New Broad-Host-Range Promoter Probe Vectors Based on the Plasmid RK2 Replicon

FEMS Microbiol Lett. 2001 Feb 5;195(1):91-6. doi: 10.1111/j.1574-6968.2001.tb10503.x.

Abstract

Broad-host-range plasmid RK2-based promoter probe vectors with a known nucleotide sequence were constructed. In the absence of an upstream promoter, the expression of two tested reporter genes (luc and lacZ) in Escherichia coli was virtually zero, while insertion of the Ptrc promoter resulted in strong inducer-dependent expression. The lacZ-based vectors were mobilized into Pseudomonas fluorescens ST, Pseudomonas putida KT2442, Sphingomonas spp. and Burkholderia spp. LB400, and expression analyses indicated that the properties observed in E. coli are maintained across the species barriers. In addition, the previously established knowledge of RK2 molecular biology allows easy manipulations of features such as plasmid copy number, further extending the application potential of the vectors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Probes
  • Genetic Vectors*
  • Gram-Negative Bacteria / genetics*
  • Molecular Sequence Data
  • Plasmids / genetics*
  • Promoter Regions, Genetic / genetics*
  • Replication Origin / genetics
  • Replicon / genetics*
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • DNA Probes
  • beta-Galactosidase

Associated data

  • GENBANK/AF187995
  • GENBANK/AF187996
  • GENBANK/AF320510