| Mef2c真核表达载体的构建及其在HEK293T细胞中的表达 |
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| 引用本文： | 温见燕,于长安,杨鹏,阴彬,李耿,柯元南,廖文强. Mef2c真核表达载体的构建及其在HEK293T细胞中的表达[J]. 山东医药, 2009, 49(4): 15-17 |
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| 作者姓名： | 温见燕 于长安 杨鹏 阴彬 李耿 柯元南 廖文强 |
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| 作者单位： | 中日友好医院,全国中西医结合心血管病中心,北京100029;中国医学科学院,北京协和医学院基础医学研究所医学分子生物学国家重点实验室 |
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| 摘 要： | 目的构建小鼠Mef2c基因真核表达质粒并转染HEK293T细胞。方法采用RT-PCR方法从小鼠心脏组织的总cDNA中扩增出小鼠的Mef2c的基因,采用基因重组技术将Mef2c的cDNA片段插入真核表达载体peD—NA3．1（＋）,构建小鼠Mef2c真核表达质粒,脂质体转染HEK293T细胞进行表达。结果酶切和测序结果证实Mef2c真核表达质粒构建成功,经脂质体转染293T细胞后,Western blot检测证明Mef2c蛋白在真核细胞中成功表达。结论成功构建真核表达质粒pcDNA3．1（＋）．Mef2c,为进一步研究小鼠Mef2c在心肌分化过程中的作用及其调控机制奠定了实验基础。
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| 关 键 词： | MEF2 基因表达 载体构建 |
Construction of eukaryotic expression vector for Mef2c and its expression in HEK293T cells |
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| WEN Jian-yan,YU Chang-an,YANG Peng,YIN Bin,LI Gen,KE Yuan-nan,LIAO Wen-qiang. Construction of eukaryotic expression vector for Mef2c and its expression in HEK293T cells[J]. Shandong Medical Journal, 2009, 49(4): 15-17 |
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| Authors: | WEN Jian-yan YU Chang-an YANG Peng YIN Bin LI Gen KE Yuan-nan LIAO Wen-qiang |
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| Affiliation: | WEN Jian-yan , YU Chang-an, YANG Peng, YIN Bin, LI Gen, KE Yuan-nan, LIAO Wen-qiang ( 1 National Integrative Medicine Center for Cardiovascular Diseases, China-Japan Friendship Hospital, Beijing 100029, P. R. China) |
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| Abstract: | Objective To construct a recombinant mouse MeE2c eukaryotic vector and transfect HEK293T in vitro. Methods The Mei2c was amplified by polymerase chain reaction （PCR） from mouse heart tissue, and the Mef2c gene was inserted into eukaryotic expression vector pcDNA3.1 （ ＋ ）. The recombinant pcDNA3.1-Mef2c plasmids were transfected into HEK293T cells using Lipofectamine. Results Mef2c eukaryotic vector was successfully constructed and confirmed by endonuclease digestion and DNA sequencing. The expression of Mef2c protein was detected by Western blot,which suggested that it could be expressed in eukaryotic cells. Coulusions These results provided a basis for further investigation of Mei2c function. |
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| Keywords: | MEF2 |
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