IL－6／IL－2融合蛋白（CH925）在大肠杆菌中高效表达  

OVEREXPRESSION OF RECOMBINANT HUMAN FUSION PROTEIN IL-6/IL-2(CH925)IN E.COLI

作　　者：赵春华[1] 王嘉玺[1] 毛宁[1] 江飞子[1] 张明伟[1] 黄碧莲 彭善云[1] 任蕴芳[1] 唐佩弦[1] 

机构地区：[1]北京军事医学科学院基础医学研究所

出　　处：《中华微生物学和免疫学杂志》1994年第3期164-167,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家青年"863"高科技资助

摘　　要：本文设计重组ＩＬ－６／ＩＬ－２的嵌合基因，选取合适的中间接头，并对ＩＬ－６及ＩＬ－２功能区基因进行修饰，运用ＰＣＲ拼接技术，克隆了中间接头与ＩＬ－２功能区基因。再与ＩＬ－６基因连接后转化Ｅ．ｃｏｌｉ，经序列分析证实获得ＩＬ－６／ＩＬ－２融合蛋白表达载体ＰＦＩＬ－６／２经诱导表达ＩＬ－６／ＩＬ－２（ＣＨ９２５）占菌体总蛋白３２％，融合蛋白分子量约为３７ｋＤ，分别以ＩＬ－６及ＩＬ－２依赖株测得融合蛋白具有双功能生物活性。An expression vector encoding the fusion protein IL-6/IL-2 had been constructed by the authors.After theflexilbe linker was synthesized and ligated with IL-2 gene fragment by PCR amplification,the IL-6 gene fragmentwas inserted into the upstream of linker-IL-2 sequence. The molecule of IL-6-linker-IL-2fusion gene was namedE.coli DH5α /pFIL-6/2 and cloned as well as identified by DNA sequencing.The expressed protein named asCH925 showed a strong band on SDS-PAGE and amounted to 32%of total cell protein.The estimated molecularweight was about 37kD. After renaturation , this fusion protein showed both IL-2 and IL-6 activity,which was as-sayed by CTLL-2 and 7TD1-dependent cell lines, respectively. The specific activity of IL-2 was 2.1×106U/mg,and for IL-6 it was up to 2. 3× 108U/mg.

关 键 词：融合蛋白 白细胞介素 大肠杆菌 

分 类 号：R392.12[医药卫生—免疫学]