大肠杆菌乙酰辅酶A羧化酶accD亚基表达载体的构建及遗传转化研究

Escherichia coli ACETYL-CoA CARBOXYLASE(ACCase) accD SUBUNIT GENE EXPRESSION VECTOR CONSTRUCTION AND GENETIC TRANSFORMATION

为探索大肠杆菌异质型乙酰辅酶A羧化酶(ACCase)的β-CT亚基accD基因对油料作物含油量的影响,将目的基因E.coli accD(eaccD)与拟南芥Rubisco小亚基(rbcS)转运肽基因融合,构建以油菜Napin启动子驱动的种子特异表达载体。本试验应用农杆菌介导法,将该表达载体转入油菜,通过PCR检测获得了7个阳性转基因株系,RT-PCR检测其中3株,证明eaccD基因在转基因株系的种子中转录,在转基因植株叶片中不转录,说明该表达载体转入油菜中能引起植株种子eaccD基因的特异表达。同时分析还表明转基因株系的含油量与对照植株相比提高约9%。

To investigate the influence of Escherichia coli accD（eaccD） gene on Brassica fatty acid content,an expression vector was constructed by fusing the Arabidopsis rubisco small subunit（rbcS） transit peptide and the Brassica seed specific promoter Napin to eaccD gene.The expression vector was transformed into Brassica napus via Agrobacterium-mediated transformation and seven transgenic plants were obtained.The eaccD transcript was confirmed by RT-PCR analysis.The data showed that eaccD was transcripted in seeds but not in leaves.This demonstrated that the expression vector can result in the specific over expression of eaccD in immatural seeds of Brassica.Seed oil contents were detected by Soxhlet extraction.The total oil content of transgenic seeds was approximately 9% higher than that of the wild plant.Therefore,expression of eaccD in Brassica could significantly increase the fatty acid content.