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As a key link between innate and adaptive immune responses, the interferon (IFN) system is the first line of defense against viral infection.IFN, and in particular, IFN-α, has been used as an effective therapeutic agent for virus infection in the clinic However, different subtypes of IFN-αdemonstrate distinct antiviral activity.Therefore, it is important to identify IFN-α subtypes with high antiviral activity to develop genetically engineered antiviral drugs.In this study, we cloned the genes forl3 IFN-α subtypes from peripheral blood lymphocytes of the mink.The homologies of the 13 mink IFN-α genes were 93.6-99.3% and 88.8-98.4% at the nucleotide and amino acid sequence levels, respectively.In contrast to human and canine IFN-α subtypes, most mink IFN-α subtypes contained two N-glycosylation sites.We expressed and purified 13 mink IFN-α subtypes in Escherichia coli.The cytopathic effect inhibition assayshowed that all the 13 recombinant mink IFN-α subtypes inhibited the propagation of VSV in WISH cells, with IFN-α2 and IFN-α12 demonstrating the highest activity.Furthermore, recombinant mink IFN-α2 and IFN-α12 significantly suppressed the propagation of canine distemper virus in Vera cells, with IFN-α2 demonstrating higher activity.In summary, we identified mink IFN-α2 subtype as a promising prototype for the development of effective antiviral drugs.