1. A genomic strategy for cloning, expressing and purifying efflux proteins of the major facilitator superfamily.
- Author
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Gerda Szakonyi, Dong Leng, Pikyee Ma, Kim E. Bettaney, Massoud Saidijam, Alison Ward, Saeid Zibaei, Alastair T. Gardiner, Richard J. Cogdell, Patrick Butaye, Anne-Brit Kolsto, John OReilly, Ryan J. Hope, Nicholas G. Rutherford, Christopher J. Hoyle, and Peter J. F. Henderson
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BIOMOLECULES , *ESCHERICHIA coli , *MEMBRANE proteins , *ENTEROBACTERIACEAE - Abstract
A genomic strategy for the overexpression of bacterial multidrug and antibiotic resistance membrane efflux proteins in Escherichia coli is described. Expression is amplified so that the encoded proteins from a range of Gram-positive and Gram-negative bacteria comprise 5% to 35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGS(His)6-tag or a Strep-tag at the C terminus. These tags facilitate the purification of the overexpressed proteins in milligram quantities for structural studies. The strategy is illustrated for the bicyclomycin resistance efflux protein, Bcr, of E. coli. [ABSTRACT FROM AUTHOR]
- Published
- 2007
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