种子特异启动子的DGAT基因植物表达载体构建

Construction of A Seed-Specific Expression Plant Vector Targeting the DGAT Gene

利用PCR技术从甘蓝型油菜(Brassica napus L.)的基因组DNA和拟南芥(Arabidopsis thaliana)cDNA中分别扩增种子特异启动子napin和二脂酰甘油酰基转移酶(DGAT)基因片段,并将它们依次插入到植物表达载体pCAMBIA1390的多克隆位点处,构建了种子特异启动子napin驱动的DGAT基因植物表达载体p1390ND,冻融法将其导入根癌农杆菌(Agrobacterium tumefaciens)LBA4404、EHA105和AGL1中,PCR检测结果证明,重组质粒p1390ND已成功导入农杆菌细胞。该载体可应用于油料能源植物种子含油量的遗传改良。

By the PCR technique, seed-specific promoter （napin） and diacylglycerol acyhransferase （DGA T） were amplified from Brassica napus L. genomic DNA and Arabidopsis thaliana cDNA respectively. The two fragments were inserted into the multiple cloning sites of the binary vector pCAMBIA1390 respectively. The resulted seed-specific expression vector was named as p1390ND. The vector was transformed into the Agrohacterium tumefaciens strains LBA4404, EHAI05 and AGLI. It could provide an excellent genetic resources for genetic improvement of seed oil content of energy plants.