Construction of human-derived consensus interferon α and verification of its antiviral activity
Objective To construct a new human-derived consensus interferon α(cIFNα)sequence and verify its antiviral effect.Methods Total 57 human-derived IFNα sequences were synthesized,and the conservative amino acid preference at each site was selected by software comparison analysis.A new cIFN sequence,hIC,was synthesized into pUC-57 vector and connected with pCK to construct eukaryotic expression vector pCK-hIC,which was transfected into 293T cells to express the target protein hIC.A549 cells were incubated with the target protein before and after infection with enhanced green fluores-cent protein vesicular stomatitis virus(VSV-EGFP).The effect of hIC on VSV-EGFP replication was analyzed by fluores-cence observation,crystal violet staining and flow cytometry in vitro,and the downstream gene expression of IFN was verified by qPCR.Results The plasmid pCK-hIC was constructed correctly as verified by double enzyme digestion and sequen-cing.The expressed hIC protein,with a relative molecular mass of about 27 000,significantly reduced the fluorescence expression of VSV-EGFP,significantly inhibited virus proliferation and activated the expression of interferon-stimulated gene 15(ISG15),2'-5'-oligoadenylate synthetase 1(OAS1)and interferon-inducible transmembrane protein(1FITM).Conclusion The hIC has good antiviral effect,which lays a foundation for the follow-up research and development.
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