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通过RT-PCR扩增,从苹果叶片cDNA中克隆了FT基因的同源基因MdFT,构建了花椰菜病毒35S启动子驱动的MdFT植物表达载体35S::MdFT,并利用根癌农杆菌介导法将其导入番茄栽培品种‘中蔬四号’;同时转化拟南芥AtFT基因作为阳性对照。从添加卡那霉素的筛选培养基上再生了抗性植株,PCR扩增证明,外源基因MdFT和AtFT已经整合到转基因番茄的基因组,半定量RT-PCR则证明它们已经在转基因番茄中得到异位过量表达。形态鉴定发现,转基因番茄植株比非转基因对照植株开花早,表明成功地从苹果中克隆了成花素基因MdFT,该基因具有通过转基因缩短苹果树童期的潜在价值。
The homology gene MdFT of FT gene was cloned from cDNA of apple leaf by RT-PCR amplification. The MdFT plant expression vector 35S :: MdFT driven by 35S promoter of cauliflower virus was constructed and transformed into Agrobacterium tumefaciens It was introduced into tomato cultivar Zhongzhong 4 and AtFT gene of Arabidopsis thaliana was also used as a positive control. Resistant plants were regenerated from kanamycin-supplemented selection medium and PCR amplification demonstrated that the exogenous genes MdFT and AtFT have been integrated into the genome of transgenic tomato and semi-quantitative RT-PCR proved that they have been obtained in transgenic tomato Ectopic overexpression. Morphological identification revealed that transgenic tomato plants flowered earlier than non-transgenic control plants, indicating that MdFT was successfully cloned from apple, which has the potential to shorten the apple tree’s childhood through transgenesis.