动物造模 z-bio 2022-06-08 216

　　Objective: To construct pEGFP-N1/IL-37b eukaryotic expression vector, and to detect its expression in THP-1 cells.

　　Methods: Total RNA was extracted from human PBMCs, the coding region sequence of IL-37b gene was amplified by RT-qPCR, cloned into pEGFP-N1 eukaryotic expression vector, and the constructed recombinant plasmid pEGFP-N1/IL-37b was transfected In THP-1 cells, the expression of IL-37 was detected by RT-qPCR and Western blot.

　　RESULTS: Double-enzyme digestion and gene sequencing showed that the IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1; RT-qPCR and Western blot results showed that the expression level of IL-37 was significantly increased after transfection into THP-1 cells ( P < 0.01).

　　Conclusion: A new eukaryotic expression vector pEGFP-N1/IL-37b for anti-inflammatory factor IL-37 was successfully constructed, laying a foundation for further research on the function of IL-37 and its relationship with related diseases.