小麦DREB基因的克隆及植物表达载体构建

Cloning of DREB Gene of Triticum aestivum and Construction of Its Plant Expression Vectors

脱水应答元件结合蛋白在高等植物应答干旱、高盐和低温胁迫中发挥重要的作用。根据GenBank中小麦(Triticum aestivum L.)DREB基因的cDNA序列设计引物,采用RT-PCR技术从小麦中克隆了DREB基因837 bp的编码区。为进一步研究小麦DREB基因的功能,以pMD18-T-DREB质粒为模板,PCR扩增DREB基因片段,构建了该基因的植物表达载体。经菌液PCR和测序鉴定后,转化到农杆菌LBA4404中,为通过转基因技术深入研究小麦DREB基因的功能奠定了基础。

Dehydration-responsive element binding protein played critical roles in the response to dehydration, salinity, and cold stresses in higher plants. Primers were designed according to the cDNA sequence of wheat DREB gene in Genbank, a length of 837 bp from cDNA of DREB gene was cloned by RT-PCR. To investigate the function of DREB gene in wheat,the plant expression vector of DREB gene was constructed. The full-length open reading frame of DREB gene was amplified by PCR using pMD18-T-DREB as template. PCR and sequencing results showed that plant expression vector of DREB gene was constructed successfully. This constructed vector was then transformed into Agrobacterium LBA4404. This constructed vector provided an effective tool for the further study of DREB gene function.