Plant Biofilm Inhibitors to Discover Biofilm Genes

植物生物膜抑制剂发现生物膜基因

• 批准号: 7098858
• 负责人: Thomas K Wood
• 金额: $ 30.34万
• 依托单位: TEXAS ENGINEERING EXPERIMENT STATION
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-19 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7098858
• 关键词: Escherichia coli; algae; antibacterial agents; bacteria infection mechanism; biofilm; cell cell interaction; confocal scanning microscopy; furans; gene induction /repression; genetic regulation; green fluorescent proteins; growth inhibitors; microarray technology; microorganism growth; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; plant extracts; transcription factor

DESCRIPTION (provided by applicant): To characterize non-toxic, effective anti-biofilm agents that do not affect bacterial growth, a research program is proposed to investigate the ability of naturally-occurring marine furanones and other plant-derived compounds to inhibit the formation of E. coli biofilms. In addition, the genetic basis of biofilm formation will be investigated using temporal DNA microarray studies, knock-outs, and NMR structure determinations of the discovered biofilm regulatory proteins. This project will discover the genes required for E. coli biofilm formation by two complementary approaches using DNA microarrays: direct identification of biofilm genes and by inhibiting biofilms with plant compounds. In addition, this project will identify how the plant-derived biofilm inhibitors influence biofilms, and will discover other plant-derived compounds which inhibit biofilms. The biofilm genes that are found will be rich targets for other classes of compounds that may be used to prevent biofilms in industry and medicine. The specific objectives of this project are: 1. synthesize natural furanones from the red alga Delisea pulchra and evaluate these furanones and other plant-derived compounds for their ability to prevent biofilm formation using confocal microscopy and 96-well rapid assays 2. evaluate the impact of the plant-derived inhibitors on the gene expression of E. coli using DNA microarrays (identify which genes are affected while the inhibitor does not affect growth) 3. discover biofilm genes using DNA microarrays for E. coli (submerged biofilm) 4. discover cell-to-cell signaling genes using DNA microarrays by inhibiting communication with furanones 5. study the importance of the genes elucidated with the gene chips in terms of biofilm formation and architecture using knock-out mutants that are labeled with the green fluorescence protein and by tagging the promoters of biofilm regulatory proteins 6. determine the structure and thereby the function of the important biofilm regulatory proteins using NMR 7. deduce a model for bacterial biofilm formation based on the genetic information (which genes induced/repressed) and protein information discerned (via NMR)

描述（由申请人提供）：为了表征不影响细菌生长的无毒、有效的抗生物膜剂，提出了一项研究计划来研究天然存在的海洋呋喃酮和其他植物源化合物抑制形成的能力大肠杆菌生物膜。此外，还将利用时间 DNA 微阵列研究、敲除和已发现的生物膜调节蛋白的 NMR 结构测定来研究生物膜形成的遗传基础。该项目将通过使用 DNA 微阵列的两种互补方法来发现大肠杆菌生物膜形成所需的基因：直接识别生物膜基因和通过植物化合物抑制生物膜。此外，该项目将确定植物源性物质如何 生物膜抑制剂影响生物膜，并将发现其他抑制生物膜的植物源化合物。 所发现的生物膜基因将成为其他类别化合物的丰富靶标，这些化合物可用于防止工业和医学中的生物膜。 该项目的具体目标是： 1. 从红藻 Delisea pulchra 中合成天然呋喃酮，并使用共焦显微镜和 96 孔快速检测评估这些呋喃酮和其他植物源性化合物防止生物膜形成的能力 2. 使用DNA微阵列评估植物源抑制剂对大肠杆菌基因表达的影响（确定哪些基因受到影响，而抑制剂不影响生长） 3. 使用大肠杆菌 DNA 微阵列发现生物膜基因（水下生物膜） 4. 使用 DNA 微阵列通过抑制与呋喃酮的通讯来发现细胞间信号传导基因 5. 研究基因芯片阐明的基因在生物膜形成和生物膜形成方面的重要性 使用绿色荧光蛋白标记的敲除突变体并标记生物膜调节蛋白的启动子的架构 6. 使用 NMR 确定重要生物膜调节蛋白的结构和功能 7. 根据遗传信息（哪些基因 诱导/抑制）和识别蛋白质信息（通过 NMR）