Expression of segment a of infectious bursal disease virus in pichia pastoris

• Main Authors: 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
• Format: Article
• Language: English; English
• Published: Journal of Pure and Applied Microbiology 2014
• Subjects: Q Science (General)
• Online Access: 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; http://irep.iium.edu.my/39365/
• Institution: Universiti Islam Antarabangsa Malaysia

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.