重组hIL-2表达载体的构建及在毕赤酵母中的表达  

Expression Vector Construction and Expression of a Modified Human Interleukin-2 in pichia pastoris

作　　者：王松[1] 苗利光[1] 李海涛[1] 刘艳环[1] 王文昊[1] 安亚雄[1] 冯卓[1] 

机构地区：[1]中国农业科学院特产研究所,吉林吉林132109

出　　处：《特产研究》2012年第1期8-11,共4页Special Wild Economic Animal and Plant Research

基　　金：吉林省科技发展计划项目(20100219)

摘　　要：本试验根据hIL-2氨基酸序列、三维结构及其与受体IL-2R结合机制,设计并合成IL-2Rα亚基结合位点修饰性hIL-2基因片段。合成的修饰性hIL-2基因序列被插入真核表达载体pPIC9K,并转化入毕赤酵母基因组中。经筛选得到阳性重组子后,通过诱导培养得到了表达产物。对表达产物进行Elisa初步测定,结果成功表达出重组hIL-2,表达水平达0.93g/L。本项研究为进一步开展IL-2Rα结合位点修饰性hIL-2的生物活性研究奠定了基础。In this experiment, based on the amino acid sequence and threedimensional structure of hIL2 and its interaction mechanism with hIL2R, sitedirected mutagenesis of gene sequence of binding sites between hIL2 and IL2R was designed and constructed. Synthetic modified hIL2 gene was inserted into pPIC9K, which was transformed into Pichia pastoris genome to express target protein. After the inducing expression, the target protein rhIL2 was identified by ELISA method. The results showed that rhIL2 was highly expressed, and the content of rhlL2 was 0.93g/L. It will lay a foundation for further study on the biological activity of rhIL2.

关 键 词：人白细胞介素2 修饰 载体构建 表达 

分 类 号：Q786[生物学—分子生物学]