【摘 要】
：
目的 在大肠杆菌中表达并纯化大肠杆菌不耐热肠毒素无毒突变体mLT63融合蛋白,并探索其免疫反应性.方法 采用不同的诱导温度,IPTG浓度,葡萄糖含量,pH值以及不同的培养基优化重
【机 构】
：
郑州大学公共卫生学院流行病学教研室,郑州,450052;河南省分子医学重点学科开放实验室
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目的 在大肠杆菌中表达并纯化大肠杆菌不耐热肠毒素无毒突变体mLT63融合蛋白,并探索其免疫反应性.方法 采用不同的诱导温度,IPTG浓度,葡萄糖含量,pH值以及不同的培养基优化重组菌TB1(pMAL-c2X-mlt)的表达条件,表达产物经超声粉碎,超声上清经初步盐析、应用直链淀粉亲和层析柱纯化,Western blot鉴定纯化产物的免疫学反应性.结果 融合蛋白在诱导表达产物中的含量达到16.11%,纯化产物的纯度达到72.58%.纯化产物能够被抗CT血清所识别.结论 成功表达并纯化出了大肠杆菌不耐热肠毒素无毒突变体mLT63,获得了纯度较高并具有良好免疫反应性的融合蛋白.
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