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中文摘要：
为了增强多肽表位的免疫原性,构建了霍乱毒素B亚单位(Cholera toxin B subunit,CTB)和p277的融合基因CTB-p277,将该融合基因克隆到pET28a(+)中,获得原核表达载体pET28a-CTB-p277;并将该质粒转入大肠杆菌菌株BL21(DE3)中;重组菌株经2%的乳糖诱导5 h后的表达产物用12%的SDS-PAGE分析表明可以表达融合蛋白,融合蛋白占全菌蛋白约40%,且主要以包含体的形式存在.包含体经洗涤、裂解,用DEAE纤维素离子交换纯化,可以获得99.1%的重组蛋白.重组蛋白的包含体经复性后,在体外可以自组装成五聚体.重组蛋白免疫KK-Ay小鼠后,ELISA的分析表明可以产生抗p277的抗体.血糖和体重的测定结果显示,给药组小鼠的血糖有明显的降低,体重也有相应的减轻.同时GM1-ELISA的检测表明,CTB-p277在体外可以和神经节苷脂GM1(Monosialoganglioside)特异结合,具有生物活性.
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