Objective: To construct the eukaryotic expression vector pEGFP N1/IL-37b of full-length and mature IL-37b, and to detect the expression of both in RAW 264.7 cells.
Methods: Using the plasmid pUBC/IL-37b containing the full-length coding region gene of IL-37b as a template, a eukaryotic expression vector capable of expressing the full-length and mature form of IL-37b was constructed. The recombinant plasmid pEGFP N1/IL-37b was constructed. Transfected into RAW 264.7 cells, the expression of full-length and mature forms of IL-37b was detected by western blot and confocal microscopy, and the effects of full-length and mature forms of IL-37b on LPS-induced IL-37b were detected by real-time PCR. 6 Inhibition of expression.
RESULTS: The constructed pEGFP N1/IL-37b could express the full-length and mature forms of IL-37b in cells after transfection, and could inhibit the expression of IL-6 induced by LPS.
Conclusion: The eukaryotic expression vector of full-length and mature form of IL-37b was successfully constructed, which lays a solid foundation for further research on the inhibitory effect and mechanism of IL-37b on inflammation.