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Microbiol."],"abstract":"<jats:p>The lipopolysaccharide (LPS) O-polysaccharide (O-PS) is the main virulence factor in <jats:italic>Brucella<\/jats:italic>. After synthesis in the cytoplasmic membrane, O-PS is exported to the periplasm by the Wzm\/Wzt system, where it is assembled into a LPS. This translocation also engages a bactoprenol carrier required for further biosynthesis pathways, such as cell wall biogenesis. Targeting O-PS export by blockage holds great potential for vaccine development, but little is known about the biological implications of each Wzm\/Wzt moiety. To improve this knowledge and to elucidate its potential application as a vaccine, we constructed and studied <jats:italic>wzm<\/jats:italic>\/<jats:italic>wzt<\/jats:italic> single- and double-deletion mutants, using the attenuated strain <jats:italic>Brucella melitensis<\/jats:italic> Rev1 as the parental strain. This allowed us to describe the composition of <jats:italic>Brucella<\/jats:italic> peptidoglycan for the first time. We observed that these mutants lack external O-PS yet trigger changes in genetic transcription and in phenotypic properties associated with the outer membrane and cell wall. The three mutants are highly attenuated; unexpectedly, Rev1\u0394<jats:italic>wzm<\/jats:italic> also excels as an immunogenic and effective vaccine against <jats:italic>B. melitensis<\/jats:italic> and <jats:italic>Brucella ovis<\/jats:italic> in mice, revealing that low persistence is not at odds with efficacy. Rev1\u0394<jats:italic>wzm<\/jats:italic> is attenuated in BeWo trophoblasts, does not infect mouse placentas, and is safe in pregnant ewes. Overall, these attributes and the minimal serological interference induced in sheep make Rev1\u0394<jats:italic>wzm<\/jats:italic> a highly promising vaccine candidate.<\/jats:p>","DOI":"10.3389\/fmicb.2022.908495","type":"journal-article","created":{"date-parts":[[2022,7,4]],"date-time":"2022-07-04T09:09:14Z","timestamp":1656925754000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Brucella melitensis Wzm\/Wzt System: Changes in the Bacterial Envelope Lead to Improved Rev1\u0394wzm Vaccine Properties"],"prefix":"10.3389","volume":"13","author":[{"given":"Sara","family":"Mena-Bueno","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Irati","family":"Poveda-Urkixo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oihane","family":"Irazoki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leyre","family":"Palacios","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Felipe","family":"Cava","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana","family":"Zabalza-Barangu\u00e1","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda Jes\u00fas","family":"Grill\u00f3","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,7,4]]},"reference":[{"key":"B1","author":"Alton","year":"1988","journal-title":"Techniques for the Brucellosis Laboratory."},{"key":"B2","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool.","volume":"215","author":"Altschul","year":"1990","journal-title":"J. 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