RT Journal Article
T1 A Versatile Molecular Tagging Method for Targeting Proteins to Avian Reovirus muNS Inclusions. Use in Protein Immobilization and Purification
A1 Brandariz Núñez, Alberto
A1 Menaya Vargas, Rebeca
A1 Benavente Martínez, Francisco Javier
A1 Martínez Costas, José Manuel
K1 Cytoplasmic inclusions
K1 Green fluorescent protein
K1 Plasmid construction
K1 Polymerase chain reaction
K1 Protein expression
K1 Baculoviruses
K1 Birds
K1 Insects
AB Background: Avian reoviruses replicate in viral factories, which are dense cytoplasmic compartments estabilished byprotein-protein interactions. The non-structural protein muNS forms the factory scaffold that attracts other viralcomponents 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 thatthe muNS region consisting of residues 477–542 was recruited with the best efficiency, and this raised the idea of using thisfragment as a molecular tag for delivering foreign proteins to muNS inclusions. By combining such tagging system with ourpreviously established method for purifying muNS inclusions from baculovirus-infected insect cells, we have developed anovel protein purification protocol.Conclusions/Significance: We show that our tagging and inclusion-targeting system can be a simple, versatile and efficientmethod for immobilizing and purifying active proteins expressed in baculovirus-infected cells. We also demonstrate thatmuNS inclusions can simultaneously recruit several tagged proteins, a finding which may be used to generate proteincomplexes and create multiepitope particulate material for immunization purposes.
PB PLOS
YR 2010
FD 2010
LK http://hdl.handle.net/10347/22844
UL http://hdl.handle.net/10347/22844
LA eng
NO Brandariz-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).
NO This work was supported by grants from the European Commission under contracts ERAS-CT-2003-980409 (as part of the European ScienceFoundation 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 apredoctoral FPI fellowship from the Spanish Ministerio de Ciencia y Tecnología
DS Minerva
RD 28 abr 2026