Objective: This is a ground work in preparation for the development of a vaccine against tumor, in which 17-1A glucoprotein, a tumor associated antigen (TAA), was cloned and expressed in Pichia pastoris.
Methods: The cDNA of 17-1A gene was cloned from nephridial tissue of mouse embryo by RT-PCR and subsequently inserted into pPICZaA plasmid after digestion with Xho I and Xba I. The linearized recombinant plasmid was transformed into Pichia pastoris strain GS115. 17-1A glucoproteins were secreted into culture supernatant through alpha-signal peptide of AOX I after inducation with methanol. The protein products were analyzed and characterized by SDS-PAGE and Western-blot.
Results: The plasmid pPICZalphaA-17-1A was constructed and verified by sequenceing. 17-1A glucoproteins were expressed stably and glycosylated in Pichia pastoris.
Conclusion: Glycosylated tumor associated antigen 17-1A can be expressed in Pichia pastoris.