Genetic complementation of the urease-negative Helicobacter pylori mutant N6ureB::TnKm.
Helicobacter pylori produces urease composed of the structural subunits UreA and UreB. Isogenic mutants produced by shuttle mutagenesis from the wild-type strain N6 are widely used in the literature. We describe the genetic complementation of the mutant N6ureB::TnKm by stable transformation with the vector pHel2 containing the cloned genes ureA and ureB and their specific promoter sequence. The orientation of the cloned insert was found to be crucial for urease expression. The majority of complemented clones functionally expressed urease at higher levels than did N6. Homologous recombination between chromosomal and cloned genes occurred at a frequency of 5%.
Department of Clinical Microbiology, Hygiene Institute, University of Vienna, Währinger Gürtel 18-20/5P, A-1090, Vienna, Austria. elmar.rokita@akh-wien.ac.at
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Makristathis
D003001:Cloning, Molecular; D018274:Electroporation; D005816:Genetic Complementation Test; D016480:Helicobacter pylori / Q000235:genetics* / Q000472:pathogenicity; D014169:Transformation, Bacterial; D014510:Urease / Q000172:deficiency* / Q000235:genetics; D014774:Virulence / Q000235:genetics
D014510:Urease