论文部分内容阅读
目的 将甲型流感病毒核蛋白(NP)基因克隆到原核表达载体进行可溶性融合表达,制备纯化的病毒核蛋白,为制备甲型流感病毒单克隆抗体提供材料。方法 提取甲型流感病毒RNA ,设计引物,RT PCR扩增NP基因,利用基因工程的手段,将甲型流感病毒的NP基因在大肠埃希菌中进行融合表达,并将表达产物进行亲和层析。结果 成功构建了甲型流感病毒NP基因原核表达载体,经亲和层析制备了较高纯度的目的表达产物。结论 通过合理控制发酵时间、生长温度和诱导物浓度,制备了较为理想的可溶性甲型流感病毒核蛋白。
Objective To clone and purify the nucleocapsid protein (NP) gene of influenza A virus into the prokaryotic expression vector for fusion expression and to prepare the purified viral nucleoprotein for the preparation of the monoclonal antibody against influenza A virus. Methods The influenza A virus DNA was extracted and the primers were designed. The NP gene was amplified by RT PCR. The NP gene of influenza A virus was fused and expressed in Escherichia coli by genetic engineering. Analysis. Results The prokaryotic expression vector of NP gene of influenza A virus was successfully constructed and the target product of higher purity was prepared by affinity chromatography. Conclusion By optimizing the fermentation time, growth temperature and the concentration of inducer, an ideal soluble nucleoprotein of influenza A virus was prepared.