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。

关键词: 小麦, TaSIM, 原核表达

Abstract: To investigate the function of TaSIMgene,the cDNA of TaSIMgene 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℃.

Key words: Wheat, TaSIM, Prokaryotic expression

中图分类号: 

• Q785