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.
Methods in molecular biology (Clifton, N.J.)
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Astex Pharmaceuticals, Cambridge, UK.
Animals, Humans, Spodoptera, Baculoviridae, Membrane Proteins, Recombinant Proteins, Genetic Vectors, Sf9 Cells