【摘要】 乙酰辅酶A羧化酶(ACCase)是脂肪酸生物合成的限速酶,在油料作物脂肪酸代谢和调控基因工程中具有重要的应用价值。本研究利用同尾酶反复酶切的方法,构建了拟南芥Rubisco SSU小亚基转运肽与大肠杆菌异质型ACCase各亚基基因的融合串联种子特异表达载体,在根癌农杆菌(Agrobacterium tumefaciens)介导的下胚轴转化甘蓝型油菜高油品种CY2,研究异质型ACCase转入油菜对含油量的影响。经过抗生素筛选获得11株转化再生植株,PCR、Southern分子检测证明四个目的基因已整合到T0代再生植株油菜基因组中。RT-PCR检测其中4株,说明在转基因株系中目标基因在籽粒中特异表达。对T2代转基因油菜含油量进行测定,研究表明转基因株系的含油量比对照提高了约5.7%。更多还原

【Abstract】 Acetyl-coenzyme A carboxylase(ACCase) was the rate-limiting enzyme in the biosynthesis of fatty acids. It played an important role in the oil crop fatty acid metabolism and regulation. Plant expression vector was constructed by adding an Arabidopsis Rubisco SSU chloroplast signal peptide at 5’ end of the target gene and hooked with the seed special expression promoter Napin. The expression vector was transformed into the high oil Brassica napus cv. CY2 via Agrobacterium-mediated transformation to investigate the influence of heteromeric ACCase on seed oil content. Eleven positive transformants were obtained. Four target genes were integrated into the T0 genome of transgenic lines by PCR and Southern blot verification. Semi-quantitative RT-PCR analysis revealed that the four target genes could be specially expressed in transgenic plants seeds. Moreover, the average seed oil content of the T2 transgenic plants was about 5.7% higher than the control.更多还原