The interferon-alpha genes from three chicken lines and its effects on H9N2 influenza viruses.
The interferon-alpha genes from three chicken lines were cloned by a direct PCR technique, and the effects of recombinant protein expressed in a prokaryotic system on highly pathogenic H9N2 influenza viruses were investigated. The cloned ChIFN-alpha gene encoded a protein of 193 amino acids with a signal sequence of 31 amino acids and mature peptides of 162 amino acids. Comparison of ChIFN-alpha sequences, detected six amino acids substitutions at positions 50, 58, 65, 81, 181, and 183. Homology analysis indicated that ChIFN-alpha genes could be subdivided into two lineages, SH-ChIFN-alpha and WJ-ChIFN-alpha. In addition, both SH-ChIFN-alpha and WJ-ChIFN-alpha were expressed with the N-terminal 6 consecutive histidine residues in a high-level prokaryotic expression system. Recombinant chicken interferon-alpha (rChIFN-alpha) protein has anti-VSV activity of more than 1 x 10(8) U/mg. Moreover, High concentration (10,000U) of rSH-ChIFN-alpha resulted in over 40% inhibition of the H9N2 virus infection in chicken embryos (Ovo), and 100% inhibition from one day-old to five day-old chickens (Vivo). The results suggested that rChIFN-alpha is a potential agent against many Chicken viral strains.
Xia
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The 211 Key Laboratory of Veterinary Molecular Biology, College of Animal Medicine, China Agricultural University, Beijing, China. xiachun@cau.edu.cn
Liu
J
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Wu
Z G
ZG
Lin
C Y
CY
Wang
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D002642:Chick Embryo; D002645:Chickens / Q000502:physiology*; D003001:Cloning, Molecular; D003588:Cytopathogenic Effect, Viral / Q000276:immunology; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D005260:Female; D007370:Interferon Type I / Q000235:genetics / Q000494:pharmacology*; D016898:Interferon-alpha / Q000235:genetics*; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000187:drug effects*; D009976:Orthomyxoviridae Infections / Q000188:drug therapy* / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000188:drug therapy* / Q000821:virology*; D011994:Recombinant Proteins; D016415:Sequence Alignment; D013047:Specific Pathogen-Free Organisms
D007370:Interferon Type I; D016898:Interferon-alpha; D011994:Recombinant Proteins; D004247:DNA