Oligonucleotide probes for the detection of TEM-1 and TEM-2 beta-lactamase genes and their transposons

Can J Microbiol. 1987 Mar;33(3):205-11. doi: 10.1139/m87-035.

Abstract

We describe the use of molecular probes to detect the TEM-type beta-lactamase genes. As a general probe, we prepared a 656 base pair restriction fragment, entirely within the TEM structural gene. This probe was specific for the TEM family, hybridizing only with TEM-1 and TEM-2. The TEM-1 and TEM-2 beta-lactamases differ by only one amino acid. We synthesized two oligonucleotides whose central bases correspond to this difference. The use of these oligonucleotides enables us to discriminate between TEM-1 and TEM-2 genes. Using oligonucleotides homologous to parts of Tn3, we also monitored the presence of TnA-like transposons in bacteria harboring different beta-lactamase genes. Only the TEM-1 and TEM-2 genes were found to be on transposons with terminal sequences identical to those of Tn3. All hybridization experiments were performed with both dot-blot and colony-hybridization techniques, and the suitability of these two methods for epidemiological studies is compared.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Transposable Elements*
  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics
  • Enterobacteriaceae / enzymology
  • Enterobacteriaceae / genetics
  • Genes, Bacterial*
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides
  • Penicillinase*
  • Pseudomonas aeruginosa / enzymology
  • Pseudomonas aeruginosa / genetics
  • Repetitive Sequences, Nucleic Acid

Substances

  • DNA Transposable Elements
  • DNA, Bacterial
  • Oligodeoxyribonucleotides
  • Penicillinase