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铜绿假单胞菌外膜蛋白I的原核表达、纯化及免疫保护作用研究

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.07.032, PP. 207-213

Keywords: 铜绿假单胞菌,OprI蛋白,免疫保护,正交试验,诱导条件

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Abstract:

铜绿假单胞菌外膜蛋白OprI具有较强的免疫原性,在疫苗上具有开发前景。利用分子方法获得OprI蛋白的表达菌株。Westernblotting验证表明抗体能与OprI蛋白特异性结合。SDS-PAGE电泳切胶纯化获得OprI蛋白。将OprI蛋白免疫小鼠并攻毒铜绿假单胞菌,发现OprI蛋白激活的特异性免疫对小鼠铜绿假单胞菌感染的保护率达到57.14%,与对照组相比较达到显著性。采用正交试验设计,获得OprI菌株最佳诱导表达条件为:加IPTG菌夜OD600值0.8,加IPTG终浓度0.3mmol/L,诱导时间8h,诱导温度28℃;菌株最佳培养条件为转速230r/min,葡萄糖浓度0%,装液量50mL。

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