Plant Biofilm Inhibitors to Discover Biofilm Genes

描述（由申请人提供）：为了表征无毒，有效的抗生物膜剂，不影响细菌生长，提出了一项研究计划，以调查天然存在的海洋呋喃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，生物膜形成的遗传基础将通过时间DNA微阵列研究、敲除和发现的生物膜调节蛋白的核磁共振结构测定来研究。该项目将通过DNA微阵列的两种互补方法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和用植物化合物抑制生物膜。此外，本项目将确定植物是如何衍生的

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)