Partial DNA sequence of a beta-lactamase produced by a Shigella flexneri strain.
A probe was constructed by radioactive labelling, and enzymatically a DNA fragment of plasmid pMAM-1, which codes for a beta-lactamase in Shigella flexneri UCSM 129, was obtained by amplification of a small part of the gene using the polymerase chain reaction technique (PCR). Since previous published work indicated that this beta-lactamase was of the TEM type, the primers used to amplify the gene were two highly conserved DNA regions in all TEM beta-lactamases. A 500 bp DNA probe was obtained which, by hybridization assays, facilitated the identification of restriction fragments of the plasmid containing the beta-lactamase gene. Two DNA fragments were sequenced by the Sanger method adapted to the PCR technique, and the sequence obtained showed a 100% homology with beta-lactamases TEM-1, TEM-2, TEM-13 and TEM-19. An intragenic restriction site, detected for Pst I, suggested that there is only one copy of the beta-lactamase gene per plasmid copy.
Department of Molecular Biology, Faculty of Biological Sciences, University of Concepción, Chile.
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D018628:Gene Dosage; D005798:Genes, Bacterial; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D012763:Shigella flexneri / Q000201:enzymology* / Q000235:genetics; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics*
D015342:DNA Probes; D004269:DNA, Bacterial; D001618:beta-Lactamases