论文部分内容阅读
本研究利用PCR技术从’赤霞珠’葡萄(Vitis vinifera L.cv Cabernet Sauvignon)基因组DNA中扩增得到与花色素3,5-O-双葡萄糖苷合成相关的Vv5GT3基因的启动子。对启动子序列的顺式作用元件进行预测分析发现,Vv5GT3基因的启动子序列除了含有植物启动子的基本结构元件外,还有多个与胁迫和激素诱导相关的顺势调控元件。以荧光素酶基因(Luc)作为为报告基因,构建了Vv5GT3基因启动子的植物表达载体p CAMBIA1300-Vv5GT3 promoter。以根癌农杆菌GV3101感受态细胞为受体,转化植物融合表达载体进行瞬时表达,并对启动子进行了功能活性分析,结果表明该启动子能够驱动Luc报告基因在烟草叶片组织中表达。用不同的光照条件处理被农杆菌侵染的烟草叶片,结果显示长时间光照有利于启动子的激活,而遮光减弱了Vv5GT3启动子的活性。
In this study, the promoter of Vv5GT3 gene related to the synthesis of anthocyanidin 3,5-O-bis-glucoside was amplified from genomic DNA of Vitis vinifera L.cv Cabernet Sauvignon by PCR. Prediction of the cis-acting elements of the promoter sequence revealed that the promoter sequence of the Vv5GT3 gene contained multiple homeopathic elements related to stress and hormone induction in addition to the basic structural elements of plant promoters. The luciferase gene (Luc) was used as the reporter gene to construct the plant expression vector p CAMBIA1300-Vv5GT3 promoter of Vv5GT3 gene promoter. The Agrobacterium tumefaciens GV3101 competent cells were transformed into plant fusion expression vector for transient expression, and the promoter activity analysis was performed. The results indicated that the promoter can drive the expression of Luc reporter gene in tobacco leaves. Agrobacterium-infected tobacco leaves were treated with different lighting conditions. The results showed that long-time light was beneficial to promoter activation, while light-shielding reduced Vv5GT3 promoter activity.