Ultracentrifuges for Molecular Biology Core

申请资金用于三个超离心机（贝克曼L7-80）和八个超离心机转子（NVT65、60Ti、SW-55Ti和SW-41Ti各两个），这些超离心机将位于分子生物学核心设施中，为卫斯理大学生物、分子生物学和生物化学以及化学三个系的实验室提供服务。这些超离心机将取代最近因使用年限问题而不能使用的两台仪器。这三台新仪器的购置将使校园内的超离心机数量达到四台，尽管第四台机器的使用将因其老化而受到限制。这些仪器将由7个主要用户和4个次要用户的实验室使用。它们还将被细胞生物学和分子生物学领域的两名新教员使用。这些仪器是分子生物学和生物化学的标准方法所必需的，包括用于膜制备的许多不同的亚细胞分离方法，以及分离蛋白质或蛋白质/RNA复合物。对于某些纯度和产量很重要的DNA制备，也需要超离心。这些方法将应用于几个不同的项目。这些研究包括：大肠杆菌蛋白定位机制；莱姆病诊断新抗原的鉴定原核生物DNA复制膜相关调节剂的拓扑结构果蝇配对蛋白功能的解剖哺乳动物视觉皮层的形成；酵母细胞分裂周期的分析鸡胚软骨细胞2(1)胶原基因的调控以及铁蛋白在抵御氧自由基毒性中的作用。卫斯理大学的配套资金将支付50%的仪器费用。

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.