{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T12:49:03Z","timestamp":1740142143578,"version":"3.37.3"},"reference-count":33,"publisher":"Wiley","license":[{"start":{"date-parts":[[2014,1,1]],"date-time":"2014-01-01T00:00:00Z","timestamp":1388534400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"funder":[{"DOI":"10.13039\/501100004386","name":"Universiti Malaya","doi-asserted-by":"publisher","award":["RG409-12HTM","FRGS FP037-2013A","UM.C\/625\/1\/HIR\/MoE\/E000044-20001"],"award-info":[{"award-number":["RG409-12HTM","FRGS FP037-2013A","UM.C\/625\/1\/HIR\/MoE\/E000044-20001"]}],"id":[{"id":"10.13039\/501100004386","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004386","name":"Universiti Malaya","doi-asserted-by":"publisher","award":["RG409-12HTM","FRGS FP037-2013A","UM.C\/625\/1\/HIR\/MoE\/E000044-20001"],"award-info":[{"award-number":["RG409-12HTM","FRGS FP037-2013A","UM.C\/625\/1\/HIR\/MoE\/E000044-20001"]}],"id":[{"id":"10.13039\/501100004386","id-type":"DOI","asserted-by":"publisher"}]},{"name":"High Impact Research MoE","award":["RG409-12HTM","FRGS FP037-2013A","UM.C\/625\/1\/HIR\/MoE\/E000044-20001"],"award-info":[{"award-number":["RG409-12HTM","FRGS FP037-2013A","UM.C\/625\/1\/HIR\/MoE\/E000044-20001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Scientific World Journal"],"published-print":{"date-parts":[[2014]]},"abstract":"<jats:p>The Gram-negative saprophyte<jats:italic>Burkholderia pseudomallei<\/jats:italic>is the causative agent of melioidosis, an infectious disease which is endemic in Southeast Asia and northern Australia. This bacterium possesses many virulence factors which are thought to contribute to its survival and pathogenicity. Using a virulent clinical isolate of<jats:italic>B. pseudomallei<\/jats:italic>and an attenuated strain of the same<jats:italic>B. pseudomallei<\/jats:italic>isolate, 6 genes<jats:italic>BPSL2033, BP1026B_I2784, BP1026B_I2780, BURPS1106A_A0094, BURPS1106A_1131<\/jats:italic>, and<jats:italic>BURPS1710A_1419<\/jats:italic>were identified earlier by PCR-based subtractive hybridization. These genes were extensively characterized at the molecular level, together with an additional gene<jats:italic>BPSL3147<\/jats:italic>that had been identified by other investigators. Through a reverse genetic approach, single-gene knockout mutants were successfully constructed by using site-specific insertion mutagenesis and were confirmed by PCR.<jats:italic>BPSL2033::Km<\/jats:italic>and<jats:italic>BURPS1710A_1419::Km<\/jats:italic>mutants showed reduced rates of survival inside macrophage RAW 264.7 cells and also low levels of virulence in the nematode infection model.<jats:italic>BPSL2033::Km<\/jats:italic>demonstrated weak statistical significance (<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M1\"><mml:mi>P<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>0.049<\/mml:mn><\/mml:math>) at 8 hours after infection in macrophage infection study but this was not seen in<jats:italic>BURPS1710A_1419::Km<\/jats:italic>. Nevertheless, complemented strains of both genes were able to partially restore the gene defects in both<jats:italic>in vitro<\/jats:italic>and<jats:italic>in vivo<\/jats:italic>studies, thus suggesting that they individually play a minor role in the virulence of<jats:italic>B. pseudomallei<\/jats:italic>.<\/jats:p>","DOI":"10.1155\/2014\/590803","type":"journal-article","created":{"date-parts":[[2014,8,11]],"date-time":"2014-08-11T17:07:39Z","timestamp":1407776859000},"page":"1-9","source":"Crossref","is-referenced-by-count":3,"title":["Molecular Characterization of Putative Virulence Determinants in<i>Burkholderia pseudomallei<\/i>"],"prefix":"10.1155","volume":"2014","author":[{"given":"Suat Moi","family":"Puah","sequence":"first","affiliation":[{"name":"Department of Biomedical Science, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S. 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