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将大豆中已克隆的一个新的ERF转录因子基因(GmERF6)构建到原核表达载体pET28上,导入大肠杆菌Rosetta(DE3)中,对其进行IPTG诱导。结果表明:在IPTG浓度为0.3 mol.L-1,诱导时间为3 h时,重组蛋白得到表达,分子量大约为30 kDa。SDS-PAGE电泳结果表明重组蛋白主要以包涵体形式存在。用8 mol.L-1尿素对其进行溶解后经Ni柱纯化,得到较好的纯化效果,Bradford法测定其蛋白浓度为0.56 mg.mL-1。
A novel ERF transcription factor gene (GmERF6) cloned in soybean was constructed into the prokaryotic expression vector pET28 and introduced into E. coli Rosetta (DE3) for IPTG induction. The results showed that the recombinant protein was expressed at an IPTG concentration of 0.3 mol·L-1 and an induction time of 3 h, with a molecular weight of about 30 kDa. SDS-PAGE electrophoresis results show that the recombinant protein is mainly in the form of inclusion bodies. The purified product was purified by Ni column after dissolving with 8 mol·L-1 urea, and its protein concentration was 0.56 mg.mL-1 by Bradford method.