A Multi-User Phosphorimaging System

此多用户组应用程序建议获得必要的仪器，以允许在俄克拉荷马州健康科学中心校园的大学的DNA，RNA和蛋白质的准确定量。 该仪器将包括一个磷光成像仪和附件，以及一台用于图像处理和数据分析的计算机和一个用于数据存档的可重写光盘驱动器。目前，我们拥有最先进的设施，支持细胞生物学，遗传学和分子生物学研究的大多数其他方面。然而，目前在俄克拉荷马州市健康科学中心校园还没有一种快速的定量和定性分析DNA、RNA和蛋白质的方法。相反，我们必须依靠放射自显影、放射自显影的密度扫描或带闪烁计数的凝胶切片进行定量。除了提供核酸和蛋白质的定量分析外，由于仪器的灵敏度增加，磷光成像仪将允许研究人员减少实验中放射性核素的量。 该机构将立即受益于以下部门的研究项目：免疫学、病理学、儿科学、眼科学和解剖科学：i）三个研究病毒转录、翻译和复制的项目（Palumbo，Kennedy和Lloyd），ii）研究原核基因表达的转录和翻译调控的四个项目（Dyer，Ferretti，Stull，Gilmore），iii）两个研究受体表达的项目（Palumbo，Dyer），iv）将确定化脓链球菌分离株的物理和遗传图谱的项目（Ferretti），v）四个项目将检查细胞因子和趋化因子水平以及mRNA水平（Murphy，Grammas，佩雷拉，坎宁安），vi）一个将检查少数群体中MHC基因座多态性的项目（Hildebrand），vii）定量掺入膜中的放射性标记蛋白水平的项目（Tweten）; viii）和两个研究细胞基因表达的项目，包括参与花生四烯酸代谢的酶、金属蛋白酶和平滑肌肌动蛋白（Palumbo，霍华德和Tomasek）。每个项目目前都得到国家校外资金的支持（NIH，USDA等）。或来自俄克拉荷马州科学技术促进中心（OCAST）或长老会健康基金会的“当地”资金。 这一科学事业的重要组成部分是基础分子生物学和蛋白质化学的重要培训。这种培训包括微生物学和免疫学系以及其他系内博士后研究员和研究生的持续研究教育。 此外，在该部门和大学的几个方案支持本科生，少数民族高中教师和少数民族高中学生的研究培训。因此，磷光成像仪将显着提高微生物学和免疫学系和健康科学中心校园内的研究和研究培训环境。 虽然磷光成像仪将被安置在微生物学和免疫学系，该设施将提供给所有研究人员在俄克拉荷马州市校园，但研究人员购买自己的磷光成像板和用户费用，以帮助支付费用旅游维护协议。因此，该仪器将在系外产生重大影响，丰富整个俄克拉荷马州健康科学中心校园的研究和研究培训能力。与这种潜力相一致，我们已经获得了重要的机构支持，用于购买磷光成像仪，因为健康科学中心已经承诺购买仪器所需资金的30%。

This multi-user group application proposes to acquire the necessary instrumentation to allow accurate quantitation of DNA, RNA, and protein at the University of Oklahoma Health Science Center Campus. This instrumentation will include one phosphorimagel and accessories with an attendant computer for image processing and data analysis, and a rewriteable optical disk drive for archiving data. Currently, we have state-of-the-art facilities that support cell biology, genetics and most other aspects of molecular biological research. However, a rapid means for quantitative and qualitative analyses of DNA, RNA, and protein are not currently available on the Oklahoma City Health Science Center Campus. Instead, we must rely on autoradiographs, densometric scans of autoradiographs, or gel slices with scintillation counting for quantitation. In addition to providing a quantitative analysis of nucleic acids and proteins, a phosphorimager will allow investigators to reduce the amount of radioactive nuclides in their experiments because of the increased sensitivity of the instrument. Research projects in the Departments ol Mlcrohlology an lmmunology, Pathology, Pediatrics, Opthamology, and Anatomical Sciences that will immediately benefit from this facility include: i) three projects that examine virus transcription, translation, and replication (Palumbo, Kennedy, and Lloyd), ii) four projects that examine transcriptional and translational regulation of procaryotic gene expression (Dyer, Ferretti, Stull, Gilmore), iii) two projects that examine receptor expression (Palumbo, Dyer), iv) a project that will determine physical and genetic map of Streptococcus pyogeMes isolates (Ferretti), v) four projects that will examine cytokine and chemokine levels and mRNA levels (Murphy, Grammas, Pereira, Cunningham), vi) a project that will examine the polymorphism of MHC loci in minority populations (Hildebrand), vii) a project to quanti tate levels of radiolabelled protein incorporated into membranes (Tweten); viii) and two projects that examine expression of cellular genes including enzymes involved in arachidonate metabolism, metalloproteinases, and smooth muscle actin (Palumbo, Howard, and Tomasek). Every project is currently supported by national extramural funding (NIH, USDA, etc.) or "local" funding from the Oklahoma Center for the Advancement of Science and Technology (OCAST) or the Presbyterian Health Foundation. Intertwined with, and an essential component of this scientific enterprise, is significant training in basic molecular biology and protein chemistry. This training includes the ongoing research education of post-doctoral fellows and graduate students within the Department of Microbiology and Immunology and the other Departments. Further, several programs in the Department and the University support research training for undergraduate students, minority high school teachers and minority high school students. Thus, the phosphorimager will significantly enhance the research and research training environment within the Department of Microbiology and Immunology and the Health Science Center Campus in general. Although the phosphorimager will be housed in the Department of Microbiology and Immunology, the facility will be available to all investigators on the Oklahoma City campus, subject to investigators purchasing their own phosphorimaging plates and user fees to help defray costs tour a maintenance agreement. Thus, this instrument will have significant impact outside the Department, enriching the research and research training capability of the entire University of Oklahoma Health Science Center campus. Consistent with this potential, we have obtained significant institutional support for the acquisition of a phosphorimager, as the Health Sciences Center has pledged 30% of the funds needed to purchase the instrumentation.