Construction and expression of eukaryotic expression vector of human IL-37b gene
Received:January 06, 2016  
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KeyWord:Interlukin-37;Gene cloning;Eukaryotic expression vector;THP-1 cells
姚静 石河子大学医学院, 新疆 石河子
程江 石河子大学第一附属医院检验科, 新疆 石河子
裴雪枫 辽宁医学院, 辽宁 锦州
王静宇 中国航天员科研训练中心, 航天医学基础与应用国家重点实验室, 北京
袁明 中国航天员科研训练中心, 航天医学基础与应用国家重点实验室, 北京
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      Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells (PBMCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eukaryotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detected by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1.RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P < 0.01) after transfection in THP-1 cells. Conclusions We successfully constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foundation for further study on IL-37 functions and its association with related diseases.
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