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dc.creatorMancilla, M. (Marcos)-
dc.creatorGrillo, M.J. (María Jesús)-
dc.creatorMiguel, M.J. (María Jesús) de-
dc.creatorLopez-Goñi, I. (Ignacio)-
dc.creatorSan-Roman, B. (Beatriz)-
dc.creatorZabalza-Barangua, A. (Ana)-
dc.creatorMoriyon, I. (Ignacio)-
dc.date.accessioned2014-03-25T11:41:57Z-
dc.date.available2014-03-25T11:41:57Z-
dc.date.issued2013-
dc.identifier.citationMancilla M, Grillo MJ, de Miguel MJ, Lopez-Goni I, San-Roman B, Zabalza-Barangua A, et al. Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine. Vet Res 2013 Oct 31;44:105-9716-44-105.es_ES
dc.identifier.issn0928-4249-
dc.identifier.urihttps://hdl.handle.net/10171/35589-
dc.description.abstractBrucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production.es_ES
dc.language.isoenges_ES
dc.publisherBioMed Centrales_ES
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/221948-
dc.rightsinfo:eu-repo/semantics/openAccesses_ES
dc.rightsinfo:eu-repo/semantics/openAccesses_ES
dc.subjectO-polysaccharide synthesises_ES
dc.subjectGram-negative bacteriaes_ES
dc.subjectSmooth lipopolysaccharidees_ES
dc.subjectDNA polymorphismes_ES
dc.subjectGenomic islandes_ES
dc.subjectPCR assayes_ES
dc.subjectAbortuses_ES
dc.subjectVirulencees_ES
dc.subjectGenees_ES
dc.subjectIdentificationes_ES
dc.titleDeletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccinees_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.identifier.doihttp://dx.doi.org/10.1186/1297-9716-44-105es_ES

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