title: Monitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors.
abstract: A family of versatile promoter-probe plasmids for gene expression analysis was developed based on a modular expression plasmid system (pZ). The vectors contain different replicons with exchangeable antibiotic cassettes to allow compatibility and expression analysis on a low-, midi- and high-copy number basis. Suicide vector variants also permit chromosomal integration of the reporter fusion and stable vector derivatives can be used for in vivo or in situ expression studies under non-selective conditions. Transcriptional and translational fusions to the reporter genes gfp(mut3.1), amCyan, dsRed2, luxCDABE, phoA or lacZ can be constructed, and presence of identical multiple cloning sites in the vector system facilitates the interchange of promoters or reporter genes between the plasmids of the series. The promoter of the constitutively expressed gapA gene of Escherichia coli was included to obtain fluorescent and bioluminescent expression constructs. A combination of the plasmids allows simultaneous detection and gene expression analysis in individual bacteria, e.g. in bacterial communities or during mouse infections. To test our vector system, we analyzed and quantified expression of Yersinia pseudotuberculosis virulence genes under laboratory conditions, in association with cells and during the infection process.
authors.affiliation: Department of Molecular Infection Biology, Helmholtz Centre for Infection Research, Braunschweig, Lower Saxony, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Uliczka
authors.forename: Fabio
authors.initials: F
authors.lastname: Pisano
authors.forename: Annika
authors.initials: A
authors.lastname: Kochut
authors.forename: Wiebke
authors.initials: W
authors.lastname: Opitz
authors.forename: Katharina
authors.initials: K
authors.lastname: Herbst
authors.forename: Tatjana
authors.initials: T
authors.lastname: Stolz
authors.forename: Petra
authors.initials: P
authors.lastname: Dersch
mesh_terms: D000818:Animals; D019677:Artificial Gene Fusion; D001419:Bacteria / Q000235:genetics* / Q000472:pathogenicity*; D001424:Bacterial Infections; D001483:Base Sequence; D002460:Cell Line; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000472:pathogenicity
chemical_list: D008164:Luminescent Proteins