{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:53:01Z","timestamp":1791247981355,"version":"4.1.0"},"reference-count":44,"publisher":"National Academy of Sciences","issue":"14","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1999,7,6]]},"abstract":"<jats:p>\n                    <jats:italic>Legionella pneumophila<\/jats:italic>\n                    is the cause of Legionnaires\u2019 disease, which is a form of potentially fatal pneumonia. To identify genes required for virulence of the bacterium, a library of 1,386\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    signature tagged transposon mutants was studied for guinea pig virulence. The mutants were screened in pools of 96 each in a guinea pig model of\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    pneumonia. Sixteen unique mutant clones were determined to have attenuated virulence after being screened twice in the animal model. All 16 mutants failed to multiply in both lungs and spleens. Four of the sixteen had no apparent defect for intracellular multiplication in macrophages. Partial DNA sequences of the interrupted genes adjacent to the transposon insertions showed that six of them had mutations in five known\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    virulence genes:\n                    <jats:italic>dotB<\/jats:italic>\n                    ,\n                    <jats:italic>dotF\/icmG<\/jats:italic>\n                    ,\n                    <jats:italic>dotO\/icmB<\/jats:italic>\n                    ,\n                    <jats:italic>icmX<\/jats:italic>\n                    , and\n                    <jats:italic>proA<\/jats:italic>\n                    . Three of the sequenced clones contained mutations in genes without known homology to other published bacterial genes, and seven clones appeared to be homologous to five different known bacterial genes but are still being characterized. With this methodology, we demonstrate the existence of\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    genes responsible for non-macrophage-related virulence. The discovery of\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    virulence genes indicates the utility of the signature tagged mutagenesis technique for pulmonary pathogens.\n                  <\/jats:p>","DOI":"10.1073\/pnas.96.14.8190","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:40:52Z","timestamp":1027680052000},"page":"8190-8195","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":114,"title":["Discovery of virulence genes of\n                    <i>Legionella pneumophila<\/i>\n                    by using signature tagged mutagenesis in a guinea pig pneumonia model"],"prefix":"10.1073","volume":"96","author":[{"given":"Paul H.","family":"Edelstein","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine and Department of Medicine, University of Pennsylvania School of Medicine, 3400 Spruce Street, Philadelphia, PA 19104; First Department of Internal Medicine, University of the Ryukyus Faculty of Medicine, Okinawa 903-01, Japan; and Department of Microbiology and Immunology, Stanford University School of Medicine, Fairchild D-039, 299 Campus Drive, Stanford, CA 94305"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martha A. 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