| 人p75NTR荧光真核表达载体的构建及其在HEK293细胞中的表达 |
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| 引用本文： | 鲁秀敏,朱枫,黄鹏,余瑛,王永堂.人p75NTR荧光真核表达载体的构建及其在HEK293细胞中的表达[J].重庆工学院学报,2008,22(12):48-52. |
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| 作者姓名： | 鲁秀敏 朱枫 黄鹏 余瑛 王永堂 |
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| 作者单位： | [1]重庆工学院化学与生物工程学院,重庆400050 [2]第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室,重庆400042 |
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| 基金项目： | 国家自然科学基金资助项目(30600665); 重庆市自然科学基金资助项目(CSTC,2008BB5107); 第三军医大学青年基金资助项目(06XG048); 创伤烧伤与复合伤国家重点实验室开放课题基金资助项目(2006A-3); 重庆工学院科研启动基金资助项目(2008ZD21) |
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| 摘 要： | 构建了人p75神经营养素受体（p75 neurotrophin receptor，p75NTR）cDNA序列的绿色荧光真核表达载体，并鉴定其在人胚肾293（human embryo kidney293，HEK293）细胞中的表达．采用PCR方法从含人p75NTR的pDC316-HP75质粒中扩增目的片段，经EcoRI和SalI双酶切，定向克隆于pEGFP-N1质粒中，构建绿色荧光真核表达载体pEGFP-N1-HP75，经酶切及测序鉴定后，通过脂质体转染HEK293细胞，激光共聚焦及Western blot法鉴定人p75NTR的表达．结果表明，酶切鉴定和序列分析证实重组质粒含有人p75NTR编码序列，转染实验表明，重组质粒能够在HEK293细胞中表达出具有活性的人p75NTR片段．
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| 关 键 词： | p75神经营养素受体 荧光 重组表达 HEK293细胞 |
Construction of Eukaryotic Fluorescence Expression Plasmid of Human P75NTR and Its Expression in HEK293 Cells |
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| LU Xiu-min,ZHU Feng,HUANG Peng,YU Ying,WANG Yong-tang.Construction of Eukaryotic Fluorescence Expression Plasmid of Human P75NTR and Its Expression in HEK293 Cells[J].Journal of Chongqing Institute of Technology,2008,22(12):48-52. |
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| Authors: | LU Xiu-min ZHU Feng HUANG Peng YU Ying WANG Yong-tang |
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| Affiliation: | LU Xiu-min1,ZHU Feng1,HUANG Peng1,YU Ying1,WANG Yong-tang2 (1.School of Chemistry , Bioengineering,Chongqing Institute of Technology,Chongqing,400050,China,2.State Key Laboratory of Trauma,Burns , Combined Injury,Institute of Surgery Research,Daping Hospital,Third Military Medical University,Chongqing 400042,China) |
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| Abstract: | 1.Objective: To establish a eukaryotic fluorescence expression plasmid of human p75 neurotrophin receptor(p75NTR) cDNA and investigate its expression in human embryo kidney 293(HEK293) cells.2.Methods: The pEGFP-N1-HP75 eukaryotic fluorescence expression vector is constructed by cloning a 835bp insertion of human p75NTR cDNA from pDC316-HP75 plasmid containing the full length open reading frame to pEGFP-N1 vector.After analysis of enzyme hydrolyze and DNA sequence,the recombinant eukaryotic expression vecto... |
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| Keywords: | p75 neurotrophin receptor fluorescence recombinant expression HEK293 cell |
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