油菜ICE1基因表达载体构建及转化烟草-兰州大学学报.PDFVIP

兰州大学学报: 自然科学版, 2016, 52(2) / 4 月 Journal of Lanzhou University ：Natural Sciences，2016 ，52(2) / April 油菜ICE 1基因表达载体构建及转化烟草 张腾国, 毛玉珊, 常 燕, 王 娟, 聂亭亭 西北师范大学 生命科学学院, 兰州 730070 摘 要：从油菜中克隆得到了ICE 1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ICE 1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. 关键词：油菜; ICE 1 基因; 转化烟草 中图分类号：Q946 文献标识码：A 文章编号: 0455-2059(2016)02-0277-06 DOI: 10.13885/j.issn.0455-2059.2016.02.019 Construction of an expression vector of ICE1 gene of Brassica campestris and its transformation into tobacco Zhang Teng-guo, Mao Yu-shan, Chang Yan, Wang Juan, Nie Ting-ting School of Life Sciences, Northwest Normal University, Lanzhou 730070, China Abstract: A complete open reading frame of ICE 1 gene was cloned from Brassica campestris (rape). Then the ICE 1 gene expression vectors containing CaMV35S promoters were constructed. The ICE 1 gene was intro- duced into tobacco genome by Agrobacterium tumefaciens mediated transformation. The PCR assay results showed that ICE 1 had been integrated into the genome of tobacco successfully. When transgenic and non-trans- genic plants were stressed by 2 ℃with seven days, the non-transgenic tobacco wilted seriously but the trans- genic 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 ℃for 1 h, APX and SOD activities were significantly higher in the transgenic lines than in the non-transgenic plants. Key words: Brassica campestris ; ICE 1 gene; transgenic tobacco