{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T09:27:52Z","timestamp":1791278872458,"version":"4.1.0"},"reference-count":33,"publisher":"American Society for Microbiology","issue":"24","license":[{"start":{"date-parts":[[2008,12,15]],"date-time":"2008-12-15T00:00:00Z","timestamp":1229299200000},"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":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2008,12,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:italic>Burkholderia psedudomallei<\/jats:italic>\n            is the etiologic agent of melioidosis, and the bacterium is listed as a potential agent of bioterrorism because of its low infectious dose, multiple infectious routes, and intrinsic antibiotic resistance. To further accelerate research with this understudied bacterium, we developed a\n            <jats:italic>Himar1<\/jats:italic>\n            -based random mutagenesis system for\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            (\n            <jats:italic>HimarBP<\/jats:italic>\n            ). The transposons contain a Flp recombinase-excisable, approved kanamycin resistance selection marker and an R6K origin of replication for transposon rescue. In vivo mutagenesis of virulent\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            strain 1026b was highly efficient, with up to 44% of cells transformed with the delivery plasmid harboring chromosomal\n            <jats:italic>HimarBP<\/jats:italic>\n            insertions. Southern analyses revealed single insertions with no evidence of delivery plasmid maintenance. Sequence analysis of rescued\n            <jats:italic>HimarBP<\/jats:italic>\n            insertions revealed random insertions on both chromosomes within open reading frames and intergenic regions and that the orientation of insertions was largely unbiased. Auxotrophic mutants were obtained at a frequency of 0.72%, and nutritional supplementation experiments supported the functional assignment of genes within the respective biosynthetic pathways.\n            <jats:italic>HimarBP<\/jats:italic>\n            insertions were stable in the absence of selection and could be readily transferred between naturally transformable strains. Experiments with\n            <jats:italic>B. thailandensis<\/jats:italic>\n            suggest that the newly developed\n            <jats:italic>HimarBP<\/jats:italic>\n            transposons can also be used for random mutagenesis of other\n            <jats:italic>Burkholderia<\/jats:italic>\n            spp., especially the closely related species\n            <jats:italic>B. mallei<\/jats:italic>\n            . Our results demonstrate that comprehensive transposon libraries of\n            <jats:italic>B. pseudomallei<\/jats:italic>\n            can be generated, providing additional tools for the study of the biology, pathogenesis, and antibiotic resistance of this pathogen.\n          <\/jats:p>","DOI":"10.1128\/aem.01973-08","type":"journal-article","created":{"date-parts":[[2008,10,25]],"date-time":"2008-10-25T01:10:51Z","timestamp":1224897051000},"page":"7529-7535","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":31,"title":["In Vivo\n            <i>Himar1<\/i>\n            Transposon Mutagenesis of\n            <i>Burkholderia pseudomallei<\/i>"],"prefix":"10.1128","volume":"74","author":[{"given":"Drew A.","family":"Rholl","sequence":"first","affiliation":[{"name":"Department of Microbiology, Immunology and Pathology, Rocky Mountain Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research, Colorado State University, Fort Collins, Colorado 80523-1690"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lily A.","family":"Trunck","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Pathology, Rocky Mountain Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research, Colorado State University, Fort Collins, Colorado 80523-1690"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Herbert P.","family":"Schweizer","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Pathology, Rocky Mountain Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research, Colorado State University, Fort Collins, Colorado 80523-1690"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.15.8927"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.012602299"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.8.2644-2652.2003"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00531-08"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1099\/00207713-48-1-317"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.18.2.383-416.2005"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02430-07"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.7.2710-2715.1998"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.7.2116-2125.1997"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.2144\/96205bm05"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0606713104"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0403302101"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2036282100"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0511100103"},{"key":"e_1_3_2_16_2","first-page":"13","volume":"9","year":"1987","unstructured":"Liss, L. 1987. 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