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

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

Research output: Contribution to journalArticle

39 Citations (Scopus)

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
Pages (from-to)419-427
Number of pages9
JournalMicrobiology and Immunology
Volume47
Issue number6
Publication statusPublished - 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

  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Microbiology

Cite this

Huda, N. M., Chen, J., Morita, Y., Kuroda, T., Mizushima, T., & Tsuchiya, T. (2003). Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1. Microbiology and Immunology, 47(6), 419-427.

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

In: Microbiology and Immunology, Vol. 47, No. 6, 2003, p. 419-427.

Research output: Contribution to journalArticle

Huda, NM, Chen, J, Morita, Y, Kuroda, T, Mizushima, T & Tsuchiya, T 2003, 'Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1', Microbiology and Immunology, vol. 47, no. 6, pp. 419-427.
Huda, Nazmul Md ; 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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