Cloning and Expression of ORF 2 Protein of PCV2 in Pichia Pastoris

Abstract

Porcine circovirus type 2 (PCV2) is a major pathogen responsible for post-weaning multisystemic wasting syndrome in pigs, causing significant economic losses in the swine industry. The ORF2 gene of PCV2 encodes the capsid (Cap) protein, the primary immunogenic component and a key target for vaccine development and serological diagnostics. In this study, the ORF2 gene was cloned and expressed in Pichia pastoris X-33 to establish a eukaryotic expression system for the recombinant Cap protein. Codon optimization was employed to enhance expression efficiency in the yeast host. Successful cloning was confirmed by restriction analysis and PCR, while expression of recombinant Cap protein was verified by SDS-PAGE and western blotting. These results highlight P. pastoris as a robust platform for the large-scale production of PCV2 Cap protein, providing a foundation for future subunit vaccine development and improved diagnostic strategies against PCV2 infection.