Construction of Recombinant pPIC9K-IFNα-2b and Expression in Pichia pastoris

Objective: A recombinanted DNA Pichia pastroris was constructed to express mature IFNα-2b with Pichia pastroris expression system. Methods: According to the synonymous codon bias of Pichia pastoris, The cDNA fragment coding for mature IFNα-2b was designed and synthesized. Then the gene was inserted into the plasmid pPIC9K of Pichia pastoris to obtain secretory plasmid pPIC9K-IFNα-2b. After electroporation of Pichia pastoris GS115 (His-Mut+),some high-copy transformants (His+Muts) were selected by G418. After 5 days induction, the result was tested by SDS-PAGE. Results: PCR and DNA sequencing results showed that the recombined plasmid was constructed successfully. SDS-PAGE result showed that IFNα-2b was expressed successfully. Conclusion: IFNα-2b was successfully expressed in Pichia pastroris.