| Domestic infective diseases which cause the tremendous economic losses to poultry industry, provoke the serious threat of poultry industry. Interferons (IFNs), a family of cytokines, have many kinds of biological activities, such as interfering with the replication of various viruses, decreasing cell proliferation and modifying immunological processes.Therefore the research on the chicken interferon has the important meaning.One pair of primers were designed according to the sequences of chicken Interferon-alpha published in GenBank. Chicken interferon-alpha gene were amplified from genome DNA extracted from liver tissue of luoman chicken, hainan chicken and gushi chicken, respectively,and were linked to pGEM-T Easy vector, then transformed to competent cells of JM109. By blue-white spot screening , idenfication of plasmid PCR and enzyme digestion,positive strain was sent to Takara for sequencing. And sequencing results showed that chicken Interferon-alpha gene of luoman chicken, hainan chicken and gushi chicken were obtained.The amplified fragment of ChIFN-α was 582bp in length, which includes one open-reading frame, encoding 162 amino acid residues..The results of homology analysis with other chicken interferon-alpha gene published previously in GenBank by DNAStar analysis reveal there was identity of 99.3% at nucleotide homology and that of 97.9% at amino acid homology.On this basis,another pair of primers were designed. Maturity protein gene of IFN-α was amplified from pGEM- ChlFN-α and was digested by SphI and SalI,then was inserted into pQE30 of the prokaryotic expression vector which was digested by SphI and SalI, the recombinant expressed plasmid pQEIFN-α was constructured ,then transformed to competent cells of JM109. pQEIFN-α was identificated by PCR, enzyme digestion and DNA sequencing conclusively. The result indicated that the gene of encoding ChIFN-α maturity protein was inserted correctly to the target site of...
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