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dc.contributor.authorBrandariz Núñez, Alberto
dc.contributor.authorMenaya Vargas, Rebeca
dc.contributor.authorBenavente Martínez, Francisco Javier
dc.contributor.authorMartínez Costas, José Manuel
dc.date.accessioned2020-06-06T10:17:44Z
dc.date.available2020-06-06T10:17:44Z
dc.date.issued2010
dc.identifier.citationBrandariz-Nuñez, A., Menaya-Vargas, R., Benavente, J., & Martinez-Costas, J. (2010). A versatile molecular tagging method for targeting proteins to avian reovirus muNS inclusions. Use in protein immobilization and purification. PLoS One, 5(11).
dc.identifier.urihttp://hdl.handle.net/10347/22844
dc.description.abstractBackground: Avian reoviruses replicate in viral factories, which are dense cytoplasmic compartments estabilished by protein-protein interactions. The non-structural protein muNS forms the factory scaffold that attracts other viral components in a controlled fashion. To create such a three-dimensional network, muNS uses several different selfinteracting domains. Methodology/Principal Findings: In this study we have devised a strategy to identify muNS regions containing selfinteracting domains, based on the capacity of muNS-derived inclusions to recruit muNS fragments. The results revealed that the muNS region consisting of residues 477–542 was recruited with the best efficiency, and this raised the idea of using this fragment as a molecular tag for delivering foreign proteins to muNS inclusions. By combining such tagging system with our previously established method for purifying muNS inclusions from baculovirus-infected insect cells, we have developed a novel protein purification protocol. Conclusions/Significance: We show that our tagging and inclusion-targeting system can be a simple, versatile and efficient method for immobilizing and purifying active proteins expressed in baculovirus-infected cells. We also demonstrate that muNS inclusions can simultaneously recruit several tagged proteins, a finding which may be used to generate protein complexes and create multiepitope particulate material for immunization purposes.
dc.description.sponsorshipThis work was supported by grants from the European Commission under contracts ERAS-CT-2003-980409 (as part of the European Science Foundation EUROCORES Programme EuroSCOPE, web: http://www.esf.org/euroscope); the Spanish Ministerio de Ciencia y Tecnología (BFU2007-61330, BFU 205- 24982-E, web: http://www.mityc.es/) and Xunta de Galicia (08CSA009203PR, web: http://www.conselleriaiei.org/ga/dxidi/index.php). ABN was the recipient of a predoctoral FPI fellowship from the Spanish Ministerio de Ciencia y Tecnología
dc.language.isoeng
dc.publisherPLOS
dc.rights© 2010 Brandariz-Nuñez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
dc.rights.urihttp://creativecommons.org/licenses/by/2.0
dc.subjectCytoplasmic inclusions
dc.subjectGreen fluorescent protein
dc.subjectPlasmid construction
dc.subjectPolymerase chain reaction
dc.subjectProtein expression
dc.subjectBaculoviruses
dc.subjectBirds
dc.subjectInsects
dc.titleA Versatile Molecular Tagging Method for Targeting Proteins to Avian Reovirus muNS Inclusions. Use in Protein Immobilization and Purification
dc.typejournal article
dc.identifier.doi10.1371/journal.pone.0013961
dc.relation.publisherversionhttps://doi.org/10.1371/journal.pone.0013961
dc.type.hasVersionVoR
dc.identifier.essn1932-6203
dc.rights.accessRightsopen access
dc.contributor.affiliationUniversidade de Santiago de Compostela. Departamento de Bioquímica e Bioloxía Molecular
dc.description.peerreviewedSI
dc.relation.projectIDinfo:eu-repo/grantAgreement/MEC/Plan Nacional de I+D+i 2004-2007/BFU2007-61330/ES/CARACTERIZACION MOLECULAR DE LOS REOVIRUS AVIARES


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© 2010 Brandariz-Nuñez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
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 © 2010 Brandariz-Nuñez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited





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