Amplification, cloning and sequencing of interferon-α gene from poodle.

One pair of primers was designed and synthesized according to the canine interferon-α(IFN-α) gene nucleic acid sequence published in the GenBank.The IFN-α gene was amplified from genome DNA of poodle liver by PCR.The purified PCR products were linked to pGEM-T Easy vector and then transformed into competent cells of DH5α.By identification of blue-white spot screening,plasmid PCR and enzyme digestion,positive strain was sent to TaKaRa for sequencing.The sequencing results showed that poodle IFN-α gene was obtained.The cloned poodle IFN-α gene span is 564 bp,which includes one open-reading frame encoding 187 amino acid residues.And there are three potential N-glycosalation sites and ten cysteines in the deduced amino acid sequence.Homology of nucleotide and amino aids of IFN-α gene between poodle and humans and other animals and the phylogenetic tree indicated that IFN-α gene varied in species,and the closer the relationship,the higher the homology.