{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T07:03:17Z","timestamp":1780383797909,"version":"3.54.1"},"reference-count":39,"publisher":"Proceedings of the National Academy of Sciences","issue":"26","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2001,12,18]]},"abstract":"<jats:p>\n            We have developed a simple and efficient procedure for adding an\n epitope-encoding tail to one or more genes of interest in the bacterial\n chromosome. The procedure is a modification of the gene\n replacement method of Datsenko and Wanner [Datsenko, K. A. &amp;\n Wanner, B. L. (2000)\n            <jats:italic>Proc. Natl. Acad. Sci. USA<\/jats:italic>\n            97,\n 6640\u20136645]. A DNA module that begins with the epitope-encoding\n sequence and includes a selectable marker is amplified by PCR with\n primers that carry extensions (as short as 36 nt) homologous to the\n last portion of the targeted gene and to a region downstream from it.\n Transformation of a strain expressing bacteriophage \u03bb\n            <jats:italic>red<\/jats:italic>\n            functions yields recombinants carrying the targeted\n gene fused to the epitope-encoding sequence. The resulting\n C-terminal-tagged protein can be identified by standard\n immuno-detection techniques. In an initial application of the method,\n we have added the sequences encoding the FLAG and 3xFLAG and influenza\n virus hemagglutinin epitopes to various genes of\n            <jats:italic>Salmonella\n enterica<\/jats:italic>\n            serovar Typhimurium, including putative and\n established pathogenic determinants present in prophage genomes.\n Epitope fusion proteins were detected in bacteria growing\n            <jats:italic>in\n vitro<\/jats:italic>\n            , tissue culture cells, and infected mouse tissues. This\n work identified a prophage locus specifically expressed in\n bacteria growing intracellularly. The procedure described here should\n be applicable to a wide variety of Gram-negative bacteria and is\n particularly suited for the study of intracellular pathogens.\n          <\/jats:p>","DOI":"10.1073\/pnas.261348198","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T14:36:44Z","timestamp":1027694204000},"page":"15264-15269","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":489,"title":["Epitope tagging of chromosomal genes in\n            <i>Salmonella<\/i>"],"prefix":"10.1073","volume":"98","author":[{"given":"Sergio","family":"Uzzau","sequence":"first","affiliation":[{"name":"Centre de G\u00e9n\u00e9tique Mol\u00e9culaire, Centre National\r de la Recherche Scientifique, 91198 Gif-sur-Yvette, France; and\r Department of Biomedical Sciences, Center for\r Biotechnology Development and Biodiversity Research, University of\r Sassari, 07100, Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nara","family":"Figueroa-Bossi","sequence":"additional","affiliation":[{"name":"Centre de G\u00e9n\u00e9tique Mol\u00e9culaire, Centre National\r de la Recherche Scientifique, 91198 Gif-sur-Yvette, France; and\r Department of Biomedical Sciences, Center for\r Biotechnology Development and Biodiversity Research, University of\r Sassari, 07100, Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Salvatore","family":"Rubino","sequence":"additional","affiliation":[{"name":"Centre de G\u00e9n\u00e9tique Mol\u00e9culaire, Centre National\r de la Recherche Scientifique, 91198 Gif-sur-Yvette, France; and\r Department of Biomedical Sciences, Center for\r Biotechnology Development and Biodiversity Research, University of\r Sassari, 07100, Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lionello","family":"Bossi","sequence":"additional","affiliation":[{"name":"Centre de G\u00e9n\u00e9tique Mol\u00e9culaire, Centre National\r de la Recherche Scientifique, 91198 Gif-sur-Yvette, France; and\r Department of Biomedical Sciences, Center for\r Biotechnology Development and Biodiversity Research, University of\r Sassari, 07100, Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2001,12,11]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/21.14.3329"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/yea.320111306"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0061(19980915)14:12<1139::AID-YEA306>3.0.CO;2-B"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0061(199907)15:10B<963::AID-YEA399>3.0.CO;2-W"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.171.5.2609-2613.1989"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/145.4.877"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.17.4750-4752.1998"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/27.5.1296"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2001.02234.x"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.8.2063-2071.1998"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/134.4.1013"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.120163297"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1119(00)00071-8"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.100127597"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(95)00193-A"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1997.4451807.x"},{"key":"e_1_3_3_17_2","first-page":"580","volume":"7","author":"Prickett K S","year":"1989","unstructured":"K S Prickett, D C Amberg, T P Hopp BioTechniques 7, 580\u2013589 (1989).","journal-title":"BioTechniques"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.117.4.1223"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1994.tb06231.x"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.2144\/00284pf01"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.170.6.2575-2583.1988"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.172.11.6557-6567.1990"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.83.14.5189"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2000.01902.x"},{"key":"e_1_3_3_25_2","volume-title":"Genetic Analysis of Pathogenic Bacteria","author":"Maloy S R","year":"1996","unstructured":"S R Maloy, W J Stewart, R K Taylor Genetic Analysis of Pathogenic Bacteria (Cold Spring Harbor Lab. 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