Plant Biofilm Inhibitors to Discover Biofilm Genes

植物生物膜抑制剂发现生物膜基因

基本信息

批准号：
7248685
负责人：
Thomas K Wood
金额：
$ 29.41万
依托单位：
TEXAS ENGINEERING EXPERIMENT STATION
依托单位国家：
美国
项目类别：
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/7248685
关键词：
Affect Architecture Bacteria Bacterial Infections Bacterial Model Biological Assay Biomolecular Nuclear Magnetic Resonance Cell Communication Cells Class Collaborations Communication Complex Confocal Microscopy DNA Microarray Chip DNA Microarray format Development Engineering Escherichia coli Figs - dietary Fluorescence Gene Chips Gene Expression Genes Genetic Gram-Negative Bacteria Green Fluorescent Proteins Growth Human Industry Knock-out Knowledge Label Laser Scanning Confocal Microscopy Lead Letters Life Marines Medicine Microbial Biofilms Modeling Molecular Plant Extracts Plants Proteins Protocols documentation Purpose Red Algae Regulator Genes Research Research Personnel Resistance Science Sequoia Signal Transduction Staging Structure System Testing Wood material Work base cell growth chemical synthesis genetic regulatory protein inhibitor/antagonist mutant novel palmitoleic acid prevent programs promoter tool ursolic acid

项目摘要

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的，有效的抗生物膜药，提出了一项研究计划，以研究自然存在的海洋富兰酮和其他植物衍生化合物抑制大肠杆菌生物膜形成的能力。此外，将使用时间DNA微阵列研究，敲除和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。该项目将通过两种互补方法使用DNA微阵列来发现大肠杆菌生物膜形成所需的基因：直接鉴定生物膜基因并抑制具有植物化合物的生物膜。此外，该项目将确定植物衍生的生物膜抑制剂会影响生物膜，并会发现抑制生物膜的其他植物衍生的化合物。发现的生物膜基因将是其他类型的化合物的丰富靶标，这些靶标可能用于预防工业和医学中的生物膜。该项目的具体目标是： 1。从红色藻类Delisea Pulchra中合成天然呋喃酮，并评估这些Furanones和其他植物衍生的化合物，以防止使用共聚焦显微镜和96孔快速测定的生物膜形成生物膜形成 2。使用DNA微阵列评估植物衍生的抑制剂对大肠杆菌基因表达的影响（确定哪些基因受影响，而抑制剂不影响生长） 3。使用DNA微阵列进行大肠杆菌（浸没生物膜）发现生物膜基因 4。使用DNA微阵列来发现细胞到细胞信号基因，通过抑制与呋喃酮的通信 5。研究用基因芯片阐明的基因的重要性使用用绿色荧光蛋白标记并通过标记生物膜调节蛋白的启动子的结构突变体的结构 6。确定结构，从而使用NMR来调节重要生物膜调节蛋白的功能 7。根据遗传信息推导细菌生物膜形成的模型（哪些基因诱导/抑制）和蛋白质信息（通过NMR）