Expression of LMP2A Gene of Epstein-Barr Virus in Pichia pastoris

Objective: To construct a recombinant plasmid pPICZαA-LMP2A for the expression of latent-infection membrane protein 2A(LMP2A) gene in Pichia pastoris. Methods: The plasmid pPCIcc containing the full length of LMP2A cDNA was digested with EcoR I and Not I and cloned into expression vector pPICZαA. The constructed plasmid pPICZαA-LMP2A was linarized with Sac I and transformed to Pichia pastoris GS115. The transformants were selected by planting cells on YPD/Zeocine plates, and then identified by PCR and expressed in BMGY and BMMY media. The expression product was analyzed by SDS-PAGE. Results: Based on the result of the 12% SDS-PAGE and western-blot, a recombinant protein about 53 KDa expressed in the yeast supernatant was identified to be recombinant LMP2A. Conclusion: The LMP2A cDNA successful expression was the foundation of further research in its fuction and constructed the suitable genetic engineering vaccine against EBV associated disease.