| WWOX基因真核表达载体的构建及其在SMMC-7721中的表达 |
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| 引用本文： | 刘峰,易继林,李兴睿,杨志芳,陈钢,沈文状,马小鹏.WWOX基因真核表达载体的构建及其在SMMC-7721中的表达[J].临床外科杂志,2008,16(10):663-665. |
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| 作者姓名： | 刘峰 易继林 李兴睿 杨志芳 陈钢 沈文状 马小鹏 |
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| 作者单位： | 华中科技大学同济医学院附属同济医院普外Ⅳ科,武汉,430030 |
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| 摘 要： | 目的克隆人WWOX基因及构建WWOX基因真核表达载体并研究其表达。方法取人新鲜正常肝组织，提取总RNA，采用逆转录-聚合酶联链反应（RT—PCR）扩增WWOX基因，将该基因定向克隆到真核表达载体pEGFP—N1中，构建真核细胞表达载体pEGFP—N1-WWOX，用限制性内切酶酶切分析，DNA序列分析鉴定重组质粒；将测序正确的WWOX基因通过脂质体介导下转染肝癌细胞SMMC-7721。结果重组质粒pEGFP—N1-WWOX经HindⅢ和BamHI双酶切后，获得4700bp片断和1234bp插入片断，序列分析表明插入的片段与Gene Bank发布的序列一致；荧光显微镜下可见转染的SMMC-7721细胞有绿色荧光蛋白的表达。结论成功构建了真核表达载体pEGFP—N1-WWOX，WWOX基因在SMMC-7721细胞内成功表达。
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| 关 键 词： | WWOX基因 克隆 真核载体 基因转染 |
Construction of WWOX eukaryotic expression vector and its expression in SMMC-7721 cell line |
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| LIU Feng,YI Ji-lin,LI Xing-rui,et al..Construction of WWOX eukaryotic expression vector and its expression in SMMC-7721 cell line[J].Journal of Clinical Surgery,2008,16(10):663-665. |
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| Authors: | LIU Feng YI Ji-lin LI Xing-rui |
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| Institution: | LIU Feng,YI Ji-lin,LI Xing-rui,et al.(Department of General Surgery,Tongji Hospital of Tongji Medical College,Huazhong University of Science , Technology,Wuhan 430030,China) |
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| Abstract: | Objective To construct a eukaryotic expression vector of human WWOX full length gene and transfect it into SMMC-7721 cell lines.Methods Total mRNA was extracted from normal human liver tissue,and then the full-length WWOX cDNA was obtained by RT-PCR.The WWOX gene was cloned into pEGFP-N1 vector and the pEGFP-N1-WWOX vector was constructed and transfected into SMMC-7721 cells.Results A fragment of 4700bp and inserted fragment of 1234bp were got by cutting positive recombinant plasmid of pEGFP-N1-WWOX with Hi... |
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| Keywords: | WWOX gene clone eukaryotic vector gene transfection |
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