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.
