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利用工程菌ＰＷＲ－１／ＪＭ１０９以融合蛋白的形式表达人工合成的恶性疟重组基因ｐｆＣＭＲ。经培养、ＩＰＴＧ诱导后收集菌体、超声波破菌、透析、电泳纯化等步骤，分离表达蛋白。结果在ＳＤＳ－ＰＡＧＥ中显示６５ｋＤ蛋白诱导ＢＡＬＢ／ｃ小鼠的体液免疫和细胞免疫反应进行了观察。结果表明，纯化后的蛋白免疫鼠血清能与表达产物粗提物产生特异的免疫反应，其抗体滴度高达１∶５１０２；Ｗｅｓｔｅｒｎｂｌｏｔ在６５ｋＤ处亦产生特异免疫反应；另在体外用ＰＨＡ和粗提物以及纯化蛋白刺激免疫鼠脾细胞，其Ｔ淋巴细胞转化率分别高达４４．２０±５．２６％、２４．００±４．７４％和３３．５０±３．８４％。提示融合蛋白中含有恶性疟原虫抗原Ｔ－／Ｂ－细胞位点。
Recombinant Plasmodium falciparum recombinant pfCMR was expressed as a fusion protein using engineered bacteria PWR-1 / JM109. After culturing, the cells were collected by IPTG induction, broken by ultrasound, dialyzed and purified by electrophoresis. Results The humoral and cellular immune responses induced by 65 kD protein in BALB / c mice were observed in SDS-PAGE. The results showed that the purified protein immunized rat serum can produce specific immune response with the crude extract of the expressed product with an antibody titer up to 1: 5102; Western blot also produced a specific immune response at 65 kD; The T lymphocyte transformation rates of splenocytes from immunized mice were 44.20 ± 5.26%, 24.00 ± 4.74% and 33.50 ± 3.84%, respectively. Suggesting that the fusion protein contains the Plasmodium falciparum antigen T- / B-cell site.