Green fluorescent protein as a novel marker and reporter system in Helicobacter sp.
In order to be able to study gene regulation in single, live Helicobacter pylori bacteria in vitro or in contact with host cells, we established the green fluorescent protein gene gfp from Aequorea victoria as a reporter gene for use with Helicobacter species. We describe here the construction of genomic transcriptional fusions of the promoterless gfp gene with the flaA and flaB promoters of H. pylori. We have also constructed a Mini-Tn3-km-gfp transposon to be used for shuttle transposon mutagenesis in H. pylori and H. mustelae. A marker strain with wild-type phenotype, carrying multiple plasmid-borne copies of gfp under the control of the H. pylori flaB promoter, was constructed for studies of bacterial distribution and transmission in animal models.
Josenhans
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Ruhr-Universität Bochum, Medizinische Mikrobiologie, Bochum, Germany.
Friedrich
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Suerbaum
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D019677:Artificial Gene Fusion / Q000379:methods; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015415:Biomarkers; D015153:Blotting, Western; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial / Q000235:genetics; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D016998:Helicobacter / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction
D001426:Bacterial Proteins; D015415:Biomarkers; D004251:DNA Transposable Elements; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins