| 人TLR4胞外段真核表达质粒的构建及其在HEK293 细胞中的表达 |
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| 引用本文： | 陈伟,史忠. 人TLR4胞外段真核表达质粒的构建及其在HEK293 细胞中的表达[J]. 中国急救医学, 2005, 25(12): 899-901 |
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| 作者姓名： | 陈伟 史忠 |
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| 作者单位： | 中国人民解放军第三军医大学新桥医院急救部,重庆,400037 |
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| 基金项目： | 国家重点基础研究发展规划973项目课题资助（No.2005CB522605） |
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| 摘 要： | 目的 构建人Toll—like receptor 4（TLR4）胞外段真核表达质粒并观察其表达。方法 通过PCR扩增人TLR4胞外段基因，将其克隆入真核表达质粒pcDNA3．1（＋），重组质粒peDNA3．1（＋）-eTLR4转染入人胚肾293细胞（HEK293 细胞）中，RT-PCR检测TLR4胞外段的表达。结果 测序结果表明，成功构建人TLR4胞外段真核表达质粒pcDNA3．1（＋）-eTLR4，并在HEK293细胞中获得表达。结论 TLR4胞外段能在HEK293细胞中获得正确表达，为TLR4信号通路的进一步研究打下了基础。
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| 关 键 词： | Toll样受体4 基因表达 内毒素 |
| 文章编号： | 1002-1949(2005)12-0899-03 |
| 收稿时间： | 2005-07-16 |
| 修稿时间： | 2005-07-16 |
Construction of eukaryotic expression plasmid of extracellular domain of human TLR4 and its expression in HEK293 cell |
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| CHEN Wei,SHI Zhong. Construction of eukaryotic expression plasmid of extracellular domain of human TLR4 and its expression in HEK293 cell[J]. Chinese Journal of Critical Care Medicine, 2005, 25(12): 899-901 |
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| Authors: | CHEN Wei SHI Zhong |
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| Affiliation: | Emergency Department, Xinqiao Hospital, the Third Military Medical University, Chongqing 400037, China |
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| Abstract: | Objective To construct eukaryotic expression plasmid of extracellular domain of human toll-like receptor 4 and determine its expression in HEK293 cell.Methods Human TLR4 extracellular cDNA codon domain was amplified by polymerase chain reaction(PCR) and cloned into pcDNA3.1(+).Recombinant plasmid pcDNA3.1(+)-eTLR4 was transfected into HEK293 cell.The expression of eTLR4 was determined by RT-PCR.Results The eTLR4 gene was proved to be correct by DNA sequencing.The eTLR4 mRNA was detected in HEK293 cell.Conclusion Extracellular domain of TLR4 could be expressed correctly in HEK293 cell,which laid the basis for the advanced study of TLR4 signaling pathway. |
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| Keywords: | Toll- like receptor 4 Gene expression Endotoxin |
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