Cloning And Prokaryotic Expression Of Chicken α And β Interferon Gene

Three primers were designed according to the sequences of chicken interferonαandβin Genebank, and the whole gene of the chicken interferonα,the mature protein gene without the signal peptide and the mature protein gene without the signal peptide of chickenβwere amplified through PCR from the DNA of chicken liver. The resulting specific sequences were cloned into the vector pMD19-T which were further transferred into the competent cell DH5α, and the plasmids were extracted, identified and sequenced. The result shows that the whole chicken interferonαgene consisted of 582bp, the mature protein gene without the signal peptide 486bp. Compared with the sequences in the Genebank, the sequence homologies were 98.5% and 98.2% respectively. Compared with the sequences of other subtypes and the subtype IFNA1, the sequence homologies were above 97.9% and 99.3% respectively. While the mature chicken interferonβgene without signal peptide consisted of 531bp with a 100% sequence homology with the referred gene.The cloned mature protein genes of the chicken interferonαandβwere subcloned into the corresponding sites of the expressed vector pGEX-2T in norientation, and the prokaryotic reconstitution expressed vectors pGEX-ChIFNα' and pGEX-ChIFNβ' were constructed, and were transformed into competent cell BL21, and the results showed that the mature protein genes of the chicken interferonαandβwere inserted correctly into the prokaryotic expressed vector pGEX-2T. The positive bacteria strains with the reconstituted expressing plasmids were induced with 1mM IPTG and were expressed , through electrophoresis, appeared both an abvious band at 44KD which showed the expression of GST-ChIFNα' and GST-ChIFNβ' fusion proteins by the reconstituted expressing vectors pGEX-ChIFNα' and pGEX-ChIFNβ'.