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| Construction and expression of eukaryotic expression vector of human IL-37b gene |
| Received:January 06, 2016 |
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| DOI：10.3969/j.issn.1005-4847.2016.03.010 |
| KeyWord:Interlukin-37;Gene cloning;Eukaryotic expression vector;THP-1 cells |
| Author | Institution |
| 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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| Abstract: |
| 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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