论文部分内容阅读
大豆油酸脱氢酶(FAD2-1B)基因是种子特异表达基因,利用PCR方法从大豆基因组DNA中分离FAD2-1B基因的启动子片段,命名为FP。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,如Skn-1 motif、AACACA、SEF4 motif、E-box、ACGT等。将克隆得到的FP片段替换pCAMBIA1301中的CaMV35S启动子,构建表达载体pCAM-FP。通过农杆菌介导法在大豆各组织中进行瞬时表达,GUS组织化学染色显示FP驱动GUS基因在大豆根、茎、叶中基本不表达,在种子中有较高的表达活性,推测FP启动子具有种子特异表达活性。
The soybean oleic dehydrogenase (FAD2-1B) gene is a seed-specific expression gene. The promoter fragment of the FAD2-1B gene was isolated from soybean genomic DNA by PCR and named FP. The analysis of PLACE online promoter prediction tools showed that there are many typical seed-specific expression elements in the sequence, such as Skn-1 motif, AACACA, SEF4 motif, E-box and ACGT. The cloned FP fragment was substituted for the CaMV35S promoter in pCAMBIA1301 to construct an expression vector pCAM-FP. Agrobacterium tumefaciens-mediated transient expression in soybean tissues, GUS histochemical staining showed that FP-driven GUS gene in soybean roots, stems and leaves were basically not expressed in the seed has a higher expression activity speculated that the FP promoter Seed-specific expression activity.