| Netrin-1慢病毒表达载体的构建及鉴定 |
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| 引用本文： | 陈景祥,王国华,任小宝,屈纪富,孙溦.Netrin-1慢病毒表达载体的构建及鉴定[J].现代生物医学进展,2010,10(4). |
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| 作者姓名： | 陈景祥 王国华 任小宝 屈纪富 孙溦 |
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| 作者单位： | 1. 第三军医大学西南医院急诊科,重庆,400038
2. 解放军第154医院内三科,河南,信阳,464250 |
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| 基金项目： | 重庆市自然科学基金计划项目(CSTC,2007BB5040) |
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| 摘 要： | 目的:构建携带有Netrin-1基因的逆转录病毒载体,为研究Netrin-1在神经发育中的作用奠定基础。方法:PCR扩增Netrin-1基因片段后,将其克隆入慢病毒表达载体pLXSN;通过PCR、酶切、测序鉴定重组质粒。重组质粒转染PA317包装细胞后获得包装的病毒颗粒。病毒颗粒感染人脑胶质瘤细胞SW038-C2,经Western blot证明重组病毒在真核细胞内表达Netrin-1的情况。结果:经PCR扩增、酶切和测序验证,重组质粒构建正确,命名为pLX-NT。Western blot证明在感染细胞泳道有一特异性条带。结论:成功构建了能表达Netrin-1的慢病毒载体。
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| 关 键 词： | Netrin-1 慢病毒载体 神经发育 |
Construction and identification of lentiviral vector with Netrin-1 gene |
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| CHEN Jing-xiang,WANG Guo-hua,REN Xiao-bao,QU Ji-fu,SUN Wei.Construction and identification of lentiviral vector with Netrin-1 gene[J].Progress in Modern Biomedicine,2010,10(4). |
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| Authors: | CHEN Jing-xiang WANG Guo-hua REN Xiao-bao QU Ji-fu SUN Wei |
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| Abstract: | Objective: To construct a lentiviral vector expressing Netrin-1 gene for studying the role of Netrin-1 in the development of neuron. Methods: The gene fragment of netrin-1 was obtained by PCR and cloned into lentiviral vector pLXSN. Then the recombinant plasmid was confirmed by PCR,restriction endonuclease digestion and sequencing. The recombinant retroviral vector pLX-NT was selected to transfect packaging cell PA317 to gain virus particles with infection ability. Human glioblastoma multiform cells SW038-C... |
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| Keywords: | Netrin-1 lentiviral vector development of neuron |
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