Large-Scale Production of Membrane Proteins in Saccharomyces cerevisiae: Using a Green Fluorescent Protein Fusion Strategy in the Production of Membra

The production of membrane proteins in the large quantities necessary for structural analysis requires many optimization steps. The GFP-fusion-based scheme described in earlier chapters (Chapters 4 , 8 , and 16 ) facilitates these steps by allowing the selection of high yielding clones that produce detergent-stable membrane proteins. Here, we describe the experimental steps required to establish the reproducible, large-scale production and purification of membrane protein–GFP fusions using S. cerevisiae .