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

Construction of eukaryotic expressing vector and transient expression of NS-1 gene of H9N2 avian influenza virus in 293 Cells

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。

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.