The goal of this proposal is to advance essential facility services in high speed automated DNA sequencing as a joint effort of the Pacific Biomedical Research Center (PBRC) and the Research Centers in Minority Institutions Program (RCMI) to increase the University's capability for health-related and biomolecular research and graduate education. Funds are requested to purchase a Perkin-Elmer ABI 377 DNA Sequencer with XL Upgrade Translation Interface Processor and associated software. The new DNA sequencer will enable the Biotechnology/Molecular Biology Instrumentation and Training Facility (BMBITF) to provide state-of-the- art instrumentation and techniques for many ongoing and new research projects, to stay current on technology and develop new supporting services. Over five years ago, the BMBITF begin supporting health- related and biomolecular research with a single ABI 373 DNA sequencer to meet the University's then modest demand for sequencing. Since then, DNA sequencing has taken a quantum leap and become an essential element in many research programs on this campus. The demand for this service is increasing at such a rate that the volume of work involved outruns the capacity of the existing nucleotide sequencer. The requested high- throughput sequencing instrument would eliminate the delays that the research community presently experiences due to facility user-overload. The instrumentation and accessories from Perkin-Elmer, which at the present time is the best instrument for the needs of the Facility, costs $150,000. The ability of research groups in biomedical areas at the University to obtain sequence data locally and rapidly is important for maintaining their competitive position for obtaining extramural funding. This proposal focuses on nine representative NIH-supported research projects in detail to demonstrate the future use of the instrument. These are as follows: I. Human Decidua and Fetal Membranes as a Paracryne system, 2. Elastin Gene in Pseudoxanthoma Elasticum; 3. Matrix: Regulation in Pre- and Postnatal Elastic Tissues; 4. Lysyl Oxydase Gene Mutations in Colon Cancer; 5. Elastic Gene Mutations in Supravascular Aortic Stenosis; 6. Colonization of Epithelium by Symbiotic Vibrio Fischeri; 7. Connexin 45 Phosphorylation in RSV- Infected Cells; 8. Cloning of the Rat Human Relax Receptor Genes; 9. The Effects of Distension on the Human Fetal Membranes. The requested instrument will also be used in academic training of qualified students, primarily those participating in the RCMI Program of PBRC.
