The Protein/DNA Sequence/Synthesis Facility of the Biotechnology Center is the resource for protein sequencing, peptide synthesis, DNA synthesis and DNA sequencing at the University of Wisconsin. Eleven users of our facility are applying for a Biomedical Research Support Shared Instrumentation Grant to purchase equipment to modernize, upgrade, and expand capabilities. We propose to add: a) microbore HPLC for purification of peptides derived from enzymatic digestions of proteins, b) multiple peptide synthesis capability to provide small quantities of short peptides economically, and c) dye labeled terminator chemistry for DNA sequencing. We propose to modernize and upgrade our existing instrumentation by: a) replacing our original gas phase sequencer with a pulsed-liquid phase sequencer, b) enhance quality control of DNA synthesis with capillary gel electrophoresis and data analysis, c) adding additional capacity to synthesize oligodeoxynucleotides by replacing our original DNA synthesizer, and d) adding a thermal cycler for DNA sequencing reactions. Projects proposed here include the study of: 1) kinases and the regulation of cell growth and differentiation, 2) genetic polymorphisms related to milk production and expression of proteins, 3) RNA polymerase and the regulation of transcription, 4) physiological ethology in primate research, 5) influenza viruses, 6) mapping quantitative trait loci in the mouse and cow, 7) structural features in the tRNA that determine acceptor identity, 8) pathways of resistance to anti-picornaviral drugs and the structure and synthesis of nodaviruses, 9) receptors and transcripts of bovine placental prolactin-related hormones, 10) high-speed automated DNA sequencing, and 11) paternity determination and genetic variability in wild, endangered primates. This improved facility will give a large number of faculty who are conducting NIH-supported research increased access to several very powerful technologies. It will extend several technologies to scientists who do not have the background to implement the protocols but have need for the data in their research. It will significantly improve economy and timeliness of delivery. this will contribute significantly to the success of much of the research and is essential for maintaining the strength and continued progress of many research programs at the University.
