从油菜中克隆得到了ICE1基因的开放阅读框序列.构建了以CaMV35S为启动子的植物表达载体,经农杆菌介导法转化烟草,获得转基因植株.通过PCR检测,目的基因成功转入烟草中.将转油菜ICE1基因的烟草和非转基因烟草置于2℃处理7 d,非转基因烟草出现萎蔫,转基因烟草没有明显变化,非转基因烟草丙二醛浓度明显高于转基因烟草.将转基因烟草与非转基因烟草分别置于0、-2、-4℃各1 h后,转基因烟草APX、SOD活性明显高于非转基因烟草.
A complete open reading frame of ICE1 gene was cloned from Brassica campestris(rape).Then the ICE1 gene expression vectors containing CaMV35 S promoters were constructed.The ICE1 gene was introduced into tobacco genome by Agrobacterium tumefaciens mediated transformation.The PCR assay results showed that ICE1 had been integrated into the genome of tobacco successfully.When transgenic and non-transgenic plants were stressed by 2 ℃ with seven days,the non-transgenic tobacco wilted seriously but the transgenic tobacco underwent little change and the content of MDA was significantly higher in the non-transgenic plants than in the transgenic lines.When transgenic and non-transgenic plants were stressed at 0,-2,-4 ℃ for1 h,APX and SOD activities were significantly higher in the transgenic lines than in the non-transgenic plants.