An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris

The glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted...

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Detalhes bibliográficos
Autores: Picotto, Leandro Daniel, Sguazza, Guillermo Hernán, Tizzano, Marco Antonio, Galosi, Cecilia Mónica, Cavalitto, Sebastián Fernando, Pecoraro, Marcelo Ricardo Ítalo
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2017
País:Argentina
Recursos:Universidad Nacional de La Plata
Repositorio:SEDICI (UNLP)
Idioma:inglés
OAI Identifier:oai:sedici.unlp.edu.ar:10915/119258
Acesso em linha:http://sedici.unlp.edu.ar/handle/10915/119258
Access Level:acceso abierto
Palavra-chave:Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
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spelling An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastorisPicotto, Leandro DanielSguazza, Guillermo HernánTizzano, Marco AntonioGalosi, Cecilia MónicaCavalitto, Sebastián FernandoPecoraro, Marcelo Ricardo ÍtaloBioquímicaPichia pastorisDO-statRabies virusGlycoproteinThe glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted protein, using a simplified DO-stat control feeding strategy. This strategy involves the addition of methanol when the dissolved oxygen (DO) level rises above the setpoint avoiding methanol accumulation and oxygen limitation. The G-protein expression was evaluated by SDS-PAGE, ELISA, and western blot assays. Like native G-protein, the recombinant G-protein was found reactive when it was challenged against specific antibodies. The data indicate that the recombinant G-protein can be easily expressed and isolated, and may be useful as a safe source in the production of diagnostic kits and subunit vaccines to prevent rabies.Facultad de Ciencias VeterinariasCentro de Investigación y Desarrollo en Fermentaciones Industriales2017info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf124-130http://sedici.unlp.edu.ar/handle/10915/119258enginfo:eu-repo/semantics/altIdentifier/issn/1046-5928info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2017.02.004info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2024-05-08T13:06:18Zoai:sedici.unlp.edu.ar:10915/119258Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292024-05-08 13:06:19.137SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
title An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
spellingShingle An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
Picotto, Leandro Daniel
Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
title_short An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
title_full An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
title_fullStr An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
title_full_unstemmed An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
title_sort An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
dc.creator.none.fl_str_mv Picotto, Leandro Daniel
Sguazza, Guillermo Hernán
Tizzano, Marco Antonio
Galosi, Cecilia Mónica
Cavalitto, Sebastián Fernando
Pecoraro, Marcelo Ricardo Ítalo
author Picotto, Leandro Daniel
author_facet Picotto, Leandro Daniel
Sguazza, Guillermo Hernán
Tizzano, Marco Antonio
Galosi, Cecilia Mónica
Cavalitto, Sebastián Fernando
Pecoraro, Marcelo Ricardo Ítalo
author_role author
author2 Sguazza, Guillermo Hernán
Tizzano, Marco Antonio
Galosi, Cecilia Mónica
Cavalitto, Sebastián Fernando
Pecoraro, Marcelo Ricardo Ítalo
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
topic Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
description The glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted protein, using a simplified DO-stat control feeding strategy. This strategy involves the addition of methanol when the dissolved oxygen (DO) level rises above the setpoint avoiding methanol accumulation and oxygen limitation. The G-protein expression was evaluated by SDS-PAGE, ELISA, and western blot assays. Like native G-protein, the recombinant G-protein was found reactive when it was challenged against specific antibodies. The data indicate that the recombinant G-protein can be easily expressed and isolated, and may be useful as a safe source in the production of diagnostic kits and subunit vaccines to prevent rabies.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://sedici.unlp.edu.ar/handle/10915/119258
url http://sedici.unlp.edu.ar/handle/10915/119258
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/1046-5928
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2017.02.004
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
124-130
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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score 15.811543

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