ArticleApplied and environmental microbiology2008
Development of inducible systems to engineer conditional mutants of essential genes of Helicobacter pylori.
Article in Applied and environmental microbiology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed, 63 citations in OpenAlex.
- Article
- Bacterial flagella hijack type IV pili proteins to control motility.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Interplay between Amoxicillin Resistance and Osmotic Stress in Helicobacter pylori.Journal of bacteriology · 2022Article
- An Overview of Autophagy inFrontiers in cellular and infection microbiology · 2022Review
- Small RNA mediated gradual control of lipopolysaccharide biosynthesis affects antibiotic resistance in Helicobacter pylori.Nature communications · 2021Article
- A peptide of a type I toxin-antitoxin system inducesProceedings of the National Academy of Sciences of the United States of America · 2020Article
- The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen Helicobacter pylori.mBio · 2020Article
- Cell morphology as a virulence determinant: lessons from Helicobacter pylori.Current opinion in microbiology · 2020Review
- HupA, the main undecaprenyl pyrophosphate and phosphatidylglycerol phosphate phosphatase in Helicobacter pylori is essential for colonization of the stomach.PLoS pathogens · 2019Article
- Maturation of atypical ribosomal RNA precursors in Helicobacter pylori.Nucleic acids research · 2019Article
- Site-directed mutagenesis of Campylobacter concisus respiratory genes provides insight into the pathogen's growth requirements.Scientific reports · 2018Article
- Three SpoA-domain proteins interact in the creation of the flagellar type III secretion system inThe Journal of biological chemistry · 2018Article
- Iron-sulfur protein maturation in Helicobacter pylori: identifying a Nfu-type cluster carrier protein and its iron-sulfur protein targets.Molecular microbiology · 2018Article
- Article
- Mechanistic insights into type I toxin antitoxin systems in Helicobacter pylori: the importance of mRNA folding in controlling toxin expression.Nucleic acids research · 2017Article
- Identification of the RNA Pyrophosphohydrolase RppH of Helicobacter pylori and Global Analysis of Its RNA Targets.The Journal of biological chemistry · 2017Article
- Overexpression of serine protease HtrA enhances disruption of adherens junctions, paracellular transmigration and type IV secretion of CagA byGut pathogens · 2017Article
- Characterization in Helicobacter pylori of a Nickel Transporter Essential for Colonization That Was Acquired during Evolution by Gastric Helicobacter Species.PLoS pathogens · 2016Article
- Role of the N-Acetylmuramoyl-l-Alanyl Amidase, AmiA, of Helicobacter pylori in Peptidoglycan Metabolism, Daughter Cell Separation, and Virulence.Microbial drug resistance (Larchmont, N.Y.) · 2016Article
- Genetic Diversity as Consequence of a Microaerobic and Neutrophilic Lifestyle.PLoS pathogens · 2016Article
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Escherichia coli-Helicobacter pylori shuttle vector pHeL2 was modified to introduce the inducible LacI(q)-pTac system of E. coli, in which the promoters were engineered to be under the control of H. pylori RNA polymerase. The amiE gene promoter of H. pylori was taken to constitutively express the LacI(q) repressor. Expression of the reporter gene lacZ was driven by either pTac (pILL2150) or a modified version of the ureI gene promoter in which one or two LacI-binding sites and/or mutated nucleotides between the ribosomal binding site and the ATG start codon (pILL2153 and pILL2157) were introduced. Promoter activity was evaluated by measuring beta-galactosidase activity. pILL2150 is a tightly regulated expression system suitable for the analysis of genes with low-level expression, while pILL2157 is well adapted for the controlled expression of genes encoding recombinant proteins in H. pylori. To exemplify the usefulness of these tools, we constructed conditional mutants of the putative essential pbp1 and ftsI genes encoding penicillin-binding proteins 1 and 3 of H. pylori, respectively. Both genes were cloned into pILL2150 and introduced in the parental H. pylori strain N6. The chromosomally harbored pbp1 and ftsI genes were then inactivated by replacing them with a nonpolar kanamycin cassette. Inactivation was strictly dependent upon addition of isopropyl-beta-d-thiogalactopyranoside. Hence, we were able to construct the first conditional mutants of H. pylori. Finally, we demonstrated that following in vitro methylation of the recombinant plasmids, these could be introduced into a large variety of H. pylori isolates with different genetic backgrounds.
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