确定的机制在感染中可能很重要 致病性大肠杆菌和其他致病细菌的循环。 一组九个 大肠杆菌蛋白在最初的初期瞬时表达 从固定阶段产生生长。 这些蛋白质的有限时间 提出，它们是由涉及调节的因素控制的 生长反应的重要方面。 直接目标 建议是确定编码生长特异性蛋白质的基因。 一旦确定，这些基因将被用作记者研究 调节生长反应。 第一个具体目的是识别编码特定生长特异性的基因 蛋白质通过从中获得氨基酸序列信息 已经描述了生长特异性蛋白。 最先进的质量 光谱法将用于分析凝胶纯化的蛋白质。 这 获得的序列信息将用于识别基因 完成的大肠杆菌基因组序列。 第二个目的是确定 使用基于PCR的差分在出生过程中专门制作的成绩单 展示。 差异表达的cDNA的序列将是 与大肠杆菌基因组序列相比，以鉴定基因。 一旦确定了生长特异性基因，它们的功能。 和 将研究调节其表达的机制。 突变将是 构造和突变体的表型。 标准分子 将使用方法，例如向记者基因的北印迹和融合。 表征表达的表达方式是生长特异性基因 受控。