{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T10:08:24Z","timestamp":1784887704665,"version":"3.55.0"},"reference-count":21,"publisher":"American Society for Microbiology","issue":"23","license":[{"start":{"date-parts":[[2014,12,1]],"date-time":"2014-12-01T00:00:00Z","timestamp":1417392000000},"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":["J Bacteriol"],"published-print":{"date-parts":[[2014,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            CTX\u03a6, a filamentous vibriophage encoding cholera toxin, uses a unique strategy for its lysogeny. The single-stranded phage genome forms intramolecular base-pairing interactions between two inversely oriented XerC and XerD binding sites (XBS) and generates a functional phage attachment site,\n            <jats:italic>attP<\/jats:italic>\n            (\n            <jats:italic>+<\/jats:italic>\n            ), for integration. The\n            <jats:italic>attP<\/jats:italic>\n            (\n            <jats:italic>+<\/jats:italic>\n            ) structure is recognized by the host-encoded tyrosine recombinases XerC and XerD (XerCD), which enables irreversible integration of CTX\u03a6 into the chromosome dimer resolution site (\n            <jats:italic>dif<\/jats:italic>\n            ) of\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Vibrio cholerae<\/jats:named-content>\n            . The\n            <jats:italic>dif<\/jats:italic>\n            site and the XerCD recombinases are widely conserved in bacteria. We took advantage of these conserved attributes to develop a broad-host-range integrative expression vector that could irreversibly integrate into the host chromosome using XerCD recombinases without altering the function of any known open reading frame (ORF). In this study, we engineered two different arabinose-inducible expression vectors, pBD62 and pBD66, using XBS of CTX\u03a6. pBD62 replicates conditionally and integrates efficiently into the\n            <jats:italic>dif<\/jats:italic>\n            of the bacterial chromosome by site-specific recombination using host-encoded XerCD recombinases. The expression level of the gene of interest could be controlled through the P\n            <jats:sub>BAD<\/jats:sub>\n            promoter by modulating the functions of the vector-encoded transcriptional factor AraC. We validated the irreversible integration of pBD62 into a wide range of pathogenic and nonpathogenic bacteria, such as\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">V. cholerae<\/jats:named-content>\n            ,\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Vibrio fluvialis<\/jats:named-content>\n            ,\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Vibrio parahaemolyticus<\/jats:named-content>\n            ,\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Escherichia coli<\/jats:named-content>\n            ,\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Salmonella enterica<\/jats:named-content>\n            , and\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Klebsiella pneumoniae<\/jats:named-content>\n            . Gene expression from the P\n            <jats:sub>BAD<\/jats:sub>\n            promoter of integrated vectors was confirmed in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">V. cholerae<\/jats:named-content>\n            using the well-studied reporter genes\n            <jats:italic>mCherry<\/jats:italic>\n            ,\n            <jats:italic>eGFP<\/jats:italic>\n            , and\n            <jats:italic>lacZ<\/jats:italic>\n            .\n          <\/jats:p>","DOI":"10.1128\/jb.01966-14","type":"journal-article","created":{"date-parts":[[2014,9,16]],"date-time":"2014-09-16T07:57:53Z","timestamp":1410854273000},"page":"4071-4080","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":14,"title":["A Novel, Broad-Range, CTX\u03a6-Derived Stable Integrative Expression Vector for Functional Studies"],"prefix":"10.1128","volume":"196","author":[{"given":"Bhabatosh","family":"Das","sequence":"first","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reena","family":"Kumari","sequence":"additional","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Archana","family":"Pant","sequence":"additional","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sourav","family":"Sen Gupta","sequence":"additional","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shruti","family":"Saxena","sequence":"additional","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ojasvi","family":"Mehta","sequence":"additional","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gopinath Balakrish","family":"Nair","sequence":"additional","affiliation":[{"name":"Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.67.2.238-276.2003"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2004.12.008"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2009.06653.x"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1017061108"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.272.5270.1910"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1997.3911758.x"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature00782"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2005.07.002"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0910212107"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0006531"},{"key":"e_1_3_3_12_2","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1002\/j.1460-2075.1995.tb07142.x","article-title":"Site-specific recombination in the replication terminus region of Escherichia coli: functional replacement of dif","volume":"14","author":"Leslie NR","year":"1995","unstructured":"LeslieNRSherrattDJ. 1995. 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