Neurturin基因的克隆及其在Vero细胞中的表达 |
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引用本文: | 马开利,孙茂盛,施锐,张莹,杨芳,奎翔,谭振国,李鸿钧. Neurturin基因的克隆及其在Vero细胞中的表达[J]. 中国生物制品学杂志, 2008, 21(4): 265-268 |
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作者姓名: | 马开利 孙茂盛 施锐 张莹 杨芳 奎翔 谭振国 李鸿钧 |
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作者单位: | 中国医学科学院北京协和医学院医学生物学研究所,昆明650118 |
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基金项目: | 云南省科技强省计划基础研究重点项目 |
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摘 要: | 目的克隆Neurturin(NTN)基因,并在Vero细胞中表达。方法用RT-PCR方法扩增hNTN cDNA,并克隆至pcDNA3真核表达载体中,经Lipofectamine2000转染Vero细胞,挑选稳定表达克隆,用RT-PCR及免疫荧光分析鉴定,并对转染后细胞的形态及生长状况进行分析。结果重组质粒pcDNA3/hNTN转染Vero细胞后获得了稳定表达克隆,且有目的蛋白表达,转染后细胞形态和生长特性发生了一定改变。结论获得了稳定表达NTN蛋白的Vero细胞株,为其移植并进行帕金森病猴基因治疗动物实验奠定了基础。
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关 键 词: | Neurturin基因 Vero细胞 克隆 表达 帕金森病 |
文章编号: | 1004-5503(2008)04-0265-04 |
修稿时间: | 2007-08-21 |
Cloning of Neurturin Gene and Its Expression in Vero Cells |
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Abstract: | Objective To clone Neurturin(NTN) gene and express in Vero cells.Methods Amplify human NTN cDNA by RT-PCR and clone to eurkaryotic expression vector pcDNA3.Transfect Vero cells with the constructed recombinant plasmid pcDNA3/hNTN in mediation of Lipofectamine 2000.Identify the expressed product by RT-PCR and IFA.Observe the morphology and growth of transfected Vero cells.Results The positive clones for stable expression of NTN was obtained after transfection of Vero cells with recombinant plasmid pcDNA3/hNTN,and the target protein was expressed.The morphology and growth of transfected Vero cells showed significant change.Conclusion The Vero cell strain for stable expression of NTN was established,which laid a foundation of gene therapy of Parkinson disease in rhesus. |
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Keywords: | Neurturin gene Vero cells Cloning Expression Parkinson disease |
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