CARBOHYDRATE METABOLISM OF E COLI

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。该项目的具体目标是：1)开发能够分解复杂碳水化合物混合物的毛细管电泳法；2)将这些分析方法应用于康威实验室提供的粘液提取物，以确定它们的碳水化合物含量。更具体地说，将确定培养了大肠杆菌的粘液提取物对碳水化合物代谢的偏好。这个项目将是俄克拉荷马大学的泰瑞尔·康韦博士和东南部俄克拉荷马州立大学的蒂姆·史密斯博士共同努力的成果。Conway对大肠杆菌的全基因组表达谱显示了一些基因，这些基因是由模拟哺乳动物肠道中营养物质供应的条件诱导的；人们发现，主要的代谢途径是打开的。小鼠定植试验中的突变分析显示，几种糖似乎对大肠杆菌的定植很重要。这些发现的一个重要扩展是确定粘液中可用的糖，并确定大肠杆菌对个别粘液衍生糖的偏好顺序。拟议的代谢组学研究将利用毛细管电泳法监测培养在合成和正宗粘液上的各种大肠杆菌菌株和特定突变株的碳水化合物代谢。康威博士的实验室将产生野生型和突变型培养物的时间进程样本。史密斯博士的实验室将开发基于毛细管电泳法的生物分析方法，以确定提取物中的碳水化合物含量。目的是确定定植的大肠杆菌的确切代谢偏好。这一合作将提供更深入的了解，了解胃肠道病原体如何获得感染宿主并开始疾病过程所需的营养，并应建立在这一模式系统的知识基础上。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The specific aim of this project is to: 1) develop capillary electrophoresis-based assays capable of resolving complex mixtures of carbohydrates and 2) apply the assays to mucus extracts supplied from Conway's laboratory to determine their carbohydrate content. More specifically, the preference for metabolism of carbohydrates from mucus extracts on which Escherichia coli (E. coli) has been culture will be determined. This project will be a collaborative effort between Dr. Tyrrell Conway of the University of Oklahoma and Dr. Tim Smith at Southeastern Oklahoma State University. Conway's whole-genome expression profiling of E. coli revealed genes which were induced by conditions designed to mimic the nutrient availability in the mammalian intestine; it was found that primarily metabolic pathways were turned on. Mutational analysis in mouse colonization assays revealed several sugars that appeared to be important for E. coli to colonize. An important extension of these findings is to identify the sugars that are available in mucus and to determine the order of E. coli preference for the individual mucus-derived sugars. The proposed metabolomics study will utilize capillary electrophoresis to monitor the carbohydrate metabolism of various E. coli strains and specific mutants cultured on synthetic and authentic mucus. Dr. Conway's laboratory will generate time-course samples of wild type and mutant cultures. Dr. Smith's laboratory will develop the bioanalytical assays based around capillary electrophoresis necessary to determine the carbohydrate content in the extracts. The goal is to determine the exact metabolic preference of colonized E. coli. This collaboration will provide a more in depth understanding of how gastrointestinal pathogens acquire the nutrients necessary to infect their host and begin the disease process and should build on the knowledge of this model system.