Objective: To construct pEGFP-N1/IL-37b eukaryotic expression vector and detect its expression in THP-1 cells.
Method: Extract human PBMC total RNA, use RT-qPCR technology to amplify IL-37b gene coding region sequence, clone into pEGFP-N1 eukaryotic expression vector, recombinant plasmid pEGFP-N1/Build IL-37b. In THP-1 cells, IL-37 expression was detected by T-qPCR and Western blot.
Result: The results of double enzyme digestion and gene sequencing showed that the IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1. The results of T-qPCR and Western blot showed that IL-37 expression level increased significantly after transfection. THP-1 cells (Pu003c0.01).
Conclusion: We have successfully constructed a new type of anti-inflammatory factor IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which laid the foundation for further research on the relationship between IL-37 function and related diseases.