Cloning and expression of VP2 and A segment genes of infectious bursal disease virus in yeast.
2011
Goudarzi, Hossein | Pourbakhsh, Seyed Ali | Shahbazzadeh, Delavar | Azizi, Mohammad
Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease, an infectious disease of global economic importance in poultry. The heterologous proteins in P.pastoris are fast, simple and inexpensive. In this study, VP2 and segment A - encoding genes of CLassical D78 IBDV were amplified using reverse transcription (RT) Polymerase chain reaction (PCR) and Cloned into pPICZ A and pPICZB vectors. Recombinant plasmid DNA were integrated into the chromosome of the transformed pichia pastoris by electroporation and expressed protein identified by SDS- PAGE and western blotting.A recombinant protein was secreted into the supernatant from the yeast and nonsecreted VP2 protein when induced with methanol. The expressed supernatant and pellets were bound with chicken anti IBDV PolyCLonal antibodies. Western blotting with antibodies against D78 IBDV indicated that the recombinant VP2 protein retained its antigenicity.The concentration of secreted VP2 protein was 0.67mg/l and nonsecreted was 6.26mg/l. The production of recombinant VP2 Protein indicated that p.pastoris was an efficient secreted and nonsecreted expression system for D78 IBDV. In this study segment A not expressed in yeast. Key words :IBDV- VP2- A Segment- Cloning- Expression- Pichia Pastoris.
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