CLONING AND EXPRESSION OF STREET RABIES VIRUS GLYCOPROTEIN GENE IN YEAST (Pichia pastoris)
| dc.contributor.advisor | RAMANJINI GOWDA, P. H. | |
| dc.contributor.author | NAGESHA, S. N. | |
| dc.date.accessioned | 2017-02-23T05:49:04Z | |
| dc.date.available | 2017-02-23T05:49:04Z | |
| dc.date.issued | 2009-05-16 | |
| dc.description.abstract | Rabies is a preventable infectious viral disease of mammals, most often transmitted through the bite of rabid animals and causes significant number of annual deaths in rural areas of Asia and Africa. Immunization with rabies vaccine is the most effective means of preventing rabies virus infection. As the presently available vaccine are expensive and involves risk of handling live virus hence inexpensive, safe and efficacious vaccines are urgently needed to address the infectious viral diseases. The yeast, Pichia pastoris, acts as an ideal system for expression of heterologous genes, as the post translational modifications like glycosylation, protein folding successfully occurs in the yeast and which secretes out the heterologous protein into media which makes protein purification simple. The glycoprotein of rabies virus is most antigenic and immunogenic determinant present in rabies virion and can serve as effective target for development of vaccine. Hence the street rabies glycoprotein gene (1.6 Kbp) was cloned into pBKS+ plasmid vector which was characterized by PCR, restriction digestion and by sequencing, further the rabies glycoprotein gene was subcloned into yeast transfer vector (pPICZαA) for expression. Linearised recombinant vector containing glycoprotein gene was introduced into yeast (Pichia pastoris, GS115). The recombinant yeast clones were confirmed by PCR analysis. The recombinant protein expressed in yeast using methanol at 0.5% (v/v) and the expression of glycoprotein gene product was confirmed by SDS-PAGE, the 70kDa protein confirmed the antigen specificity by western blot and ELISA. The expressed recombinant rabies glycoprotein could be used as vaccine after the immunization trials in animals. | en_US |
| dc.identifier.other | Th-9273 | |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810002297 | |
| dc.language.iso | en | en_US |
| dc.pages | 137 | en_US |
| dc.publisher | University of Agricultural Sciences GKVK, Bangalore | en_US |
| dc.sub | Plant Biotechnology | en_US |
| dc.subject | null | en_US |
| dc.theme | EXPRESSION OF STREET RABIES | en_US |
| dc.these.type | Ph.D | en_US |
| dc.title | CLONING AND EXPRESSION OF STREET RABIES VIRUS GLYCOPROTEIN GENE IN YEAST (Pichia pastoris) | en_US |
| dc.type | Thesis | en_US |