Acquisition of Molecular Imaging and Plate Reading Systems

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Abstract BullerjahnTo enhance ongoing research activities in the departments of Biological Sciences and Chemistry at Bowling Green State University, a molecular imaging system, and a multifunctionplate reader will be placed in common lab facilities shared by the two departments. The laboratories of seven major users will use the instruments. Molecular imagers analyze radioactive, fluorescent and luminescent samples, yielding linear data over a wide dynamic range. The imager will be used to visualize and quantify electrophoretic mobility shift assays, western blots, RNase protection assays, northern blots, sequencing gels, and DNA footprints. A multifunction plate reader capable of photometric, fluorescence and luminescence modes with adequate software for data analysis and storage will be used on similar projects. The molecular imaging system will be used in several very diverse projects, such as quantitation of ferredoxin and flavodoxin levels in metal-limited phytoplankton by western blotting; functional analyses of Fur-type repressor-operator interactions; northern/western analyses of mRNA and protein levels in brain cells comprising the circadian pacemaker;quantitiation of western blots of hybrid Nif proteins involved in nitrogenase expression and assembly; assessing protein phosphorylation of ErbB3 receptors in; characterizing transcription of genes involved in crayfish monoamine synthesis; and quantifying by gel electrophoresis RNA-RNA interactions in model systems optimized for determining binding constants. The projects employing the plate reader are studies parallel to those employing the molecular imager, and such work includes screening cyanobacterial promoter fusions by luciferase-dependent luminescence; using such promoter fusions in a cyanobacterial-based metal biosensor assay; determining phosphate nutrient status in phytoplankton by assaying alkaline phosphatase activity; measuring interacting Nif proteins by a beta-galactosidase assay; and quantifying cAMP and cGMP levels following activation of oocytes bearing the AT2 receptor by angiotensin II. Given the diversity of the research projects described above, the award will provide direct benefits to the widest possible spectrum of research projects at BGSU. The instrumentation will provide analytical tools currently not available on campus, thus the immediate impact will beImproved data acquisition for the PI, co-PIs and major users. Such benefits will allow these faculty to make more rapid progress on their funded research projects. The instrumentation will also be available to other scientists on campus, thus yielding an improved environment forresearch and training of graduate and undergraduate students.