Protein Expression and Purification

蛋白质表达和纯化

• 批准号: 6940587
• 负责人: ANDREW D MESECAR
• 金额: $ 19.34万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6940587
• 关键词: SARS virus; biomedical facility; endopeptidases; protein purification; transfection; virus protein

The major objective of this Core is to supply the various Projects and Cores with a continuous supply of highly purified protein in amounts necessary to complete the individual specific aims for each Project and Core, and therefore the long-term objectives of the overall Program Project. The specific aims are to (1) collaborate with Projects 1 & 2 in the cloning of SCLpro and PLpro into over-expression vectors, and to then determine their optimum expression conditions for each enzyme; (2) develop rapid and efficient purification procedures for SCLpro and PLpro enzyme constructs and determine the optimum conditions for their stabilization and storage; and (3) produce large amounts of highly purified enzyme that will be utilized by Projects 1-3, and by computational Core. The ultimate goal is to provide Projects 1-3, and the computational Core with a continuous supply of protein for the purposes of biochemical, kinetic, and biophysical characterization, x-ray structure elucidation, and high-throughput enzymatic and ligand-binding and bioassays.

该核心的主要目的是为各种项目和核心提供连续的 供应高度纯化的蛋白质，以完成每个项目和核心的各个特定目标所需的量，因此供应整体计划项目的长期目标。具体目的是（1）与SCLPRO和PLPRO的克隆中的项目1和2合作 进入过表达的载体，然后确定其每个酶的最佳表达条件； （2）为SCLPRO和PLPRO酶构建体开发快速有效的纯化程序，并确定其稳定和储存的最佳条件； （3）生产大量高纯化的酶，这些酶将由项目1-3和计算核心使用。最终目标是提供项目1-3，并为生化，动力学和生物物理表征，X射线结构阐明以及高通量酶促酶促和配体结合和生物分子而提供蛋白质连续供应。