Construction of a Pastoris expression vector for a fusion protein consisting of anti-HBsAg scfv and interferon-α

PurposeTo construct an Pastoris expression vector for anti-HBsAg ScFv and Interferon-а fusion protein.MethodsThe target protein gene was constructed using DNA overlap PCR technology, and then was cloned to an expression vector pPICZаB.After DNA sequencing,the resulting plasmid pPICZaB-HBScFv-IFN-а was linearized and transformed into P.Pastoris GS115.The recombinant Pichia strains,identified by direct PCR and zeocin-resistant screening of Pichia transformants,were cultured and induced with methanol,andthe expression production was detected by SDS-PAGE.ResultsDNA sequencing confirmed that the target gene has correctly cloned to the expression vector.The results of direct PCR showed that the target gene had correctly transformed into GS115,and a 44 000 recombinant protein was observed by SDS-PAGE after being induced with methanol.ConclusionA Pastoris expression vector for anti-HBsAg ScFv and Interferon-а fusion protein has been constructed successfully.