腐蹄病节瘤拟杆菌纤毛蛋白与绵羊IL-2融合基因的分泌型表达

Abstract

用改进的 Mg Cl2 法制备宿主菌株PAK/ 2 pfs感受态细胞 ;由 JM10 5工程菌中提取纤毛蛋白与绵羊白细胞介素 2 ( Pilin-IL-2 )融合基因表达载体 ,转化宿主细胞 PAK/ 2 pfs,筛选出具有 Pilin-IL-2融合基因表达载体的阳性克隆菌株 ;在营养肉汤中进行 Pilin-IL-2融合基因的表达。培养 16～ 18h后 ,离心 ,向上清液中加入饱和 ( NH4) 2 SO4提取 Pilin-IL-2融合基因表达蛋白。用羊抗兔 E型节瘤拟杆菌抗血清与提取的融合蛋白进行对流免疫电泳 ,证明该融合蛋白具有特异性

The pmPilin IL-2 was transformed into the host competent cell PAK/rpfs and the recombinant pmPilin IL-2 was expressed in the supernatant of the culture of the transformed cell PAK/rpfs.The recombinant pmPilin IL-2 was purified by (NH_4)_2_SO_4 from the supernatant of the culture of the transformed PAK/rpfs. The specific reaction of the recombinant pmPilin IL-2 and antiserum of B.nodosus serotype E pilin was demonstrated by cross electrophoresis.