屋尘螨重组变应原Der p 2的表达、纯化及免疫学活性鉴定

Expression,purification and identification of the recombinant allergen Der p2 from Dermatophagoides pteronyssinus and investigation on its immunological activities

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。

The recombinant plasmid p ET24a Der-p2 was firstly transformed to E. coil 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.