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Mechanisms of Enhanced Resistance of E. coli Biofilms

大肠杆菌生物膜增强耐药性的机制

基本信息

批准号：
7246623
负责人：
MICHAEL R BENOIT
金额：
$ 4.6万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-01 至 2008-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7246623
关键词：
3-Dimensional Address Affect Animals Antibiotic Resistance Antibiotics Antimicrobial Resistance Architecture Bacteria Bacteria sigma factor KatF protein Biochemical Biological Process Bioreactors Brush Border Cell physiology Cells Characteristics Chronic Communicable Diseases Communities Condition DNA Microarray Chip DNA Microarray format Development Devices Disease Environment Escherichia coli Food Gene Expression Gene Expression Profile Gene Proteins Genes Genetic Transcription Genitourinary system Hour Human In Vitro Individual Infection Knock-out Knowledge Laboratories Laboratory Organism Liquid substance Location Mass Spectrum Analysis Mechanical Stress Mechanics Microbe Microbial Biofilms Microbiology Microgravity Modification Molecular Nutrient Operative Surgical Procedures Personal Satisfaction Phase Physiology Polysaccharides Process Proteins Proteome Research Resistance Resistance development Robin bird Role Route Safety Sigma Factor Simulate Space Flight Staging Stimulus Stress Surface System Texas Tissue-Specific Gene Expression Training Translations Two-Dimensional Gel Electrophoresis Virulence Virulent Water Supply Work bacterial resistance base cellular microvillus denitrification drinking water experience gastrointestinal gastrointestinal microvillus interest microbial multidisciplinary mutant novel prevent protein degradation protein protein interaction repository research study respiratory response shear stress skills tissue culture wasting water treatment yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Bacteria within a biofilm are more effective in phenomena that are detrimental to human well being. Based on the work of Dr. Matin and others, it is hypothesized that due to low-shear fluid effects, such as potentially altered macromolecular folding, and documented changes in gene expression, biofilms under low-shear conditions are different from their conventional counterparts. Using novel adaptations to a rotating wall vessel (RWV) bioreactor, devised in the Matin lab, E. coli biofilms will be cultivated in low-shear fluid environments. What unique genes and proteins are expressed at different developmental stages of the low-shear biofilms will be determined, using DNA microarrays, 2-D gel electrophoresis, and mass spectrometry. Mutants unable to form low-shear biofilms or unable to enhance resistance will be isolated. Use of knockouts in selected genes will further explore the role of individual genes in low-shear biofilm formation and antibiotic resistance. What effect RWVs may have on protein-protein interactions will be determined, using two hybrid and TAP-tagging systems, with suitable proteins. Fundamental and enabling information will result in better control of biofilms for preventing infectious disease.

描述(申请人提供)：生物膜内的细菌在对人类健康有害的现象中更有效。根据Matin博士和其他人的工作，假设由于低剪切流体效应，例如可能改变的大分子折叠，以及已知的基因表达变化，低剪切条件下的生物膜与传统条件下的生物膜不同。利用Matin实验室设计的旋转壁容器(RWV)生物反应器的新改型，大肠杆菌生物膜将在低剪切流体环境中培养。利用DNA微阵列、双向凝胶电泳法和质谱仪，将确定在低剪切生物膜的不同发育阶段表达哪些独特的基因和蛋白质。不能形成低剪切生物膜或不能增强抗性的突变株将被隔离。在选定的基因中使用敲除将进一步探索单个基因在低剪切生物膜形成和抗生素耐药性中的作用。RWV可能对蛋白质-蛋白质相互作用有什么影响将通过使用两种杂交和TAP标签系统以及合适的蛋白质来确定。基本和有利的信息将导致更好地控制生物膜以预防传染病。