Acquisition of Molecular Imaging and Plate Reading Systems

为了加强鲍林格林州立大学生物科学和化学系正在进行的研究活动，分子成像系统和多功能平板阅读器将放置在两个系共享的实验室设施中。七个主要用户的实验室将使用这些仪器。分子成像仪分析放射性、荧光和发光样品，在很大的动态范围内产生线性数据。该成像仪将用于可视化和量化电泳迁移率改变分析、蛋白质印迹、核糖核酸酶保护分析、Northern印迹、测序凝胶和DNA足迹。将在类似项目中使用具有光度、荧光和发光模式的多功能板阅读器，并配有适当的数据分析和存储软件。该分子成像系统将用于几个非常不同的项目，如通过Western blotting定量测定金属受限浮游植物中的铁氧还蛋白和黄还蛋白水平；对Fur-型阻遏子-操纵子相互作用的功能分析；对包括昼夜节律起搏器的脑细胞中的mRNA和蛋白质水平的Northern/West分析；对参与固氮酶表达和组装的杂交NIF蛋白的蛋白质印迹进行定量；评估ErbB3受体的蛋白质磷酸化；鉴定参与小龙虾单胺合成的基因的转录；以及在优化用于确定结合常数的模型系统中用凝胶电泳定量RNA-RNA的相互作用。使用平板阅读器的项目是与使用分子成像仪的项目平行的研究，这类工作包括通过依赖荧光素酶的发光来筛选蓝藻启动子的融合；在基于蓝藻的金属生物传感器分析中使用这种启动子融合；通过检测碱性磷酸酶的活性来确定浮游植物中的磷酸盐营养状态；通过β-半乳糖苷酶检测相互作用的NIF蛋白；以及在携带AT2受体的卵细胞被血管紧张素II激活后量化cAMP和cGMP的水平。鉴于上述研究项目的多样性，该奖项将为北卡罗来纳州立大学最广泛的研究项目提供直接好处。该仪器将提供目前在校园内没有的分析工具，因此，直接的影响将是改善PI、共同PI和主要用户的数据采集。这样的福利将使这些教员在他们资助的研究项目上取得更快的进展。该仪器还将向校园内的其他科学家开放，从而为研究生和本科生的研究和培训创造一个更好的环境。

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.