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An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris
- Source :
- SEDICI (UNLP), Universidad Nacional de La Plata, instacron:UNLP
- Publication Year :
- 2017
- Publisher :
- Academic Press Inc Elsevier Science, 2017.
-
Abstract
- 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.<br />Facultad de Ciencias Veterinarias<br />Centro de Investigación y Desarrollo en Fermentaciones Industriales
- Subjects :
- 0301 basic medicine
Bioquímica
DO-STAT
PICHIA PASTORIS
030106 microbiology
medicine.disease_cause
Pichia
law.invention
Pichia pastoris
03 medical and health sciences
Viral Envelope Proteins
Western blot
law
DO-stat
Otras Ciencias Veterinarias
medicine
Host cell surface
chemistry.chemical_classification
biology
medicine.diagnostic_test
Ciencias Veterinarias
Rabies virus
GLYCOPROTEIN
RABIES VIRUS
medicine.disease
biology.organism_classification
Virology
Recombinant Proteins
030104 developmental biology
chemistry
CIENCIAS AGRÍCOLAS
biology.protein
Recombinant DNA
Rabies
Antibody
Glycoprotein
Biotechnology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- SEDICI (UNLP), Universidad Nacional de La Plata, instacron:UNLP
- Accession number :
- edsair.doi.dedup.....c455ddf86960c2d1bc562df5b0560258