农杆菌介导Ds转座因子的黄瓜遗传转化

Agrobacterium-mediated Ds Transposon Transformation in Cucumber

以黄瓜品种"中农15"子叶外植体为试料,绿色荧光蛋白GFP基因为报告基因,利用农杆菌介导的遗传转化法将构建的激活表达标签pDsBar导入黄瓜,获得转基因黄瓜群体,对转化群体进行GFP基因活性及分子检测。结果表明,GFP已经整合到黄瓜基因组中;以Bar基因设计引物进行PCR检测表明,阳性率为72.3%;通过Southern杂交分析,发现26.0%为1个插入位点,51.0%为2个插入位点,T-DNA在基因组中的平均拷贝数为2.2个。

The activation tagging pDsBar was transformed into cucumber via Agrobacterium-mediated with GFP as reporter genes. The transformed cucumber population was gained. GFP gene activation and molecular detection were conducted with the transformed plants. The results showed that GFP gene was interknitted into tomato genome. The positive rate was 72.3 % by PCR detection. Analyzed with southern blotting, about 26.0 % among T1 population with one inserted site and 51.0 % with two inserted sites were detected. The average inserted T-DNA sites was 2.2.