| IL18-IL2融合基因的克隆及其在大肠埃氏菌(E.coli)中的表达 |
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| 引用本文： | 杜广营,张国营,刘昆,栾金灵,段慧英.IL18-IL2融合基因的克隆及其在大肠埃氏菌(E.coli)中的表达[J].烟台大学学报(自然科学与工程版),2011,24(2). |
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| 作者姓名： | 杜广营 张国营 刘昆 栾金灵 段慧英 |
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| 作者单位： | 烟台大学药学院,山东烟台,264005 |
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| 摘 要： | 构建IL-18和IL-2成熟区的融合基因IL18-IL2,并实现其在大肠埃氏菌中的大量表达.抽提经PHA刺激增殖的人淋巴细胞总RNA,RT-PCR法获得IL-18和IL-2基因的成熟区序列,并通过一段Linker相连得到融合基因IL18-IL2;将IL18-IL2克隆至原核表达载体pBV220,并转化至大肠埃氏菌BL21(DE3);阳性克隆经42℃温度诱导表达后用SDS-PAGE和Western blot分析重组蛋白的表达情况及正确性.经SDS-PAGE分析,融合基因IL18-IL2在宿主菌中能够表达,其相对表观分子质量约为34.5 kD;Western blot进一步证明重组蛋白正确.实验结果表明,已成功构建了表达人IL18-IL2的菌株,为进一步研究融合蛋白IL18-IL2体内外抗肿瘤活性提供了基础.
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| 关 键 词： | 白细胞介素-18 白细胞介素-2 pBV220载体 原核表达 融合蛋白 |
Cloning and Expression of a Fusion Protein IL18-IL2 in Escherichia Coli |
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| DU Guang-ying,ZHANG Guo-ying,LIU Kun,LUAN Jin-ling,DUAN Hui-ying.Cloning and Expression of a Fusion Protein IL18-IL2 in Escherichia Coli[J].Journal of Yantai University(Natural Science and Engineering edirion),2011,24(2). |
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| Authors: | DU Guang-ying ZHANG Guo-ying LIU Kun LUAN Jin-ling DUAN Hui-ying |
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| Institution: | DU Guang-ying,ZHANG Guo-ying,LIU Kun,LUAN Jin-ling,DUAN Hui-ying(School of Pharmacy,Yantai University,Yantai 264005,China) |
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| Abstract: | We report here the cloning and expressing of an fusion protein IL18-IL2 containing the mature hu-man interleukin-18 peptide and interleukin-2 peptide in Escherichia Coli.The mature human IL-18 and IL-2cDNAs are cloned by RT-PCR with the total RNAs,extracted from human PBMC stimulated by PHA,as tem-plates.The fusion gene IL18-IL2 connected by a flexible linker is inserted into the pBV220 expression vector and sequenced.Then the recombinant plasmid pBV-IL18-IL2 is transformed into BL21(DE3) and induced at 42 ... |
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| Keywords: | interleukin-18 interleukin-2 pBV220 vector prokaryotic expression fusion protein |
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