We propose to establish a DNA analysis facility with the purchase of several key instruments for the automated analysis of DNA molecules. The instrumentation requested is commercially available and represents the core of many genome and DNA sequencing centers at other major research universities. This type of facility is essential for research projects that require experiments to be done on a genome-wide scale. There are currently three such research programs at the University of Oregon; all of these absolutely require the establishment of the proposed DNA analysis facility if they are to achieve their stated goals. In addition, the acquisition of the requested instrumentation is likely to have significant ancillary benefits. Many other users of the instruments will be able to complete their projects more efficiently and economically by incorporating the automated methods into their research programs. The establishment of the DNA analysis facility should also greatly aid our recruitment of new faculty with research interests in developmental genetics, molecular neuroscience and molecular evolution. These are all areas in which we plan to hire new faculty in the near future. Specifically, funds are requested for: 1) an Integrated Separation Systems AutoGen 740 that can prepare up to 200 DNA samples of viral DNA, bacterial plasmid DNA, yeast artificial chromosomal (YAC) DNA or genomic DNA in a 24 hour period; 2) a Biomek 2000 robotic workstation and sideloader for robotic manipulations of clones and nucleic acids; and 3) an ABI 377 DNA sequencer and the necessary accessories to perform high throughput automated DNA sequence and microsatellite analysis. In addition, several small equipment items (including Macintosh PowerPC 8100, two GeneAmp 9600 thermocyclers, and two microfuges) are requested to equip the DNA analysis facility. Presently, there is no facility in the State of Oregon that can provide the research support of the proposed DNA analysis facility. Fundin g this proposal will allow the faculty to fully develop recently initiated projects that utilize emerging intellectual and technical advances in the genetic analysis of complex (quantitative) characters, and that take advantage of map-based cloning strategies to isolate and characterize genes of major effect. These projects all aim to elucidate the genetic and molecular basis of fundamental biological processes, and encompass a broad spectrum of biology, from evolution and population genetics, to developmental genetics, to behavioral genetics. For example, the plant genetics project that will supported by these instruments will identify genes within the plant genome that control the evolution of different strategies of plant reproduction within a single species in which individuals have been geographically isolated for many generations. The dog project uses the same experimental logic being applied in the plant project, but in this case, the plan is to take advantage of the striking morphological and behavioral differences between dog breeds to gain insight into the genetic and molecular basis for these differences. The zebrafish research efforts are concentrated on identifying genes important for embryonic development of the organism, including genes required for the proper development and functioning of the nervous system. An important approach which is being pursued by these investigators is to identify new mutations that effect developmental processes in question. The ability to place these new mutations onto a high quality genetic map will facilitate the cloning of the genes and eventual molecular understanding of how these genes function. It should be noted that many of the projects described in this proposal are collaborative in nature. This reflects the efforts of our faculty to combine our individual expertise in order to advance our fields of interest more rapidly. Establishing the DNA analysis facility with the acquisition of the requested instrumentati on will represent a major advance in these collaborative efforts, and will provide the infrastructure necessary for the success of our research programs.
