| 人SET基因RNA干扰重组质粒载体的构建及其在肝细胞中的表达 |
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| 引用本文： | 黄新凤,刘建军,蒋英芝,席仁荣,杨亮.人SET基因RNA干扰重组质粒载体的构建及其在肝细胞中的表达[J].热带医学杂志,2011,11(3):291-294. |
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| 作者姓名： | 黄新凤 刘建军 蒋英芝 席仁荣 杨亮 |
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| 作者单位： | 深圳市疾病预防控制中心毒理检验科,广东深圳,518020 |
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| 基金项目： | 国家自然科学基金，深圳市科技计划重点项目 |
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| 摘 要： | 目的建立真核质粒载体介导的人SET基因RNA干扰表达体系,并检测其在肝细胞中对SET蛋白表达的影响,为研究SET蛋白在三氯乙烯致肝细胞毒性的作用打下基础。方法设计并合成2对人SET基因特异性短发夹RNA(short hair-pin RNA,shRNA)寡核苷酸链,退火形成双链DNA后插入到真核表达质粒psiRNA-hH1 neo中产生shRNA重组质粒载体,经酶切和测序鉴定成功后,转染至L-02肝细胞中,48h后收集细胞提取总蛋白,Western blotting检测干扰后SET蛋白的表达。结果 shRNA重组质粒的酶切和测序鉴定均正确,Western blotting结果显示重组质粒psiRNA1组的干扰效果较好,对SET蛋白表达的抑制率达60%左右,与正常对照组相比,差异具有统计学意义(P<0.05)。结论成功构建针对人SET基因的RNA干扰真核质粒载体,其能有效地抑制肝细胞内SET蛋白的表达,为深入研究SET蛋白在三氯乙烯致肝细胞毒性中的作用奠定了基础。
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| 关 键 词： | RNA干扰 SET基因 真核质粒载体 Western Blotting |
Construction of human SET gene RNA interference recombinant plasmid vector and its expression in L-02 liver cells |
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| HUANG Xin-feng,LIU Jian-jun,JIANG Ying-zhi,XI Ren-rong,YANG Liang.Construction of human SET gene RNA interference recombinant plasmid vector and its expression in L-02 liver cells[J].Journal Of Tropical Medicine,2011,11(3):291-294. |
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| Authors: | HUANG Xin-feng LIU Jian-jun JIANG Ying-zhi XI Ren-rong YANG Liang |
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| Institution: | HUANG Xin-feng,LIU Jian-jun,JIANG Ying-zhi,XI Ren-rong,YANG Liang(Shenzhen Municipal Center for Disease Control and Prevention,Guangdong,Shenzhen 518020,China) |
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| Abstract: | Objective To construct human patient SE transloction(SET) gene RNA interference recombinant plasmid vector and investigate SET protein's expression in the liver cells.The aim of this study was to lay a good basis for the further study of effect of SET protein on trichloroethylene-induced liver cells.Methods The 2 pairs of short hair-pin RNA oligonucleotides targeting human SET gene were designed and synthesized.After annealing,these double-strand DNA were cloned to eukaryotic expression plasmid Vector psiRN... |
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| Keywords: | Western Blotting |
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