CAJ | 학술논문

[目的]为了提高植原体膜蛋白Imp基因在E．coliBL21（DE3）中的表达量，优化Imp基因的原核表达条件。[方法]通过设计正交试验，考察不同的培养条件对工程菌EcoliBL21（DE3）-pET．28a（＋）-Imp的影响。在获得最佳培养条件的基础上考察不同诱导条件对Imp蛋白表达量的影响。利用SDS．PAGE和Gene Tools凝胶分析软件分析融合蛋白Imp的表达量。[结果]表达条件优化结果表明，最佳培养条件为：温度37℃，pH7．0，装液量20％，振荡速度200n/min。最佳诱导条件为：温度37℃，起始OD600≈1．5，IPTG终浓度0．1mmol/L，诱导培养时间6h。[结论]在最佳条件下Imp表达量达到70．98mg／L，确定了Imp融合蛋白在大肠杆菌的优化表达条件。
[목적]위료제고식원체막단백Imp기인재E．coliBL21（DE3）중적표체량，우화Imp기인적원핵표체조건。[방법]통과설계정교시험，고찰불동적배양조건대공정균EcoliBL21（DE3）-pET．28a（＋）-Imp적영향。재획득최가배양조건적기출상고찰불동유도조건대Imp단백표체량적영향。이용SDS．PAGE화Gene Tools응효분석연건분석융합단백Imp적표체량。[결과]표체조건우화결과표명，최가배양조건위：온도37℃，pH7．0，장액량20％，진탕속도200n/min。최가유도조건위：온도37℃，기시OD600≈1．5，IPTG종농도0．1mmol/L，유도배양시간6h。[결론]재최가조건하Imp표체량체도70．98mg／L，학정료Imp융합단백재대장간균적우화표체조건。
[Objective] This study aimed to increase the expression level of immun- odominant membrane protein gene （Imp） of phytoplasmas in E. coil BL21 （DE3）. [Method] On the basis of orthogonal experiment, effects of different culture conditions on recombinant bacteria E. coil BL21-pET-28a（＋）-Imp were investigated. Based on the obtained optimal culture condition, effects of different induction conditions on the ex- pression level of Imp protein were explored. The expression level of Imp fusion pro- tein was analyzed by using SDS-PAGE and Gene Tools software. [Result] The re- sults showed that the optimal conditions for culture were at 37℃, pH 7.0, with liq- uid volume of 20% and oscillation speed of 200 r/min, for induction were at 37℃ for 6 h, with initial OD600 of about 1.5 and IPTG final concentration of 0.1 mmol/L. [Conclusion] The expression level of Imp achieved 70.98 mg/L under the optimal conditions. Optimized conditions for expression of Imp fusion protein in E. coil were determined.