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  • Title: [Expression and characterization of mouse 17-1A antigen from Pichia pastoris].
    Author: Wang YS, Tian L, Diao P, Hu M, Du XB, Wei YQ.
    Journal: Sichuan Da Xue Xue Bao Yi Xue Ban; 2006 Jul; 37(4):506-9. PubMed ID: 16909589.
    Abstract:
    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.
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