【摘要】 乙酰辅酶A羧化酶(ACCase)催化脂肪酸合成的第一步,是脂肪酸合成的限速酶。采用PCR方法分别从陆地棉和拟南芥基因组中扩增出ACCase羧基转移酶β-CT亚基编码基因accD,ACCase羧基转移酶α-CT亚基编码基因CAC3的定位于叶绿体的转运肽序列。将CAC3基因转运肽序列与accD基因进行体外重组,构建融合植物表达载体pBI-CAC3tp-accD。重组质粒通过冻融法转化根癌农杆菌GV3101。渗透法转化拟南芥,收种子,在含卡那霉素的MS培养基中发芽筛选。用叶盘法转化烟草,经不定芽诱导、生根培养,获转基因烟草植株。T1代转基因拟南芥和转基因烟草植株经卡那霉素检测、PCR、RT-PCR检测后,初步表明目的基因已在植株中转化成功,并可以正常转录。更多还原

【Abstract】 The acetyl-CoA carboxylase(ACCase) is the rate-limiting enzyme of fatty acids synthesis. The accD Gene and transit peptide of CAC3 Gene was amplified from Gossypuum hirsutum Genome and Arabidopsis thaliana Geneome. Plant Expression Vector of Fusion Transit Peptide of CAC3 Gene and accD Gene was constructed.The Vector of pBI-CAC3tp-accD were transferred into Agrobacterium tumefaciens GV3101. Using infiltration and leaf discs method,the gene were transferred into Arabidopsis thaliana and tobacco cell. The seeds of Arabidopsis thaliana were selected on solid medium containing Kanamycin. Transferred leafs of tobacco were selected on solid medium containing Kanamycin. Transgenic Arabidopsis thaliana(T1) and tobacco plants were found containing purpose gene by PCR. After RT-PCR,it shows that the CAC3tp-accD gene could transcript normally.更多还原