Hyper-IL-6融合基因的构建及在真核细胞中的表达  

Construction and expression of fusion gene encoding Hyper-IL-6 in mammalian cells

作　　者：郄春花[1] 秦波[1] 黄爱龙[1] 刘杞[1] 陈国民[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所感染性疾病分子生物学教育部重点实验室,重庆400010

出　　处：《细胞与分子免疫学杂志》2006年第2期141-143,147,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：重庆市科委科研基金项目(No.2004-54号)

摘　　要：目的:构建融合基因Hyper-IL-6并在真核细胞中进行表达。方法:采用基因拼接(GeneSOEing)法将人类可溶性白细胞介素6受体和白细胞介素6的编码基因用一富含甘氨酸序列的接头经PCR扩增融合,并定向克隆至pIRES2-EGFP,构建与绿色荧光蛋白(GFP)共表达的融合重组质粒,转染哺乳动物细胞,通过绿色荧光蛋白、RT-PCR和免疫组化检测Hyper-IL-6蛋白的表达。结果:PCR扩增出1224bp的Hyper-IL-6编码序列并成功构建重组质粒pIRES-HIL-6,重组质粒转染真核细胞后经检测证实Hyper-IL-6能在真核细胞中表达。结论:人类Hyper-IL-6重组表达质粒的成功构建与真核表达为以后对其进行活性分析和生物学功能的研究提供了基础。AIM： To construct and express fusion gene encoding Hyper-lL-6 in mammalian cells. METHODS： Using geneSOEing method, human slL-6R cDNA was fused with IL-6 cDNA by a linker rich in glycine through PCR technique and cloned into plRES2-EGFP. The constructed plasmid was transfected into mammalian cells. The expression of fu- sion gene encoding Hyper-lL-6 was detected by GFP and RT-PCR and immunocytochemistry. RESULTS： The recombinant plasmid plRES-HIL-6 was successfully constructed and expressed in mammalian cells, CONCLUSION： The success in construction and expression of human Hyper-lL-6 makes it possible to further study its biological functions.

关 键 词：HYPER-IL-6 绿色荧光蛋白 真核细胞 表达 

分 类 号：R392.11[医药卫生—免疫学]