Mechanisms of Enhanced Resistance of E. coli Biofilms

大肠杆菌生物膜增强耐药性的机制

• 批准号: 7110644
• 负责人: MICHAEL R BENOIT
• 金额: $ 4.4万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 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/7110644
• 关键词: Escherichia coli; bacterial genetics; bacterial proteins; biofilm; bioreactors; drug resistance; gene mutation; gene targeting; mass spectrometry; pharmacogenetics; postdoctoral investigator; protein protein interaction; shear stress; two dimensional gel electrophoresis; virulence; yeast two hybrid system

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（由申请人提供）：生物膜内的细菌在对人类健康有害的现象中更有效。基于马丁博士和其他人的工作，假设由于低剪切流体效应，例如潜在改变的大分子折叠，以及记录的基因表达变化，低剪切条件下的生物膜与传统的生物膜不同。采用新的适应旋转壁容器（RWV）生物反应器，设计在马丁实验室，E。大肠杆菌生物膜将在低剪切流体环境中培养。什么独特的基因和蛋白质表达在不同的发展阶段的低剪切生物膜将被确定，使用DNA微阵列，2-D凝胶电泳和质谱。将分离不能形成低剪切生物膜或不能增强抗性的突变体。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抗性中的作用。RWVs可能对蛋白质-蛋白质相互作用有什么影响将使用两个杂交和TAP标记系统与合适的蛋白质来确定。基本的和有利的信息将导致更好地控制生物膜，以预防传染病。