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Development of a replicating plasmid based on the native oriC in <em>Mycoplasma pneumoniae</em>.

hal.structure.identifierDepartment of General Microbiology, Institute of Microbiology and Genetics
dc.contributor.authorBLÖTZ, Cedric
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorLARTIGUE, Carole
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorVALVERDE TIMANA, Yanina
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorRUIZ, Estelle
hal.structure.identifierBarcelona Institute of Science and Technology [BIST]
dc.contributor.authorPAETZOLD, Bernhard
hal.structure.identifierDepartment of General Microbiology, Institute of Microbiology and Genetics
dc.contributor.authorBUSSE, Julia
hal.structure.identifierDepartment of General Microbiology, Institute of Microbiology and Genetics
dc.contributor.authorSTÜLKE, Jörg
dc.date.issued2018
dc.identifier.issn1350-0872
dc.description.abstractEnBacteria of the genus Mycoplasma have recently attracted considerable interest as model organisms in synthetic and systems biology. In particular, Mycoplasma pneumoniae is one of the most intensively studied organisms in the field of systems biology. However, the genetic manipulation of these bacteria is often difficult due to the lack of efficient genetic systems and some intrinsic peculiarities such as an aberrant genetic code. One major disadvantage in working with M. pneumoniae is the lack of replicating plasmids that can be used for the complementation of mutants and the expression of proteins. In this study, we have analysed the genomic region around the gene encoding the replication initiation protein, DnaA, and detected putative binding sites for DnaA (DnaA boxes) that are, however, less conserved than in other bacteria. The construction of several plasmids encompassing this region allowed the selection of plasmid pGP2756 that is stably inherited and that can be used for genetic experiments, as shown by the complementation assays with the glpQ gene encoding the glycerophosphoryl diester phosphodiesterase. Plasmid-borne complementation of the glpQ mutant restored the formation of hydrogen peroxide when bacteria were cultivated in the presence of glycerol phosphocholine. Interestingly, the replicating plasmid can also be used in the close relative, Mycoplasma genitalium but not in more distantly related members of the genus Mycoplasma. Thus, plasmid pGP2756 is a valuable tool for the genetic analysis of M. pneumoniae and M. genitalium.
dc.description.sponsorshipA model-driven approach to minimal cell engineering for medical therapy - ANR-15-SYNB-0001
dc.language.isoen
dc.publisherMicrobiology Society
dc.subjectoriC
dc.subjectcomplementation
dc.subjectmollicutes
dc.subject.enreplicating plasmid
dc.subject.ensystems biology
dc.title.enDevelopment of a replicating plasmid based on the native oriC in <em>Mycoplasma pneumoniae</em>.
dc.typeArticle de revue
dc.identifier.doi10.1099/mic.0.000711
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.page1372-1382
bordeaux.volume164
bordeaux.issue11
bordeaux.peerReviewedoui
hal.identifierhal-02628284
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-02628284v1
bordeaux.COinSctx_ver=Z39.88-2004&amp;rft_val_fmt=info:ofi/fmt:kev:mtx:journal&amp;rft.jtitle=Microbiology&amp;rft.date=2018&amp;rft.volume=164&amp;rft.issue=11&amp;rft.spage=1372-1382&amp;rft.epage=1372-1382&amp;rft.eissn=1350-0872&amp;rft.issn=1350-0872&amp;rft.au=BL%C3%96TZ,%20Cedric&amp;LARTIGUE,%20Carole&amp;VALVERDE%20TIMANA,%20Yanina&amp;RUIZ,%20Estelle&amp;PAETZOLD,%20Bernhard&amp;rft.genre=article


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