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流感病毒属于正黏病毒科,为有包膜包裹的单股负链RNA病毒。它的8个基因片段编码至少16种病毒蛋白,其中3个蛋白组成流感病毒的聚合酶复合体。流感病毒聚合酶碱性蛋白1(PB1)是该复合体的组分之一,在病毒的转录、复制及重配中发挥重要的作用。为研究其功能,构建His-PB1(aa 550–755)融合蛋白原核表达质粒,IPTG诱导融合蛋白表达,通过镍柱亲和将其纯化,然后作为抗原免疫家兔。对抗血清进行间接ELISA检测,表明抗体效价可达1∶100 000。兔源PB1蛋白抗血清经亲和纯化后,用于免疫印迹检测流感病毒WSN毒株PB1蛋白以及外源转染的FLAG-PB1蛋白,结果表明该PB1抗体具有良好的特异性,同时也能特异性识别其他亚型的A型流感病毒的PB1蛋白,为进一步研究流感病毒PB1蛋白的功能奠定了基础。
Influenza virus belongs to orthomyxoviridae, which is an enveloped single-stranded minus-strand RNA virus. Its eight gene segments encode at least 16 viral proteins, three of which make up the polymerase complex of the influenza virus. Influenza Virus Polymerase Basic protein 1 (PB1) is one of the components of this complex that plays an important role in the transcription, replication and reassortment of viruses. To investigate its function, a prokaryotic expression plasmid containing His-PB1 (aa 550-755) fusion protein was constructed. IPTG induced the expression of the fusion protein, purified by nickel affinity chromatography, and then used as antigen to immunize rabbits. Anti-sera were tested by indirect ELISA, indicating that the antibody titers reached 1: 100,000. The purified PB1 antiserum was used to detect the PB1 protein of influenza virus WSN strain and FLAG-PB1 exogenously transfected by PB1 protein. The results showed that the PB1 antibody has good specificity and specificity The PB1 protein that recognizes other subtypes of type A influenza virus, which lays the foundation for further study on the function of influenza virus PB1 protein.