ubi启动子驱动的花生芪合酶基因表达载体的构建及玉米遗传转化初报

研究克隆了玉米的ubi启动子,以pBI 121为基本载体,构建了含有ubi启动子的中间载体pBI UB及ubi启动子驱动的GFP基因的植物表达载体pBI UG;采用基因枪法将载体pBI UG转入洋葱表皮细胞对启动子活性进行检测,结果表明GFP基因在洋葱表皮细胞中得到了表达。本实验同时采用RT-PCR技术克隆了花生的芪合酶基因(Res),该基因编码的蛋白质的氨基酸序列与已公布的芪合酶的氨基酸序列的同源性为99.49 %,并且164位的活性中心Cys没有发生突变;将该Res基因插入载体pBI UB构建了ubi启动子控制下的花生芪合酶基因植物表达载体pBI UA,并对玉米进行了遗传转化,得到了转化苗。

Construction of ubi promoter-based plant expression vector of stilbene synthase gene and transformation of type ii callus of maize by particle bombardment

ubi promoter of maize was cloned in our research. pBIUB vector containing this promoter and plant expression vector pBIUG with GFP gene driven by ubi promoter were constructed on the basis of pBI121. pBIUG was introduced into onion epidermis cells by particle bombardment and the results indicated that GFP gene expressed. RT-PCR was employed to clone Stilbene synthase gene of peanut in our experiment. The concensus between the amino acid sequence of the protein the cloned gene encoded and that of Stilbene synthase published was 99.49 % while no mutation occurred at the site of Cys164. Plant expression vector pBIUA, constructed by inserting Res gene into vector pBIUB, was transferred into type II callus of maize and transforments were obtained.