CORE--PROTEIN PURIFICATION

该核心将产生用于几种不同目的的蛋白质：使用x射线晶体学和核磁共振测定结构；金属结合分析；金属对翻译后修饰的响应分析使用脂质体重构和注射到动物体内产生抗体的金属运输测量。蛋白质纯化核心将从大规模培养的细菌中纯化蛋白质；杆状病毒感染昆虫细胞和酵母菌。虽然CEMH的大多数成员在他们的实验室中使用蛋白质表达，但有几个原因使蛋白质表达集中。首先，中心将集中专家和试剂。该核心将包含细菌表达系统目录，允许大量生产和纯化表位标记蛋白（HIS， GST，麦芽糖，TAP）。其次，将核心的规模设计到

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).