Cloning, expression and antiviral bioactivity of red-crowned crane interferon-α

Abstract

Interferon-α (IFN-α) genes have been cloned from a variety of animals, but information regarding crane IFN-α has not been reported to date. In this study, we cloned a full-length Red-crowned Crane interferon-α (crIFN-α) gene sequence consisting of a 486bp partial 5' UTR, 741bp complete ORF and 559bp partial 3' UTR. This gene encodes a protein of 246 amino acids and shares 60 to 80% identity with avian IFN-α and less than 45% identity with mammalian IFN-α. The expression of crIFN-α with an N-terminal His-tag was investigated in Escherichia coli, and the protein was purified on a nickel column. To obtain activated proteins, crIFN-α inclusion bodies were renatured by dialysis. In vitro cytopathic inhibition assays indicated that the recombinant crIFN-α could inhibit the replication of vesicular stomatitis virus in chicken fibroblasts. These antiviral activities were abrogated by rabbit anti-crIFN-α antibodies in vitro. In addition, an immunofluorescence assay indicated that crIFN-α could be expressed in chicken fibroblasts and was primarily located in the cytoplasm. Taken together, our results suggest that the crIFN-α gene may play an important role in inhibiting the replication of viruses.

Keywords: Antiviral activity; Gene clone; Immunofluorescence; Interferon-α; Red-crowned Crane.