【摘要】 单核增生性李斯特杆菌(L.monocytogenes)的主要毒力基因hly与融合表达载体pGEX-6P-1相连,转化大肠杆菌BL-21。在摇床条件下,利用异丙基硫代-βD-半乳糖苷(IPTG)为诱导剂,分析比较不同IPTG浓度对表达产物的影响,以确定最佳的IPTG浓度。结果表明,在装有LB培养基的三角瓶中,在37℃情况下,当IPTG终浓度为0.6mmol.L-1时,hly基因的表达量最大。当再增加IPTG的浓度时,抑制重组蛋白的表达。Western-blot分析,所表达的蛋白具有抗原特异性。实验为重组李氏溶血素的工业化生产提供依据。更多还原

【Abstract】 The recombinant of pGEX-6P-hly was transformed into E.coli BL-21,induced and expressed by IPTG,to analyze the effect of different concentration of IPTG on the expression of recombinant Listeriolysin O,and to confirm the best induced concentration of IPTG.Experimental results indicated that under shake incubator,LB medium and inducing 2.4 hours with 37 ℃,hly expression quantity was the most when ultima concentration of IPTG was 0.6 mmol·L-1.Western-blot analysis showed that recombinant fusion protein could react with a specific anti-serum.This provided dependable experimental accordance for industry production of Listeriolysin O when IPTG was inducer.更多还原