Cloning, expression and antiviral bioactivity of red-crowned crane interferon-α.
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.
College of Wildlife Resources, Northeast Forestry University, Harbin 150040, PR China. Electronic address: tianli_1988@yahoo.com.
Li
L
Tian
College of Wildlife Resources, Northeast Forestry University, Harbin 150040, PR China. Electronic address: zpp13796033057@163.com.
Panpan
P
Zhao
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, PR China. Electronic address: maboLHT@163.com.
Bo
B
Ma
College of Wildlife Resources, Northeast Forestry University, Harbin 150040, PR China. Electronic address: guowanyang2009@163.com.
Guangyang
G
Guo
College of Wildlife Resources, Northeast Forestry University, Harbin 150040, PR China. Electronic address: sunyingdongzi@163.com.
Ying
Y
Sun
College of Wildlife Resources, Northeast Forestry University, Harbin 150040, PR China. Electronic address: xingmingweinefu@sina.com.
Mingwei
M
Xing
D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology; D030161:Avian Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001717:Birds / Q000235:genetics* / Q000378:metabolism; D002645:Chickens; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D015870:Gene Expression; D007106:Immune Sera / Q000276:immunology / Q000494:pharmacology; D016898:Interferon-alpha / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000502:physiology
D000998:Antiviral Agents; D030161:Avian Proteins; D007106:Immune Sera; D016898:Interferon-alpha; D011994:Recombinant Proteins
Antiviral activity; Gene clone; Immunofluorescence; Interferon-α; Red-crowned Crane