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dc.contributor.author | Corral, Teresa | |
dc.contributor.author | Ver, Lorena S | |
dc.contributor.author | Mottet, Geneviève | |
dc.contributor.author | Cano, Olga | |
dc.contributor.author | Garcia-Barreno, Blanca | |
dc.contributor.author | Calder, Lesley J | |
dc.contributor.author | Skehel, John J | |
dc.contributor.author | Roux, Laurent | |
dc.contributor.author | Melero, Jose Antonio | |
dc.date.accessioned | 2019-02-20T11:59:45Z | |
dc.date.available | 2019-02-20T11:59:45Z | |
dc.date.issued | 2007-04-05 | |
dc.identifier.citation | BMC Biotechnol. 2007 Apr 5;7:17. | es_ES |
dc.identifier.issn | 1472-6750 | es_ES |
dc.identifier.uri | http://hdl.handle.net/20.500.12105/7195 | |
dc.description.abstract | BACKGROUND: Embryonated chicken eggs have been used since the mid-20th century to grow a wide range of animal viruses to high titers. However, eggs have found so far only limited use in the production of recombinant proteins. We now describe a system, based on a Sendai virus minigenome, to produce large amounts of heterologous viral glycoproteins in the allantoic cavity of embryonated eggs. RESULTS: Soluble forms of human respiratory syncytial virus (HRSV) and human metapneumovirus (HMPV) fusion (F) proteins, devoid of their transmembrane and cytoplasmic domains, were produced in allantoic fluids using the Sendai minigenome system. The first step was rescuing in cell cultures Sendai virus minigenomes encoding the proteins of interest, with the help of wild type Sendai virus. The second step was propagating such recombinant defective viruses, together with the helper virus, in the allantoic cavity of chicken embryonated eggs, and passage to optimize protein production. When compared with the production of the same proteins in the culture supernatant of cells infected with vaccinia recombinants, the yield in the allantoic fluid was 5-10 fold higher. Mutant forms of these soluble proteins were easily constructed by site-directed mutagenesis and expressed in eggs using the same approach. CONCLUSION: The simplicity and economy of the Sendai minigenome system, together with the high yield achieved in the allantoic fluid of eggs, makes it an attractive method to express soluble glycoproteins aimed for structural studies. | es_ES |
dc.description.sponsorship | This work was supported in part by grants PM99-9014 and SAF2006-07805 from Ministerio de Educación y Ciencia and 01/24 from ISCIII to J.A.M. and by the Medical Research Council to J.J.S and L.J.C. G.M. and L.R. have been supported by grants from the Swiss National Foundation for the Scientific Research, from the Académique de Geneva and from the Swisslife Stiftung. L.S.V. was recipient of a predoctoral fellowship from Ministerio de Educación y Ciencia (Spain). | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | BioMed Central (BMC) | es_ES |
dc.type.hasVersion | VoR | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | * |
dc.subject.mesh | Allantois | es_ES |
dc.subject.mesh | Animals | es_ES |
dc.subject.mesh | Body Fluids | es_ES |
dc.subject.mesh | Chick Embryo | es_ES |
dc.subject.mesh | Chickens | es_ES |
dc.subject.mesh | Cricetinae | es_ES |
dc.subject.mesh | Genome, Viral | es_ES |
dc.subject.mesh | Glycoproteins | es_ES |
dc.subject.mesh | Humans | es_ES |
dc.subject.mesh | Ovum | es_ES |
dc.subject.mesh | Sendai virus | es_ES |
dc.subject.mesh | Solubility | es_ES |
dc.subject.mesh | Viral Proteins | es_ES |
dc.title | High level expression of soluble glycoproteins in the allantoic fluid of embryonated chicken eggs using a Sendai virus minigenome system | es_ES |
dc.type | journal article | es_ES |
dc.rights.license | Atribución 4.0 Internacional | * |
dc.identifier.pubmedID | 17411439 | es_ES |
dc.format.volume | 7 | es_ES |
dc.format.number | 1 | es_ES |
dc.format.page | 17 | es_ES |
dc.identifier.doi | 10.1186/1472-6750-7-17 | es_ES |
dc.contributor.funder | Ministerio de Educación y Ciencia (España) | |
dc.contributor.funder | Instituto de Salud Carlos III | |
dc.description.peerreviewed | Sí | es_ES |
dc.relation.publisherversion | https://doi.org/10.1186/1472-6750-7-17 | es_ES |
dc.identifier.journal | BMC biotechnology | es_ES |
dc.repisalud.centro | ISCIII::Centro Nacional de Microbiología | es_ES |
dc.repisalud.institucion | ISCIII | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/ES/PM99-9014 | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/ES/SAF2006-07805 | es_ES |
dc.rights.accessRights | open access | es_ES |