| 可诱导稳定表达DLX4细胞系的构建及其表达分析 |
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| 引用本文： | 张连美,傅俊江.可诱导稳定表达DLX4细胞系的构建及其表达分析[J].泸州医学院学报,2011,34(5):480-482. |
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| 作者姓名： | 张连美 傅俊江 |
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| 作者单位： | 泸州医学院医学基础研究中心,四川泸州,646000 |
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| 基金项目： | 国家自然基金项目(81172049); 四川省教育厅重点项目(10ZA038); 泸州医学院启动基金资助项目 |
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| 摘 要： | 目的：建立一种可诱导稳定表达方法,并构建DLX4细胞系。分析DLX4细胞系的表达调控。方法：将DLX4全长基因克隆入pcDNA5/FRT/TO表达载体,将其转染入宿主细胞Flp-InTMT-RExTM-293细胞系,潮霉素B筛选或者阳性克隆。采用不同剂量的DOX或者不同作用时间对pcDNA5/DLX4细胞系进行诱导表达,用Western blotting检测DLX4表达情况。结果：构建了可被Doxycline（DOX）诱导的稳定表达pcDNA5/DLX4的细胞系。0.001μg/ml的DOX即可诱导pcDNA5/DLX4细胞系的表达,0.1μg/ml DOX即可诱导该细胞系表达达到高峰;DOX对该细胞系作用1h后即可诱导该细胞系表达,对其作用16h后该细胞系表达明显增强。结论：构建了可被DOX诱导稳定表达的DLX4细胞系,不同剂量的DOX和不同作用时间对DLX4细胞系表达有不同的影响。该细胞系的建立为深入探索DLX4的功能提供了一种新的有效方法和实验材料。
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| 关 键 词： | Doxycline DLX4 稳定细胞系 蛋白质表达 诱导 |
ESTABLISHMENT OF INDUCIBLE DLX4 CELL LINE WITH STABLE EXPRESSION AND IT'S EXPRESSIVE ANALYSIS |
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| Zhang Lianmei,et al The Research Center for Preclinical Medicine,Luzhou Medical College.ESTABLISHMENT OF INDUCIBLE DLX4 CELL LINE WITH STABLE EXPRESSION AND IT'S EXPRESSIVE ANALYSIS[J].Journal of Luzhou Medical College,2011,34(5):480-482. |
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| Authors: | Zhang Lianmei The Research Center for Preclinical Medicine Luzhou Medical College |
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| Institution: | Zhang Lianmei,et al The Research Center for Preclinical Medicine,Luzhou Medical College |
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| Abstract: | Objective: To establish an inducible and stable expression method to make DLX4 cell line,and to analyze the expressive regulation of DLX4 cell line by Doxycline.Methods: The DLX4 ORF was cloned into pcDNA5/FRT/TO expression vector,then transfected into the host cell line Flp-InTMT-RExTM-293,and the positive clones were selected using hygromycin B as selection drug.The expression of pcDNA5/DLX4 cell line was induced with different dosage of DOX or different action time with certain DOX dosage.Western blottin... |
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| Keywords: | Doxycline DLX4 Stable cell line Protein expression Induction |
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