论文部分内容阅读
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 glycinin promoter G1p was cloned by PCR from the genomic DNA of soybean “Jidou 2” with a length of about 686 bp. The PLACE online promoter predictive tool analysis showed that the sequence contained many typical seeds Specific expression element. The cloned G1p was substituted for the CaMV35S promoter in pCAMBIA1301 to construct the vector pCAM-G1p for fusion expression of G1p and GUS gene. The results of Agrobacterium tumefaciens-mediated transient expression in soybean root, stem, leaf and seed GUS activity was detected only in seeds, whereas GUS activity was not detectable in other tissues of roots, stems and leaves. The result shows that the 686 bp fragment upstream of G1 gene has the function of seed-specific promoter, and G1p is a more efficient seed-specific promoter.