Astex Pharmaceuticals, Cambridge, UK.
Diamond Light Source Ltd., Didcot, Oxfordshire, UK.
Methods Mol Biol. 2021;2199:67-94. doi: 10.1007/978-1-0716-0892-0_5.
This chapter describes the step-by-step methods employed by the Structural Genomics Consortium (SGC) for screening and producing proteins in the baculovirus expression vector system (BEVS). This eukaryotic expression system was selected and a screening process established in 2007 as a measure to tackle the more challenging kinase, RNA-DNA processing, and integral membrane protein families on our target list. Here, we discuss our platform for identifying soluble proteins from 3 mL of insect cell culture and describe the procedures involved in producing protein from liter-scale cultures.
本章介绍了结构基因组学联盟(SGC)在杆状病毒表达载体系统(BEVS)中筛选和生产蛋白质的分步方法。这个真核表达系统于 2007 年被选中并建立了一个筛选流程,作为解决我们目标列表中更具挑战性的激酶、RNA-DNA 处理和整合膜蛋白家族的一种手段。在这里,我们讨论了从 3 毫升昆虫细胞培养物中鉴定可溶性蛋白质的平台,并描述了从大规模培养物中生产蛋白质的过程。