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

发布时间：2018-05-13 08:48

  本文选题：铜绿假单胞菌 + Opr　； 参考：《生物技术通报》2015年07期

【摘要】：铜绿假单胞菌外膜蛋白Opr I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Opr I蛋白的表达菌株。Western blotting验证表明抗体能与Opr I蛋白特异性结合。SDS-PAGE电泳切胶纯化获得Opr I蛋白。将Opr I蛋白免疫小鼠并攻毒铜绿假单胞菌,发现Opr I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Opr I菌株最佳诱导表达条件为:加IPTG菌夜OD600值0.8,加IPTG终浓度0.3 mmol/L,诱导时间8 h,诱导温度28℃;菌株最佳培养条件为转速230 r/min,葡萄糖浓度0%,装液量50 m L。
[Abstract]:Pseudomonas aeruginosa outer membrane protein (Opr I) has strong immunogenicity and is promising in vaccine development. The expression strain of Opr I protein was obtained by molecular method. Western blotting analysis showed that the antibody could bind to Opr I protein specifically. SDS-PAGE gel electrophoresis was used to purify Opr I protein. Opr I protein was immunized against Pseudomonas aeruginosa in mice. It was found that the protective rate of specific immunity activated by Opr I protein against Pseudomonas aeruginosa infection was 57.14%, which was significantly higher than that in control group. By orthogonal design, the optimal expression conditions of Opr I were obtained as follows: the OD600 value of IPTG strain was 0.8, the final concentration of IPTG was 0.3 mmol / L, the induction time was 8 h, and the induction temperature was 28 鈩，

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