This is a shared equipment proposal for the acquisition of a high performance liquid chromatography system (HPLC). HPLC provides a major improvement over conventional chromatography by increasing resolution, improving yields, and decreasing elution times and volumes. HPLC separations are applicable to virtually any class of compound, including small organic molecules, lipids, nucleic acids or macromolecules such as proteins, due to the diversity of available columns. The requested HPLC consists of the basic components required to provide a fully functional and highly versatile semi-preparative system. The unit includes an IBM 486 computer with the appropriate software for control of the individual HPLC components and data analysis; a single two solvent pump system capable of gradient formation; a manual sample injector; a Diode array detector for monitoring elution of a wide variety of molecules the Diode array permits the user to scan multiple wavelengths during an experiment, which will allow the researcher to determine if the eluted fraction is pure or not ; the Quick Res software for analysis of co-eluting compounds; and an initial assortment of HPLC columns to allow separations based on different physical properties. The requested instrument will enhance and modernize facilities available for research and teaching in the department of Biochemistry, Microbiology and Molecular Biology in addition to being available to other faculty and students outside of the department. The HPLC system will directly benefit several faculty research projects such as: 1) the metabolism and degradation of tetracycline and hydrocarbons by bacteria; 2) mechanisms regulating mammalian proteolytic enzymes, in vivo and in vitro; 4) post-translational modification as a regulatory mechanism in signal transduction processes in Dictyostelium; 5) purification of phenolics and hormones; 6) studies of the intermediate and slow blocks against polyspermy and embryonic stress tolerance in f ucoid algae and 7) the regulation of membrane lipid and fatty acid composition and the metabolism of these molecules in a variety of eubacteria, eukaryotes and plants. The graduate and undergraduate curricula in Biochemistry, Microbiology and Molecular Biology at the University of Maine include several laboratory courses that will also be enhanced by the availability of a HPLC system. Proposed uses include purification of monoclonal antibodies for Immunology; separation and isolation of DNA and polymerase chain reaction products for Molecular Biology and Bacterial Genetics; purification of proteins and peptide mapping in Biochemistry; and characterization of membrane lipids from several microorganisms for Bacterial Physiology. The inclusion of HPLC within these courses will provide our students with an important experience in using current methodologies. Such skills are commonly required and are greatly beneficial for ensuring their competitiveness for post-graduate studies, research jobs or careers.
