Abstract:A pair of PCR primers, with EcoR I and Not I restricted enzyme sites in the 5， and 3， end respectively, was designed according to the major capsid protein (MCP) gene of turbot viral reddish body iridovirus (TRBIV). After PCR amplification, a recombinant expression vector including α factor peptide and 6×His tag was constructed by inserting TRBIV MCP gene into GAP promoter downstream of pGAPZαA directly. The constructed vector pGAPZαA MCP was linearized by Avr Ⅱ, and then transformed and intergrated into the host Pichia pastoris X 33 genome by electroporation. The recombinant yeasts with high level secreted expression of MCP were screened by ELISA Dot. The expressed proteins were identified by SDS PAGE and Western blot. The results revealed that a recombinant protein with the molecular weight of approximately 51kDa was secreted into the supernatant from the recombinant yeast cells successfully. The expressional amount of TRBIV MCP could reach to 60.2 μg/ml in the supernatant secreted from recombinant yeast after being fermented at 28 °C for 72 h.
