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Nucleotide sequence of the Streptococcus faecalis plasmid gene encoding the 3'5"-aminoglycoside phosphotransferase type III
Authors:P Trieu-Cuot  P Courvalin
Institution:Laboratoire de Biochimie, L.A. CNRS 271, Unité de Bactériologie Médicale, Institut Pasteur, F-7 5724 ParisFrance Tel. 306 19 19 ext. 3516
Abstract:We have cloned in Escherichia coli and sequenced a 1489-bp DNA fragment conferring resistance to kanamycin and originating from the streptococcal plasmid pJH1. The resistance gene was located by analysis of the initiation and termination codons in an open reading frame (ORF) of 792 bp. The deduced gene product, a 3'5'-aminoglycoside phosphotransferase of type III, has an Mr of 29,200. Comparison of its amino acid sequence with those of type I (Oka et al., 1981) and type II (Beck et al., 1982) 3' phosphotransferase, from transposable elements Tn903 and Tn5, respectively, indicated a statistically significant structural relationship between these enzymes from phylogenetically remote bacterial genera. The degree of homology observed indicate that phosphotransferase type III and type I genes have diverged from a common ancestor and that the phosphotransferase type II gene has emerged more recently from the type I evolutionary pathway.
Keywords:Recombinant DNA  gene cloning  chemical and dideoxy sequencing  amino acid sequence comparison  Ami  amikacin  Amp  ampicillin  APH-I  3′5″-aminoglycoside phosphotransferase type I  APH-II  3′-aminoglycoside phosphotransferase type II  APH-III  3′5″-aminoglycoside phosphotransferase type III  But  butirosin  bp  base pairs  EtBr  ethidium bromide  Kan  kanamycin  Liv  lividomycin  MIC  minimum inhibitory concentration  Neo  neomycin  ORF  open reading frame  Par  paromomycin  p  plasmid  RBS  ribosome-binding site (Shine-Dalgarno sequence)  Rib  ribostamycin>  R  resistance plasmid  RF  double-stranded replicative form  SDS  sodium dodecyl sulfate  Spc  spectinomycin  Str  streptomycin  Tn  transposon  Tob  tobramycin  Resistance phenotypes include Em  erythromycin  Km  kanamycin  Sm  streptomycin  Tc  tetracycline
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