| mIL-21／PcDNA3．1真核表达载体的构建及鉴定 |
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| 引用本文： | 王丽娜,林志娟,鞠吉雨,李芳娜,冯永堂,孙萍,苗乃发.mIL-21／PcDNA3．1真核表达载体的构建及鉴定[J].潍坊医学院学报,2009,31(3):179-180,192. |
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| 作者姓名： | 王丽娜 林志娟 鞠吉雨 李芳娜 冯永堂 孙萍 苗乃发 |
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| 作者单位： | 潍坊医学院免疫学教研室,山东省高校免疫学重点实验室,山东,潍坊,261053 |
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| 基金项目： | 潍坊市科技局计划项目 |
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| 摘 要： | 目的构建mIL-21／PcDNA3．1真核表达载体。方法从ConA活化的BALB／c小鼠脾细胞中抽提总RNA，RT—PCR方法扩增小鼠IL-21基因，XhoⅠ和HindⅢ双酶切后克隆入真核细胞高效表达载体pcDNA3．1中，构建小鼠IL-21真核表达载体mIL-21／PcDNA3．1，并经PCR、限制性酶谱分析和DNA序列分析对重组子的正确性进行鉴定。结果构建的小鼠IL-21真核表达载体mIL-21／PcDNA3．1经PCR、限制性酶谱分析和DNA序列分析证明获得了目的基因，其序列与Genbank中登录的小鼠IL-21基因序列完全一致。结论成功构建了。mIL-21／PcDNA3．1真核表达载体，为IL-21分子的进一步研究奠定了基础。
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| 关 键 词： | IL-21 IL-21/PcDNA3.1 真核表达 |
Construction and Verification of Eukaryotic Expression Vector of mIL-21/PcDNA3.1 |
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| WANG Li-na,LIN Zhi-juan,JU Ji-yu,LI Fang-na,FENG Yong-tang,SUN Ping,MIAO Nai-fa.Construction and Verification of Eukaryotic Expression Vector of mIL-21/PcDNA3.1[J].Journal of Weifang Medical College,2009,31(3):179-180,192. |
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| Authors: | WANG Li-na LIN Zhi-juan JU Ji-yu LI Fang-na FENG Yong-tang SUN Ping MIAO Nai-fa |
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| Institution: | ( Department of Key Laboratory of Immunology, Weifang Medical College, Weifang 261053, China ) |
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| Abstract: | Objective To clone murine interleukin 21 （mIL-21）gene,and construct it＇s eukaryotic expression plasmid IL-21/ PcDNA3.1. Methods Extracting total RNA from ConA activated murine T cells by RT-PCR, and then the gene is cloned into eukaryocytic expression plasmid pcDNA3. 1 to form recombinant plasmid mIL-21/pcDNA3. 1. The correctness of recombinant plasmid is identified by PCR,restrict digestion analysis and DNA sequencing. Results The recombinant plasmid mlL-21/PcDNA3.1 was confirmed by PCR,restrict digestion analysis and DNA sequencing. Conclusion An eukaryotic expression plasmid mIL-21/PcDNA3. 1 was constructed successfully, and to lay the foundation for further study. |
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| Keywords: | IL-21 IL-21/PcDNA3 1 |
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