A scalable, GFP‐based pipeline for membrane protein overexpression screening and purification

Abstract

We describe a generic, GFP‐based pipeline for membrane protein overexpression and purification in Escherichia coli . We exemplify the use of the pipeline by the identification and characterization of E. coli YedZ, a new, membrane‐integral flavocytochrome. The approach is scalable and suitable for high‐throughput applications. The GFP‐based pipeline will facilitate the characterization of the E. coli membrane proteome and serves as an important reference for the characterization of other membrane proteomes.