ArticleApplied and environmental microbiology2015
A replicative plasmid vector allows efficient complementation of pathogenic Leptospira strains.
Article in Applied and environmental microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 37 papers.
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Who cites it
37 citing papers in PubMed.
- In vivo dual RNA-Seq uncovers key effectors of epithelial barrier disruption by an extracellular pathogen.Nature communications · 2026Article
- Creating a Library of Random Transposon Mutants in Leptospira.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Gene Knockout in Leptospira spp. by CRISPR/Cas9-NHEJ (Non-Homologous End-Joining) and CRISPR-Prime Editing.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Bioinformatic Identification of CRISPR-Cas Systems inPathogens (Basel, Switzerland) · 2025Article
- Article
- Linking genomic evolutionary transitions to ecological phenotypic adaptations inbioRxiv : the preprint server for biology · 2025Article
- Induced protein expression in Leptospira spp. and its application to CRISPR/Cas9 mutant generation.Scientific reports · 2025Article
- Inter-species Transcriptomic Analysis Reveals a Constitutive Adaptation Against Oxidative Stress for the Highly Virulent Leptospira Species.Molecular biology and evolution · 2024Article
- Leptospiral adhesins: from identification to future perspectives.Frontiers in microbiology · 2024Review
- Signal-sensing triggers the shutdown of HemKR, regulating heme and iron metabolism in the spirochete Leptospira biflexa.PloS one · 2024Article
- LipL41 and LigA/LigB Gene Silencing on a LipL32 KnockoutPathogens (Basel, Switzerland) · 2023Article
- Hematogenous dissemination of pathogenic and non-pathogenicFrontiers in cellular and infection microbiology · 2022Article
- Heterologous production of the adhesin LIC13411 from pathogenicFrontiers in cellular and infection microbiology · 2022Article
- Transcriptional analysis of CRISPR I-B arrays ofFrontiers in microbiology · 2022Article
- A Novel Breakthrough inFrontiers in microbiology · 2022Article
- The CRISPR-associated Cas4 protein fromCurrent research in microbial sciences · 2021Article
- The oxidative stress response of pathogenic Leptospira is controlled by two peroxide stress regulators which putatively cooperate in controlling virulence.PLoS pathogens · 2021Article
- Role of the major determinant of polar flagellation FlhG in the endoflagella-containing spirochete Leptospira.Molecular microbiology · 2021Article
- Genetic manipulation of pathogenic Leptospira: CRISPR interference (CRISPRi)-mediated gene silencing and rapid mutant recovery at 37 °C.Scientific reports · 2021Article
- Article
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3 authors.
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Abstract
Leptospirosis, an emerging zoonotic disease, remains poorly understood because of a lack of genetic manipulation tools available for pathogenic leptospires. Current genetic manipulation techniques include insertion of DNA by random transposon mutagenesis and homologous recombination via suicide vectors. This study describes the construction of a shuttle vector, pMaORI, that replicates within saprophytic, intermediate, and pathogenic leptospires. The shuttle vector was constructed by the insertion of a 2.9-kb DNA segment including the parA, parB, and rep genes into pMAT, a plasmid that cannot replicate in Leptospira spp. and contains a backbone consisting of an aadA cassette, ori R6K, and oriT RK2/RP4. The inserted DNA segment was isolated from a 52-kb region within Leptospira mayottensis strain 200901116 that is not found in the closely related strain L. mayottensis 200901122. Because of the size of this region and the presence of bacteriophage-like proteins, it is possible that this region is a result of a phage-related genomic island. The stability of the pMaORI plasmid within pathogenic strains was tested by passaging cultures 10 times without selection and confirming the presence of pMaORI. Concordantly, we report the use of trans complementation in the pathogen Leptospira interrogans. Transformation of a pMaORI vector carrying a functional copy of the perR gene in a null mutant background restores the expression of PerR and susceptibility to hydrogen peroxide comparable to that of wild-type cells. In conclusion, we demonstrate the replication of a stable plasmid vector in a large panel of Leptospira strains, including pathogens. The shuttle vector described will expand our ability to perform genetic manipulation of Leptospira spp.
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