| Smyd1干扰真核表达质粒及其稳定转染H9c2细胞系的构建 |
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| 引用本文： | 李帆,廖四芳,刘华友,鲁建鑫,刘宪楚,刘明,吴秀山,李永青. Smyd1干扰真核表达质粒及其稳定转染H9c2细胞系的构建[J]. 激光生物学报, 2011, 20(5). DOI: 10.3969/j.issn.1007-7146.2011.05.016 |
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| 作者姓名： | 李帆 廖四芳 刘华友 鲁建鑫 刘宪楚 刘明 吴秀山 李永青 |
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| 作者单位： | 湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室,心脏发育研究中心,湖南长沙410081 |
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| 基金项目： | 国家然科学基金项目(30930054、30871417、30971663); 国家重点基础研究发展计划项目(2005CB522505); 湖南省自然科学基金项目(09JJ5014) |
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| 摘 要： | 运用RNA干扰技术(RNA interference RNAi)构建pSUPER.retro-Smyd1真核表达质粒,经鉴定后用脂质体法转染H9c2细胞,通过G418筛选出稳定表达pSUPER.retro-Smyd1的细胞系,最后经Western blot及RT-PCR实验鉴定其干扰效果。经鉴定,通过H9c2细胞系构建的pSUPER.retro-Smyd1干扰细胞系的干扰效果显著。因此,本实验成功构建了Smyd1干扰真核表达质粒及其稳定转染的H9c2细胞系,为进一步研究Smyd1基因在心脏发育中的作用奠定了良好的实验基础。
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| 关 键 词： | Smydl基因 RNAi技术 siRNA H9c2细胞系 稳定转染 |
Construction of RNAi Recombinant Eukaryotic Expression Vectors of Smyd1 and Establishment of Its Stably Transfected H9c2 Cell Lines |
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| LI Fan,LIAO Sifang,LIU Huayou,LU Jianxin,LIU Xianchu,LIU Ming,WU Xiushan,Li Yongqing. Construction of RNAi Recombinant Eukaryotic Expression Vectors of Smyd1 and Establishment of Its Stably Transfected H9c2 Cell Lines[J]. ACTA Laser Biology Sinica, 2011, 20(5). DOI: 10.3969/j.issn.1007-7146.2011.05.016 |
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| Authors: | LI Fan LIAO Sifang LIU Huayou LU Jianxin LIU Xianchu LIU Ming WU Xiushan Li Yongqing |
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| Affiliation: | LI Fan,LIAO Sifang,LIU Huayou,LU Jianxin,LIU Xianchu,LIU Ming,WU Xiushan~*,Li Yongqing~* (The Center for Heart Development,Key Lab of MOE for Development Biology and Protein Chemistry,Hunan Normal University,Changsha 410081,Hunan,China) |
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| Abstract: | The eukaryotic expression vector of pSUPER-Smyd1 was constructed for RNA interference.After verification, we transfected the plasmids into H9c2 cells by Lipofectamine 2000.After screening by G418,stable transfected cell lines were established to express pSUPER-Smyd1.Finally,the interferential effect was demonstrated by Western blot and RT-PCR.In a word,RNAi recombinant eukaryotic expression vector of Smyd1 was successfully Constructed and the establishment of its stably transfected H9c2 cell lines laid a so... |
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| Keywords: | Smyd1 gene RNAi siRNA H9c2 cell lines stable transfection |
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