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dc.contributor.author
Serena, Maria Soledad
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dc.contributor.author
Geisler, Christoph
dc.contributor.author
Metz, German Ernesto
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dc.contributor.author
Corva, Santiago Gerardo
dc.contributor.author
Mórtola, Eduardo Carlos
dc.contributor.author
Larsen, Alejandra
dc.contributor.author
Jarvis, Donald L.
dc.contributor.author
Echeverria, Maria Gabriela
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dc.date.available
2017-01-18T20:27:47Z
dc.date.issued
2013-07
dc.identifier.citation
Serena, Maria Soledad; Geisler, Christoph; Metz, German Ernesto; Corva, Santiago Gerardo; Mórtola, Eduardo Carlos; et al.; Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs; Elsevier; Protein Expression And Purification; 90; 1; 7-2013; 1-8
dc.identifier.issn
1046-5928
dc.identifier.uri
http://hdl.handle.net/11336/11596
dc.description.abstract
Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.subject
Suid Herpesvirus 1
dc.subject
Baculovirus
dc.subject
Insect Cell
dc.subject
Glycoprotein
dc.subject
Elisa
dc.subject
Gc Content
dc.subject.classification
Ciencias Veterinarias
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dc.subject.classification
Ciencias Veterinarias
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dc.subject.classification
CIENCIAS AGRÍCOLAS
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dc.title
Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-01-17T19:04:26Z
dc.journal.volume
90
dc.journal.number
1
dc.journal.pagination
1-8
dc.journal.pais
Países Bajos
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dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: Serena, Maria Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.fil
Fil: Geisler, Christoph. University of Wyoming. Department of Molecular Biology; Estados Unidos
dc.description.fil
Fil: Metz, German Ernesto. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.fil
Fil: Corva, Santiago Gerardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
dc.description.fil
Fil: Mórtola, Eduardo Carlos. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
dc.description.fil
Fil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
dc.description.fil
Fil: Jarvis, Donald L.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
dc.description.fil
Fil: Echeverria, Maria Gabriela. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.journal.title
Protein Expression And Purification
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.pep.2013.04.008
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592813000752
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://pubmed.ncbi.nlm.nih.gov/23631926/
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