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将克隆的烟草 β-1 ,3 -葡聚糖酶基因插入双子叶植物高效表达载体 p Bin4 3 8中 ,构建了植物表达载体 p BLG。利用农杆菌介导法 ,将此抗真菌基因导入甘蓝型双低油菜品种 H1 65中 ,得到了抗卡那霉素的再生植株 ,并探讨了影响油菜分化的几个要素。结果表明 :乙酰丁香酮及硝酸银等物质对油菜分化及绿苗的形成具有重要作用 ;油菜对卡那霉素十分敏感 ,提高卡那霉素浓度有可能将一部分转化体淘汰。对 8株再生植株进行了 PCR检测 ,其中 4株表现为阳性。进一步的点杂交分析表明 ,有 3株表现出较强的杂交信号 ,初步说明外源基因已整合到油菜基因组中
The cloned tobacco β-1, 3-glucanase gene was inserted into dicotyledonous high efficient expression vector p Bin4 3 8 to construct plant expression vector p BLG. Using Agrobacterium - mediated method, the antifungal gene was introduced into Brassica napus variety H165, and the regenerated plants resistant to kanamycin were obtained. Several factors influencing rape differentiation were also discussed. The results showed that acetosyringone and silver nitrate played an important role in the differentiation of rapeseed and the formation of green shoots. Rapeseed was very sensitive to kanamycin, and it was possible to eliminate some of the transformants by increasing the concentration of kanamycin. Eight regenerated plants were tested by PCR, of which 4 were positive. Further point hybridization analysis showed that 3 of them showed strong hybridization signals, which indicated that foreign genes had been integrated into the genome of rape