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Expression of Porcine Epizootic Diarrhea Virus S1 Gene in Pichia pastoris

TAN Bo-min, YU Xi-yao, ZHONG Ze-min, JIANG Zhi-qiong, HUANG Yu-mao   

  1.  (College of Veterinary MedicineSouth China Agricultural UniversityGuangzhou 510642,China)

  • Revised:2014-01-17 Online:2014-06-20 Published:2014-07-24

Abstract: To develop a new vaccines can efficiently express the porcine epizootic diarrhea virus S1 antigen epitope genes mainly,the recombinant plasmid pPICZαC-S1 was constructed by integrat S1 antigen epitope genes mainly into yeast expression vector pPICZαC and transformed into Pichia pastoris X33 by electroporation after linearized by Sac Ⅰ restriction enzymes. The positive transformants (pPICZαC-S1-X33) were screened by ZeocinTM. The transformants were inoculated into culture medium and induced with methanol. The result was demonstrated by SDS-PAGE and Western blotting, and showed the mainly antigen epitope of spike protein was secreted successfully expression in Pichia pastoris. Expression of the protein molecular weight for 42 ku, and the ability to combine with PEDV positive serum specificity.

Key words:

porcine epizootic diarrhea virus (PEDV); antigenic epitope; secreted expression; Pichia pastoris