Acquisition of Equipment for Structural Studies of Macromolecular Assemblies Using Cryo-EM

采购使用冷冻电镜进行大分子组装体结构研究的设备

• 批准号: 10635738
• 负责人: JOACHIM FRANK
• 金额: $ 12.24万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10635738
• 关键词: Administrative Supplement; Aging; Bacteria; Biological Process; Communication; Cryoelectron Microscopy; Ensure; Equipment; Eukaryota; Frequencies; Grant; In Vitro; Manufacturer Name; Modeling; Nature; Parents; Preparation; Publishing; Research; Ribonucleases; Ribosomal Proteins; Ribosomes; Sampling; Structure; System; Technology; Time; Translations; equipment acquisition; experimental study; macromolecular assembly; remote location

PROJECT SUMMARY The experimental research under the parent R35 grant focuses on time-resolved (TR) cryo-electron microscopy (cryo-EM) to study short-lived states during translation in eukaryotes. TR cryo-EM is a promising new way to obtain information about the dynamics of biological processes that can be carried out in vitro. Our successful applications of this technology to translation in bacteria have been published in Structure, Nature and Nature Communications. This research requires large-scale high-quality purifications of ribosomes and protein factors -- twenty to fifty times more sample quantity than in normal blotting cryo-EM experiments. Our present AKTA purification system is aging and shows numerous problems, which are detailed in the Equipment section of this proposal. We do not have access to core equipment or other groups’ equipment that is both in reach (i.e., on the same campus) and RNAse-free, a stringent requirement for any preparations involving ribosomes. Moreover, the manufacturer has recently decided to discontinue support of this model within less than a year. Thus, to ensure uninterrupted pursuit of our research we request an equipment supplement allowing us to purchase an AKTA pure 25 M (Cytiva Product Number: 29018226) purification system ($122,412.70).

项目概要 母公司 R35 资助下的实验研究重点是时间分辨 (TR) 冷冻电子显微镜 （冷冻电镜）用于研究真核生物翻译过程中的短暂状态。 TR 冷冻电镜是一种很有前途的新方法 获得有关可在体外进行的生物过程动态的信息。我们成功的 该技术在细菌翻译中的应用已发表在《Structure》、《Nature》和《Nature》杂志上 通讯。 这项研究需要对核糖体和蛋白质因子进行大规模高质量纯化——二十到五十个 样品数量是正常印迹冷冻电镜实验的两倍。 我们现有的 AKTA 净化系统已经老化并显示出许多问题，详细信息请参见 本提案的设备部分。我们无法获得核心设备或其他团体的设备 既可触及（即在同一校园内）又不含 RNAse，这是对任何准备工作的严格要求 涉及核糖体。此外，制造商最近决定停止对该型号的支持 不到一年的时间。 因此，为了确保我们的研究不间断地进行，我们要求提供设备补充，使我们能够 购买 AKTA pure 25 M（Cytiva 产品编号：29018226）纯化系统（122,412.70 美元）。