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. 1994 Aug;62(8):3586–3589. doi: 10.1128/iai.62.8.3586-3589.1994

A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach.

M Tsuda 1, M Karita 1, M G Morshed 1, K Okita 1, T Nakazawa 1
PMCID: PMC303000  PMID: 8039935

Abstract

The urease of Helicobacter pylori has been proposed to be one of its pathogenic factors. A kanamycin resistance determinant was inserted in a cloned urease gene, and transformation-mediated allelic exchange mutagenesis was carried out to introduce the disrupted gene into the corresponding wild-type chromosomal region of a clinical isolate of H. pylori, CPY3401. The resulting mutant, HPT73, had the null activity of urease. Nude mouse stomachs were challenged with these two isogenic strains to examine the role of urease in pathogenesis. Gastritis was found in the CPY3401-challenged stomachs, from which bacteria indistinguishable from CPY3401 were recovered. There was no gastritis in the HPT73-challenged stomachs, and we could not recover H. pylori from them. These results indicated that H. pylori urease is essential for colonizing the nude mouse stomach.

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Selected References

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  1. Cussac V., Ferrero R. L., Labigne A. Expression of Helicobacter pylori urease genes in Escherichia coli grown under nitrogen-limiting conditions. J Bacteriol. 1992 Apr;174(8):2466–2473. doi: 10.1128/jb.174.8.2466-2473.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Eaton K. A., Brooks C. L., Morgan D. R., Krakowka S. Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun. 1991 Jul;59(7):2470–2475. doi: 10.1128/iai.59.7.2470-2475.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ferrero R. L., Cussac V., Courcoux P., Labigne A. Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J Bacteriol. 1992 Jul;174(13):4212–4217. doi: 10.1128/jb.174.13.4212-4217.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Haas R., Meyer T. F., van Putten J. P. Aflagellated mutants of Helicobacter pylori generated by genetic transformation of naturally competent strains using transposon shuttle mutagenesis. Mol Microbiol. 1993 May;8(4):753–760. doi: 10.1111/j.1365-2958.1993.tb01618.x. [DOI] [PubMed] [Google Scholar]
  5. Hu L. T., Foxall P. A., Russell R., Mobley H. L. Purification of recombinant Helicobacter pylori urease apoenzyme encoded by ureA and ureB. Infect Immun. 1992 Jul;60(7):2657–2666. doi: 10.1128/iai.60.7.2657-2666.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Karita M., Kouchiyama T., Okita K., Nakazawa T. New small animal model for human gastric Helicobacter pylori infection: success in both nude and euthymic mice. Am J Gastroenterol. 1991 Nov;86(11):1596–1603. [PubMed] [Google Scholar]
  7. Lee A., Fox J., Hazell S. Pathogenicity of Helicobacter pylori: a perspective. Infect Immun. 1993 May;61(5):1601–1610. doi: 10.1128/iai.61.5.1601-1610.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Pérez-Pérez G. I., Olivares A. Z., Cover T. L., Blaser M. J. Characteristics of Helicobacter pylori variants selected for urease deficiency. Infect Immun. 1992 Sep;60(9):3658–3663. doi: 10.1128/iai.60.9.3658-3663.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rose R. E. The nucleotide sequence of pACYC184. Nucleic Acids Res. 1988 Jan 11;16(1):355–355. doi: 10.1093/nar/16.1.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Segal E. D., Shon J., Tompkins L. S. Characterization of Helicobacter pylori urease mutants. Infect Immun. 1992 May;60(5):1883–1889. doi: 10.1128/iai.60.5.1883-1889.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Segal E. D., Tompkins L. S. Transformation of Helicobacter pylori by electroporation. Biotechniques. 1993 Feb;14(2):225–226. [PubMed] [Google Scholar]
  12. Suerbaum S., Josenhans C., Labigne A. Cloning and genetic characterization of the Helicobacter pylori and Helicobacter mustelae flaB flagellin genes and construction of H. pylori flaA- and flaB-negative mutants by electroporation-mediated allelic exchange. J Bacteriol. 1993 Jun;175(11):3278–3288. doi: 10.1128/jb.175.11.3278-3288.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Taylor L. A., Rose R. E. A correction in the nucleotide sequence of the Tn903 kanamycin resistance determinant in pUC4K. Nucleic Acids Res. 1988 Jan 11;16(1):358–358. doi: 10.1093/nar/16.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tsuda M., Karita M., Nakazawa T. Genetic transformation in Helicobacter pylori. Microbiol Immunol. 1993;37(1):85–89. doi: 10.1111/j.1348-0421.1993.tb03184.x. [DOI] [PubMed] [Google Scholar]
  15. Wang Y., Roos K. P., Taylor D. E. Transformation of Helicobacter pylori by chromosomal metronidazole resistance and by a plasmid with a selectable chloramphenicol resistance marker. J Gen Microbiol. 1993 Oct;139(10):2485–2493. doi: 10.1099/00221287-139-10-2485. [DOI] [PubMed] [Google Scholar]

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