Journal:

Article Title: Expression of the Helicobacter pylori ureI Gene Is Required for Acidic pH Activation of Cytoplasmic Urease

doi:

Figure Lengend Snippet: PCR analysis confirming the replacement of the ureI gene with the kanamycin cassette. Genomic DNA was isolated from the H. pylori wild type and H. pylori ureI mutant strains and used as a template for PCR. PCR primers corresponding to regions of the ureB and ureE gene were used as described in Materials and Methods. A PCR product of 1,281 bp was detected for the H. pylori wild type strains (ATCC 43504, lane 1; ATCC 49503, lane 3). A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4).

Article Snippet: A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 7 caption a7 Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies.

Techniques: Isolation, Mutagenesis

Journal:

Article Title: Expression of the Helicobacter pylori ureI Gene Is Required for Acidic pH Activation of Cytoplasmic Urease

doi:

Figure Lengend Snippet: PCR analysis of ureI from genomic DNA of gastric and nongastric Helicobacter species. Genomic DNA was isolated from the different bacterial species and used as template for PCR of ureI as described in Materials and Methods. A single PCR product of about 587 bp was detected in the lanes containing genomic DNA from H. pylori ATCC 43504 and ATCC 49503 and from H. nemestrinae. No PCR product was detected in the lanes containing genomic DNA from the ureI deletion mutants, H. mustelae or the nongastric Helicobacter species.

Article Snippet: A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 7 caption a7 Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies.

Techniques: Isolation

Journal:

Article Title: Expression of the Helicobacter pylori ureI Gene Is Required for Acidic pH Activation of Cytoplasmic Urease

doi:

Figure Lengend Snippet: Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies. Membrane protein extracts were size fractionated by SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with anti-UreI (UP2/3 and UP4/5 mixture) as described in Materials and Methods. A single band corresponding to a molecular size of about 21 kDa was detected in the lanes containing membrane proteins from H. pylori ATCC 43504 and ATCC 49503 and from H. nemestrinae. No immunoreactivity was detected in the lanes containing membrane proteins from the ureI deletion mutants or the nongastric Helicobacter species.

Article Snippet: A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 7 caption a7 Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies.

Techniques: Western Blot, Membrane, Polyacrylamide Gel Electrophoresis

Journal:

Article Title: Expression of the Helicobacter pylori ureI Gene Is Required for Acidic pH Activation of Cytoplasmic Urease

doi:

Figure Lengend Snippet: pH dependence of urease activity of H. pylori and H. pylori ureI mutant. The urease activity of the intact H. pylori wild type (■) increased rapidly as the medium pH decreased. The urease activity remained high down to a medium pH of 2.5. In contrast, there was no increase of urease activity in the H. pylori ureI mutant (●). The urease activity of the H. pylori ureI mutant lysates displayed a single pH optimum (○). Urease activity was measured radiometrically under strong buffering conditions in the presence of 5 mM urea as described in Materials and Methods (micromoles of CO2 liberated per minute per milligram of protein; n = 3, ± the standard error of the mean [SEM]).

Article Snippet: A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 7 caption a7 Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies.

Techniques: Activity Assay, Mutagenesis

Journal:

Article Title: Expression of the Helicobacter pylori ureI Gene Is Required for Acidic pH Activation of Cytoplasmic Urease

doi:

Figure Lengend Snippet: Effect of detergent permeabilization on the urease activity of intact H. pylori and H. pylori ureI mutant. Addition of 0.01% C12E8 to intact H. pylori and H. pylori ureI mutant increases urease activity at neutral pH. Urease activity was measured radiometrically under strong buffering conditions in the presence of 5 mM urea as described in Materials and Methods (micromoles of CO2 liberated per minute per milligram of protein; n = 3, ±SEM).

Article Snippet: A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 7 caption a7 Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies.

Techniques: Activity Assay, Mutagenesis

Journal:

Article Title: Expression of the Helicobacter pylori ureI Gene Is Required for Acidic pH Activation of Cytoplasmic Urease

doi:

Figure Lengend Snippet: Effect of urea on the membrane potential of H. pylori wild type and ureI mutants. Membrane potential was measured by using DiSC3(5) at pH 4.5 under pH clamp conditions as described in Materials and Methods. Urea was added as indicated by the arrow and resulted in an increase of membrane potential in H. pylori (A) but not in the H. pylori ureI mutant (B). The horizontal arrow at the point of equilibration of the dye indicates bacterial membrane potential prior to urea addition.

Article Snippet: A PCR product of 1,325 bp was detected for the H. pylori ureI mutant strains (ATCC 43504-I, lane 2; ATCC 49503-I, lane 4). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 7 caption a7 Western blot analysis of membrane protein extracts from gastric and nongastric Helicobacter species by using UreI antibodies.

Techniques: Membrane, Mutagenesis