Optimization of GFP levels for analyzing Salmonella gene expression during an infection

Green fluorescent protein (GFP) is an attractive reporter for Salmonella gene expression analysis but might interfere with virulence when expressed at high levels. To identify suitable GFP levels, we constructed a series of Salmonella strains expressing different amounts of GFP and measured their fl...

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Bibliographic Details
Published in:FEBS letters 2002-06, Vol.521 (1), p.105-108
Main Authors: Wendland, Meike, Bumann, Dirk
Format: Article
Language:English
Subjects:
Animals
Disease Models, Animal
Female
Flow Cytometry
Gene Expression
Genes, Reporter
Green fluorescent protein
Green Fluorescent Proteins
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Mice
Mice, Inbred BALB C
Salmonella enterica
Salmonella enterica - genetics
Salmonella enterica - growth & development
Salmonella enterica - pathogenicity
Salmonella Infections - microbiology
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Summary:Green fluorescent protein (GFP) is an attractive reporter for Salmonella gene expression analysis but might interfere with virulence when expressed at high levels. To identify suitable GFP levels, we constructed a series of Salmonella strains expressing different amounts of GFP and measured their fluorescence and colonization levels in infected mice. The results show that GFP concentrations in the range of 7000–200 000 molecules per Salmonella cell are detectable in ex vivo samples using flow cytometry, and cause no major Salmonella virulence defect. Appropriate GFP levels can be obtained with weak promoters and stable GFP, or strong promoters and destabilized GFP.
ISSN:0014-5793
1873-3468
DOI:10.1016/S0014-5793(02)02834-X