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A key step in the production of recombinant membrane proteins for structural studies is the optimization of protein yield and quality. One commonly used approach is to fuse the protein to green fluorescent protein (GFP), enabling expression to be tracked without the need to purify the protein. Combining fusion to green fluorescent protein with the baculovirus expression system provides a useful platform for both screening and production of eukaryotic membrane proteins. In this chapter we describe our protocol for the expression screening of membrane proteins in insect cells using fusion to GFP as a reporter. We use both SDS-PAGE with in-gel fluorescence imaging and fluorescence-detection size-exclusion chromatography (FSEC) to screen for expression.

Original publication

DOI

10.1007/978-1-4939-2230-7_11

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2015

Volume

1261

Pages

197 - 209

Keywords

Animals, Baculoviridae, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Genetic Vectors, Green Fluorescent Proteins, Membrane Proteins, Recombinant Fusion Proteins, Sf9 Cells, Spodoptera