| β-防御素-2荧光素酶报告基因载体及克罗恩突变体的构建 |
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| 引用本文： | 姚国鹏,智发朝,张迎春,陈正彦,智佳,林勇,关婧,王继德,姜泊.β-防御素-2荧光素酶报告基因载体及克罗恩突变体的构建[J].第四军医大学学报,2009(20):2077-2080. |
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| 作者姓名： | 姚国鹏 智发朝 张迎春 陈正彦 智佳 林勇 关婧 王继德 姜泊 |
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| 作者单位： | 南方医科大学南方医院消化内科研究所; |
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| 基金项目： | 广东省自然科学基金(03004770) |
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| 摘 要： | 目的：构建人β-防御素-2（hBD-2）荧光素酶报告基因质粒及其克罗恩病（Crohn’s，CD）相关突变体，并检测其在人胚胎肾T细胞（HEK-293T）中的活化．方法：通过基因重组构建包含hBD-2启动子序列的报告基因质粒hBD-2—780-luc，再通过定点突变技术在启动子区（-233位点）引入变异点（G变为C），构建突变体hBD-2-mut-luc；经酶切及测序验证后，将报告基因质粒和野生型NOD2／CARD15蛋白真核表达载体共转染到HEK一293T中，TNF—α刺激后测定双荧光素酶的表达．结果：酶切和测序证实，构建了重组体hBD-2-780-luc及其突变体hBD-2-mut—luc，两者均能表达荧光素酶活性，且明显高于空载体．结论：hBD-2报告基因质粒hBD-2-780-luc及其CD相关突变体hBD-2-mut—luc构建成功，并在细胞内活化表达荧光素酶．
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| 关 键 词： | β-防御素-2 克罗恩病 荧光素酶报告基因质粒 定点突变 |
Construction of luciferase reporter gene plasmid containing hBD-2 and mutant correlated with Crohn's disease |
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| YAO Guo-Peng,ZHI Fa-Chao,ZHANG Ying-Chun,CHEN Zheng-Yan,ZHI Jia,LIN Yong,GUAN Jing,WANG Ji-De,JIANG BoInstitute of Gastroenterology,Nanfang Hospital,Southern Medical University,Guangzhou ,China.Construction of luciferase reporter gene plasmid containing hBD-2 and mutant correlated with Crohn's disease[J].Journal of the Fourth Military Medical University,2009(20):2077-2080. |
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| Authors: | YAO Guo-Peng ZHI Fa-Chao ZHANG Ying-Chun CHEN Zheng-Yan ZHI Jia LIN Yong GUAN Jing WANG Ji-De JIANG BoInstitute of Gastroenterology Nanfang Hospital Southern Medical University Guangzhou China |
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| Institution: | YAO Guo-Peng,ZHI Fa-Chao,ZHANG Ying-Chun,CHEN Zheng-Yan,ZHI Jia,LIN Yong,GUAN Jing,WANG Ji-De,JIANG BoInstitute of Gastroenterology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China |
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| Abstract: | AIM:To construct and identify luciferase reporter gene plasmid containing hBD-2 and its mutant correlated with Crohn's disease.METHODS:The reporter gene plasmid hBD-2-780-luc was constructed by genetic recombination and hBD-2-mut-luc by site-directed mutagenesis to induce the mutation of the promoter for the-233 site(G)in hBD-2 into C.After lipofectin-mediated transient transfection of HEK-293T with plasmids including one of the promoter gene plasmids and NOD2/CARD15 eukaryotic expression vector,luciferase ... |
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| Keywords: | human beta-defensin 2 Crohn's disease luciferase reporter gene plasmid site-directed mutagenesis |
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