{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T14:16:49Z","timestamp":1792678609853,"version":"4.5.0"},"reference-count":55,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2003,4,1]],"date-time":"2003-04-01T00:00:00Z","timestamp":1049155200000},"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 Virol"],"published-print":{"date-parts":[[2003,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The genome of the coronavirus transmissible gastroenteritis virus (TGEV) has been engineered as an expression vector with an infectious cDNA. The vector led to the efficient (&gt;40 \u03bcg\/10\n            <jats:sup>6<\/jats:sup>\n            cells) and stable (&gt;20 passages) expression of a heterologous gene (green fluorescent protein [GFP]), driven by the transcription-regulating sequences (TRS) of open reading frame (ORF) 3a inserted in the site previously occupied by the nonessential ORFs 3a and 3b. Expression levels driven by this TRS were higher than those of an expression cassette under the control of regulating sequences engineered with the N gene TRS. The recombinant TGEV including the GFP gene was still enteropathogenic, albeit with a 10- to 10\n            <jats:sup>2<\/jats:sup>\n            -fold reduction in enteric tissue growth. Interestingly, a specific lactogenic immune response against the heterologous protein has been elicited in sows and their progeny. The engineering of an additional insertion site for the heterologous gene between viral genes N and 7 led to instability and to a new genetic organization of the 3\u2032 end of the recombinant viruses. As a consequence, a major species of subgenomic mRNA was generated from a TRS with the noncanonical core sequence 5\u2032-CUAAAA-3\u2032. Extension of the complementarity between the TRS and sequences at the 3\u2032 end of the viral leader was associated with transcriptional activation of noncanonical core sequences. The engineered vector led to expression levels as high as those of well-established vectors and seems very promising for the development of vaccines and, possibly, for gene therapy.\n          <\/jats:p>","DOI":"10.1128\/jvi.77.7.4357-4369.2003","type":"journal-article","created":{"date-parts":[[2003,3,12]],"date-time":"2003-03-12T18:46:25Z","timestamp":1047494785000},"page":"4357-4369","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":90,"title":["Engineering the Transmissible Gastroenteritis Virus Genome as an Expression Vector Inducing Lactogenic Immunity"],"prefix":"10.1128","volume":"77","author":[{"given":"Isabel","family":"Sola","sequence":"first","affiliation":[{"name":"Centro Nacional de Biotecnologi\u0301a, CSIC, Department of Molecular and Cell Biology, Campus Universidad Auto\u0301noma, Cantoblanco, Madrid"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Alonso","sequence":"additional","affiliation":[{"name":"Centro Nacional de Biotecnologi\u0301a, CSIC, Department of Molecular and Cell Biology, Campus Universidad Auto\u0301noma, Cantoblanco, Madrid"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonia","family":"Zu\u0301n\u0303iga","sequence":"additional","affiliation":[{"name":"Centro Nacional de Biotecnologi\u0301a, CSIC, Department of Molecular and Cell Biology, Campus Universidad Auto\u0301noma, Cantoblanco, Madrid"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mo\u0301nica","family":"Balasch","sequence":"additional","affiliation":[{"name":"Fort-Dodge Veterinaria, Department of Research and Development, Girona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Plana-Dura\u0301n","sequence":"additional","affiliation":[{"name":"Fort-Dodge Veterinaria, Department of Research and Development, Girona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Enjuanes","sequence":"additional","affiliation":[{"name":"Centro Nacional de Biotecnologi\u0301a, CSIC, Department of Molecular and Cell Biology, Campus Universidad Auto\u0301noma, Cantoblanco, Madrid"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.22.12989"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.10.5516"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.3.1293-1308.2002"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-83-3-567"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1998.9059"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1006\/smvy.1997.0109"},{"key":"e_1_3_2_8_2","unstructured":"Nidoviruses 2000"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-002-0847-x"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.3.1422-1434.2002"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2001.0908"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4615-2996-5_45"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.7.4291-4298.1996"},{"key":"e_1_3_2_14_2","first-page":"835","year":"2000","unstructured":"Enjuanes, L., D. 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