Prokaryotic Expression of Tianfu Goose IFN-α Mature Peptide and its Activity Against Virus

A primer was designed reference to the complete ORF of Tianfu goose IFN-α,the mature peptide was cloned and connected into pET32a to construct the prokaryotic expression vector.Then the IFN-α gene was expressed in Rosetta and identified by SDS-PAGE.After the expressed protein was purified by Ni-IMAC affinity chromatography and dialyzed to renaturation,its antiviral activity was determinated by micro cytopathic effect inhibition.The results revealed that IFN-α mature peptide gene was 486 bp and encoded a polypeptide of 161 amino acid.The prokaryotic expression vector pET32a-mGoIFN-α was constructed successfully and the gene was expressed.SDS-PAGE and Western-blotting assays demonstrated the expressed protein was about 38 ku.After purifying and renaturating,the antiviral activity of IFN was evaluated by using vesicular stomatitis virus(VSV) and had an antiviral specific activity of 5.52×103 U/mg,which Showed that the recombinant protein had a good antivital activity.