【摘要】 目的　流感病毒NS1基因敲除毒株(delNS1毒株)的获得可以使我们建立更有效的生物学关系,主要表现在流感病毒繁殖周期和致病的过程中,流感病毒NS1蛋白和双链RNA活性蛋白激酶(PKR)之间的生物学关系.资料表明,缺少功能性的PKR可以使delNS1毒株在其他非允许宿主中繁殖,这表明流感病毒NS1蛋白的主要功能是对抗或阻止PKR调解抗病毒效应.因此,本实验为了证明流感病毒NS1蛋白对其复制和毒力的影响.方法　应用定点突变技术进行流感病毒NS1基因的第38位和第41位氨基酸残基位点的定点突变;然后进行细胞转染包装成新的缺陷病毒;再对其进行转染后产量的测定,如TCID50和PFU.结果　包装成的缺陷病毒与包装成的非缺陷病毒相比,缺陷病毒的TCID50和PFU明显下降.结论　本实验证明了流感病毒NS1蛋白在流感病毒复制过程中,使其产量和毒力下降.更多还原

【Abstract】 ObjectiveThe availability of an influenza virus NS1 gene knockout virus（delNS1 virus） allowed us to establish the significance of biological relationship between the influenza virus NS1 protein and double-stranded-RNA-activated protein kinase（PKR） in the life cycle and pathogenicity of influenza virus. Results show that the lack of functional PKR permits the delNS1 virus to replicate in other nonpermissive hosts, suggesting that the major function of the influenza virus NS1 protein is to counteract or prevent the PKR-mediated antiviral response. Therefore, the result can prove that an influenza virus NS1 protein may affect its copy and virulence.MethodsDelNS1 virus was mutated by ribonucleoprotein transfection of an altered viral influenza NS gene. An NS1 protein was encoded with mutations at amino acid peptide positions 38（R→A） and 41（K→A）. DelNS1 virus was packed by eukaryotic translation. The production of DelNS1 virus was tested by TCID₅₀ and PFU.ResultsThe production of DelNS1 protein receded, such as TCID₅₀ and PFU.ConclusionThe results show that the production and virulence of DelNS1 virus receded in NS1 protein synthesis.更多还原