{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T13:47:42Z","timestamp":1791294462708,"version":"4.1.0"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2005,4,1]],"date-time":"2005-04-01T00:00:00Z","timestamp":1112313600000},"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":["J Bacteriol"],"published-print":{"date-parts":[[2005,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>The unusual and complex cell wall of pathogenic mycobacteria plays a major role in pathogenesis, with specific complex lipids acting as defensive, offensive, or adaptive effectors of virulence. The phthiocerol and phthiodiolone dimycocerosate esters (PDIMs) comprise one such category of virulence-enhancing lipids. Recent work in several laboratories has established that the<jats:italic>Mycobacterium tuberculosis fadD26-mmpL7<\/jats:italic>(Rv2930-Rv2942) locus plays a major role in PDIM biosynthesis and secretion and that PDIM is required for virulence. Here we describe two independent transposon mutants (WAg533 and WAg537) of<jats:italic>Mycobacterium bovis<\/jats:italic>, both of which carry an insertion in Mb0100 (=<jats:italic>M. tuberculosis<\/jats:italic>Rv0097) to reveal a new locus involved in PDIM biosynthesis. The mutations have a polar effect on expression of the downstream genes Mb0101, Mb0102 (<jats:italic>fadD10<\/jats:italic>), Mb0103, and Mb0104 (<jats:italic>nrp<\/jats:italic>), and Mb0100 is shown to be in an operon comprising these genes and Mb0099. Reverse transcription-PCR analysis shows elevated transcription of genes in the operon upstream from the transposon insertion sites in both mutants. Both mutants have altered colony morphology and do not synthesize PDIMs or glycosylphenol-PDIM. Both mutants are avirulent in a guinea pig model of tuberculosis, and when tested as a vaccine, WAg533 conferred protective immunity against<jats:italic>M. bovis<\/jats:italic>infection at least equal to that afforded by<jats:italic>M. bovis<\/jats:italic>bacillus Calmette-Gue\u0301rin.<\/jats:p>","DOI":"10.1128\/jb.187.7.2267-2277.2005","type":"journal-article","created":{"date-parts":[[2005,3,17]],"date-time":"2005-03-17T21:41:33Z","timestamp":1111095693000},"page":"2267-2277","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":58,"title":["Transposon Mutagenesis of Mb0100 at the<i>ppe1<\/i>-<i>nrp<\/i>Locus in<i>Mycobacterium bovis<\/i>Disrupts Phthiocerol Dimycocerosate (PDIM) and Glycosylphenol-PDIM Biosynthesis, Producing an Avirulent Strain with Vaccine Properties At Least Equal to Those of<i>M. bovis<\/i>BCG"],"prefix":"10.1128","volume":"187","author":[{"given":"Grant S.","family":"Hotter","sequence":"first","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barry J.","family":"Wards","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pania","family":"Mouat","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gurdyal S.","family":"Besra","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Birmingham, Edgbaston, Birmingham"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Gomes","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Birmingham, Edgbaston, Birmingham"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"Singh","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shalome","family":"Bassett","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pamela","family":"Kawakami","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul R.","family":"Wheeler","sequence":"additional","affiliation":[{"name":"Veterinary Laboratories Agency, New Haw, Addlestone, Surrey, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geoffrey W.","family":"de Lisle","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Desmond M.","family":"Collins","sequence":"additional","affiliation":[{"name":"AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"16741","DOI":"10.1074\/jbc.272.27.16741","volume":"272","year":"1997","unstructured":"Azad, A. K., T. D. Sirakova, N. D. Fernandes, and P. E. Kolattukudy. 1997. Gene knockout reveals a novel gene cluster for the synthesis of a class of cell wall lipids unique to pathogenic mycobacteria. J. Biol. Chem.272:16741-16745.","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"4787","DOI":"10.1073\/pnas.93.10.4787","volume":"93","year":"1996","unstructured":"Azad, A. K., T. D. Sirakova, L. M. Rogers, and P. E. Kolattukudy. 1996. Targeted replacement of the mycocerosic acid synthase gene in Mycobacterium bovis BCG produces a mutant that lacks mycosides. Proc. Natl. Acad. Sci. USA93:4787-4792.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_4_2","first-page":"91","volume":"101","year":"1998","unstructured":"Besra, G. S. 1998. Preparation of cell-wall fractions from mycobacteria. Methods Mol. Biol.101:91-107.","journal-title":"Methods Mol. 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