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

Pinheiro, Leonardo B., Gibbs, Moreland D., Vesey, Graham, Smith, James J., & Berquist, Peter L. (2008) Fluorescent reference strains of bacteria by chromosomal integration of a modified green fluorescent protein gene. Applied Microbiology and Biotechnology, 77(6), pp. 1287-1295.

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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.

Impact and interest:

7 citations in Scopus
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6 citations in Web of Science®

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ID Code: 12661
Item Type: Journal Article
Additional Information: For more information, please refer to the journal’s website (see hypertext link) or contact the author.
Keywords: Fluorescence, reference, bacteria, GFP, strain, indicator
DOI: 10.1007/s00253-007-1253-9
ISSN: 1432-0614
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > MICROBIOLOGY (060500) > Microbial Genetics (060503)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > MICROBIOLOGY (060500) > Infectious Agents (060502)
Divisions: Past > QUT Faculties & Divisions > Faculty of Science and Technology
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright 2008 Springer
Copyright Statement: The original publication is available at SpringerLink http://www.springerlink.com
Deposited On: 26 Feb 2008
Last Modified: 29 Feb 2012 23:40

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