An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
- Autores
- Picotto, Leandro Daniel; Sguazza, Guillermo Hernán; Tizzano, Marco Antonio; Galosi, Cecilia Mónica; Cavalitto, Sebastián Fernando; Pecoraro, Marcelo Ricardo Ítalo
- Año de publicación
- 2017
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- 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.
Facultad de Ciencias Veterinarias
Centro de Investigación y Desarrollo en Fermentaciones Industriales - Materia
-
Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by-nc-sa/4.0/
- Repositorio
.jpg)
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/119258
Ver los metadatos del registro completo
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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:UNLP2025-09-03T11:00:15Zoai: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:13292025-09-03 11:00:16.15SEDICI (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 |
| dc.description.none.fl_txt_mv |
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. Facultad de Ciencias Veterinarias Centro de Investigación y Desarrollo en Fermentaciones Industriales |
| 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 |
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info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Articulo http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
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article |
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publishedVersion |
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http://sedici.unlp.edu.ar/handle/10915/119258 |
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eng |
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eng |
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info:eu-repo/semantics/altIdentifier/issn/1046-5928 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2017.02.004 |
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