CORE--PROTEIN EXPRESSION

利用分离蛋白的研究，从抗体的产生到酶的生化特性，以及对分泌信号因子的动物注射研究，对于了解它们的生物学作用至关重要。为了促进这类研究，我们计划实施蛋白质表达核心设施(PE核心)。PE核心将为DERC成员生产和表征纯化的蛋白质。具体地说，该中心将提供以下服务：(1)在大肠杆菌中表达细菌蛋白重组蛋白。一般来说，在这个系统中只能产生结构简单的蛋白质。(2)用细胞分选法快速富含高表达细胞片段的哺乳动物表达。(3)用标准亲和层析、离子交换和凝胶过滤层析技术进行纯化。(4)用标准生化方法和质谱学方法表征纯度、合适的折叠和齐聚状态。

Studies making use of isolated proteins, ranging from production of antibodies to biochemical characterization of enzymes and animal injection studies of secreted signaling factors, are critical to understand their biological roles. To facilitate such studies, we plan to implement a Protein Expression Core Facility (the PE core). The PE core will produce and characterize purified proteins for members of the DERC. Specifically, the core will provide the following services: (1) Bacterial protein expression of recombinant proteins in E. coli. In general, only proteins with simple architecture will be produced in this system. (2) Mammalian expression using a cellsorting method to quickly enrich the fraction of highly expressing cells. (3) Purification using standard affinity, ion exchange, and gel filtration chromatography techniques. (4) Characterization of purity, appropriate folding, and oligomerization state, using standard biochemical methods and mass spectrometry.