Construction and Secreted Expression of Recombinant Procarboxypeptidase B in Pichia pastoris

Objective To construct the recombinant procarboxypeptidase B in Pichia pastoris.Methods The yeast plasmid containing procarboxypeptidase B coding gene was constructed,and then transferred to Pichia pastoris.The positive clone was obtained by G418 resistance selection in His-medium.The rabbit anti-rat recombinant procarboxypeptidase B serum was prepared with recombinant procarboxypeptidase B expressed in E.coli.Western-blotting analysis was used to determine the expressed protein with the prepared antiserum after fermentation and methanol induction of recombinant yeast.Results The rabbit anti-rat recombinant procarboxypeptidase B serum with higher antibody titer was got.The result of Western-blotting analysis showed that the supernatant was positive with the antiserum after methanol induction of recombinant yeast.Conclusion The recombinant procarboxypeptidase B in Pichia pastoris can be constructed with the successfulsecretion of recombinant procarboxypeptidase B.