{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T06:16:48Z","timestamp":1775888208751,"version":"3.50.1"},"reference-count":48,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2006,4,1]],"date-time":"2006-04-01T00:00:00Z","timestamp":1143849600000},"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":[[2006,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>In this paper we describe construction of a luciferase-based vector, pPL2<jats:italic>lux<\/jats:italic>, and use of this vector to study gene expression in<jats:italic>Listeria monocytogenes<\/jats:italic>. pPL2<jats:italic>lux<\/jats:italic>is a derivative of the listerial integration vector pPL2 and harbors a synthetic<jats:italic>luxABCDE<\/jats:italic>operon encoding a fatty acid reductase complex (LuxCDE) involved in synthesis of the fatty aldehyde substrate for the bioluminescence reaction catalyzed by the LuxAB luciferase. We constructed pPL2<jats:italic>lux<\/jats:italic>derivatives in which the<jats:italic>secA<\/jats:italic>and<jats:italic>hlyA<\/jats:italic>promoters were translationally fused to<jats:italic>luxABCDE<\/jats:italic>and integrated as a single copy into the chromosome of<jats:italic>L. monocytogenes<\/jats:italic>EGD-e. Growth experiments revealed that<jats:italic>hlyA<\/jats:italic>was expressed predominantly in the stationary phase in LB medium buffered at pH 7.4, whereas<jats:italic>secA<\/jats:italic>expression could be detected in the exponential growth phase. Moreover, the correlation between luciferase activity and transcription levels, as determined by reverse transcriptase PCR, was confirmed using conditions known to lead to repression and activation of hemolysin expression (addition of cellobiose and activated charcoal, respectively). Furthermore, hemolysin expression could be monitored in real time during invasion of an intact monolayer of C2Bbe1 (Caco-2-derived) cells. Finally, hemolysin expression could be detected in the livers, spleens, and kidneys of mice 3 days postinfection. These experiments clearly established the effectiveness of pPL2<jats:italic>lux<\/jats:italic>as a quantitative reporter system for real-time, noninvasive evaluation of gene expression in<jats:italic>L. monocytogenes<\/jats:italic>.<\/jats:p>","DOI":"10.1128\/aem.72.4.2876-2884.2006","type":"journal-article","created":{"date-parts":[[2006,4,6]],"date-time":"2006-04-06T13:43:49Z","timestamp":1144331029000},"page":"2876-2884","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":66,"title":["Novel Luciferase Reporter System for In Vitro and Organ-Specific Monitoring of Differential Gene Expression in<i>Listeria monocytogenes<\/i>"],"prefix":"10.1128","volume":"72","author":[{"given":"Peter A.","family":"Bron","sequence":"first","affiliation":[{"name":"Alimentary Pharmabiotic Centre"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ian R.","family":"Monk","sequence":"additional","affiliation":[{"name":"Alimentary Pharmabiotic Centre"},{"name":"Department of Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sine\u0301ad C.","family":"Corr","sequence":"additional","affiliation":[{"name":"Alimentary Pharmabiotic Centre"},{"name":"Department of Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Colin","family":"Hill","sequence":"additional","affiliation":[{"name":"Alimentary Pharmabiotic Centre"},{"name":"Department of Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cormac G. M.","family":"Gahan","sequence":"additional","affiliation":[{"name":"Alimentary Pharmabiotic Centre"},{"name":"Department of Microbiology"},{"name":"School of Pharmacy, University College Cork, Cork, Ireland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(86)90130-7"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-64.12.2058"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.13.3450-3456.2002"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.2.894-904.2005"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.8.3635-3642.1998"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.62.7.2252-2256.1996"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.1.310-317.2004"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.23.7829-7835.2004"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.57.030502.090934"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2004.04003.x"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.6.3594-3600.2000"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.5.3350-3358.2001"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2672.2005.02559.x"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.6.3924-3932.2001"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.55.7.1641-1646.1987"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1063447"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.57.8.2255-2259.1991"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1092712"},{"key":"e_1_3_2_20_2","first-page":"443","volume":"85","year":"1995","unstructured":"Israelsen, H., S. M. Madsen, E. Johansen, A. Vrang, and E. B. Hansen. 1995. Environmentally regulated promoters in lactococci. Dev. Biol. Stand.85:443-448.","journal-title":"Dev. Biol. Stand."},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.19.5722-5734.2003"},{"key":"e_1_3_2_22_2","first-page":"7374","volume":"72","year":"2005","unstructured":"Kim, H., K. J. Boor, and H. Marquis. 2005. Listeria monocytogenes \u03c3B contributes to invasion of human intestinal epithelial cells. Infect. Immun.72:7374-7378.","journal-title":"Infect. Immun."},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.15.4177-4186.2002"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2002.03072.x"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0505.990502"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/mr.55.1.123-142.1991"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2003.03413.x"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.1.131-139.1999"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.71.3.1356-1363.2005"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(90)90479-B"},{"key":"e_1_3_2_31_2","first-page":"29","volume":"188","year":"2000","unstructured":"Phillips-Jones, M. K. 2000. Use of a lux reporter system for monitoring rapid changes in alpha-toxin gene expression in Clostridium perfringens during growth. FEMS Microbiol. Lett.188:29-33.","journal-title":"FEMS Microbiol. Lett."},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2005.04.035"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.60.2.587-593.1994"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1993.tb06391.x"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.11.7074-7082.2001"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002480000091"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.2.717-727.2004"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/0923-2508(96)84712-7"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.8.4759-4766.2001"},{"key":"e_1_3_2_40_2","unstructured":"Sambrook J. E. F. Fritsch and T. Maniatis. 1989. Molecular cloning: a laboratory manual 2nd ed. Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y."},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7012(01)00362-1"},{"key":"e_1_3_2_42_2","doi-asserted-by":"crossref","first-page":"1183","DOI":"10.1111\/j.1365-2958.2004.04454.x","volume":"55","year":"2005","unstructured":"Sleator, R. D., H. H. Wemekamp-Kamphuis, C. G. Gahan, T. Abee, and C. Hill. 2005. A PrfA-regulated bile exclusion system (BilE) is a novel virulence factor in Listeria monocytogenes. Mol. Microbiol.55:1183-1195.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.50.2.540-542.1985"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.11.4124-4127.1995"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.14.3.584-640.2001"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.63.7.2566-2571.1997"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2004.00414.x"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.5.3152-3159.2005"},{"key":"e_1_3_2_49_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1998.tb13045.x"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.72.4.2876-2884.2006","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.72.4.2876-2884.2006","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,3]],"date-time":"2024-02-03T16:22:53Z","timestamp":1706977373000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.72.4.2876-2884.2006"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,4]]},"references-count":48,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2006,4]]}},"alternative-id":["10.1128\/AEM.72.4.2876-2884.2006"],"URL":"https:\/\/doi.org\/10.1128\/aem.72.4.2876-2884.2006","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,4]]},"assertion":[{"value":"2005-08-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2006-01-16","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2006-04-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}