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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Autores principales: Picotto, Leandro Daniel, Sguazza, Guillermo Hernán, Tizzano, Marco Antonio, Galosi, Cecilia Mónica, Cavalitto, Sebastián Fernando, Pecoraro, Marcelo Ricardo Ítalo
Formato: Articulo
Lenguaje:Inglés
Publicado: 2017
Materias:
Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/119258
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-119258
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
spellingShingle Bioquímica
Pichia pastoris
DO-stat
Rabies virus
Glycoprotein
Picotto, Leandro Daniel
Sguazza, Guillermo Hernán
Tizzano, Marco Antonio
Galosi, Cecilia Mónica
Cavalitto, Sebastián Fernando
Pecoraro, Marcelo Ricardo Ítalo
An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
topic_facet 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.
format Articulo
Articulo
author Picotto, Leandro Daniel
Sguazza, Guillermo Hernán
Tizzano, Marco Antonio
Galosi, Cecilia Mónica
Cavalitto, Sebastián Fernando
Pecoraro, Marcelo Ricardo Ítalo
author_facet Picotto, Leandro Daniel
Sguazza, Guillermo Hernán
Tizzano, Marco Antonio
Galosi, Cecilia Mónica
Cavalitto, Sebastián Fernando
Pecoraro, Marcelo Ricardo Ítalo
author_sort Picotto, Leandro Daniel
title An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
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 effective and simplified do-stat control strategy for production of rabies glycoprotein in pichia pastoris
publishDate 2017
url http://sedici.unlp.edu.ar/handle/10915/119258
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