人参SQS基因的干扰载体构建及转化人参愈伤组织

摘要

According to the SQS gene of Panax ginseng, the authors designed a sense primer and an antisense primer to construct a SQS interference expression vector. The ginseng callus was transformed by agrobacterium-mediated methods, and the selected resistant callus was assayed by PCR and real-time PCR, and the ginsenosides of resistant callus was analyzed by HPLC. Compared with the non-transformation ginseng callus, the SQS gene' s expression of positive callus decreased, and the ginsenosides also had certain changes. SQS is a critical biosynthetic enzyme of ginsenoside. To inhibit the expression of SQS can mediate the change of the content of the ginsenosides of ginseng.%以5年生人参为材料,根据人参SQS基因设计正义及反义片段引物,构建了SQS基因干扰表达载体; 利用农杆菌转化法转化人参愈伤组织,对获得的抗性愈伤组织进行PCR检测及实时定量PCR检测,并对抗性愈伤组织的皂苷进行HPLC分析.实验结果表明,与非转化人参愈伤组织相比,转化后的愈伤组织SQS基因表达量降低,皂苷含量有所变化.SQS是人参皂苷生物合成途径的关键酶,抑制SQS基因的表达,可调控人参皂苷含量变化.