Expression of IFN-λ1 from Congjiang pigs and its effect on anti-PRRSV proliferation.

Abstract

An eukaryotic expression system of Congjiang pigs IFN-λ1 was constructed to obtain its expression in CHO-K1 cells and the inhibition effect of Congjiang pig IFN-λ1 on PRRSV proliferation was verified. The eukaryotic expression plasmid pEGFP-PoIFN-λ1 was constructed from the pig IFN-λ1 gene fragment and transfected into CHO-K1 cells. Expression was detected by fluorescence microscopy and Western blotting. The influence on the proliferation of PRRSV was assessed. The results of the study showed that the recombinant plasmid pEGFP-PoIFN-λ1 was constructed correctly. After transfection, green fluorescent signal was detected in CHO-K1 cells by fluorescence microscopy. Western blot analysis revealed that in cells at different time periods after transfection, porcine IFN-λ1 was expressed, with the highest expression observed 36 h after transfection. The antiviral activity of the supernatant after 36 h of transfection was determined by the micro cytopathic inhibition method, and the biological activity was 2.1×103 U/mL. Quantitative PCR was used to detect the proliferation of PRRSV, and the results showed that Congjiang pigs IFN-λ1 significantly inhibited the mRNA expression of PRRSV and viral proliferation in a dose- and time-dependent manner. This study established a Congjiang pig IFN-λ1 eukaryotic expression system, and the quantitative PCR method showed that it has a significant inhibitory effect on the proliferation of PRRSV, which lays a foundation for the future production of antiviral drugs and clinical application.