小菜蛾类钙粘蛋白片段的原核融合表达载体构建及其条件优化

Prokaryotic Expression Vector Construction of Plutella xylostella Cadherin Fragments and Optimization of Its Conditions

构建了小菜蛾钙粘蛋白片段基因的重组载体,为提高其在大肠杆菌中的表达量,研究了时间、温度、IPTG浓度对融合蛋白表达量以及可溶性的影响。结果表明,使用LB培养基37℃培养1.5h后,采用终浓度为0.1mmol·L-1的IPTG,在25℃180r·min-1诱导培养36h融合蛋白的表达量最大,并且温度的改变对融合蛋白的可溶性几乎没有影响,SDS-PAGE电泳检测结果表明,融合蛋白的分子质量与预计大小一致,约为16kDa。

In this study,we constructed a recombinant of Plutella xylostella cadherin gene fragment.In order to enhance its expression in Escherichia coli,we studied the effect of time, temperature and IPTG concentration on fusion protein expression and its solubility.The results showed that the fusion protein gave rise to the maximum expression when host cells were cultured in LB at 37℃for 1.5 h and then induced by 0.1 mmol·L-1 IPTG at 25℃180 r·min-1 for 36 h.And the temperature change had little impact on the solubility of fusion proteins. SDS-PAGE test also indicated that the molecular weight of fusion proteins was the same as expected,which is about 16 kDa.