| 人源性中性内肽酶稳定转染细胞株的建立 |
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| 引用本文： | 孔峰,胡晓燕,杨玲玲,丁岩,王丽娜,崔行.人源性中性内肽酶稳定转染细胞株的建立[J].山东大学学报(医学版),2006,44(2):118-121. |
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| 作者姓名： | 孔峰 胡晓燕 杨玲玲 丁岩 王丽娜 崔行 |
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| 作者单位： | 山东大学医学院生物化学与分子生物学研究所,山东,济南,250012 |
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| 摘 要： | 目的：获得中性内肽酶（NEP）及失活中性内肽酶（inNEP）稳定表达的细胞株。方法：用酶切及基因测序鉴定质粒pcDNA3．1-hNEP及pcDNA3．1-inNEP（E585V）；以脂质体Ljpofectamjne^TM2000介导pcDNA3．1-hNEP及pcDNA3．1-inNEP（E585V）转染神经母细胞瘤细胞SK-N-SH，分别获得两者的稳定表达细胞株；用RT-PCR及Western blot分析NEP的表达。结果：转染4周后，可见G418单克隆抗性细胞株形成，转染pcDNA3．1．hNEP及pcDNA3．1-inNEP（E585V）的细胞NEP表达均增高。结论：获得稳定转染pcDNA3．1-hNEP及ocDNA3．1-inNEP（E585V）的细胞株。
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| 关 键 词： | 中性内肽酶 阿尔茨海默病 基因治疗 转染 细胞转化 肿瘤 |
| 文章编号： | 1670-7554（2006）02-0018-04 |
| 收稿时间： | 2005-03-09 |
| 修稿时间： | 2005-03-09 |
Building of stably transfected cell strain on human neprilysin |
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| KONG Feng,HU Xiao-yan,YANG Ling-ling,DING Yan,WANG Li-na,CUI Xing.Building of stably transfected cell strain on human neprilysin[J].Journal of Shandong University:Health Sciences,2006,44(2):118-121. |
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| Authors: | KONG Feng HU Xiao-yan YANG Ling-ling DING Yan WANG Li-na CUI Xing |
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| Institution: | Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan 250012, Shandong, China |
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| Abstract: | Objective: To obtain human neprilysin stably transfected neuroblastoma cell strain. Methods: The plasmid pcDNA3.1-hNEP and pcDNA3.1-inNEP(E585V) were identified with restriction endonuclease digesting and gene sequencing, then were transfected into SK-N-SH cells with LipofectamineTM2000, G418 was used for screening. The stably transfected cells were confirmed with RT-PCR and Western blot. Results: The monoclonal cell strains resisting to G418 were formed 4 weeks after transfected with pcDNA3.1-hNEP and pcDNA3.1-inNEP(E585V)respectively. The results of RT-PCR and Western blot showed that the stably transfected cells expressed much more NEPs than control cells. Conclusion:Human neprilysin transfected neuroblstoma cell strain has been obtained. |
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| Keywords: | Neutral endopeptidase Alzheimer disease Gene thearapy Transfection Cell transformation neoplastic |
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