Abstract:

IL-1β plays crucial roles in mediating immune regulatory and inflammatory responses. In order to investigate the biological function of swamp eel IL-1β, gene-specific primers were synthesized and used for amplifying the cDNA fragments. Subsequently, the purified fragments were subcloned into pET-28a. The recombinant vector was sequenced and transformed into BL21(DE3). In addition, a recombinant strain was induced with IPTG and then Ni-NTA affinity chromatography was performed to isolate the rIL-1β protein. Moreover, SDS-PAGE and Western blotting were carried out to analyze the recombinant protein. Finally, qRT-PCR was performed to investigate the effects on expression of several genes in liver after intraperitoneal injection of rIL-1β at a dose of 2 μg/kg. Results displayed that the prokaryotic expression vector was successfully constructed as well as rIL-1β protein was efficiently isolated with obvious activity. Intraperitoneal injection of rIL-1β upgraded the expression of IL-1β and NF-κB and inhibited the expression of AKR and Nrf2 in liver of swamp eel. All above results provided us a good opportunity to further our study on the function of fish IL-1β in immune system.

Key words: IL-1β, swamp eel, bioactivity analysis, recombinant expression