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

许多病毒感染的一个特征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。这一识别事件的核心是蛋白质-核酸复合物的分子特征，它允许成功的相互作用发生。McKenna实验室主要关注了解病毒核酸分子与2‘-5’寡聚腺苷酸合成酶（OAS）酶之间的相互作用，OAS酶是一种核酸结合蛋白家族，通过阻断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项目，在分子水平上了解自然产生的病毒核酸分子的特征，这些分子被这种酶家族专门识别。生物物理和结构生物学方法将用于表征oas核酸复合物，并帮助了解这些酶如何区分病毒和宿主细胞核酸分子。本研究计划的一个关键起点是开发高产、高纯度的蛋白质和核酸材料的生产和分离策略。本提案所要求的快速高效液相色谱系统将是用于纯化大量这些试剂的中心设备，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，麦肯纳实验室将处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所起的重要作用，将快速高效液相色谱系统纳入本已多学科的培训环境中，将培养出高素质的人才，为科学事业做好充分准备。

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.