{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T01:46:16Z","timestamp":1791942376914,"version":"4.3.4"},"reference-count":62,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2000,3,1]],"date-time":"2000-03-01T00:00:00Z","timestamp":951868800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Infect Immun"],"published-print":{"date-parts":[[2000,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Bacteria of the genus\n            <jats:italic>Brucella<\/jats:italic>\n            are facultative intracellular pathogens which have developed the capacity to survive and multiply in professional and nonprofessional phagocytes. The genetic basis of this aspect of\n            <jats:italic>Brucella<\/jats:italic>\n            virulence is still poorly understood. To identify new virulence factors, we have adapted signature-tagged transposon mutagenesis, which has been used essentially in animal models, to an in vitro human macrophage infection model. A library of 1,152\n            <jats:italic>Brucella suis<\/jats:italic>\n            1330 tagged mini-Tn\n            <jats:italic>5<\/jats:italic>\n            Km2 mutants, in 12 pools, was screened for intracellular survival and multiplication in vitamin D\n            <jats:sub>3<\/jats:sub>\n            -differentiated THP1 cells. Eighteen mutants were identified, and their attenuation was confirmed in THP1 macrophages and HeLa cells. For each avirulent mutant, a genomic fragment containing the transposon was cloned. The genomic DNA sequence flanking the transposon allowed us to assign functions to all of the inactivated genes. Transposon integration had occurred in 14 different genes, some of which were known virulence genes involved in intracellular survival or biosynthesis of smooth lipopolysaccharide (the\n            <jats:italic>virB<\/jats:italic>\n            operon and\n            <jats:italic>manB<\/jats:italic>\n            ), thus validating the model. Other genes identified encoded factors involved in the regulation of gene expression and enzymes involved in biosynthetic or metabolic pathways. Possible roles in the virulence of\n            <jats:italic>Brucella<\/jats:italic>\n            for the different factors identified are discussed.\n          <\/jats:p>","DOI":"10.1128\/iai.68.3.1297-1303.2000","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:01:19Z","timestamp":1027764079000},"page":"1297-1303","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":160,"title":["Identification of\n            <i>Brucella suis<\/i>\n            Genes Affecting Intracellular Survival in an In Vitro Human Macrophage Infection Model by Signature-Tagged Transposon Mutagenesis"],"prefix":"10.1128","volume":"68","author":[{"given":"Vincent","family":"Foulongne","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->INSERM U431, Faculte\u0301 de Me\u0301decine, 30900 N\u0131\u0302mes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gise\u0301le","family":"Bourg","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->INSERM U431, Faculte\u0301 de Me\u0301decine, 30900 N\u0131\u0302mes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chantal","family":"Cazevieille","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->INSERM U431, Faculte\u0301 de Me\u0301decine, 30900 N\u0131\u0302mes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvie","family":"Michaux-Charachon","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->INSERM U431, Faculte\u0301 de Me\u0301decine, 30900 N\u0131\u0302mes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"O'Callaghan","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->INSERM U431, Faculte\u0301 de Me\u0301decine, 30900 N\u0131\u0302mes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.3.1008-1016.1998"},{"key":"e_1_3_2_3_2","first-page":"1","article-title":"Laboratory techniques in brucellosis","volume":"55","author":"Alton G. 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