【摘要】 本研究应用酸脱毛法 ,将供试工程菌培养物进行脱毛、提取制备表达蛋白 ,并对酸脱毛法的最适pH值、最佳温度、最佳时间进行了探讨 ,SDS_PAGE分析结果证明 ,酸脱毛法脱毛pH值为 2、脱毛温度为 6 0℃、脱毛时间为 30分钟时 ,表达蛋白分离提取的效果最佳。对提取蛋白进行琼扩 ,SDS_PAGE、Western_blot分析 ,结果表明所提取蛋白具有较高的抗原活性 ,并能被K88K99单克隆抗体所识别。更多还原

【Abstract】 In the study,K 88 K 99 expression proteins were effectively extracted by the acid detached fimbriae method from engineered Strain.The optimum conditions(i.e.pH value, temperature and time)of the acid detached fimbrae method had been investigated.In SDS_PAGE analyses,the results showed that protein bands are only K 88 K 99 fimbriae proteins obtained from E. coli by detaching fimbriae at 60℃ for 30 minutes in the pH 2 acid solution.The fimbriae antigens activities were demonstrated by double immunodiffusion test and western blotting analyses. The expression proteins could be recognized by monoclonal antibodies against K 88 K 99 fimbriae更多还原