腐蹄病节瘤拟杆菌纤毛蛋白与绵羊白细胞介素-2融合基因在PAK/2pfs细胞中的表达与鉴定

Expression and Identification of the fused Dichelobacter Pili and Ovine Interleukin 2 Gene in Pseudomonasaeruginus

将前期工作中筛选出的含有腐蹄病节瘤拟杆菌纤毛蛋白与绵羊白细胞介素-2(Pilin-IL-2)融合基因表达载体的PAK/2pfs工程菌[1]接种于营养肉汤,40℃培养16～18h,离心,向上清液中加入饱和(NH4)2SO4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,且有较高的表达量。经SDS-PAGE电泳和Westernblot确证其为Pilin-IL-2融合蛋白。

The pmPilinIL2 was transformed into the host competent cell PAK/rpfs and the recombinant pmPilinIL2 was expressed in the supernatant of the coltures of the transformant cell PAK/rpfs.The recombinant pmPilinIL2 was purified by (NH_4)_2SO_4 from the supernatant of the colture of the transformed PAK/rpfs. The specificreaction of the recombinant pmPilinIL2 and antiserum of D.nodosus serotype E pilin was demonstrated by cross electrophoresis.33000 Expression protein was found by SDS-PAGE.Through Western-biotting,the expression protein was recognized by antiserum of D.nodosus serotype E pilin.