| 牛白细胞介素-18基因原核表达载体的构建 |
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| 引用本文： | 倪雪,宋佰芬,李迎春,邹珊,崔玉东.牛白细胞介素-18基因原核表达载体的构建[J].黑龙江八一农垦大学学报,2009,21(6):51-53. |
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| 作者姓名： | 倪雪 宋佰芬 李迎春 邹珊 崔玉东 |
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| 作者单位： | 黑龙江八一农垦大学生命科学技术学院,大庆,163319 |
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| 摘 要： | 根据已发表的IL-18蛋白cDNA序列保守区设计一对特异性引物,应用RT-PCR技术从ConA活化的牛外周血单核细胞中扩增到编码牛IL-18蛋白基因,其大小为582 bp。将该基因克隆到pMD18-T载体中,经序列测定表明该基因与gengbank上发表的牛白细胞介素-18基因序列同源性为98%。将该基因从重组pMD-gIL-18质粒中亚克隆到表达载体pET-32 a（＋）质粒中,构建原核表达重组质粒,经酶切和PCR鉴定后表明构建的重组质粒为阳性。
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| 关 键 词： | 白细胞介素 原核表达 载体构建 |
Prokaryotic Expression Vector Construction of Cow IL-18 gene |
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| Ni Xue,Song Baifen,Li Yingchun,Zou Shan,Cui Yudong.Prokaryotic Expression Vector Construction of Cow IL-18 gene[J].Journal of Heilongjiang August First Land Reclamation University,2009,21(6):51-53. |
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| Authors: | Ni Xue Song Baifen Li Yingchun Zou Shan Cui Yudong |
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| Institution: | (College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319) |
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| Abstract: | According to the encoding sequence of the IL-18 protein and the multiple cloning sites characteristic of the prokaryotic expression vector pET-32 a （＋） in Genebank, appropriate primers specific to IL-18 gene were designed. The target DNA fragments were obtained by RT-PCR amplification and then were cloned into the pMD18-T vector. After identification by restriction enzyme digestion and sequencing analysis, the specific DNA fragments were cloned into the prokaryotic expression vector pET-32 a （＋）. The positive recombinant plasmids were named pET-32a （＋）-IL-18 after identification by restriction enzyme digestion and PCR amplification. Results showed that recombinant expression plasmids pET-32a（＋）-IL-18 were successfully constructed. |
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| Keywords: | IL-18 prokaryotic expression vector construction |
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