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Volume 154, Issue 4

Application of unstable Gfp variants to the kinetic study of gene expression during infection Free

Abstract

An unstable type of green fluorescent protein (Gfp) tagged with a C-terminal extension, which is a target for tail-specific protease, was used as a reporter gene in . To analyse Gfp expression in legionellae, transcriptional fusions of unstable with the specific (ntraellular ultiplication) promoters (P, P and P) were constructed. Infection studies using J774.1 macrophages as the host, and strains carrying P-, P- and P- fusions, indicated that the , and genes could be expressed intracellularly. Expression of , and genes in infected cells was examined by flow cytometry. Furthermore, fluorescent intracellular legionellae were detected directly by confocal microscopy. Real-time quantitative RT-PCR revealed the differences in the gene expression of , and that of and , during infection. Expression of was high in the late stage of infection, while that of and was high in the early phase only. We show that unstable is a useful reporter gene whose expression in legionellae can be followed in real-time, and that it allows analysis of promoter activities in legionellae and monitoring of the infection process.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
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2008-04-01
2024-04-25
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Application of unstable Gfp variants to the kinetic study of Legionella pneumophila icm gene expression during infection
Microbiology 154, 1015 (2008); https://doi.org/10.1099/mic.0.2007/013144-0
/content/journal/micro/10.1099/mic.0.2007/013144-0
/content/journal/micro/10.1099/mic.0.2007/013144-0
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