MiniTn7-transposon delivery vectors for inducible or constitutive fluorescent protein expression in Enterobacteriaceae

FEMS Microbiol Lett. 2016 Aug;363(16):fnw178. doi: 10.1093/femsle/fnw178. Epub 2016 Jul 20.

Abstract

Here we present the generation and function of two sets of bacterial plasmids that harbor fluorescent genes encoding either blue, cyan, yellow or red fluorescent proteins. In the first set, protein expression is controlled by the strong and constitutive nptII promoter whereas in the second set, the strong tac promoter was chosen that underlies LacI(q) regulation. Furthermore, the plasmids are mobilizable, contain Tn7 transposons and a temperature-sensitive origin of replication. Using Escherichia coli S17-1 as donor strain, the plasmids allow fast and convenient Tn7-transposon delivery into many enterobacterial hosts, such as the here-used E. coli O157:H7. This procedure omits the need of preparing competent recipient cells and antibiotic resistances are only transiently conferred to the recipients. As the fluorescence proteins show little to no overlap in fluorescence emission, the constructs are well suited for the study of multicolored synthetic bacterial communities during biofilm production or in host colonization studies, e.g. of plant surfaces. Furthermore, tac promoter-reporter constructs allow the generation of so-called reproductive success reporters, which allow to estimate past doublings of bacterial individuals after introduction into environments, emphasizing the role of individual cells during colonization.

Keywords: CUSPER; Escherichia coli O157:H7; fluorescent proteins; reproductive success.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biofilms
  • Chromosomes, Bacterial
  • Cloning, Molecular / methods
  • DNA Transposable Elements*
  • DNA, Bacterial
  • Drug Resistance, Microbial / genetics
  • Escherichia coli O157 / genetics*
  • Genetic Vectors*
  • Luminescent Proteins / genetics*
  • Plasmids*
  • Promoter Regions, Genetic

Substances

  • DNA Transposable Elements
  • DNA, Bacterial
  • Luminescent Proteins
  • red fluorescent protein