【摘要】 将蛇毒类凝血酶基因(TLE)亚克隆到原核表达载体pMAL-p2X上,对重组表达质粒鉴定正确后,转化大肠埃希菌BL21进行诱导表达;同时采用不同的OD值、诱导时间I、PTG浓度和诱导温度对尖吻蝮蛇蛇毒类凝血酶基因重组表达质粒pMAL-p2X进行了条件优化。研究发现,不同的诱导时间、IPTG诱导浓度和诱导温度与融合蛋白表达量有很大关系。SDS-PAGE结果显示,当OD值为0.5,IPTG终浓度为0.3 mmol/L,诱导温度为37℃时,诱导3 h后蛋白表达量最高,超出此范围可使表达量显著减少。在优化诱导条件后,融合蛋白的表达量可达全菌总蛋白量的66%。更多还原

【Abstract】 TLE gene was subcloned into prokaryotic expression vector pMAL-p2X and transformed into E.coli BL21 after identification of the recombinant vector.The expression of recombinant vector pMAL-p2X which contained TLE gene of Agkistrodon acutus in E.coli BL21 was induced at a variety of OD value,temperature,density of IPTG,and induction time.The results showed that the expression level was correlated with OD value,temperature,density of IPTG and induction time.SDS-PAGE results also confirmed that the expression level was highest in the conditions of OD value 0.5,0.3 mmol/L IPTG,under 37 ℃ and 3 hours of induction time.The amount of recombinant gene expression products could reach as high as 66% total proteins of the host.更多还原