{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T21:59:17Z","timestamp":1790546357013,"version":"4.1.0"},"reference-count":30,"publisher":"American Society for Microbiology","issue":"22","license":[{"start":{"date-parts":[[2015,11,15]],"date-time":"2015-11-15T00:00:00Z","timestamp":1447545600000},"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":[[2015,11,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Leptospirosis is a zoonotic disease that affects \u223c1 million people annually, with a mortality rate of &gt;10%. Currently, there is an absence of effective genetic manipulation tools for targeted mutagenesis in pathogenic leptospires. Transcription activator-like effectors (TALEs) are a recently described group of repressors that modify transcriptional activity in prokaryotic and eukaryotic cells by directly binding to a targeted sequence within the host genome. To determine the applicability of TALEs within\n            <jats:named-content content-type=\"genus-species\">Leptospira<\/jats:named-content>\n            spp., two TALE constructs were designed. First, a constitutively expressed TALE gene specific for the\n            <jats:italic>lacO<\/jats:italic>\n            -like region upstream of\n            <jats:italic>bgaL<\/jats:italic>\n            was\n            <jats:italic>trans<\/jats:italic>\n            inserted in the saprophyte\n            <jats:named-content content-type=\"genus-species\">Leptospira biflexa<\/jats:named-content>\n            (the TALE\n            <jats:sub>\u03b2gal<\/jats:sub>\n            strain). Reverse transcriptase PCR (RT-PCR) analysis and enzymatic assays demonstrated that BgaL was not expressed in the TALE\n            <jats:sub>\u03b2gal<\/jats:sub>\n            strain. Second, to study the role of LigA and LigB in pathogenesis, a constitutively expressed TALE gene with specificity for the homologous promoter regions of\n            <jats:italic>ligA<\/jats:italic>\n            and\n            <jats:italic>ligB<\/jats:italic>\n            was\n            <jats:italic>cis<\/jats:italic>\n            inserted into the pathogen\n            <jats:named-content content-type=\"genus-species\">Leptospira interrogans<\/jats:named-content>\n            (TALE\n            <jats:sub>\n              <jats:italic>lig<\/jats:italic>\n            <\/jats:sub>\n            ). LigA and LigB expression was studied by using three independent clones: TALE\n            <jats:sub>\n              <jats:italic>lig1<\/jats:italic>\n            <\/jats:sub>\n            , TALE\n            <jats:sub>\n              <jats:italic>lig2<\/jats:italic>\n            <\/jats:sub>\n            , and TALE\n            <jats:sub>\n              <jats:italic>lig3<\/jats:italic>\n            <\/jats:sub>\n            . Immunoblot analysis of osmotically induced TALE\n            <jats:sub>\n              <jats:italic>lig<\/jats:italic>\n            <\/jats:sub>\n            clones demonstrated 2- to 9-fold reductions in the expression levels of LigA and LigB, with the highest reductions being noted for TALE\n            <jats:sub>\n              <jats:italic>lig1<\/jats:italic>\n            <\/jats:sub>\n            and TALE\n            <jats:sub>\n              <jats:italic>lig2<\/jats:italic>\n            <\/jats:sub>\n            , which were avirulent\n            <jats:italic>in vivo<\/jats:italic>\n            and nonrecoverable from animal tissues. This study reconfirms galactosidase activity in the saprophyte and suggests a role for LigA and LigB in pathogenesis. Collectively, this study demonstrates that TALEs are effective at reducing the expression of targeted genes within saprophytic and pathogenic strains of\n            <jats:named-content content-type=\"genus-species\">Leptospira<\/jats:named-content>\n            spp., providing an additional genetic manipulation tool for this genus.\n          <\/jats:p>","DOI":"10.1128\/aem.02202-15","type":"journal-article","created":{"date-parts":[[2015,9,5]],"date-time":"2015-09-05T02:39:35Z","timestamp":1441420775000},"page":"7888-7892","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":38,"title":["Control of Gene Expression in Leptospira spp. by Transcription Activator-Like Effectors Demonstrates a Potential Role for LigA and LigB in Leptospira interrogans Virulence"],"prefix":"10.1128","volume":"81","author":[{"given":"Christopher J.","family":"Pappas","sequence":"first","affiliation":[{"name":"Institut Pasteur, Unit\u00e9 de Biologie des Spiroch\u00e8tes, Paris, France"},{"name":"Manhattanville College, Department of Biology, Purchase, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathieu","family":"Picardeau","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Unit\u00e9 de Biologie des Spiroch\u00e8tes, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"66","volume-title":"Abstr 8th Int Leptospirosis Soc Sci Meet","author":"Hagan JE","year":"2013","unstructured":"Hagan JE, Costa F, Calcagno J, Kane M, Torgerson P, Martinez-Silveira MS, Stein C, Abela-Ridder B, Ko A. 2013. Global morbidity and mortality of leptospirosis: a systematic review, p 66. Abstr 8th Int Leptospirosis Soc Sci Meet, Fukuoka, Japan, 8 to 11 October 2013."},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-662-45059-8_4"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-662-45059-8_7"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1178811"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1178817"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks404"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1039\/C2CC37107C"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.resmic.2004.09.007"},{"key":"e_1_3_2_10_2","first-page":"45","article-title":"Nutrition of Leptospira pomona and growth of 13 other serotypes: fractionation of oleic albumin complex and a medium of bovine albumin and polysorbate 80","volume":"26","author":"Ellinghausen HC","year":"1965","unstructured":"Ellinghausen HC, McCullough WG. 1965. Nutrition of Leptospira pomona and growth of 13 other serotypes: fractionation of oleic albumin complex and a medium of bovine albumin and polysorbate 80. Am J Vet Res 26:45\u201351.","journal-title":"Am J Vet Res"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.94.1.27-31.1967"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00173-15"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1194"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-11-129"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01619-14"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00663-13"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01803-14"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02172-07"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.05695-11"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0030097"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00531-13"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00713-12"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2003.03619.x"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01635-06"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jir875"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00989-08"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00427-15"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.009175-0"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.meegid.2008.10.012"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0016879"},{"key":"e_1_3_2_31_2","volume-title":"Experiments in molecular genetics","author":"Miller JH","year":"1972","unstructured":"Miller JH. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY."}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02202-15","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02202-15","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T02:55:30Z","timestamp":1645584930000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.02202-15"}},"subtitle":[],"editor":[{"given":"M. A.","family":"Elliot","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2015,11,15]]},"references-count":30,"journal-issue":{"issue":"22","published-print":{"date-parts":[[2015,11,15]]}},"alternative-id":["10.1128\/AEM.02202-15"],"URL":"https:\/\/doi.org\/10.1128\/aem.02202-15","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,11,15]]},"assertion":[{"value":"2015-07-06","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-09-01","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-10-21","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}