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| 线粒体抗病毒信号蛋白MAVS真核表达质粒的构建及表达 | |
| 引用本文： | 倪才飞,赵帆,郑子瑞,黄蓓,马红芳,李力,宋婷,魏从文,何湘,张艳红,钟辉. 线粒体抗病毒信号蛋白MAVS真核表达质粒的构建及表达[J]. 军事医学科学院院刊, 2008, 32(6) |
| 作者姓名： | 倪才飞 赵帆 郑子瑞 黄蓓 马红芳 李力 宋婷 魏从文 何湘 张艳红 钟辉 |
| 作者单位： | 1. 安徽大学生命科学院,合肥,230039;军事医学科学院生物工程研究所,北京,100850 2. 军事医学科学院生物工程研究所,北京,100850 3. 安徽大学生命科学院,合肥,230039 4. 军事医学科学院生物工程研究所,北京,100850;军事医学科学院野战输血研究所,北京,100850 5. 军事医学科学院生物工程研究所,北京,100850;军事医学科学院疾病预防控制所,北京,100850 |
| 摘 要： | 目的:构建线粒体抗病毒信号蛋白(mitoehondrial antiviral signaling protein,MAVS)真核表达质粒.方法:从人胚肾细胞系HEK293T细胞的总RNA中经过逆转录和PCR获得MAVS的全长cDNA双链片段.经过酶切后连接到真核表达载体pcDNA3/flag中,挑选出转化生成的阳性克隆进行测序,将序列正确的重组质粒命名为pcDNA3/flag-MAVS.借助脂质体转染peDNA3/flag-MAVS质粒到HEK293T细胞中,用Westem印迹检测目的蛋白的表达.同时用β干扰素的荧光素酶报告基因(IFN-β-luc)检测MAVS蛋白活性.结果:Western印迹实验证明pcDNA3/flag-MAVS可以在HEK293T细胞中表达MAVS,并且能使IFN-β报告基因活性明显增强.结论:构建了重组质粒peDNA3/flag-MAVS,在细胞中表达MAVS后具有刺激IFN-β转录的生物活性. |
| 关 键 词： | IFN-β 荧光素酶报告基因 |
Construction and expression of mitochondrial antiviral signaling protein(MAVS)in eukaryotic expression vectors |
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| NI Cai-Fei,ZHAO Fan,ZHENG Zi-Rui,HUANG Bei,MA Hong-Fang,LI Li,SONG Ting,WEI Cong-Wen,HE Xiang,ZHANG Yan-Hong,ZHONG Hui. Construction and expression of mitochondrial antiviral signaling protein(MAVS)in eukaryotic expression vectors[J]. Bulletin of the Academy of Military Medical Sciences, 2008, 32(6) | |
| Authors: | NI Cai-Fei ZHAO Fan ZHENG Zi-Rui HUANG Bei MA Hong-Fang LI Li SONG Ting WEI Cong-Wen HE Xiang ZHANG Yan-Hong ZHONG Hui |
| Affiliation: | NI Cai-Fei~(1,2) ZHAO Fan~2 ZHENG Zi-Rui~(1,2) HUANG Bei~1 MA Hong-Fang~(2,3) LI Li~2 SONG Ting~2 WEI Cong-Wen~2 HE Xiang~(2,4) ZHANG Yan-Hong~2 ZHONG Hui~(2*) (1.School of Life Science,Anhui University,Hefei 230039,China,2.Institute of Biotechnology,Academy of Military Medical Sciences,Beijing 100850,3.Institute of Transfusion Medicine,4.Institute of Disease Prevention & Control,China) |
| Abstract: | Objective:To construct the eukaryotic expression plasmid of mitochondrial antiviral signaling protein(MAVS). Methods:The gene encoding MAVS protein was amplified by RT-PCR from total RNA of HEK293T cell line.After cleav- ase with restriction endonucleases the gene fragment was ligated to pcDNA3/flag vector.The transformed clones were se- quenced and the right one was named pcDNA3/flag-MAVS,pcDNA3/flag-MAVS was transfected into HEK293T cell line to test the expression and activity of MAVS by Western blot and... |
| Keywords: | MAVS |
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