{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T01:53:25Z","timestamp":1770429205008,"version":"3.49.0"},"reference-count":24,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2003,1,1]],"date-time":"2003-01-01T00:00:00Z","timestamp":1041379200000},"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":[[2003,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Controlled expression of cloned \u03c6X174 gene\n            <jats:italic>E<\/jats:italic>\n            in gram-negative bacteria results in lysis of the bacteria by the formation of a transmembrane tunnel structure built through the cell envelope complex. Production of bacterial ghosts is routinely monitored by classical microbiological procedures. These include determination of the turbidity of the culture and the total number of cells and the number of reproductive cells present during the time course of growth and lysis. Although conceptually simple, these methods are labor intensive and time consuming, providing a complete set of results after the determination of viable cell counts. To avoid culturing methods for bacterial growth, an alternative flow cytometric procedure is presented for the quantification of ghosts and polarized, as well as depolarized, nonlysed cells within a culture. For this method, which is based on the discriminatory power of the membrane potential-sensitive dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol, a staining protocol was developed and optimized for the maximum discrepancy in fluorescence between bacterial ghosts and viable cells. The total quantitative analysis procedure takes less than 2 min. The results derived from classical or cytometric analyses correlate with respect to the total cell numbers and the viability of the culture.\n          <\/jats:p>","DOI":"10.1128\/aem.69.1.468-474.2003","type":"journal-article","created":{"date-parts":[[2003,1,3]],"date-time":"2003-01-03T22:02:44Z","timestamp":1041631364000},"page":"468-474","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":40,"title":["Online Monitoring of\n            <i>Escherichia coli<\/i>\n            Ghost Production"],"prefix":"10.1128","volume":"69","author":[{"given":"W.","family":"Haidinger","sequence":"first","affiliation":[{"name":"Apovia AG, D-82152 Martinsried, Germany"},{"name":"Institute of Microbiology and Genetics, University of Vienna, UZAI, A-1090 Vienna"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. P.","family":"Szostak","sequence":"additional","affiliation":[{"name":"Apovia AG, D-82152 Martinsried, Germany"},{"name":"BIRD-C GmbH &amp; Co. KEG, A-1080 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W.","family":"Jechlinger","sequence":"additional","affiliation":[{"name":"Apovia AG, D-82152 Martinsried, Germany"},{"name":"Institute of Microbiology and Genetics, University of Vienna, UZAI, A-1090 Vienna"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W.","family":"Lubitz","sequence":"additional","affiliation":[{"name":"Institute of Microbiology and Genetics, University of Vienna, UZAI, A-1090 Vienna"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.65.8.3681-3689.1999"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0320(19970901)29:1<58::AID-CYTO6>3.0.CO;2-9"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(98)00423-X"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1656(00)00315-1"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0009-3084(94)90035-3"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.175.21.7105-7108.1993"},{"key":"e_1_3_2_8_2","unstructured":"Gilstrap M. J. G. Kleyn and E. W. Nester. 1983. Experiments in microbiology 2nd ed. CBS College Publishing Philadelphia Pa."},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0320(20010601)44:2<106::AID-CYTO1088>3.0.CO;2-5"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(00)00107-9"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-3659(99)00099-1"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.7.2696-2701.1995"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.9.2001"},{"key":"e_1_3_2_14_2","first-page":"359","volume":"3","year":"1980","unstructured":"Lam, J. S., D. M. Granoff, J. R. Gilsdorf, and W. Costerton. 1980. Immunogenicity of outer membrane derivatives of Haemophilus influenzae type B. Curr. Microbiol.3:359-364.","journal-title":"Immunogenicity of outer membrane derivatives of Haemophilus influenzae type B. Curr. Microbiol."},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0320(19971201)29:4<298::AID-CYTO6>3.0.CO;2-6"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2672.1995.tb05031.x"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.12.2752"},{"key":"e_1_3_2_18_2","unstructured":"Miller J. H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y."},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2672.1998.00436.x"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.1995.tb00203.x"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/0168-1656(95)00123-9"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.169.4.1750-1752.1987"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1989.tb14661.x"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00248685"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1997.5781941.x"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.69.1.468-474.2003","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.69.1.468-474.2003","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T13:57:13Z","timestamp":1645624633000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.69.1.468-474.2003"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,1]]},"references-count":24,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2003,1]]}},"alternative-id":["10.1128\/AEM.69.1.468-474.2003"],"URL":"https:\/\/doi.org\/10.1128\/aem.69.1.468-474.2003","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2003,1]]},"assertion":[{"value":"2002-06-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2002-10-31","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2003-01-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}