{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T08:14:45Z","timestamp":1777364085225,"version":"3.51.4"},"reference-count":55,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2008,5,1]],"date-time":"2008-05-01T00:00:00Z","timestamp":1209600000000},"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":[[2008,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:italic>Francisella tularensis<\/jats:italic>\n            is the etiologic agent of an intracellular systemic infection of the lymphatic system in humans called tularemia. The organism has become the subject of considerable research interest due to its classification as a category A select agent by the CDC. To aid genetic analysis of this pathogen, we have constructed a temperature-sensitive Tn\n            <jats:italic>5<\/jats:italic>\n            -based transposon delivery system that is capable of generating chromosomal reporter fusions with\n            <jats:italic>lacZ<\/jats:italic>\n            or\n            <jats:italic>luxCDABE<\/jats:italic>\n            , enabling us to monitor gene expression. Transposition is catalyzed by the hyperactive Tn\n            <jats:italic>5<\/jats:italic>\n            transposase, whose expression is driven by the\n            <jats:italic>Francisella groES<\/jats:italic>\n            promoter. When high-temperature selection (42\u00b0C) is applied to a bacterial culture carrying the transposon delivery plasmid, \u223c0.1% of the population is recovered with Tn\n            <jats:italic>5<\/jats:italic>\n            insertions in the chromosome. Nucleotide sequence analysis of a sample of mutants revealed that the insertions occur randomly throughout the chromosome. The kanamycin-selectable marker of the transposon is also flanked by FLP recombination target sequences that allow deletion of the antibiotic resistance gene when desired. This system has been used to generate transposon mutant libraries for the\n            <jats:italic>F. tularensis<\/jats:italic>\n            live vaccine strain as well as two different virulent\n            <jats:italic>F. tularensis<\/jats:italic>\n            strains. Chromosomal reporters delivered with the transposon were used to identify genes upregulated by growth in Chamberlain's defined medium. Genes in the\n            <jats:italic>fsl<\/jats:italic>\n            operon, reported to be involved in iron acquisition, as well as genes in the\n            <jats:italic>igl<\/jats:italic>\n            gene cluster were among those identified by the screen. Further experiments implicate the ferric uptake regulator (Fur) protein in the negative regulation of\n            <jats:italic>fsl<\/jats:italic>\n            but not\n            <jats:italic>igl<\/jats:italic>\n            reporters, which occurs in an iron-dependent manner. Our results indicate that we have created a valuable new transposon that can be used to identify and characterize virulence genes in\n            <jats:italic>F. tularensis<\/jats:italic>\n            strains.\n          <\/jats:p>","DOI":"10.1128\/aem.02882-07","type":"journal-article","created":{"date-parts":[[2008,3,15]],"date-time":"2008-03-15T00:59:48Z","timestamp":1205542788000},"page":"2637-2645","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":27,"title":["Identification of Differentially Regulated\n            <i>Francisella tularensis<\/i>\n            Genes by Use of a Newly Developed Tn\n            <i>5<\/i>\n            -Based Transposon Delivery System"],"prefix":"10.1128","volume":"74","author":[{"given":"Blake W.","family":"Buchan","sequence":"first","affiliation":[{"name":"Department of Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Molly K.","family":"McLendon","sequence":"additional","affiliation":[{"name":"Department of Microbiology"},{"name":"Inflammation Program, Department of Medicine, Roy J. and Lucille A. Carver School of Medicine, University of Iowa, Iowa City, Iowa 52242"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bradley D.","family":"Jones","sequence":"additional","affiliation":[{"name":"Department of Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.69.1.600-606.2003"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.59.9.3291-3296.1991"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-137-12-2697"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1998.00926.x"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1999.tb13646.x"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.60.2.690-693.1992"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/170.4.841"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/am.13.2.232-235.1965"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0264-410X(94)90284-4"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.6.3204-3217.2004"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.60.12.5164-5171.1992"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2000.tb08874.x"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00247a016"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(84)90135-5"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2001.02696.x"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.169.6.2624-2630.1987"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01975-05"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.69.4.2209-2216.2003"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.181.20.6223-6229.1999"},{"key":"e_1_3_2_21_2","first-page":"349","volume":"60","year":"1994","unstructured":"Fortier, A. H., S. J. Green, T. Polsinelli, T. R. Jones, R. M. Crawford, D. A. Leiby, K. L. Elkins, M. S. Meltzer, and C. A. Nacy. 1994. Life and death of an intracellular pathogen: Francisella tularensis and the macrophage. Immunol. Ser.60:349-361.","journal-title":"Immunol. Ser."},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0606713104"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.5.3042-3047.2004"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.71.10.5940-5950.2003"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1038\/72017"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2002.tb11369.x"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.11.6901-6904.2004"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2001.tb10928.x"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00203-002-0442-2"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1097(03)00820-6"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2006.00595.x"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01811-06"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.45685-0"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.7.3134-3141.1998"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.6.2.137"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.29121-0"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.12.7511-7519.2004"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.72.3.1878-1885.2006"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0406287"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.24.7638-7643.1997"},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1099\/13500872-140-12-3309"},{"key":"e_1_3_2_42_2","unstructured":"Experiments in molecular genetics. 1972"},{"key":"e_1_3_2_43_2","first-page":"186","volume":"4","year":"1994","unstructured":"Mokrievich, A. N., V. V. Fursov, and V. M. Pavlov. 1994. Plasmid cryotransformation. Problemy Osobo Opasnyh Infektsy Saratov4:186-190. (In Russian.)","journal-title":"Problemy Osobo Opasnyh Infektsy Saratov"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-695X.1996.tb00248.x"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-695X.1996.tb00247.x"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.13.4426-4428.1997"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1991.tb01991.x"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-6-69"},{"key":"e_1_3_2_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2007.03.002"},{"key":"e_1_3_2_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.56.5.1194-1202.1988"},{"key":"e_1_3_2_51_2","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0306219"},{"key":"e_1_3_2_52_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01865-06"},{"key":"e_1_3_2_53_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00027-06"},{"key":"e_1_3_2_54_2","doi-asserted-by":"publisher","DOI":"10.1183\/09031936.03.00088903"},{"key":"e_1_3_2_55_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0609675104"},{"key":"e_1_3_2_56_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.5.2644-2654.2005"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02882-07","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02882-07","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T16:56:06Z","timestamp":1645635366000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.02882-07"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,5]]},"references-count":55,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2008,5]]}},"alternative-id":["10.1128\/AEM.02882-07"],"URL":"https:\/\/doi.org\/10.1128\/aem.02882-07","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,5]]},"assertion":[{"value":"2007-12-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-03-05","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-05-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}