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Identification, cloning, and functional characterization of EmrD-3, a putative multidrug efflux pump of the major facilitator superfamily from Vibrio cholerae O395. Archives of microbiology [Arch Microbiol] Journal article

 
TitleIdentification, cloning, and functional characterization of EmrD-3, a putative multidrug efflux pump of the major facilitator superfamily from Vibrio cholerae O395.
Author(s)Smith KP, Kumar S, Varela MF 
InstitutionDepartment of Biology, Eastern New Mexico University, Roosevelt Hall, Room 101, Station 33, Portales, NM, 88130, USA.
SourceArch Microbiol 2009 Oct 30.
AbstractA putative multidrug efflux pump, EmrD-3, belonging to the major facilitator superfamily (MFS) of transporters and sharing homology with the Bcr/CflA subfamily, was identified in Vibrio cholerae O395. We cloned the emrD-3 gene and evaluated its role in antimicrobial efflux in a hypersensitive Escherichia coli strain. The efflux activity of this membrane protein resulted in lowering the intracellular concentration of ethidium. The recombinant plasmid carrying emrD-3 conferred enhanced resistance to several antimicrobials. Among the antimicrobials tested, the highest relative increase in minimum inhibitory concentration (MIC) of 102-fold was observed for linezolid (MIC = 256 mug/ml), followed by an 80.1-fold increase for tetraphenylphosphonium chloride (TPCL) (156.2 mug/ml), 62.5-fold for rifampin (MIC = 50 mug/ml), >30-fold for erythromycin (MIC = 50 mug/ml) and minocycline (MIC = 2 mug/ml), 20-fold for trimethoprim (MIC = 0.12 mug/ml), and 18.7-fold for chloramphenicol (MIC = 18.7 mug/ml). Among the fluorescent DNA-binding dyes, the highest relative increase in MIC of 41.7-fold was observed for ethidium bromide (125 mug/ml) followed by a 17.2-fold increase for rhodamine 6G (100 mug/ml). Thus, we demonstrate that EmrD-3 is a multidrug efflux pump of V. cholerae, the homologues of which are present in several Vibrio spp., some members of Enterobacteriaceae family, and Gram-positive Bacillus spp.
LanguageENG
Pub Type(s)JOURNAL ARTICLE
PubMed ID19876617
  
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