Quorum sensing regulation of bacterial development

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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.