Construction and use of a new vector/transposon, pLBT::mini-Tn10:lac:kan, to identify environmentally responsive genes in a marine bacterium

FEMS Microbiol Lett. 1996 Jul 1;140(2-3):287-94. doi: 10.1016/0378-1097(96)00196-6.

Abstract

The previously described pLOFKm transposon delivery plasmid (J.Bacteriol. (1990) 172, 6557-6567) was engineered such that a promotorless lacZ gene was cloned within the transposon cassette, generating the vector pLBT. Using pLBT, stable insertion mutations were generated at high frequencies in Vibrio sp. S141 and Pseudomonas sp. S91, and the interrupted genes could be monitored for their pattern of regulation. Genetic screens isolated mutants defective in a variety of activities. We describe the construction and use of pLBT as a tool for reporter gene mutant analysis in bacteria other than well-characterized laboratory strains.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteria / drug effects
  • Bacteria / genetics*
  • Bacteria / isolation & purification
  • Carbon Dioxide / pharmacology
  • Conjugation, Genetic
  • DNA Transposable Elements*
  • Genes, Bacterial
  • Genes, Reporter
  • Genetic Vectors*
  • Lac Operon
  • Marine Biology
  • Mutagenesis, Insertional
  • Pseudomonas / genetics
  • Vibrio / genetics

Substances

  • DNA Transposable Elements
  • Carbon Dioxide