| 凋亡蛋白抑制因子Livin shRNA慢病毒载体的构建与鉴定 |
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| 引用本文： | 焦伊胜,王永来,张淑兰,王桂玲.凋亡蛋白抑制因子Livin shRNA慢病毒载体的构建与鉴定[J].中国现代医学杂志,2008,18(24):3601-3605. |
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| 作者姓名： | 焦伊胜 王永来 张淑兰 王桂玲 |
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| 作者单位： | 1. 中国医科大学附属盛京医院,妇产科,辽宁,沈阳,110004
2. 中国医科大学,细胞生物教研室,辽宁,沈阳,110004 |
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| 基金项目： | 辽宁省教育厅高等学校科研项目(05L475)资助 |
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| 摘 要： | 目的 构建并鉴定凋亡蛋白抑制因子Livin基因短发夹状干扰RNA(shRNA)慢病毒我体,以便进一步研究Livin的功能.方法 针对已经筛选确定的Livin基因RNAi有效靶序列,合成靶序列的Oligo DNA,退火形成双链DNA,与经Hpa I和Xho I酶切后的pGCL-GFP载体含U6启动子和绿色荧光蛋白(GFP)]连接、转化,产生pLV-shLivin慢病毒表达载体,PCR筛选阳性克隆,测序鉴定.用pLV-shLivin,pHelper 1.0和pHelper 2.0 3种质粒共转染包装细胞人胚肾293T细胞,包装产生重组慢病毒,以293T细胞GFP蛋白的表达水平测定病毒滴度.结果 PCR和测序证实,成功构建Livin shRNA的慢病毒栽体LV-shLivin.包装慢病毒,浓缩病毒悬液的滴度为3×108TU/ML.结论 成功构建Livin基因sbRNA慢病毒栽体,为应用RNAi研究Livin功能奠定基础.
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| 关 键 词： | 细胞凋亡 Livin 慢病毒载体 |
Construction and identification of lentivira l vector expressing of shRNA of Livin gene |
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| JIAO Yi-sheng
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WANG Yong-lai
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ZHANG Shu-lan
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WANG Gui-ling.Construction and identification of lentivira l vector expressing of shRNA of Livin gene[J].China Journal of Modern Medicine,2008,18(24):3601-3605. |
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| Authors: | JIAO Yi-sheng WANG Yong-lai ZHANG Shu-lan WANG Gui-ling |
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| Institution: | JIAO Yi-sheng1,WANG Yong-lai1,ZHANG Shu-lan1,WANG Gui-ling2(1.Department of Obstetrics , Gynecology,Shengjing Hospital of China Medical University,Shenyang,Liaoning 110004,P.R.China,2.Department of Cell Biology,China Medical University,Liaoning 110001,P.R.China) |
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| Abstract: | Objective] To construct and identify a lentiviral vector expressing short hairpin interference RNA(shRNA) of Livin gene to study the function of Livin.Methods] The effective sequence of siRNA targeting Livin gene was confirmed in our previous study.The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pGCL-GFP vector,which contained U6 promoter and green fluorescent protein(GFP).The resulting lentiviral vector containing ... |
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| Keywords: | apoptosis Livin Lentivirus vector |
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