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为获得大量具天然抗原活性的恶性疟原虫HRP-Ⅱ抗原,用摇瓶发酵工程菌,诱导β-半乳糖苷酶-HRP-Ⅱ融合蛋白表达;裂菌后,洗涤沉淀2～3次,用6M脲溶解;取上清经Sephacryl-200柱层析分离纯化目的蛋白,然后复性。结果重组蛋白以包涵体的形式高效表达;Sephacryl-200柱层析后,目的蛋白纯度达86％,91.48％被复性,被Dipstick即 ParsSight-F识别,表明该重组抗原纯度高,复性效果好,可用于恶性疟原虫 HRP-Ⅱ抗体的制备等研究。
In order to obtain a large number of Plasmodium falciparum HRP-Ⅱ antigen with natural antigen activity, the β-galactosidase-HRP-Ⅱ fusion protein is induced by shake flask fermentation engineering bacteria; after the split bacteria, the precipitate is washed for 2 to 3 times, 6M urea dissolved; the supernatant by Sephacryl-200 column chromatography purification of the target protein, and then renaturation. Results The recombinant protein was highly expressed in the form of inclusion bodies. After purification by Sephacryl-200 column, the purity of the recombinant protein was 86% and 91.48%, respectively. The recombinant protein was identified by Dipstick, ParsSight-F, Good, can be used for the preparation of antibodies against Plasmodium falciparum HRP-Ⅱ.