title: Functional gene cloning and characterization of the SsmE multidrug efflux pump from Serratia marcescens.
abstract: We cloned a gene responsible for multidrug resistance from chromosomal DNA of Serratia marcescens, and determined the nucleotide sequence. We designated the gene as ssmE. The deduced amino acid sequence of SsmE showed high similarity with the small multidrug resistance (SMR)-type multidrug efflux pumps. Cells of Escherichia coli KAM32, a drug hyper-susceptible mutant, transformed with a plasmid pESM437 carrying the ssmE gene showed elevated minimum inhibitory concentrations of structurally unrelated antimicrobial agents. E. coli KAM32/pESM437 showed elevated energy dependent efflux of ethidium. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis revealed that ssmE was expressed in cells of S. marcescens.
authors.affiliation: Department of Molecular Microbiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Tsushima, Okayama 700-8530, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Minato
authors.forename: Fereshteh
authors.initials: F
authors.lastname: Shahcheraghi
authors.forename: Wakano
authors.initials: W
authors.lastname: Ogawa
authors.forename: Teruo
authors.initials: T
authors.lastname: Kuroda
authors.forename: Tomofusa
authors.initials: T
authors.lastname: Tsuchiya
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D004996:Ethidium / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012706:Serratia marcescens / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004996:Ethidium