Construction of mini-Tn10luxABcam/Ptac-ATS and its use for developing a bacteriophage that transduces bioluminescence to Escherichia coli O157:H7

FEMS Microbiol Lett. 2000 Jan 15;182(2):285-9. doi: 10.1111/j.1574-6968.2000.tb08909.x.

Abstract

Mini-Tn10luxABcam/Ptac-ATS was constructed in order to develop a luciferase-transducing bacteriophage for detecting Escherichia coli O157:H7. The transposon was designed to deliver a 3.6-kb insertion that confers n-decanal-dependent bioluminescence and resistance to chloramphenicol and was constructed using mini-Tn10cam/Ptac-ATS in the plasmid pNK2884 and luxAB from Vibrio harveyi. PhiV10, a temperate bacteriophage infecting common phage types of Escherichia coli O157:H7, was mutagenized as a prophage in E. coli O157:H7 strain R508. PhiV10::luxABcamA1-23 was rescued from the strain by propagating it on a strain lacking the bacteriophage and the vector containing the transposon. The bacteriophage transduced n-decanal-dependent bioluminescence to E. coli O157:H7 strain R508 that was measurable approximately 1 h post infection.

MeSH terms

  • Coliphages / genetics*
  • Coliphages / physiology
  • DNA Transposable Elements*
  • Escherichia coli O157 / genetics*
  • Escherichia coli O157 / physiology
  • Escherichia coli O157 / virology
  • Luciferases / genetics
  • Luminescent Measurements*
  • Microscopy, Electron
  • Mutagenesis, Insertional
  • Transduction, Genetic*

Substances

  • DNA Transposable Elements
  • Luciferases