Expression of Human Cytomegalovirus pp65 Gene in Pichia pastoris

Objective To construct a recombinant Pichia pastoris for expression of human cytomegalovirus (HCMV) pp65336-507 gene. Methods Amplify HCMV pp65336-507 gene from the genome of HCMV by PCR using the designed primers to construct recombinant plasmid pPIC9/pp65336-507. Transform P. pastoris GS115 with the constructed recombinant plasmid, screen positive clones by PCR, and identify the expressed product by SDS-PAGE and Western blot. Results Positive recombinants were successfully screened. The expressed pp65336-507 protein, with a relative molecular mass of about 26 000, reacted with the specific antibody against pp65. Conclusion The recombinant Pichia pastoris for secretory expression of pp65336-507 protein was successfully constructed, and the expressed product showed good reactogenicity.