摘要
通过分子克隆,获得了Omp T蛋白的表达菌株;利用切胶纯化,获得了Omp T蛋白。用Omp T蛋白免疫小鼠,制备多克隆抗体,采用ELISA法检测抗体滴度为1∶1 600,用Western Blotting法检测抗血清特异性较好。采用正交试验,获得Omp T菌株的适宜表达条件为诱导菌液OD600 nm为1.0,IPTG添加终浓度为0.3 mmol/L,诱导时间为8 h,诱导温度32℃;适宜培养条件为葡萄糖质量分数0%,转速230 r/min,装液量50 m L。
OmpT, a major outer membrane protein of Escherichia coli, had a good prospect in vaccine development for its better immune detection. The expression strain of OmpT was obtained employing molecular clone technology and purified using gel slices approach, and the polyclonal antibody was prepared from immunized mice. The antibody titer tested by ELISA approach was 1 ： 1 600, and the antiserum tested by Western Blotting method had good specificity. From the results of orthogonal experiment, the optimal condition for inducing expression of OmpT protein were 1.0 OD600 nm value of strain, 0.3 mmol/L final concentration of IPTG, 8 h of inducing time and maintaining temperature in 32 ℃; The optimal cultural condition for glucose concentration, rotation rate, and medium volume were 0%, 230 r/min and 50 mL, respectively. These work provided a ground for industrial fermentation, protein function and vaccine development of OmpT.
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2015年第4期350-355,共6页
Journal of Hunan Agricultural University(Natural Sciences)
基金
陕西省教育厅科学研究计划项目(2013JK0723)
陕西理工学院人才启动项目(SLGQD13–15)
