| 小鼠白细胞介素21 cDNA的克隆及真核表达质粒的构建 |
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| 引用本文： | 陈国兵,窦骏,陈峻崧,洪晓武.小鼠白细胞介素21 cDNA的克隆及真核表达质粒的构建[J].细胞与分子免疫学杂志,2004,20(1):49-52. |
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| 作者姓名： | 陈国兵 窦骏 陈峻崧 洪晓武 |
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| 作者单位： | 东南大学医学院病原生物学和免疫学系,江苏,南京,210009 |
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| 摘 要： | 目的：克隆小鼠自细胞介素2l(IL-21)基因，构建真核表达质粒，用以进行肿瘤的基因治疗。方法：用RT-PCR法。从ConA活化的小鼠T细胞中扩增IL-21 cDNA。克隆人哺乳动物细胞高效表达质粒pcDNA3．1中，构建重组mIL-21真核表达质粒。重组体用载体上的通用引物和PCR下游引物为测序引物，鉴定克隆的正确性。将已鉴定的重组质粒用脂质体法转染Sp2／0细胞，用RT-PCR法鉴定转染细胞中IL-21基因的表达，用MTT比色法检测表达的mIL-21诱导的NK细胞杀伤活性的增强。结果：正确构建了重组真核表达质粒pcDNA3．1／mIL-21，并在转染的细胞中检测出IL-2l的表达，表达的mIL-21可在体外增强NK细胞的杀伤活性：结论：成功地构建了重组真核表达质粒pcDNA3．1／mIL-21，为进一步在肿瘤动物模型中进行IL-21基因治疗及疗效观察奠定了基础.
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| 关 键 词： | 白细胞介素21 RT-PCR 重组质粒 |
| 文章编号： | 1007-8738(2004)01-0049-04 |
| 修稿时间： | 2003年3月12日 |
Cloning and eukaryotic expression of mouse interleukin 21 |
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| CHEN Guo-bing,DOU Jun ,CHEN Jun-song,HONG Xiao-wu.Cloning and eukaryotic expression of mouse interleukin 21[J].Journal of Cellular and Molecular Immunology,2004,20(1):49-52. |
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| Authors: | CHEN Guo-bing DOU Jun CHEN Jun-song HONG Xiao-wu |
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| Institution: | Department of Microbiology and Immunology, Medical College, Southeast University, Nanjing 210009, China. pawnking@vip.sina.com |
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| Abstract: | AIM: To clone mouse interleukin 21 (mIL-21) gene and construct its eukaryotic expression vector. METHODS: mIL-21 cDNA was amplified from ConA-activated mouse T cells by RT-PCR. And then the cDNA was cloned into eukaryotic expression plasmid pcDNA3.1 to construct recombinant plasmid pcDNA3.1/mIL-21 which was introduced into Sp2/0 cells by lipofectin. Expression of mIL-21 gene was detected by RT-PCR and NK cytotoxicity assay. RESULTS: Recombinant plasmid pcDNA3.1/mIL-21 was constructed correctly and mIL-21 gene was expressed in transfected Sp2/0 cells. CONCLUSION: mIL-21 gene was cloned and expressed successfully, which lays the foundation for further studying anti-tumor effect of mIL-21 gene therapy in animal models. |
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| Keywords: | interleukin 21 RT-PCR recombinant plasmid |
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