| 小鼠骨骼肌Desmin启动子的克隆及表达活性分析 |
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| 引用本文： | 薛超,安星兰,王春生,朴善花,苗向阳,安铁洙. 小鼠骨骼肌Desmin启动子的克隆及表达活性分析[J]. 黑龙江畜牧兽医, 2011, 0(7) |
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| 作者姓名： | 薛超 安星兰 王春生 朴善花 苗向阳 安铁洙 |
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| 作者单位： | 东北林业大学生命科学学院;中国农业科学院北京畜牧兽医研究所; |
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| 基金项目： | 国家转基因生物新品种培育科技重大专项(2009ZX08008-004B) |
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| 摘 要： | 为了研究在体细胞中小鼠Desmin基因启动子是否具有表达活性,试验根据NCBI中小鼠Desmin启动子的序列,利用生物软件Primer Premier 6.0设计序列上、下游引物,克隆小鼠基因组中Desmin启动子片段,将克隆到的Desmin启动子片段与pAcGFP1-N1载体中的CMV启动子进行置换以构建真核表达载体pAcGFP1-N1-Des,然后将pAcGFP1-N1-Des依次转染CHO-K1、COS-7、293GP和NIH-3T3等细胞后,观察绿色荧光蛋白表达活性。结果表明:克隆得到的Desmin基因上游启动子(841 bp)与NCBI序列(NT_039173.7)比对同源性为99.76%;克隆得到的启动子中含有CAAT-box、GC-box核心调控区以及MyoD等多种转录因子结合位点;所构建的pAcGFP1-N1-Des经转染COS-7、CHO-K1和Hela细胞后可观察到呈现表达活性的绿色荧光。说明克隆获得了在COS-7、CHO-K1以及Hela细胞中的具有表达活性的小鼠Desmin基因启动子。
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| 关 键 词： | 小鼠 Desmin基因 启动子 活性分析 |
Cloning mouse Desmin promoter and detect the expressive activity |
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| XUE Chao,AN Xing-lan,WANG Chun-sheng,PIAO Shan-hua,MIAO Xiang-yang,AN Tie-zhu. Cloning mouse Desmin promoter and detect the expressive activity[J]. Heilongjiang Animal Science And veterinary Medicine, 2011, 0(7) |
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| Authors: | XUE Chao AN Xing-lan WANG Chun-sheng PIAO Shan-hua MIAO Xiang-yang AN Tie-zhu |
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| Affiliation: | XUE Chao1,AN Xing-lan1,WANG Chun-sheng1,PIAO Shan-hua1,MIAO Xiang-yang2,AN Tie-zhu1(1.College of Life Science,Northeast Forestry University,Harbin 150040,China,2.Insitute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China) |
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| Abstract: | In order to study whether the mouse Desmin gene promoter has started downstream gene expression activity in somatic cells,experiments used PCR amplification to clone mouse Desmin gene promoter and then lay a foundation for construct the vector which can express shRNA in the muscle-derived cells.Experiments were performed as follows.Primers were designed primers by Primer Premier 6.0 and amplified Desmin promoter that was used to replaced the CMV promoter in pAcGFP1-N1 vector,and then the recombinant plasmid... |
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| Keywords: | Mouse Desmin gene promoter activity analysis |
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