Funds are requested for three ultracentrifuges (Beckman L7-80) and eight ultracentrifuge rotors (two each of NVT65, 60Ti, SW-55Ti, and SW-41Ti) to be located in the molecular biology core facility that serves laboratories in the three departments of Biology, Molecular Biology and Biochemistry, and Chemistry at Wesleyan University. The ultracentrifuges will replace two instruments that have recently become non-operational due to age related circumstances. Acquisition of the three new instruments will bring to four the number of ultracentrifuges on campus although use of the fourth machine will be limited due to its age. The instruments will be used by the laboratories of seven major and four minor users. They will also be used by two new faculty to be hired in the areas of cell biology and molecular biology. The instruments are required for standard methods in molecular biology and biochemistry including a number of different subcellular fractionation methods for membrane preparations, and to separate protein or protein/RNA complexes. Ultracentrifugation is also required for certain DNA preparations where purity and yield are important. These methods will be applied to several different projects. These include studies of: Mechanisms of protein localization in E. coli; Identification of novel antigens for Lyme disease diagnosis; Topology of a membrane associated regulator of prokaryotic DNA replication; Dissection of Drosophila Paired protein function; Formation of the mammalian visual cortex; Analysis of the cell division cycle in yeast; Regulation of the 2(1) collagen gene in chick embryonic cartilage cells; and the role of ferritin in defense against oxygen radical toxicity. Matching funds from Wesleyan University will pay for 50% of the cost of the instruments.
