His-prescission蛋白酶基因在大肠杆菌中表达条件的优化

Optimization of Culture Conditions for His-prescission Protease Gene

以生物量和SDS-PAGE电泳表达量为判断标准进行单因素实验,结果表明:分析比较不同的大肠杆菌感受态细胞BL21(DE3)-T1R、Rosetta2(DE3)、Origami2(DE3)对于表达His-prescission蛋白的影响,确定最佳的感受态细胞为BL21(DE3)-T1R。正交实验结果表明:培养基初始pH为7.0,摇床转速130 r/min,摇瓶装液量1 200 mL,接种量20 m L,培养温度37℃,OD值达到0.7时,加入IPTG 0.1 m M,诱导表达时间6 h时,蛋白表达量最大。

The paper based on biomass and SDS-PAGE electrophoresis expression shows by the single factor experiment firstly that BL21(DE3)-T1R are identified as the best competent cells through the analysis and comparison of the influence on express His-prescission protein from different E.coli competent cells BL21(DE3)-T1R,Rosetta2(DE3)and Origami2(DE3).At the same time,the expression of a protein has touched the maximum level after adding IPTG 0.1 mM into the medium for the 6 h-inducing-expression time in the condition of the initial value pH 7.0 for medium,the speed of 130 r/min for shaking,the 1200 mL of shaking-bottled liquid,the 20 mL volume of inoculation,the culture temperature at 37℃ and OD value reached 0.7.The results of orthogonal experiment show the largest amount of protein expression,that the medium initial pH 7.0,table speed 130 r/min,bottled liquid volume 1200 mL,quantity of 20 mL,culture temperature of 37℃,the OD value of 0.7,join IPTG 0.1 mM,inducing expression time of 6 h.