| 重组人白细胞介素21真核表达载体的构建及体外药效学研究 |
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| 引用本文： | 饶桂荣,朱梅,杨妙青,杨富强,王洪敏,吴庆洲,陈光明.重组人白细胞介素21真核表达载体的构建及体外药效学研究[J].华南国防医学杂志,2010,24(6):431-434. |
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| 作者姓名： | 饶桂荣 朱梅 杨妙青 杨富强 王洪敏 吴庆洲 陈光明 |
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| 作者单位： | 解放军458医院全军肝病中心;广东省人民医院药学部; |
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| 摘 要： | 目的构建人的真核表达载体pVAX/IL-21,并进行体外细胞转染实验的鉴定研究。方法采用PCR扩增技术扩增含有白细胞介素21（interleukin21,IL-21）的基因,亚克隆于真核表达载体pVAX1,并进行酶切和测序分析;将所构建的质粒转染COS-7细胞检测其表达情况。结果酶谱和测序分析表明,构建的质粒pVAX/IL-21（pIL-21）基因片段的方向、序列完全正确;ELISA法定量检测双质粒转染COS-7细胞后24h、48h、72h、96h均检测到目的基因的蛋白表达,其中转染48h的表达水平较高,达到45ng/ml。结论成功构建了人IL-21的整合表达质粒pIL-21,并具有较强的体外细胞转染活性,为进一步研究IL-21的生物学功能如抗肿瘤活性等提供了实验基础。
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| 关 键 词： | 人白细胞介素21 构建 转染 基因表达 |
Construction and in vitro Pharmacodynamics of Human Interleukin 21 Eukaryotic Expression Plasmid |
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| RAO Gui-rong,ZHU Mei,YANG Miao-qing,YANG Fu-qiang,WANG Hong-min,WU Qing-zhou,CHEN Guang-ming.Construction and in vitro Pharmacodynamics of Human Interleukin 21 Eukaryotic Expression Plasmid[J].Military Medical Journal of South China,2010,24(6):431-434. |
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| Authors: | RAO Gui-rong ZHU Mei YANG Miao-qing YANG Fu-qiang WANG Hong-min WU Qing-zhou CHEN Guang-ming |
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| Institution: | .(Research Center of Hepatic Disease,No.458 Hospital of the People’s Liberation Army,Guangzhou Guangdong 510600,China ) |
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| Abstract: | Objective To develop effective eukaryotic expression plasmids,pVAX/IL-21,and perform the evaluation study on in vitro transfection experiment.Methods Interleukin 21(IL-21) genes were amplified by PCR technique,and then subcloned into eukaryotic expression palsmid pVAX1 by double enzymes and ligase.The new plasmids were analyzed by endonuclease restriction and DNA sequencing.The constructed plasmids were transfected into COS-7 cells in vitro with reagent LipofectamineTM 2000.The expression was quantified by ... |
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| Keywords: | Human interleukin 21 Construction Transfection Gene expression |
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