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A scalable, GFP-based pipeline for membrane protein overexpression screening and purification
Authors:Drew David  Slotboom Dirk-Jan  Friso Giulia  Reda Torsten  Genevaux Pierre  Rapp Mikaela  Meindl-Beinker Nadja M  Lambert Wietske  Lerch Mirjam  Daley Daniel O  Van Wijk Klaas-Jan  Hirst Judy  Kunji Edmund  De Gier Jan-Willem
Institution:Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
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.
Keywords:Escherichia coli  membrane protein overexpression  membrane protein isolation  membrane protein characterization  GFP
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