| 重组小鼠白介素-2基因在真核细胞中的转染表达 |
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| 引用本文： | 路晓辉,杜军,曾军,李晓玲,徐珍霞,蔡绍晖.重组小鼠白介素-2基因在真核细胞中的转染表达[J].广西医科大学学报,2006,23(3):352-354. |
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| 作者姓名： | 路晓辉 杜军 曾军 李晓玲 徐珍霞 蔡绍晖 |
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| 作者单位： | 暨南大学药学院,广州,510632;中山大学药学院 |
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| 摘 要： | 目的：在构建重组真核表达载体pIRESneo2／mIL-2的基础上，建立能够持续稳定表达mIL-2的哺乳类工程细胞。方法：运用分子克隆技术，将由RT-PCR获得的mIL-2cDNA片断插入真核表达质粒pIRESneo2构建成mIL-2重组表达载体pIRESneo2／mIL-2。通过脂质体转染法将pIRESneo2／mIL-2导入C2C12细胞。转染后第30天，用Western blots检测mIL-2表达情况。结果：经DNA测序证明mIL-2cDNA片断插入方向和碱基组成顺序均准确无误，Western blots检测转染真核重组表达载体pIRESneo2／mIL-2的C2C12细胞系表达mIL-2。结论：利用pIRESneo2／mIL-2构建的真核表达载体在C2C12细胞系中能够持续稳定表达mIL-2。
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| 关 键 词： | 小鼠白细胞介素-2 真核表达 基因转染 |
| 收稿时间： | 2006-01-20 |
| 修稿时间： | 2006年1月20日 |
THE EXPRESSION OF RECOMBINANT mIL-2 GENE IN TRANSFECTED EUKARYOTIC CELLS |
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| Lu Xiaohui, Du Jun, Zeng Jun,et al..THE EXPRESSION OF RECOMBINANT mIL-2 GENE IN TRANSFECTED EUKARYOTIC CELLS[J].Journal of Guangxi Medical University,2006,23(3):352-354. |
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| Authors: | Lu Xiaohui Du Jun Zeng Jun |
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| Institution: | Pharmacy School of Jinan University, Guangzhou 510632 China |
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| Abstract: | Objective:To construct a eukaryotic expressive plasmid that codes mIL-2 gene and test its expression in C_ 2 C_ 12 cells.Methods:mIL-2 gene was inserted into eukaryotic expression vector named pIRESneo2 by enzymolysis and ligation, and at the same time, the stop code of mIL-2 cDNA was taken away. It was confirmed by restrictive enzymes (Age I/BamH I) digestion and by analysis of DNA sequencing. The C_ 2 C_ 12 cells were transfected with the recombinant plasmid and the expression of recombinant mIL-2 was examined by Western blot.Result:DNA sequencing shows that the recombinant plasmid pIRESneo2/mIL-2 has been successfully constructed. And the expression of recombinant mIL-2 was testified in C_ 2 C_ 12 cells by Western blot. Conclusion:A eukaryotic engineering cell line expressing mIL-2 steadily has been established successfully. |
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| Keywords: | mouse interleukin-2 eukaryotic expression gene transfection |
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