高效表达重组猪IL-10的基因工程菌株的构建  

High-level Expression of Recombinant Porcine Interleukin-10 in E.coli

作　　者：仇华吉[1] 金吉东[1] 兰文升[1] 郭海龙[1] 周艳君[1] 王云峰[1] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《畜牧兽医学报》2003年第4期385-388,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点基础研究发展规划项目(199011902)

摘　　要：白细胞介素 10(IL 10)是一种Th2细胞等产生的能抑制Th1细胞释放细胞因子的免疫调节因子。从经脂多糖(LPS)活化的猪外周血单核细胞(PBMC)中提取总RNA,用RT PCR扩增了猪IL 10(poIL 10)编码基因,克隆并测序验证后,将其亚克隆到表达载体pPROEXTMHTb中,转化DH5α后,用IPTG诱导培养,经免疫印迹和生物活性检测显示,重组菌株表达了具有活性的重组poIL 10,重组poIL 10表达水平达3mg/ml培养液,占菌体细胞总蛋白的30 2%,本研究为poIL 10的功能研究和临床应用奠定了基础。Interleukin10(IL10)is an antiinflammatory cytokine that is produced by T cells,B cells,monocytes,and macrophages.The porcine IL10(plIL10)cDNA was amplified from the total RNA of peripheral blood mononuclear cells stimulated by LPS using RTPCR,and cloned and sequenced.The poIL10 was then subcloned into a prokaryotic expression vector.Highlevel expression of recombinant poIL10 was obtained after induction by IPTG of the recombinant bacterium containing the recombinant plasmid harboring the poIL10 gene.The expressed Histagged recombinant poIL10 showed high activity of inhibition of the production of interferongamma by porcine macrophages.The recombinant poIL10 can be a valuable therapeutic agent for the diseases with overproduction of inflammatory cytokines.

关 键 词：基因表达 基因重组 猪 IL—10 基因工程 菌株构建 

分 类 号：S828[农业科学—畜牧学]