大豆球蛋白G1启动子的克隆及表达活性分析

Cloning and Activity Analysis of Soybean G1 Promoter

利用PCR的方法从大豆品种"吉豆2号"基因组DNA中克隆得到大豆球蛋白启动子G1p,长度约为686 bp,PLACE在线启动子预测工具分析表明:序列中含有多种典型的种子特异性表达元件。将克隆得到的G1p取代pCAMBIA1301中的CaMV35S启动子,构建于G1p与GUS基因融合表达的载体pCAM-G1p,通过农杆菌介导的方法在大豆根、茎、叶和种子中进行瞬时表达分析结果显示,仅能在种子中检测到GUS活性,而在根、茎和叶其他组织中基本检测不到GUS活性。说明G1基因上游686 bp片段具有种子特异性启动子的功能,G1p是一个比较高效的种子特异性启动子。

The G1p promoter of soybean G1 was isolated from the genomic DNA of soybean Jidou2 by PCR method.Promoter sequence analysis by PLACE showed that the cloned fragment contained a lot of the motifs than constituted the seed-specific cis-elements.Replacing CaMV35S promoter of pCAMBIA1301 with the G1p fragment.Transient expression by Agrobacterium tumefaciens mediated method,the histochemical GUS analysis and fluorometric GUS analysis the expression of the GUS activity.GUS activity assays indicated that expression of GUS was active only in seeds,but not in roots stems and leaves,which suggests the G1p is a efficient seed-specific promoter.