ISSN: 0973-7510

E-ISSN: 2581-690X

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.
J Pure Appl Microbiol. 2014;8(Spl. Edn. 1):875-879
© The Author(s). 2014
Received: 08/01/2014 | Accepted: 24/03/2014 | Published: 31/05/2014
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

Article Metrics

Article View: 0

Share This Article

© The Author(s) 2014. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.