| 刘德勇，张庆丽，龙晓兰，彭和人，谢海龙.S100A11慢病毒表达载体的构建及其在肝癌细胞中的表达.[J].中南医学科学杂志.,2014,42(6):554-557. |
| S100A11慢病毒表达载体的构建及其在肝癌细胞中的表达 |
| Construction of Lentivirus Vector S100A11 andits Expression in Hepatoma Cells |
| 投稿时间：2014-06-22 |
| DOI： |
| 中文关键词: S100A11 Huh-7 细胞 慢病毒载体 |
| 英文关键词:S100A11 Huh-7 cells lentiviral vectors |
| 基金项目:湖南省教育厅湖南省高校科研项目（No.12C1051），衡阳市科技局计划项目（2012KJ34）. |
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| 中文摘要: |
| 目的构建S100A11基因慢病毒表达载体，建立Huh-7 S100A11稳定细胞株，并鉴定其表达。方法RT-PCR扩增S100A11基因片段，与pWPT载体经MluⅠ和NotⅠ同时酶切后连接，构建慢病毒表达载体pWPT-S100A11；将表达载体pWPT-GFP和pWPT-S100A11与慢病毒包装质粒 pMD2.G和psPAX2共同感染293T细胞，获得携带GFP和S100A11基因的慢病毒，将两种慢病毒分别感染Huh-7细胞，获得Huh-7 S100A11和Huh-7GFP稳定细胞株；利用Real-Time PCR和Western Blotting实验方法检测感染后S100A11的过表达情况。结果成功构建慢病毒表达载体pWPT-S100A11；慢病毒感染Huh-7细胞株后，阳性对照Huh-7GFP经荧光镜检测传染率达95%；感染Huh-7细胞后S100A11的mRNA和蛋白表达量均增加。结论成功构建慢病毒表达载体pWPT-S100A11，并建立了Huh-7 S100A11稳定细胞株，为进一步研究S100A11基因在肝癌中的生物学功能和机制奠定了基础。 |
| 英文摘要: |
| ObjectiveTo construct lentiviral vector carrying S100A11 gene and infected Huh-7 cells,and detect the expression of S100A11.MethodsS100A11 gene was obtained by RT-PCR,and then cloned into the lentiviral vector pWPT digested by MluⅠand NotⅠ to construct lentiviral vector pWPT-S100A11.The vector pWPT-GFP and pWPT-S100A11 with pMD2.G and psPAX2 co-infected 293T cells to get the recombinant lentivirus carrying GFP and S100A11 genes.The recombinant lentivirus carrying GFP and S100A11 were used to infect Huh-7 cells respectively.The expression of S100A11 in Huh-7 cells was detected by Real-Time PCR and Western blotting.ResultsThe recombinant lentivirus expression vector pWPT-S100A11 has been successfully constructed.The efficiency of recombinant lentivirus transfection reached to 95% under the green fluorescent.The mRNA and protein of S100A11 expression were all increased in Huh-7 cells after infection.ConclusionpWPT-S100A11 and Huh-7 S100A11 cells will be used in studying the function and mechanism of S100A11 in hepatic carcinomar. |
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