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 |
| Published: |
Journal of Pure and Applied Microbiology
2014
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/39365/ 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 |
| Summary: | 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. |
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