Construction and Expression of Eukaryotic Expression Vector for Interferon α2b Gene

Objective To construct a eukaryotic expression vector for interferon α2b(IFNα2b)gene and express in CHOdhfr-cells.Methods IFNα2b gene fragment was amplified from recombinant E.coli DH5α-pbv220-IFNα2b and cloned into plasmid psv-dhfr.The constructed recombinant plasmid psv2-dhfr-IFNα2b was transfected to CHO-dhfr cells,and the monoclonal cell strains growing stably were obtained by MTX pressure screening.The antiviral activity of IFNα2b in transfected cells was determined by Wish cytopathic inhibition test.The genomic DNA of transfected cells was extracted and identified by PCR.Results Both PCR and restriction analysis proved that recombinant plasmid psv2-dhfr-IFNα2b was constructed correctly.The IFNα2b expressed in transfected cells showed high antiviral activity.IFNα2b gene band was amplified using the genomic DNA of transfected cells as a template.Conclusions A eukaryotic expression vector for IFNα2b gene was successfully constructed,and the IFNα2b with biological activity was expressed in CHO-dhfr-cells,which laid a foundation of further eukaryotic expression of IFNα2b.