| H9N2亚型禽流感病毒NS1基因真核表达载体的构建及其在293细胞瞬时表达 |
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| 引用本文： | 董永军,;王丽荣,;贺会利.H9N2亚型禽流感病毒NS1基因真核表达载体的构建及其在293细胞瞬时表达[J].兽医大学学报,2014(8):1235-1238. |
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| 作者姓名： | 董永军 ;王丽荣 ;贺会利 |
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| 作者单位： | [1]河南科技学院动物科学学院,河南新乡453003; [2]河南科技学院抗体工程重点实验室,河南新乡453003 |
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| 基金项目： | 河南省教育厅科学技术研究重点资助项目（12A230003）; 河南省新乡市重点科技攻关资助项目（ZG13009） |
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| 摘 要： | 在293细胞中瞬时表达A/Chicken/Henan/1001/2010（H9N2）NS1基因蛋白,并对表达蛋白活性进行测定。试验利用PCR技术从NS1-T载体质粒上扩增NS1基因,将其克隆至pCAGGS载体,构建重组质粒NS1-pCAGGS;经酶切和测序鉴定正确后,重组质粒NS1-pCAGGS与脂质体按照一定比例混合后转染293细胞,用间接免疫荧光方法对瞬时表达细胞进行荧光信号反应。结果显示,禽流感NS1基因可以很好地在293细胞中瞬时表达,具有良好的反应原性。
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| 关 键 词： | H9N2亚型禽流感病毒 NS1基因 真核表达载体 瞬时表达 |
Construction of eukaryotic expressing vector and transient expression of NS-1 gene of H9N2 avian influenza virus in 293 Cells |
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| Institution: | DONG Yun-jun ,WANG Li-rong , HE Hui-li (1. College of Animal Science, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China ; 2. Antibody Engineering Laboratory, Henan Institute of Science and Technology ,Xinxiang,Henan 453003 ,China) |
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| Abstract: | For transient expression of NS1 gene of H9N2 avian influenza virus in 293 cells, the NS1 gene was amplified from NS1-T vector plasmid in vitro and cloned into pCAGGS vector to con struct a recombinant plasmid NSI-pCAGGS. The NSI-pCAGGS plasmid was extracted, and then was mixed with liposome in accordance with a certain proportion to transfect the 293 cells. The indirect immunofluorescence assay was used to detect the fluorescence signal response from transient expression of cells. The result showed that the NS1 gene of H9N2 avian influenza virus could be transiently expressed in 293 cells. |
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| Keywords: | H9N2 avian influenza virus NS1 gene eukaryotic expression vector transient expres sion |
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