A genomic strategy for cloning, expressing and purifying efflux proteins of the major facilitator superfamily
| Autor: | Nicholas G. Rutherford, Alison Ward, Gerda Szakonyi, Peter J. F. Henderson, Dong Leng, Pikyee Ma, Richard J. Cogdell, Kim E. Bettaney, Patrick Butaye, Ryan J. Hope, John O'Reilly, Saeid Zibaei, Christopher J. Hoyle, Anne-Brit Kolstø, Alastair T. Gardiner, Massoud Saidijam |
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| Rok vydání: | 2007 |
| Předmět: |
Microbiology (medical)
ATP Binding Cassette Transporter Subfamily B Biology medicine.disease_cause Microbiology Bicyclomycin Drug Resistance Bacterial medicine Escherichia coli Inner membrane Pharmacology (medical) Histidine Cloning Molecular Pharmacology Membrane transport protein Circular Dichroism Gene Amplification Membrane transport Bridged Bicyclo Compounds Heterocyclic Major facilitator superfamily Anti-Bacterial Agents Infectious Diseases Biochemistry Membrane protein Genes Bacterial biology.protein Spectrophotometry Ultraviolet Efflux Crystallization Plasmids |
| Zdroj: | The Journal of antimicrobial chemotherapy. 59(6) |
| ISSN: | 0305-7453 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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