Journal of Pure and Applied MicrobiologyVol. 8 No. May 2014 Special Edition

Expression of Segment a of Infectious Bursal Disease Virus in Pichia pastoris

Mohd S. Ghazali5, Raha A. Raus1*, Noriha M. Amin2, Syamsiah A. Shohaimi3, Farah D. Abu Bakar4, Norafiza Zainuddin5, Dzun N. Jimat1 and Nor Azlin Alia Nor Muhammad5

1Department of Biotechnology Engineering, Kulliyyah of Engineering, International Islamic University Malaysia, P.O. Box 10, 50728 Kuala Lumpur, Malaysia. 2Pusat PenyelidikanBioteknologi, IbuPejabat MARDI, Serdang, Selangor, Malaysia. 3Veterinary Research Institute, Jalan Sultan Azlan Shah, Ipoh, Perak. 4Fakulti SainsdanTeknologi, UniversitiKebangsaan Malaysia, Bangi, Selangor, Malaysia. 5Faculty of Health Sciences, International Islamic University of Malaysia, Kuantan, Pahang, Malaysia.

Received on 08 January 2014 and accepted on 24 March 2014

 

ABSTRACT

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.

Keywords : Segment A, IBDV Expression, Pichia pastoris, Vaccine.