The interferon-alpha genes from three chicken lines and its effects on H9N2 influenza viruses

摘要

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.