| 重组hIL-2表达载体的构建及在毕赤酵母中的表达 |
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| 引用本文： | 王松,苗利光,李海涛,刘艳环,王文昊,安亚雄,冯卓.重组hIL-2表达载体的构建及在毕赤酵母中的表达[J].特产研究,2012,34(1):8-11. |
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| 作者姓名： | 王松 苗利光 李海涛 刘艳环 王文昊 安亚雄 冯卓 |
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| 作者单位： | 中国农业科学院特产研究所,吉林吉林,132109 |
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| 基金项目： | 吉林省科技发展计划项目(20100219) |
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| 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。
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| 关 键 词： | 人白细胞介素2 修饰 载体构建 表达 |
Expression Vector Construction and Expression of a Modified Human Interleukin -2 in pichia pastoris |
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| WANG Song
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MIAO Li-guang
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LI Hai-tao
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LIU Yan-huan
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WANG Wen-hao
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AN Ya-xiong
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FENG Zhuo.Expression Vector Construction and Expression of a Modified Human Interleukin -2 in pichia pastoris[J].Special Wild Economic Animal and Plant Research,2012,34(1):8-11. |
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| Authors: | WANG Song MIAO Li-guang LI Hai-tao LIU Yan-huan WANG Wen-hao AN Ya-xiong FENG Zhuo |
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| Institution: | (Institute of Special Wild Economic Animals and Plants,CAAS,Jilin 132109,China) |
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| Abstract: | In this experiment,based on the amino acid sequence and three-dimensional structure of hIL-2 and its interaction mechanism with hIL-2R,site-directed mutagenesis of gene sequence of binding sites between hIL-2 and IL-2R was designed and constructed.Synthetic modified hIL-2 gene was inserted into pPIC9K,which was transformed into Pichia pastoris genome to express target protein.After the inducing expression,the target protein rhIL-2 was identified by ELISA method.The results showed that rhIL-2 was highly expressed,and the content of rhIL-2 was 0.93g/L.It will lay a foundation for further study on the biological activity of rhIL-2. |
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| Keywords: | HIL-2 Modification Vector construction Expression |
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