A replicative plasmid vector allows efficient complementation of pathogenic Leptospira strains

Applied and Environmental Microbiology
Christopher J PappasMathieu Picardeau

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 o...Continue Reading

References

Jul 1, 1967·Journal of Bacteriology·R C Johnson, V G Harris
Nov 23, 2006·Infection and Immunity·Pascale BourhyMathieu Picardeau
Nov 13, 2007·Applied and Environmental Microbiology·Mathieu Picardeau
Sep 17, 2009·Nature Reviews. Microbiology·Albert I KoMathieu Picardeau
Jun 1, 2010·Applied and Environmental Microbiology·Dante PoggiMathieu Picardeau
Sep 25, 2014·International Journal of Systematic and Evolutionary Microbiology·Pascale BourhyMathieu Picardeau

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Citations

Jun 27, 2018·The American Journal of Tropical Medicine and Hygiene·Carla FernandezMichael A Matthias
Nov 12, 2015·PLoS Neglected Tropical Diseases·Yanzhuo WangJinhong Qin
Mar 7, 2017·Nature Reviews. Microbiology·Mathieu Picardeau
Mar 9, 2017·Frontiers in Immunology·Maria Gomes-SoleckiCatherine Werts
Aug 8, 2018·Scientific Reports·Olivier SchiettekattePascale Bourhy
Dec 14, 2021·PloS One·Theerapat PhokaKanitha Patarakul

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