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Thus, homologous recombination between introduced DNA and the corresponding chromosomal locus has never been demonstrated for this pathogen. Leptospiral immunoglobulin-like repeat (Lig) proteins were previously identified as putative<jats:italic>Leptospira<\/jats:italic>virulence factors. In this study, a<jats:italic>ligB<\/jats:italic>mutant was constructed by allelic exchange in<jats:italic>L. interrogans<\/jats:italic>; in this mutant a spectinomycin resistance (Spc<jats:sup>r<\/jats:sup>) gene replaced a portion of the<jats:italic>ligB<\/jats:italic>coding sequence. Gene disruption was confirmed by PCR, immunoblot analysis, and immunofluorescence studies. The<jats:italic>ligB<\/jats:italic>mutant did not show decrease virulence compared to the wild-type strain in the hamster model of leptospirosis. In addition, inoculation of rats with the<jats:italic>ligB<\/jats:italic>mutant induced persistent colonization of the kidneys. Finally, LigB was not required to mediate bacterial adherence to cultured cells. Taken together, our data provide the first evidence of site-directed homologous recombination in pathogenic<jats:italic>Leptospira<\/jats:italic>species. Furthermore, our data suggest that LigB does not play a major role in dissemination of the pathogen in the host and in the development of acute disease manifestations or persistent renal colonization.<\/jats:p>","DOI":"10.1128\/iai.00989-08","type":"journal-article","created":{"date-parts":[[2008,9,23]],"date-time":"2008-09-23T00:25:42Z","timestamp":1222129542000},"page":"5826-5833","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":103,"title":["Targeted Mutagenesis in Pathogenic<i>Leptospira<\/i>Species: Disruption of the LigB Gene Does Not Affect Virulence in Animal Models of Leptospirosis"],"prefix":"10.1128","volume":"76","author":[{"given":"Julio","family":"Croda","sequence":"first","affiliation":[{"name":"Gonc\u0327alo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil"},{"name":"Laboratory of Pathology of Transmissible Diseases, Department of Pathology, Medicine School, Sa\u0303o Paulo University, Sa\u0303o Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudio Pereira","family":"Figueira","sequence":"additional","affiliation":[{"name":"Gonc\u0327alo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"Elsio A.","family":"Wunder","sequence":"additional","affiliation":[{"name":"Gonc\u0327alo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cleiton S.","family":"Santos","sequence":"additional","affiliation":[{"name":"Gonc\u0327alo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitermayer G.","family":"Reis","sequence":"additional","affiliation":[{"name":"Gonc\u0327alo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert I.","family":"Ko","sequence":"additional","affiliation":[{"name":"Gonc\u0327alo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil"},{"name":"Division of International Medicine and Infectious Disease, Weill Medical College of Cornell University, New York, New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathieu","family":"Picardeau","sequence":"additional","affiliation":[{"name":"Unite\u0301 de Biologie des Spiroche\u0300tes, Institut Pasteur, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2007.10.012"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-8-70"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00460-06"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.70.12.6926-6932.2002"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.26065-0"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(03)00830-2"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.9.3255-3258.2005"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0603979103"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01635-06"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.02344-06"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsre.2003.10.004"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.8.4853-4863.2005"},{"key":"e_1_3_2_14_2","first-page":"45","volume":"26","year":"1965","unstructured":"Ellinghausen, H. 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