Acquisition of Molecular Imaging and Plate Reading Systems

获取分子成像和读板系统

• 批准号: 0070334
• 负责人: George Bullerjahn
• 金额: $ 8.43万
• 依托单位: Bowling Green State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0070334&HistoricalAwards=false
• 关键词: Acquisition; Molecular; Imaging; Plate; Reading

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.

为了加强绿色州立大学的生物科学和化学系正在进行的研究活动，一个分子成像系统和一个多功能酶标仪将被放置在两个系共享的公共实验室设施中。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，在很宽的动态范围内产生线性数据。成像仪将用于可视化和定量电泳迁移率变动分析、蛋白质印迹、RNA酶保护分析、北方印迹、测序凝胶和DNA足迹。在类似项目中，将使用一台能够进行光度测定、荧光和发光模式的多功能读板器，并配备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多样化的项目，例如通过蛋白质印迹法定量金属限制性浮游植物中的铁氧还蛋白和黄氧还蛋白水平; Fur型阻遏物-操纵基因相互作用的功能分析; mRNA和蛋白质水平的北方/西方分析构成昼夜节律起搏器的脑细胞;参与固氮酶表达和组装的杂合Nif蛋白的蛋白质印迹定量;评估ErbB 3受体的蛋白磷酸化;表征涉及小龙虾单胺合成的基因的转录;以及通过凝胶电泳在优化用于确定结合常数的模型系统中定量RNA-RNA相互作用。 采用平板读数器的项目是与采用分子成像仪的项目平行的研究，这类工作包括：通过依赖荧光酶的发光筛选蓝细菌启动子融合体;在蓝细菌金属生物传感器测定中使用这种启动子融合体;通过测定碱性磷酸酶活性确定浮游植物中的磷酸盐营养状态;通过β-半乳糖苷酶测定测量相互作用的Nif蛋白;以及在通过血管紧张素II活化携带AT 2受体的卵母细胞后定量cAMP和cGMP水平。 鉴于上述研究项目的多样性，该奖项将为BGSU尽可能广泛的研究项目提供直接利益。 仪器将提供目前校园内没有的分析工具，因此，直接影响将是改善PI，共同PI和主要用户的数据采集。 这些好处将使这些教师在他们资助的研究项目上取得更快的进展。 该仪器也将提供给校园里的其他科学家，从而为研究生和本科生的研究和培训提供更好的环境。