Large-scale production of membrane proteins in Saccharomyces cerevisiae: using a green fluorescent protein fusion strategy in the production of membrane proteins.

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.