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

Mohd S. Ghazali, Raha A. Raus, Noriha M. Amin, Syamsiah A. Shohaimi, Farah Diba Abu Bakar, Norafiza Zainuddin, Dzun N. Jimat, Nor Azlin Alia Nor Muhammad

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)875-879
Number of pages5
JournalJournal of Pure and Applied Microbiology
Volume8
Issue numberSPEC. ISS. 1
Publication statusPublished - 2014

Fingerprint

Infectious bursal disease virus
Polyproteins
Pichia
Western Blotting
Electroporation
Open Reading Frames
Genes
Disease Outbreaks
Methanol
Epitopes
Plasmids
Proteins

Keywords

  • IBDV expression
  • Pichia pastoris
  • Segment A
  • Vaccine

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Biotechnology
  • Microbiology

Cite this

Ghazali, M. S., Raus, R. A., Amin, N. M., Shohaimi, S. A., Abu Bakar, F. D., Zainuddin, N., ... Nor Muhammad, N. A. A. (2014). Expression of segment a of infectious bursal disease virus in Pichia pastoris. Journal of Pure and Applied Microbiology, 8(SPEC. ISS. 1), 875-879.

Expression of segment a of infectious bursal disease virus in Pichia pastoris. / Ghazali, Mohd S.; Raus, Raha A.; Amin, Noriha M.; Shohaimi, Syamsiah A.; Abu Bakar, Farah Diba; Zainuddin, Norafiza; Jimat, Dzun N.; Nor Muhammad, Nor Azlin Alia.

In: Journal of Pure and Applied Microbiology, Vol. 8, No. SPEC. ISS. 1, 2014, p. 875-879.

Research output: Contribution to journalArticle

Ghazali, MS, Raus, RA, Amin, NM, Shohaimi, SA, Abu Bakar, FD, Zainuddin, N, Jimat, DN & Nor Muhammad, NAA 2014, 'Expression of segment a of infectious bursal disease virus in Pichia pastoris', Journal of Pure and Applied Microbiology, vol. 8, no. SPEC. ISS. 1, pp. 875-879.
Ghazali MS, Raus RA, Amin NM, Shohaimi SA, Abu Bakar FD, Zainuddin N et al. Expression of segment a of infectious bursal disease virus in Pichia pastoris. Journal of Pure and Applied Microbiology. 2014;8(SPEC. ISS. 1):875-879.
Ghazali, Mohd S. ; Raus, Raha A. ; Amin, Noriha M. ; Shohaimi, Syamsiah A. ; Abu Bakar, Farah Diba ; Zainuddin, Norafiza ; Jimat, Dzun N. ; Nor Muhammad, Nor Azlin Alia. / Expression of segment a of infectious bursal disease virus in Pichia pastoris. In: Journal of Pure and Applied Microbiology. 2014 ; Vol. 8, No. SPEC. ISS. 1. pp. 875-879.
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