Gene cloning and characterization of VcrM, a Na + -coupled multidrug efflux pump, from Vibrio cholerae non-O1

Nazmul Huda, Jing Chen, Yuji Morita, Teruo Kuroda, Tohru Mizushima, Tomofusa Tsuchiya

Research output: Contribution to journalArticle

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Abstract

We cloned a DNA fragment responsible for drug resistance from chromosome of Vibrio cholerae non-O1. Nucleotide sequence analysis of this fragment revealed the presence of a single open reading frame encoding a protein consisting of 445 amino acid residues. We designated the gene as vcrM. Hydropathy analysis of the deduced amino acid sequence of VcrM suggests the presence of 12 trans-membrane segments. A dendrogram showed that VcrM is a member of the DinF-subfamily within the MATE family of multidrug efflux pumps. Expression of the cloned vcrM gene in drug-hypersensitive Escherichia coli KAM32 cells made them resistant to acriflavine, 4′, 6-diamidino-2-phenylindole, Hoechst 33342, rhodamine 6G, tetraphenylphosphonium chloride (TPPCI) and ethidium bromide. Efflux of acriflavine due to VcrM was dependent on Na + or Li + . Moreover, Na + efflux was observed with VcrM when TPPCI was added to Na + -loaded cells. Therefore, we conclude that VcrM is a Na + /drug antiporter-type multidrug efflux pump.

Original languageEnglish (US)
Pages (from-to)419-427
Number of pages9
JournalMicrobiology and Immunology
Volume47
Issue number6
DOIs
StatePublished - Jan 1 2003
Externally publishedYes

Fingerprint

Vibrio cholerae non-O1
Acriflavine
Organism Cloning
Genes
Antiporters
Ethidium
Protein Sequence Analysis
Drug Resistance
Pharmaceutical Preparations
Open Reading Frames
Sequence Analysis
Chromosomes
Escherichia coli
Amino Acids
Membranes
DNA
tetraphenylphosphonium
Proteins

Keywords

  • Drug efflux pump
  • Multidrug resistance
  • VcrM
  • Vibrio cholerae non-O1

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

Cite this

Gene cloning and characterization of VcrM, a Na + -coupled multidrug efflux pump, from Vibrio cholerae non-O1 . / Huda, Nazmul; Chen, Jing; Morita, Yuji; Kuroda, Teruo; Mizushima, Tohru; Tsuchiya, Tomofusa.

In: Microbiology and Immunology, Vol. 47, No. 6, 01.01.2003, p. 419-427.

Research output: Contribution to journalArticle

Huda, Nazmul ; Chen, Jing ; Morita, Yuji ; Kuroda, Teruo ; Mizushima, Tohru ; Tsuchiya, Tomofusa. / Gene cloning and characterization of VcrM, a Na + -coupled multidrug efflux pump, from Vibrio cholerae non-O1 In: Microbiology and Immunology. 2003 ; Vol. 47, No. 6. pp. 419-427.
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