Chromatography sysem to purify nucleic acid molecules and proteins for biophysical analysis

许多病毒感染的一个特点是在病毒生命周期中产生外来核酸分子。通过识别病毒核酸分子来检测入侵的细胞蛋白允许宿主细胞发起免疫反应。这一认知事件的核心是蛋白质-核酸复合体的分子特征，这些分子特征允许成功的相互作用发生。麦肯纳实验室主要致力于了解病毒核酸分子和2‘-5’寡腺苷合成酶(OAS)酶之间的相互作用，OAS酶是一类核酸结合蛋白，通过阻止必要的病毒蛋白生产来阻止病毒复制。麦肯纳实验室将建立一个研究计划，在分子水平上了解这一酶家族特别识别的自然产生的病毒核酸分子的特征。将使用生物物理和结构生物学方法来表征OAS-核酸复合体，并帮助理解这些酶如何区分病毒和宿主细胞核酸分子。这项研究计划的一个关键起点是开发生产和分离高产率和高纯度的蛋白质和核酸材料的策略。本提案中所要求的快速高效液相色谱系统将是用于提纯大量此类试剂的中心设备，也是现代结构生物学实验室所需的设备。随着所需设备的获得，麦肯纳实验室将把自己置于核酸分离领域的前沿，用于结构生物学应用。鉴于核酸分子在生物系统和生物技术领域中发挥的重要作用，将快速性能液相色谱系统纳入本已多学科的培训环境将导致培训为科学事业提供良好装备的高素质人员。

A hallmark of many viral infections is the production of foreign nucleic acid molecules generated during the viral life cycle. Cellular proteins that detect invasion through the recognition of viral nucleic acid molecules allow host cells to mount an immune response. At the heart of this recognition event are the molecular features of the protein-nucleic acid complexes that allow for successful interaction to occur. The McKenna laboratory is principally concerned with understanding the interactions between viral nucleic acid molecules and 2'-5' oligoadenylate synthetase (OAS) enzymes, a family of nucleic-acid binding proteins that prevent viral replication by blocking necessary viral protein production. The McKenna laboratory will establish a research program to understand, at the molecular level, the features of naturally occurring viral nucleic acid molecules that are specifically recognized by this family of enzymes. Biophysical and structural biology approaches will be used to characterize OAS-nucleic acid complexes, and help understand how these enzymes distinguish between viral and host-cell nucleic acid molecules. A crucial starting point for this research program is the development of strategies to produce and isolate protein and nucleic acid materials of high yield and purity. The requested Fast Performance Liquid Chromatography system outlined in this proposal will be the central device used to purify large quantities of these reagents, and is required equipment in modern structural biology laboratories. With the acquisition of the requested equipment, the McKenna laboratory will place itself at the cutting edge of the nucleic acid isolation field for use in structural biology applications. Given the important roles that nucleic acid molecules play in biological systems and the biotechnology field, the inclusion of a Fast Performance Liquid Chromatography system into an already multidisciplinary training environment will result in the training of highly qualified personnel who are well equipped for careers in science.