Expression, purification and glycosylation analysis of chicken infectious bursal disease virus VP2 in yeast

Document Type : Full paper (Original article)

Authors

1 State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, PR, China

2 MSc Student in Biology, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, PR, China

3 Ph.D. Student in Biology, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, PR, China

Abstract

Infectious bursal disease (IBD), a highly contagious and devastating disease in young chicken, is caused
by infectious bursal disease virus (IBDV). To improve the immunogenicity of recombinant IBDV subunit
vaccine, an attempt was made to find a new way to prepare IBD vaccine containing glycosylated mVP2
antigen. Firstly, IBDV mVP2 gene (with a nucleic acid sequence encoding B cell epitope of IBDV
(KFDQML) in the 5′-end of the VP2, with a nucleic acid sequence encoding B cell epitope of IBDV (LASP)
and (His) 6-tag in the 3′-end of the VP2) was cloned. Secondly, IBDV mVP2 protein was expressed in the
methylotrophic yeast Pichia pastoris which can secret glycosylated protein. The recombinant mVP2 protein
could be stained pink with periodic acid-schiff reagents (PAS), which showed that mVP2 was glycosylated.
Finally, IBDV mVP2 protein was purified with His-Trap (1 mL) affinity chromatography. These results
indicate that glycosylated IBDV VP2 protein modified with epitope peptides can be expressed in Pichia
pastoris, which lay the groundwork for the development of a recombinant infectious bursal disease vaccine
with high immunogenicity.

Keywords

Files

Share

How to cite

Statistics