托斯卡那病毒核蛋白的表达及纯化

Study on the expression and purification of Toscana virus N protein

采用聚合酶链反应方法从含托斯卡那病毒 S 片段全序列的 cDNA 质粒 pGEM3ZS 中获得核蛋白基因片段,通过多克隆接头与5′端含有6个组氨酸残基的 pQE 30表达载体连接,在大肠杆菌中高效表达核蛋白。经纯化后免疫小鼠,产生的多克隆抗体腹水经免疫印迹法检测,与托斯卡那病毒核蛋白发生特异性反应,其酶联免疫吸附试验滴度高达1:3 200。

We obtained Toscana virus N gene segment by polymerase chain reactiont from plasmid pGEM 3Z- S which contained entire eDNA of Toscana virus S segment and then cloned into express vector pQE30 which con- tained 6 His tag in 5' end of polylinker.We also expressed and purified recombinant N protein in E.coll.The immu- nized mice serum and polyclone ascitic fluid reactecd specifically with Toscana virus N protein by Western-blot and the ELISA titer was 1:3200.