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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. Although not discussed in this chapter, the same process can also be applied to integral membrane proteins (IMPs) with slight adaptations to the purification procedure.

Original publication

DOI

10.1007/978-1-62703-691-7_6

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2014

Volume

1091

Pages

95 - 121

Keywords

Animals, Baculoviridae, Batch Cell Culture Techniques, Cell Culture Techniques, Gene Expression, Genetic Vectors, Humans, Proteins, Quality Control, Recombinant Proteins, Sf9 Cells, Transfection