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A toolbox for manipulating the genome of the major goat pathogen, Mycoplasma capricolum subsp. capripneumoniae

hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorGOURGUES, Géraldine
hal.structure.identifierAnimal, Santé, Territoires, Risques et Ecosystèmes [UMR ASTRE]
hal.structure.identifierDépartement Systèmes Biologiques [Cirad-BIOS]
dc.contributor.authorMANSO-SILVÁN, Lucía
hal.structure.identifierDépartement de biologie [Sherbrooke] [UdeS]
dc.contributor.authorCHAMBERLAND, Catherine
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorSIRAND-PUGNET, Pascal
hal.structure.identifierAnimal, Santé, Territoires, Risques et Ecosystèmes [UMR ASTRE]
dc.contributor.authorTHIAUCOURT, François
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorBLANCHARD, Alain
hal.structure.identifierUniversité de Montréal [UdeM]
dc.contributor.authorBABY, Vincent
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorLARTIGUE, Carole
dc.date.accessioned2024-04-11T10:05:50Z
dc.date.available2024-04-11T10:05:50Z
dc.date.issued2024-01-09
dc.identifier.issn1350-0872
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/197532
dc.description.abstractEnMycoplasma capricolum subspecies capripneumoniae ( Mccp ) is the causative agent of contagious caprine pleuropneumonia (CCPP), a devastating disease listed by the World Organisation for Animal Health (WOAH) as a notifiable disease and threatening goat production in Africa and Asia. Although a few commercial inactivated vaccines are available, they do not comply with WOAH standards and there are serious doubts regarding their efficacy. One of the limiting factors to comprehend the molecular pathogenesis of CCPP and develop improved vaccines has been the lack of tools for Mccp genome engineering. In this work, key synthetic biology techniques recently developed for closely related mycoplasmas were adapted to Mccp . CReasPy-Cloning was used to simultaneously clone and engineer the Mccp genome in yeast, prior to whole-genome transplantation into M. capricolum subsp. capricolum recipient cells. This approach was used to knock out an S41 serine protease gene recently identified as a potential virulence factor, leading to the generation of the first site-specific Mccp mutants. The Cre–lox recombination system was then applied to remove all DNA sequences added during genome engineering. Finally, the resulting unmarked S41 serine protease mutants were validated by whole-genome sequencing and their non-caseinolytic phenotype was confirmed by casein digestion assay on milk agar. The synthetic biology tools that have been successfully implemented in Mccp allow the addition and removal of genes and other genetic features for the construction of seamless targeted mutants at ease, which will pave the way for both the identification of key pathogenicity determinants of Mccp and the rational design of novel, improved vaccines for the control of CCPP.
dc.language.isoen
dc.publisherMicrobiology Society
dc.rights.urihttp://creativecommons.org/licenses/by/
dc.subject.enwhole-genome transplantation.
dc.subject.enCre–lox recombination system
dc.subject.enMycoplasma capricolum subsp. capripneumoniae
dc.subject.enS41 serine protease
dc.subject.enin-yeast genome cloning/engineering
dc.subject.enoriC plasmid
dc.title.enA toolbox for manipulating the genome of the major goat pathogen, Mycoplasma capricolum subsp. capripneumoniae
dc.typeArticle de revue
dc.identifier.doi10.1099/mic.0.001423
dc.subject.halSciences du Vivant [q-bio]
dc.subject.halSciences du Vivant [q-bio]/Biologie animale
dc.subject.halSciences du Vivant [q-bio]/Biologie animale/Médecine vétérinaire et santé animal
dc.subject.halSciences du Vivant [q-bio]/Microbiologie et Parasitologie
bordeaux.journalMicrobiology
bordeaux.volume170
bordeaux.hal.laboratoriesBiologie du Fruit & Pathologie (BFP) - UMR 1332*
bordeaux.issue1
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionINRAE
bordeaux.peerReviewedoui
hal.identifierhal-04512951
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-04512951v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Microbiology&rft.date=2024-01-09&rft.volume=170&rft.issue=1&rft.eissn=1350-0872&rft.issn=1350-0872&rft.au=GOURGUES,%20G%C3%A9raldine&MANSO-SILV%C3%81N,%20Luc%C3%ADa&CHAMBERLAND,%20Catherine&SIRAND-PUGNET,%20Pascal&THIAUCOURT,%20Fran%C3%A7ois&rft.genre=article


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