大豆gma-miR169c启动子的克隆及植物表达载体构建

Isolation of gma-miR169c Promoter form Glycine max and Construction of Its Plant Expression Vectors

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。

Based on the predicted sequence of gma-mi R169 c promoter region,primers was designed based on promoter sequence.A 1 935 bp fragment was cloned from the genomic DNA of the soybean Williams 82 by PCR method.Plant CARE analysis showed that the promoter sequence contains basic elements TATA-box,CAAT-box and some cis-acting elements involved in response to abiotic stresses and plant hormones.After that,the genomic DNA fragments with partial deletion in gma-mi R169 c promoter and GUS fusion expression vector were constructed by replacing Ca MV35 S promoter of p BI121.These results would be helpful for the further functional characterization of cis-acting elements and regulation mechanism in gma-mi R169 c promoter.