ArticleApplied and environmental microbiology2006
Generating tetracycline-inducible auxotrophy in Escherichia coli and Salmonella enterica serovar Typhimurium by using an insertion element and a hyperactive transposase.
Article in Applied and environmental microbiology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 15 citations in OpenAlex.
- Tet-ON and Tet-OFF regulation in hypervirulentMicrobiology spectrum · 2026Article
- A versatile regulatory toolkit of arabinose-inducible artificial transcription factors for Enterobacteriaceae.Communications biology · 2023Article
- Article
- Status quo of tet regulation in bacteria.Microbial biotechnology · 2022Review
- Characterization and potential mechanisms of highly antibiotic tolerant VBNC Escherichia coli induced by low level chlorination.Scientific reports · 2020Article
- The fitness landscape of the African Salmonella Typhimurium ST313 strain D23580 reveals unique properties of the pBT1 plasmid.PLoS pathogens · 2019Article
- Generation of a platform strain for ionic liquid tolerance using adaptive laboratory evolution.Microbial cell factories · 2017Article
- Article
- The application of Tet repressor in prokaryotic gene regulation and expression.Microbial biotechnology · 2008Review
- Random insertion of a TetR-inducing peptide tag into Escherichia coli proteins allows analysis of protein levels by induction of reporter gene expression.Applied and environmental microbiology · 2006Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We report the construction and application of a novel insertion element for transposase-mediated mutagenesis in gram-negative bacteria. Besides Km(r) as a selectable marker, the insertion element InsTet(G-)1 carries the anhydrotetracycline (atc)-regulated outward-directed PA promoter so that atc-dependent conditional gene knockouts or knockdowns are generated. The complex formed between the purified hyperactive transposase and InsTet(G-)1 was electroporated into Escherichia coli or Salmonella enterica serovar Typhimurium, and mutant pools were collected. We used E. coli strains with either TetR or the reverse variant revTetR(r2), while only TetR was employed in Salmonella. Screening of the InsTet(G-)1 insertion mutant pools revealed 15 atc-regulatable auxotrophic mutants for E. coli and 4 atc-regulatable auxotrophic mutants for Salmonella. We have also screened one Salmonella mutant pool in murine macrophage-like J774-A.1 cells using ampicillin enrichment. Two mutants with the InsTet(G-)1 insertion in the gene pyrE or argA survived this procedure, indicating a reduced intracellular growth rate in J774-A.1 cells. The nature of the mutants and the modes of their regulation are discussed.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.