This application request funding to purchase the following instruments: 1) A DNA synthesizer with three column synthesis capability and full programability (Applied Biosystems model 380B-0s). 2) A peptide systhesizer with three reaction vessels and a single resin sampler (Applied Biosystems model 430A). 3) High pressure liquid chromatography system (Beckman model 344HPLC), variable wave-length detector (model 165), and integrator recorder (model 427). These instruments will be installed in the core molecular and cell biology laboratories which were constructed recently with NHLBI funding (ICO 6 HL 35669) and are going to serve the needs of the investigators of the Cardiovascular Institute, the Biophysics Institute, and the Pulmonary and Arthritis Center. The growing utilization of the techniques of molecular biology in biomedical research necessitated the recruitment of new faculty with expertise in molecualr biology and the development of post doctoral training programs in several clinical departments. There is a daily need to utilize the nucleotide synthesizer for studies involving the structure, function and the regulation of expression of important human genes (apolipoproteins, contractile proteins, acute phase proteins, and fatty acid binding proteins). Other proteins and genes are currently being characterized. Among these are alpha, adrenergic receptor, lipoprotein lipase, glutamate dehydrogenase, sodium potassium ATPase. Synthetic peptides will also be used extensively as inhibitors of proteolytic enzymes, renin converting enzymes, kallikrein and kinin, vassopressin analogs, etc. The HPLC units will be used for purification and analysis of the synthetic oligonucleotides and peptides. There is an institutional commitment for personnel and materials to operate the instruments for the first two years. Dr. James Burton, an expert in solid phase peptide synthesis, and Drs. Zannis, Ruiz-Opazo, and Roghani will supervise the initial installation and operation of these instruments. After the initial support from Boston University, most of the financial support for the operation of the instruments will be obtained from existing or new research grants. The costs will be prorated to take into consideration the extent of utilization of each scientist and research program. A similar mechanism for dividing costs between users is already in place for other core instruments at Boston University.
