【摘要】 目的大量诱导表达含pET24a-Derp2质粒的BL21工程菌,表达产物以重组蛋白包涵体的形式存在,经包涵体洗涤与溶解后,使用结合6组氨酸的镍柱进行亲和层析纯化蛋白质,用梯度复性方法进行重组蛋白的复性,再用屋尘螨过敏性哮喘患者阳性血清经Western blot方法分析Derp2重组蛋白的免疫学特性。纯化后的融合蛋白Derp2具有较高的纯度及较强的免疫活性,可望作为有效的屋尘螨变应原诊断试剂和疫苗的候选分子。更多还原

【Abstract】 The recombinant plasmid p-ET24a-Der-p2 was firstly transformed to E.coli JM109 and purified. Then it was induced to express in large amount with IPTG. After centrifugation of the bacterial cultures,the bacterial pellet was re-suspended and lysed by freezing-thawing treatment and ultrasonication. The inclusion bodies were purified by gel-filtration and chromatography. This highly purified allergen was shown to possess good IgE-binding activity as demonstrated by Western blotting. It suggests that this recombinant allergen can be used as a candidate in vaccine development for mite allergy.更多还原