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本研究以大豆Williams 82基因组DNA为材料,根据预测的gma-mi R169c启动子序列设计引物,采用PCR方法克隆得到gma-mi R169c上游一段长度为1 935 bp的启动子序列。采用Plant CARE启动子在线预测工具分析表明,gma-mi R169c启动子序列具有TATA-box、CAAT-box基本的顺式作用元件和一些参与非生物胁迫和植物激素应答相关的顺式作用元件。将gma-mi R169c启动子部分缺失序列替代p BI121载体中的Ca MV35S启动子,构建了与GUS融合的启动子元件缺失表达载体。研究结果为进一步分析gma-mi R169c启动子元件的功能和调控机制奠定了基础。
In this study, the Williams 82 genomic DNA was used as a material to design a promoter based on the predicted gma-mi R169c promoter sequence. A 1935 bp upstream of gma-mi R169c was cloned by PCR. Analysis using the Plant CARE promoter online prediction tool showed that the gma-mi R169c promoter sequence has the TATA-box, CAAT-box basic cis-acting elements and some cis-acting elements involved in abiotic stress and plant hormone response. A partial deletion sequence of the gma-mi R169c promoter was substituted for the CaMV35S promoter in the pBI121 vector to construct a promoter-deficient expression vector fused with GUS. The results laid the foundation for further analysis of the function and regulatory mechanism of gma-mi R169c promoter element.