This proposal is to develop a DNA sequencing facility to support basic research in molecular biology, genetics, medicine, and related areas. Newly emerging instrumentation which automates many of the routine steps in dideoxy-sequencing will soon be available. The University of Wisconsin Biotechnology Center's Protein/DNA Sequence/Synthesis Facility has the opportunity to evaluate one such instrument in their efforts to bring the research community a resource facility utilizing equipment which is able to provide the best sensitivity and economy available in a shared instrumentation core laboratory. The ability to sequence DNA is central to the success of many research projects and continues to increase in importance. Mastering manual techniques is no longer essential to graduate education and thus can be provided as a service. Laboratories doing no, little, or a modest amount of DNA sequencing will benefit enormously from a central facility with automation and established expertize; those sequencing vast amounts of DNA will benefit from economies of scale and improvements due to automation including reduction of tedium. Nine major users are applying for a Biomedical Research Support Shared Instrumentation Grant to purchase an automated DNA sequencer to expand the capabilities of the Biotechnology Centers research resources. Projects proposed here include the study of: 1) regulation of gene expression, transposition (Burgess, Reznikoll); 2) molecular genetics of RNA viruses (Ahlquist); 3) molecular biology of apolipoprotein and artherogenesis (Attie); 4) mutants of E. coli H+-ATPase (Fillingame); 5) biochemistry and genetics of formation of bacterial secondary metabolites (Hutchinson); 6) structure and evolution of eukaryotic genes (Smithies); 7) retrovirus replication, variation and relation to cancer (Temin); and 8) genetic regulation of antibiotic resistance (Weisblum). This facility will give a large number of faculty who are conducting NIH-supported research increased access to a very powerful new technology. This technology will contribute significantly to the success of much of this research and is essential for maintaining the strength and continued progrrss of many research programs at this University.
