Preparation and antiviral activity study of high-activity porcine IFN-a17m protein
The aim of this study is to obtain the high antiviral activity of porcine interferon α(poIFN-α).Porcine IFN-α17(poIFN-α17)gene was mutated and synthesized,and mutated poIFN-α17 was named as poIFN-α17m,then the gene was further digested and cloned into the pVB220 plasmid to construct E.coli expression vectors pVB220-IFN-α17 and pVB220-IFN-α17m.The ex-pression vectors were transformed into E.coli DH5α and recombinant poIFN-α17 and poIFN-α17m were expressed with temperature change.The recombinant protein was purified using affinity chro-matography and identified by Western blot.The antiviral activity of recombinant poIFN-α17 and poIFN-α17m was tested in PK-15 cells against PRV and VSV.The expression of interferon stimu-lating genes(ISGs)induced by recombinant poIFN-α17 and poIFN-α17m was further detected and compared.The results demonstrated that the pVB220-IFN-α17 and pVB220-IFN-α17m expression vector were successfully constructed,and inclusion expression of poIFN-α17 and poIFN-α17m protein in E.coli DH5α was obtained after the induction with temperature change,and the recom-binant poIFN-α17 and poIFN-α17m could react with porcine interferon polyclonal antibody.Fur-thermore,the result demonstrated that the antiviral activity of poIFN-α17m against PRV and VSV was superior to poIFN-α17.Moreover,recombinant poIFN-α17m could effectively induce the ex-pression of ISGs.
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