Prokaryotic Expression Of IFN-? And Inhibits The Replication Of ALV-J In Vitro

Avian leukemia in subgroup J is a neoplastic disease caused by retroviruses and has not yet been controlled by drugs and vaccines.The reduction of productivity caused by eradication cost and tumor has caused great economic loss to the global poultry industry.IFN-? is a new type of interferon with similar antiviral and antitumor activities as type I interferon.The effect of IFN-? on ALV-J replication was studied by prokaryotic expression,which laid a foundation for the prevention and treatment of ALV-J infection.IFN-? gene was linked into pET-32 a prokaryotic expression vector to construct recombinant plasmid for double digestion validation.IFN-? protein expression was induced at the optimal expression time,purified and renatured,and verified by Western Blot.ELISA and fluorescence quantitative PCR were used to detect the inhibition of ALV-J replication on DF-1 cells by IFN-? alone and in combination with IFN-?.DF-1 cells were induced with IFN-?,and the effects on the expression of ISGs and type I interferon were examined.When IFN-? was used alone,the relative expression of the virus decreased by 52% at the final concentration of 5000 ng/mL(P < 0.001),inhibiting the replication of ALV-J in a dose-dependent manner.At the concentration of 10 ng/mL,the relative expression of IFN-? and IFN-? decreased by 21%(P < 0.01)compared with IFN-? alone;at the concentration of 100 ng/mL,the relative expression of IFN-? decreased by 32%(P < 0.001)compared with IFN-? alone,indicating that IFN-? and IFN-? have a more obvious synergistic effect.After induction of cells by IFN-?,the expression of six genes,RSAD,ISG15,MX2,IFIT3,OASL and ISG12-2,was significantly up-regulated,with the highest expression at 12 h;the expression of IFN-? was up-regulated by 2 times,and that of IFN-beta was up-regulated by 4.3 times,with the highest expression at 12 h.In conclusion,prokaryotic expression of IFN-? on DF-1 cells can inhibit ALV-J replication.The combination of IFN-? and IFN-? was more effective.IFN-? induced the highest expression of ISGs and type I interferon at 12 h.