Reliable use of green fluorescent protein in fluorescent pseudomonads

J Microbiol Methods. 2001 Jul 30;46(1):77-80. doi: 10.1016/s0167-7012(01)00262-7.

Abstract

When fluorescent pseudomonads are cultured on standard solid media under iron limiting conditions, they produce fluorescent, pigmented iron collating agents (siderophores). Siderophores can be readily identified by strong fluorescence seen under UV/blue light. The application of the eukaryotic green fluorescent protein (GFP) as a bacterial marker in microbial ecology is increasingly being used, particularly as it is a powerful method for non-destructive monitoring in situ. As gfp expressing bacteria have to be detected under UV/blue light, the fluorescence of siderophore-producing Pseudomonas spp. masks normal levels of GFP fluorescence when colonies are viewed on standard bacterial agar. Here, we describe a simple but effective way of identifying gfp-expressing Pseudomonas fluorescens using media supplemented with 0.45 mM FeSO(4).7H(2)O. This is of relevance for the screening of insertion libraries and in the application of GFP transposons as promoter probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Culture Media
  • DNA Transposable Elements
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Ferric Compounds / pharmacology
  • Fluorescence
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Mutagenesis, Insertional
  • Plasmids
  • Pseudomonas fluorescens / genetics
  • Pseudomonas fluorescens / isolation & purification
  • Pseudomonas fluorescens / metabolism*
  • Siderophores / metabolism

Substances

  • Culture Media
  • DNA Transposable Elements
  • Ferric Compounds
  • Luminescent Proteins
  • Siderophores
  • Green Fluorescent Proteins
  • ferric sulfate