A scalable, GFP-based pipeline for membrane protein overexpression screening and purification.
David Drew,
Dirk-Jan Slotboom,
Giulia Friso,
Torsten Reda,
Pierre Genevaux,
Mikaela Rapp,
Nadja M Meindl-Beinker,
Wietske Lambert,
Mirjam Lerch,
Daniel O Daley,
Klaas-Jan Van Wijk,
Judy Hirst,
Edmund Kunji and
Jan-Willem De Gier
Protein Sci 14(8):2011-7 (2005)
PMID 15987891
PMCID:
2279312
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.
DOI: 10.1110/ps.051466205
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