{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T06:25:36Z","timestamp":1789971936733,"version":"4.0.1"},"reference-count":38,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2002,5,21]],"date-time":"2002-05-21T00:00:00Z","timestamp":1021939200000},"content-version":"vor","delay-in-days":780,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2000,4]]},"abstract":"<jats:p> <jats:italic>Listeria monocytogenes<\/jats:italic> is capable of growth within the cytoplasm of infected host cells. Escape from the host cell phagosome is mediated primarily through secretion of listeriolysin, a haemolytic factor which functions to actively lyse the phagosomal membrane. Listeriolysin negative mutants of <jats:italic>L. monocytogenes<\/jats:italic> are non\u2010haemolytic on blood agar plates and demonstrate a significant reduction of virulence in the mouse model of infection. We have developed a system for the identification of <jats:italic>in vivo<\/jats:italic> induced genes in <jats:italic>L. monocytogenes<\/jats:italic> which utilizes the listeriolysin gene, <jats:italic>hly<\/jats:italic>, as both a reporter of gene expression and as a means of selection of promoter elements expressed <jats:italic>in vivo<\/jats:italic>. The system is analogous to <jats:italic>in vivo<\/jats:italic> expression technology (IVET) first reported for <jats:italic>Salmonella<\/jats:italic>, however, as listeriolysin functions in the environment of the host phagosome the loci identified in this study are most likely expressed during residence in the phagosome. The system was successfully tested using the promoter of the inducible virulence gene <jats:italic>plcA<\/jats:italic>. A bank was created by fusing a promoterless copy of <jats:italic>hly<\/jats:italic> to random promoter elements in a listeriolysin negative IVET host. Sequential inoculations of mice with this bank resulted in the isolation of clones with increased survival potential in the mouse model relative to a negative control, but which remained haemolysin negative on blood agar plates. Nine <jats:italic>in vivo<\/jats:italic> induced loci were identified including genes encoding a DNA topoisomerase III, a cellobiose transporter and a fumarase. Two isolates represented fusions to proteins of unknown function and three isolates contained no significant homologues in the database. A mutant in the fumarase gene demonstrated reduced virulence for mice and an inability to grow in cultured mouse phagocytes.<\/jats:p>","DOI":"10.1046\/j.1365-2958.2000.01869.x","type":"journal-article","created":{"date-parts":[[2003,3,12]],"date-time":"2003-03-12T11:29:48Z","timestamp":1047468588000},"page":"498-507","source":"Crossref","is-referenced-by-count":48,"title":["The use of listeriolysin to identify <i>in vivo<\/i> induced genes in the Gram\u2010positive intracellular pathogen <i>Listeria monocytogenes<\/i>"],"prefix":"10.1111","volume":"36","author":[{"given":"Cormac G. 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