甲型流感病毒核蛋白基因的克隆表达及纯化  

Clone expression and purification of influenza A virus nucleoprotein gene

作　　者：房师松[1] 程小雯[2] 刘涛[2] 刘建军[2] 赵树进[3] 周丽[2] 

机构地区：[1]华南理工大学食品与生物工程学院,广州现在510642 [2]深圳市疾病预防控制中心微生物检验科 [3]广州军区广州总医院药学部

出　　处：《中华实验和临床病毒学杂志》2005年第2期165-168,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：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 the influenza A virus NP gene into expression vector and to purify the target protein, which was used to study the preparation of monoclonal antibody. Methods The RNA of influenza A virus was extracted and primers were designed according the NP gene sequence, then the NP gene of influenza A virus was expressed in E.coli DH5α and the NP protein was purified by affinity chromatography. Results The recombinant expression vector-pGEX-4T-2-NP was successfully constructed and relatively pure target protein was obtained. Conclusion Through reasonably controlling the fermentation time, growth temperature and induction concentration, satisfactory soluble target product was obtained.

关 键 词：甲型流感病毒 核蛋白基因 纯化 表达及 RT-PCR扩增 基因原核表达载体 融合表达 亲和层析 表达产物 单克隆抗体 病毒RNA 大肠埃希菌 基因克隆 设计引物 基因工程 发酵时间 合理控制 可溶性 P基因 制备 高纯度 诱导物 

分 类 号：R373[医药卫生—病原生物学]