CORE--PROTEIN PURIFICATION

该核心将产生将用于几个不同目的的蛋白质：使用X射线晶体学和NMR进行结构测定;金属结合分析;响应于金属的翻译后修饰分析;使用脂质体重构和注射到动物中以产生抗体的金属转运测量。蛋白质纯化核心将从大规模培养的细菌、杆状病毒感染的昆虫细胞和酵母中纯化蛋白质。虽然CEMH的大多数成员在他们的实验室中使用蛋白质表达，但集中蛋白质表达有几个原因。首先，核心将集中专业知识和试剂。核心将包含细菌表达系统的目录，允许大量生产和纯化表位标记的蛋白质（HIS，GST，麦芽糖，TAP）。第二，核心的规模将被设计为 适应从相对少量的蛋白质生产不同水平的蛋白质的需要， 抗体生产所需的量（mg）至金属结合测定（10 mg）至X射线晶体学（100 mg）。

This core will produce proteins that will be used for several different purposes: structure determination using both X-ray crystallography and NMR; metal binding analyses; analysis of posttranslational modification in response to metals; metal transport measurements using liposome reconstitution and for injection into animals to generate antibodies. The Protein Purification Core will purify proteins from large-scale cultures of bacteria; bacculovirus infected insect cells and yeast. While most members of the CEMH use protein expression in their laboratories, there are several reasons to centralize protein expression. First, the Core will centralize both expertise and reagents. The Core will contain a catalogue of bacterial expressions systems, permitting the production and purification of epitope tagged proteins (HIS, GST, Maltose, TAP) in large amounts. Second, the scale of the Core will be designed to accommodate the need to produce different levels of proteins from the relatively small amounts required for antibody production (mg) to metal binding assay (10 mg) to X-Ray crystallography (100's mg).