小麦转录因子 TaSIM 的原核表达分析

摘要

为了进一步研究TaSIM基因的功能，以pJET1．2-TaSIM质粒为模板，PCR扩增TaSIM基因的cDNA编码区，构建了该基因的原核表达载体pET-28a-TaSIM，经菌液PCR和测序鉴定后转化到大肠杆菌BL21（DE3）中。结果表明，在大肠杆菌BL21（DE3）菌株中成功表达了与标签蛋白融合的TaSIM蛋白，大小约为33 kDa。 SDS-PAGE 电泳分析表明，最佳诱导表达条件为0．5 mmol /L IPTG在37℃下诱导2 h。%To investigate the function of TaSIM gene,the cDNA of TaSIM gene was amplified by PCR using pJET1.2-TaSIM as template and the full-length open reading frame was fused into a prokaryotic expression vector pET-28a.PCR and sequencing showed that the recombinant vector pET-28a-TaSIM was successfully constructed and transformed into E.coli BL21 (DE3) cells.The results indicated that the pET-28a-TaSIM with the predicted molecular weight of about 33 kDa was successfully expressed in E.coli BL21 (DE3) strain.SDS-PAGE indicated that the best expression quantity was induced with 0.5 mmol/L IPTG treatment for 2 h at 37 ℃.