The sequences of 27 chicken interferon-alpha (ChIFN-α) genes were obtained from GenBank. The gene sequences were compared and homology between them was determined by using a bio-software. On the basis of these results, a new rChIFN-α peptide sequence with 194 amino acids was assembled. Thereafter, on the basis of the new amino acid sequences and by using the most frequently occurring codes of Pichia pastoris, and a 582 bp gene sequence was formed. In order to amplify this non-templated gene, 16 primers were designed, and their gene sequences were synthesized, and amplified. This amplified gene sequence was cloned into the expression vector pPICZα-A to construct a recombination plasmid named pPICZ-rChIFN-α. Then, the recombination plasmid was induced to express the rChIFN-α protein. The results demonstrated that the recombinant plasmid pPICZ-rChIFN-α was successfully expressed in P. pastoris. Furthermore, rChIFN-α had a considerable antiviral activity against both Newcastle disease virus (NDV) and vesicular stomatitis virus (VSV). Therefore, this method of gene engineering could give direction to research on the key amino acids in the interferon or analogous proteins and enable the construction of proteins with high antiviral activity, which can be used both for research and industrial purposes.
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