Expression of segment a of infectious bursal disease virus in pichia pastoris
Recombinant plasmid containing segment A open reading frame 2 (ORF2) gene of infectious bursal disease virus (IBDV) of a very virulent subtype from local outbreak (strain 3529/92) was constructed. The gene encoding the IBDV structural polyprotein (N-VP2-VP3-VP4-C) was inserted into an expression vec...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English English |
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Journal of Pure and Applied Microbiology
2014
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| Online Access: | http://irep.iium.edu.my/39365/ http://irep.iium.edu.my/39365/2/Expression_of_Segment_A_gene.pdf http://irep.iium.edu.my/39365/4/39365_Expression%20of%20segment_Scopus.pdf |
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| author | Ghazali, Mohd. Shahrin Ahmad Raus, Raha Amin, Noriha M. Shohaimi, Syamsiah A. Abu Bakar, Farah D. Zainuddin, Norafiza Jimat, Dzun Noraini Nor Muhammad, Nor Azlin Alia |
| author_facet | Ghazali, Mohd. Shahrin Ahmad Raus, Raha Amin, Noriha M. Shohaimi, Syamsiah A. Abu Bakar, Farah D. Zainuddin, Norafiza Jimat, Dzun Noraini Nor Muhammad, Nor Azlin Alia |
| author_sort | Ghazali, Mohd. Shahrin |
| building | IIUM Repository |
| collection | Online Access |
| description | Recombinant plasmid containing segment A open reading frame 2 (ORF2) gene of infectious bursal disease virus (IBDV) of a very virulent subtype from local outbreak (strain 3529/92) was constructed. The gene encoding the IBDV structural polyprotein (N-VP2-VP3-VP4-C) was inserted into an expression vector, pPICZ prior to its transformation into Pichia pastoris by electroporation. After the induction of P. pastoris transformant with 0.5% methanol, the production of IBDV polyprotein was observed using Western blot. In P. pastoris, co- or post-translational processing of the large polyprotein occurred, generating a stable C-terminal product (VP3) of correct size, but without any detectable N-terminal product (VP2). The failure to observe the VP2 protein in Western blot analysis was probably due to the conformational epitope problem. |
| first_indexed | 2025-11-14T15:54:51Z |
| format | Article |
| id | iium-39365 |
| institution | International Islamic University Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-14T15:54:51Z |
| publishDate | 2014 |
| publisher | Journal of Pure and Applied Microbiology |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | iium-393652024-06-21T01:44:02Z http://irep.iium.edu.my/39365/ Expression of segment a of infectious bursal disease virus in pichia pastoris Ghazali, Mohd. Shahrin Ahmad Raus, Raha Amin, Noriha M. Shohaimi, Syamsiah A. Abu Bakar, Farah D. Zainuddin, Norafiza Jimat, Dzun Noraini Nor Muhammad, Nor Azlin Alia Q Science (General) Recombinant plasmid containing segment A open reading frame 2 (ORF2) gene of infectious bursal disease virus (IBDV) of a very virulent subtype from local outbreak (strain 3529/92) was constructed. The gene encoding the IBDV structural polyprotein (N-VP2-VP3-VP4-C) was inserted into an expression vector, pPICZ prior to its transformation into Pichia pastoris by electroporation. After the induction of P. pastoris transformant with 0.5% methanol, the production of IBDV polyprotein was observed using Western blot. In P. pastoris, co- or post-translational processing of the large polyprotein occurred, generating a stable C-terminal product (VP3) of correct size, but without any detectable N-terminal product (VP2). The failure to observe the VP2 protein in Western blot analysis was probably due to the conformational epitope problem. Journal of Pure and Applied Microbiology 2014-05 Article PeerReviewed application/pdf en http://irep.iium.edu.my/39365/2/Expression_of_Segment_A_gene.pdf application/pdf en http://irep.iium.edu.my/39365/4/39365_Expression%20of%20segment_Scopus.pdf Ghazali, Mohd. Shahrin and Ahmad Raus, Raha and Amin, Noriha M. and Shohaimi, Syamsiah A. and Abu Bakar, Farah D. and Zainuddin, Norafiza and Jimat, Dzun Noraini and Nor Muhammad, Nor Azlin Alia (2014) Expression of segment a of infectious bursal disease virus in pichia pastoris. Journal of Pure and Applied Microbiology, 8 spl. pp. 875-879. ISSN 0973-7510 |
| spellingShingle | Q Science (General) Ghazali, Mohd. Shahrin Ahmad Raus, Raha Amin, Noriha M. Shohaimi, Syamsiah A. Abu Bakar, Farah D. Zainuddin, Norafiza Jimat, Dzun Noraini Nor Muhammad, Nor Azlin Alia Expression of segment a of infectious bursal disease virus in pichia pastoris |
| title | Expression of segment a of infectious bursal disease virus in pichia pastoris |
| title_full | Expression of segment a of infectious bursal disease virus in pichia pastoris |
| title_fullStr | Expression of segment a of infectious bursal disease virus in pichia pastoris |
| title_full_unstemmed | Expression of segment a of infectious bursal disease virus in pichia pastoris |
| title_short | Expression of segment a of infectious bursal disease virus in pichia pastoris |
| title_sort | expression of segment a of infectious bursal disease virus in pichia pastoris |
| topic | Q Science (General) |
| url | http://irep.iium.edu.my/39365/ http://irep.iium.edu.my/39365/2/Expression_of_Segment_A_gene.pdf http://irep.iium.edu.my/39365/4/39365_Expression%20of%20segment_Scopus.pdf |