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J. Mol. Micro. Biotechnol. 3: 193-200
Prokaryote Multidrug Efflux Proteins of the Major Facilitator Superfamily: Amplified Expression, Purification and Characterisation
Alison Ward, Chris Hoyle, Sarah Palmer, John O'Reilly, Jeff Griffith, Martin Pos, Scott Morrison, Bert Poolman, Mick Gwynne,
and Peter Henderson
In bacterial genomes 3-12% of open reading frames are predicted to encode membrane transport
proteins. These proteins can be vital for antibiotic efflux, protein/toxin secretion, cell nutrition, environmental
sensing, ATP synthesis, and other functions. Some, such as the multidrug efflux proteins, are potential targets for
the development of new antibacterials and also for applications in biotechnology. In general membrane
transport proteins are poorly understood, because of the technical difficulties involved in isolating sufficient protein
for elucidation of their structure-activity relationships. We describe a general strategy for the amplified
expression, purification and characterisation of prokaryotic multidrug efflux proteins of the 'Major facilitator superfamily'
of transport proteins, using the Bacillus
subtilis multidrug resistance protein, 'Bmr', as example.
Full article [pdf]
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