Essential role of Helicobacter pylori urease in gastric colonization: definite proof using a urease-negative mutant constructed by gene replacement.

OBJECTIVE

To investigate the involvement of urease in Helicobacter pylori colonization in the nude mouse stomach by using a genetically defined urease-negative mutant.

METHODS AND

RESULTS

Through electroporation-mediated gene replacement, one of the urease genes of an H. pylori strain, CYP3401, was disrupted by insertion of a kanamycin-resistance determinant to construct a stable urease-negative mutant, HPT73. Southern analysis confirmed that gene replacement was achieved. The two isogenic strains were introduced into the stomachs of nude mice, and the number of H. pylori and the histological changes in the stomachs were investigated 1 or 4 weeks after the challenge. Gastritis was present in the CPY3401-challenged stomach, from which bacteria indistinguishable from CPY3401 were successfully recovered. In contrast, no gastritis was found in the HPT73-challenged stomach, and H. pylori was not recovered from these stomachs.

CONCLUSION

H. pylori urease is essential for colonization of the nude mouse stomach.