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Influenza Virus Propagation in Embryonated Chicken Eggs
Published on: March 19, 2015
The interferon-alpha genes from three chicken lines and its effects on H9N2 influenza viruses
1The 211 Key Laboratory of Veterinary Molecular Biology, College of Animal Medicine, China Agricultural University, Beijing, China. xiachun@cau.edu.cn
Abstract:
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.
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