CORE--PROTEIN EXPRESSION

利用分离蛋白的研究，从抗体的生产到酶的生化表征以及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，我们计划实施一个蛋白表达核心设施（PE核心）。PE核心将为DERC成员生产和表征纯化蛋白。具体而言，该核心将提供以下服务：(1)重组蛋白在大肠杆菌中的细菌蛋白表达。一般来说，只有结构简单的蛋白质才会在这个系统中产生。(2)利用细胞分选法快速富集高表达细胞部分的哺乳动物表达。(3)使用标准亲和、离子交换和凝胶过滤色谱技术进行纯化。(4)使用标准的生化方法和质谱法对纯度、适当折叠和寡聚化状态进行表征。

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.