Fluorescent reference strains of bacteria by chromosomal integration of a modified green fluorescent protein gene

L. B. Pinheiro, M. D. Gibbs, G. Vesey, J. J. Smith, Peter Bergquist

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    20 Citations (Scopus)

    Abstract

    Fluorescent reference strains of bacteria carrying a stable chromosomally integrated single copy of the gfp gene have been developed. A modified version of the gfp gene has been generated by mutagenesis and expressed under the control of the bacteriophage lambda promoter P-L. A cassette comprising bacteriophage Mu transposon arms flanking the modified gfp gene and regulatory regions was irreversibly integrated as an in-vitro-assembled transposition complex into the genomes of Escherichia coli and Salmonella spp. The modified green fluorescent protein (GFP) protein retained the fluorescence excitation and emission wavelengths of wild-type GFP. However, it fluoresced more brightly in E. coli and Salmonella compared to wild-type GFP, presumably due to improved protein maturation. Fluorescent E. coli and Salmonella strains carrying the gfp gene cassette were easily differentiated from their respective non-fluorescent parental strains on various growth media by visualization under UV light. The bacterial strains produced by this method remained viable and stably fluorescent when incorporated into a matrix for delivery of exact numbers of viable bacterial cells for use as quality control agents in microbiological procedures.

    Original languageEnglish
    Pages (from-to)1287-1295
    Number of pages9
    JournalApplied Microbiology and Biotechnology
    Volume77
    Issue number6
    DOIs
    Publication statusPublished - Jan 2008

    Keywords

    • GRAM-NEGATIVE BACTERIA
    • LACTIC-ACID BACTERIA
    • ESCHERICHIA-COLI
    • TRANSPOSITION COMPLEXES
    • MARKER
    • GFP
    • SURVIVAL
    • MUTAGENESIS
    • EXPRESSION
    • VECTORS

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