Expression Purificationand Bioactivity of Recombinant Porcine Interferon-α(IFNα)
In this study,the sequence of porcine interferon α(No.P49879)in the UniProt database was op-timized according to the codon preference of Escherichia coli,the corresponding gene fragment was synthesized,and the pET-28a-INF-α prokaryotic expression vector was constructed.The recombinant vector was trans-formed into Escherichia coli BL21(DE3),and expression was induced by IPTG.After incubation in a 20 L fer-menter for 24 h,the cells were harvested with an OD600of 108.The expressed porcine interferon α mainly existed in the form of inclusion bodies,and the label-free porcine interferon α protein was obtained through the steps of de-naturation,repletion,nickel-column purification,and enzymatic digestion.SDS-PAGE electrophoresis showed that the The molecular weight of the protein was about 19 kD and the purity was up to 95％.The cytopathic inhibi-tion assay showed that the antiviral activity of the protein was 1.28 x 107 IU/mg.In this study,the high purity and high activity of porcine interferon α protein was successfully obtained,which lays the foundation for further advan-cing the clinical application of the protein drug,and provides the basis for the commercial production.
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