| CD59真核表达载体的构建及其对Jurkat细胞增殖的影响 |
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| 引用本文： | 钟丹丹,高美华,张蓓.CD59真核表达载体的构建及其对Jurkat细胞增殖的影响[J].免疫学杂志,2011(3):228-231. |
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| 作者姓名： | 钟丹丹 高美华 张蓓 |
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| 作者单位： | 青岛大学医学院免疫学教研室; |
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| 基金项目： | 国家自然科学基金资助项目(30972677) |
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| 摘 要： | 目的构建pIRES-CD59融合基因真核表达载体,探讨重组CD59在Jurkat细胞增殖中的作用。方法 RT-PCR法选择性扩增编码CD59的基因片段,克隆入pIRES真核表达质粒。脂质体法将重组载体转染Jurkat细胞,通过G418筛选获得稳定表达CD59的细胞克隆,利用RT-PCR和Western blot检测细胞中CD59的表达,通过MTT法检测Jurkat细胞的增殖。结果经酶切和测序鉴定表明携带CD59基因的重组质粒构建成功,RT-PCR和Western blot结果显示转染Jurkat细胞的CD59基因高表达,MTT实验显示转染CD59重组质粒的Jurkat细胞增殖速度明显快于对照组(P<0.05)。结论 pIRES-CD59真核表达载体在转染细胞Jurkat中可高表达CD59分子,并可影响Jurkat细胞的增殖,为研究CD59的生物学作用奠定了基础。
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| 关 键 词： | CD59基因 Jurkat细胞 细胞增殖 |
Construction of a eukaryotic expression vector containing CD59 gene and its role in Jurkat cell proliferation |
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| ZHONG D,an,GAO Meihua,ZHANG Bei.Construction of a eukaryotic expression vector containing CD59 gene and its role in Jurkat cell proliferation[J].Immunological Journal,2011(3):228-231. |
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| Authors: | ZHONG D an GAO Meihua ZHANG Bei |
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| Institution: | ZHONG Dandan,GAO Meihua,ZHANG BeiDepartment of Immunology,Medical College of Qingdao University,Qingdao 266071,China |
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| Abstract: | pIRES-CD59 fusion gene eukaryotic expression vector was constructed in order to discuss therole of CD59 gene in Jurkat cell proliferation.CD59 gene was amplified by RT-PCR and inserted into theeukaryotic expression plasmid-pIRES.The vector containing the object gene was transfected into Jurkat cells byliposome,while steady cells cloned could get through G418 screening.CD59 mRNA and protein levels weredetected by RT-PCR and Western blot;Jurkat cell proliferation was analyzed by MTT.The eukaryotic expressionv... |
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| Keywords: | CD59 gene Jurkat cell Cell proliferation |
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