重组小鼠IL-38蛋白制备及其生物学活性分析  

Preparation of Recombinant Mouse IL-38 Protein and Its Bio-Activity Analysis

作　　者：朱俊丰[1] 赵鹏燕 刘帅 赵健[1] 李佳增 阿斯姆古丽·马木提 华子雯 赵艺淑 刘宏生[1] ZHU Jun-feng;ZHAO Peng-yan;LIU Shuai;ZHAO Jian;LI Jia-zeng;Asimuguli·Mamuti;HUA Zi-wen;ZHAO Yi-shu;LIU Hong-sheng(School of life Sciences,Liaoning Uni.,Shenyang 110036)

机构地区：[1]辽宁大学生命科学院,辽宁沈阳110036

出　　处：《微生物学杂志》2020年第1期76-80,共5页Journal of Microbiology

基　　金：辽宁省教育厅项目(LQN201711);辽宁大学博士启动项目;辽宁大学预申报基金项目(LDGY2019018)。

摘　　要：制备小鼠IL-38蛋白,并检验其生物学活性。对IL-38的基因进行密码子优化并化学合成优化后的基因,将其插入原核表达载体pET28a(+),构建pET28a-IL-38质粒,转化至大肠埃希菌BL21(DE3),经IPTG诱导表达,镍亲和层析法纯化制备IL-38蛋白;分别用IL-36γ和IL-38干预小鼠肠系膜淋巴结(mesenteric lymph node,MLN)细胞,ELISA检测各组Th1细胞因子的分泌水平。经SDS-PAGE分析获得纯度高达95%的IL-38蛋白。相对于对照组,IL-38处理组Th1细胞因子分泌水平明显降低(P<0.05)。成功制备小鼠IL-38蛋白,证实了IL-38对IL-36γ刺激的MLN细胞炎性反应具有负向调控作用,为进一步研究IL-38在炎症性疾病中的免疫调节功能提供参考。A recombinant mouse IL-38 protein was prepared and tested for bio-activity.The gene of IL-38 was optimized for its codon and chemically synthesized the gene after the optimization,the gene was inserted into the prokaryotic epression vector pET28a(+)to construct pET28a-IL-38 plasmid and transformed into E.coli BL21(DE3),and prepared IL-38 adopting IPTG induced expression,nickel affinity chromatography,and purification.Then the mesenteric lymph nodes(MLN)cells of mouse were respectively interfered with IL-36γand IL-38,and detected the excretion level of Th1 cytokines in each intervention group by ELISA.The results showed that:the IL-38 with purity as high as 95%was obtained adopting SDS-PAGE analysis.As compared with the control group,the secretion of Th1 cytokines in IL-38 treated group was significantly lower(P<0.01).Therefore,the mouse IL-38 protein was successfully prepared,and confirmed that it has negative regulation effect on MLN cell inflammatory response of MLN cells stimulated by IL-36γ,and that laid the foundation for further study on the immune regulation function in IL-38 ininflammatory diseases.

关 键 词：白细胞介素38 原核表达 纯化 生物学活性 

分 类 号：Q939.93[生物学—微生物学]