| 真核表达Hsa-miR-1基因及其检测方法的建立 |
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| 引用本文： | 胡徐庞,李伟,曹跃芬,张璇,黄青红,徐科,杨耿兵,魏旭斌,钱程,刘立.真核表达Hsa-miR-1基因及其检测方法的建立[J].浙江理工大学学报,2011(5). |
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| 作者姓名： | 胡徐庞 李伟 曹跃芬 张璇 黄青红 徐科 杨耿兵 魏旭斌 钱程 刘立 |
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| 作者单位： | 浙江理工大学生命科学学院新元医药研究所;浙江理工大学生物工程研究所; |
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| 基金项目： | 国家重点基础发展计划资助项目(973)(2010CB529406) |
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| 摘 要： | 获得Hsa-miR-1的方法主要由人工合成,需耗费很高的成本与时间。文章通过将Hsa-miR-1前体部分连接至pcDNA3.0,可实现Hsa-miR-1基因的高表达。构建含有Hsa-miR-1基因表达前体的表达载体pcDNA-Hsa-miR-1和含有与Hsa-miR-1基因完全互补序列的检测载体pEGFP-C1-CM1。通过共转染pcDNA-Hsa-miR-1和pEGFP-C1-CM1检测Hsa-miR-1基因的表达,并采用Taqman探针法定量检测Hsa-miR-1基因表达水平,成功构建了可高效表达Hsa-miR-1基因的真核生物质粒pcDNA-Hsa-miR-1,并首次建立起Hsa-miR-1基因的检测系统pEGFP-C1-CM1,该检测系统的建立将为今后Hsa-miR-1在肿瘤细胞中的研究提供准确、快捷的检测方法。
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| 关 键 词： | Hsa-miR-1载体 Taqman探针检测 pcDNA-Hsa-miR-1 pEGFP-C1-CM1 |
Eukaryotic Expression of Hsa-miR-1 Gene and Establishment of Its Detection Method |
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| HU Xu-panga,LI Weia,CAO Yue-fenb,ZHANG Xuana,HUANG Qing-honga,XU Kea,YANG Geng-bina,WEI Xu-bina,QIAN Chenga,LIU Lia.Eukaryotic Expression of Hsa-miR-1 Gene and Establishment of Its Detection Method[J].Journal of Zhejiang Sci-tech University,2011(5). |
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| Authors: | HU Xu-panga LI Weia CAO Yue-fenb ZHANG Xuana HUANG Qing-honga XU Kea YANG Geng-bina WEI Xu-bina QIAN Chenga LIU Lia |
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| Affiliation: | HU Xu-panga,LI Weia,CAO Yue-fenb,ZHANG Xuana,HUANG Qing-honga,XU Kea,YANG Geng-bina,WEI Xu-bina,QIAN Chenga,LIU Lia (Zhejiang Sci-Tech University,a.Xinyuan Institute of Medicine and Biotechnology,b.Institute of Bioengineering,Hangzhou 310018,China) |
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| Abstract: | Currently available Hsa-miR-1 mainly by the synthetic methods,is very costly and time-consuming.By part precursor of the Hsa-miR-1 connecting to the pcDNA3.0,can be realized Hsa-miR-1 genes were highly expressed.Constructing containing Hsa-miR-1 precursor gene expression vector pcDNA-Hsa-miR-1 and containing Hsa-miR-1 gene completely complementary sequence detection vector pEGFP-C1-CM1,and using Taqman probes for quantification of Hsa-miR-1 gene expression level,can successfully constructe and highly expres... |
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| Keywords: | Hsa-miR-1 vector taqman probe pcDNA-Hsa-miR-1 pEGFP-C1-CM1 |
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