Quorum sensing regulation of bacterial development

项目总结 群体感应是细菌用来检测和检测细胞间通讯的一种方法。 应对人口密度的变化并控制有利于该群体的行为 在其环境中的适应和生存(例如，共生和致病)。我的研究 该计划的重点是细菌如何使用群体感应信号来控制 毒力基因。为此，我们研究了弧菌的群体感应基因调控， 既作为相关病原体，又作为已建立的群体感应模型系统。我们专注于 群弧菌主要感应转录因子LuxR，在所有弧菌中都是保守的 是物种，是数百个基因的核心调节者。我的研究小组使用生物化学， 生物物理、分子生物学和生理学分析，以剖析LUXR的机制 对多种弧菌的调控。这些实验是我的 研究计划。 我正在申请资金，以购买Li-COR奥德赛M成像系统。购买 奥德赛M将扩大我实验室的检测范围，提高分子检测的灵敏度 分析、更换较旧的共享设备，并消除放射性材料和 影像设施费。我们将改进我们现有的生化检测方法，增加 通过荧光标记底物提高灵敏度，并消除不断增加的放射性成本 材料。此外，奥德赛M相机将消除我们目前使用的旧的、共享的 标准分子生物学分析的成像系统(DNA分析、蛋白质分析、 蛋白质印迹和平板分析)。我们将进一步为我的团队添加新技术 因为奥德赛M将使我们能够想象受弧菌感染的现场细菌感染情况 使用高分辨率荧光抗体的组织。最后，所包含的Empiria软件 包括对EMSA和蛋白质印迹分析至关重要的量化能力。

Project summary Quorum sensing is a method of cell-cell communication used by bacteria to detect and respond to changes in population density and control behaviors that benefit the group for adaption and survival in their environment (e.g., symbiosis and pathogenesis). My research program is focused on how bacteria use quorum sensing signaling to control expression of virulence genes. Toward this goal, we study quorum sensing gene regulation in Vibrio species, both as relevant pathogens and as established quorum sensing model systems. We focus on the master Vibrio quorum sensing transcription factor LuxR, which is conserved in all Vibrio species and is the core regulator of hundreds of genes. My research group uses biochemical, biophysical, molecular biology, and physiological assays to dissect the LuxR mechanisms of regulation in multiple Vibrio species. These experiments are the foundational components of my research program. I am requesting funds to purchase a LI-COR Odyssey M Imaging System. Purchase of the Odyssey M will expand my lab’s repertoire of assays, increase sensitivity in molecular analyses, replace older shared equipment, and eliminate costs for radioactive materials and facilities fees for imaging. We will improve our current biochemical assays with increased sensitivity via fluorescence-tagged substrates and eliminate the increasing costs of radioactive materials. In addition, the Odyssey M Imager will eliminate our current use of older, shared imaging systems for standard molecular biology assays (DNA analysis, protein analysis, western blots, and plate-based assays). We will further add new technology to my group because the Odyssey M will enable us to image bacterial infection in situ in Vibrio-infected tissues using fluorescent antibodies at high resolution. Finally, the included Empiria software comprises quantification capabilities that are crucial for EMSA and western blot analyses.