{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T17:41:20Z","timestamp":1793814080183,"version":"4.6.1"},"reference-count":62,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2001,1,1]],"date-time":"2001-01-01T00:00:00Z","timestamp":978307200000},"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":[[2001,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:italic>Burkholderia pseudomallei<\/jats:italic>\n            , the etiologic agent of melioidosis, is responsible for a broad spectrum of illnesses in humans and animals particularly in Southeast Asia and northern Australia, where it is endemic.\n            <jats:italic>Burkholderia thailandensis<\/jats:italic>\n            is a nonpathogenic environmental organism closely related to\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            . Subtractive hybridization was carried out between these two species to identify genes encoding virulence determinants in\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            . Screening of the subtraction library revealed A-T-rich DNA sequences unique to\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            , suggesting they may have been acquired by horizontal transfer. One of the subtraction clones, pDD1015, encoded a protein with homology to a glycosyltransferase from\n            <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n            . This gene was insertionally inactivated in wild-type\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            to create SR1015. It was determined by enzyme-linked immunosorbent assay and immunoelectron microscopy that the inactivated gene was involved in the production of a major surface polysaccharide. The 50% lethal dose (LD\n            <jats:sub>50<\/jats:sub>\n            ) for wild-type\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            is &lt;10 CFU; the LD\n            <jats:sub>50<\/jats:sub>\n            for SR1015 was determined to be 3.5 \u00d7 10\n            <jats:sup>5<\/jats:sup>\n            CFU, similar to that of\n            <jats:italic>B. thailandensis<\/jats:italic>\n            (6.8 \u00d7 10\n            <jats:sup>5<\/jats:sup>\n            CFU). DNA sequencing of the region flanking the glycosyltransferase gene revealed open reading frames similar to capsular polysaccharide genes in\n            <jats:italic>Haemophilus influenzae<\/jats:italic>\n            ,\n            <jats:italic>Escherichia coli<\/jats:italic>\n            , and\n            <jats:italic>Neisseria meningitidis<\/jats:italic>\n            . In addition, DNA from\n            <jats:italic>Burkholderia mallei<\/jats:italic>\n            and\n            <jats:italic>Burkholderia stabilis<\/jats:italic>\n            hybridized to a glycosyltransferase fragment probe, and a capsular structure was identified on the surface of\n            <jats:italic>B. stabilis<\/jats:italic>\n            via immunoelectron microscopy. Thus, the combination of PCR-based subtractive hybridization, insertional inactivation, and animal virulence studies has facilitated the identification of an important virulence determinant in\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            .\n          <\/jats:p>","DOI":"10.1128\/iai.69.1.34-44.2001","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:01:19Z","timestamp":1027764079000},"page":"34-44","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":221,"title":["Detection of Bacterial Virulence Genes by Subtractive Hybridization: Identification of Capsular Polysaccharide of\n            <i>Burkholderia pseudomallei<\/i>\n            as a Major Virulence Determinant"],"prefix":"10.1128","volume":"69","author":[{"given":"Shauna L.","family":"Reckseidler","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Center, Calgary, Alberta, Canada T2N 4N1,1 and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"DeShazer","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Bacteriology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 217022"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pamela A.","family":"Sokol","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Center, Calgary, Alberta, Canada T2N 4N1,1 and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donald E.","family":"Woods","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Center, Calgary, Alberta, Canada T2N 4N1,1 and"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1992.tb01351.x"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5331.1453"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.2144\/97225bm04"},{"key":"e_1_3_2_5_2","first-page":"1","article-title":"Genetics of capsular polysaccharide production in bacteria","volume":"150","author":"Boulnois G. 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