Construction of recombinant plasmids of VCA protein P23 of Epstein-Barr virus and its expression in Pichia pastoris

【Objective】 To construct the recombinant plasmid pPICZαA-BLRF2 for secretory expression of the BLRF2 recombinant protein efficiently in Pichia pastoris,provide the foundation for the development of the subunit vaccine and the diagnose reagent for the Epstein-Barr virus.【Methods】 EBV-BLRF2 gene was amplified from the B95-8 cell complete genomic DNA by PCR and cloned into plasmid pPICZαA.The recombinant plasmids were transformed into GSl15 yeast by electroporation.The yeast transformants were induced by methanol to express recombinant proteins.The recombinant proteins were identified by Western blotting.【Results】 The recombinant plasmid pPICZαA-BLRF2 was constructed successfully,and the recombinant protein was expressed in the Pichia pastoris prosperously.【Conclusion】 The EBV encoding gene BLRF2 of viral capsid antigens protein P23 were secretory expressed efficiently.The pPICZαA-BLRF2 yeast strain which was valuable in screening for NPC patients was obtained and can be used for further research.