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 ℃.
出处
《华北农学报》
CSCD
北大核心
2013年第2期60-62,共3页
Acta Agriculturae Boreali-Sinica
基金
新疆农业大学校前期资助课题(XJAU201019)
新疆维吾尔自治区高技术研究发展项目(201011109)
新疆维吾尔自治区高校科研启动项目(XJEDU2011S20)
作物遗传育种重点学科资助项目
