Structure and Specificity of Restriction-Modification (R-M) Systems

限制性修饰（R-M）系统的结构和特异性

• 批准号: 10797690
• 负责人: ANEEL K. AGGARWAL
• 金额: $ 11.79万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10797690
• 关键词: Cryoelectron Microscopy; DNA Binding; DNA Restriction-Modification Enzymes; Goals; Monitor; National Institute of General Medical Sciences; RNA-Binding Proteins; Services; Specificity; Structure; System; Vendor; X-Ray Crystallography; aged; fast protein liquid chromatography; instrument; parent grant; protein purification

Abstract Under the parent grant NIGMS R35-GM131780, titled “Structure and specificity of restriction-modification (R-M) systems”, this application is a supplemental request to replace an aged FPLC system that has been discontinued and is no longer supported and serviced by the vendor. All of the structural studies performed under the parent grant are heavily reliant on protein purification – a necessary prerequisite for X-ray crystallography and cryo-EM. The request is for an AKTA pure 25 M system with a Micro kit that will be 1) fully supported and serviced by the vendor, 2) allow the purification of proteins to be monitored at more than one wavelength, which is important because the aims of the parent grant are centered on DNA and RNA binding proteins, and 3) permit the purification of proteins on a microscale, which is essential, especially as the aims of the parent grant continue to grow in the direction of cryo-EM.

摘要 根据母授权NIGMS R35-GM 131780，标题为“限制修饰的结构和特异性”， (R-M)系统”，此申请是一个补充请求，以更换已老化的FPLC系统， 停止使用，供应商不再提供支持和服务。进行的所有结构研究 在父母的资助下，严重依赖于蛋白质纯化-这是X射线的必要先决条件 晶体学和冷冻电镜。该请求是一个AKTA纯25 M系统与微型套件，将1） 完全支持和服务的供应商，2）允许纯化的蛋白质进行监测， 一个波长，这是重要的，因为父母补助金的目标是集中在DNA和RNA 结合蛋白，和3）允许在微尺度上纯化蛋白，这是必需的，特别是当 母公司赠款的目标继续朝着冷冻EM的方向发展。