期刊论文

中文

生物工程学报

1000-3061

1999

15

4

482-488

10.13345/j.cjb.1999.04.013

国家自然科学基金 湖北省自然科学基金

本校为第一机构

带有1 ~2 m m 子叶柄的芥菜型油菜子叶经农杆菌感染后,培养在附加10 ~20 mg/ L卡那霉素的 M S 选择培养基上筛选转化愈伤组织及不定芽。卡那霉素抗性苗相继在含30 ~50 m g/ L 卡那霉素的选择培养基上继代培养,再转移到含20 mg/ L 卡那霉素的生根培养基上诱导生根。以苏云金杆菌杀虫晶体蛋白基因为探针,进行 Southern blot 分子杂交,得到阳性结果。 P C R 分析也证明外源基因整合到油菜基因组并稳定传递到后代。转基因植株的抗虫性和卡那霉素抗性在自交后代中得到保持,筛选得到纯合的转基因植株后代株系

Cotyledons with the petioles were cut from axenic seedlings and infected with Agrobacterium tumefaciens . After 2 days of cultivation, the explants were transferred to MS selection medium with 10～20mg/L kanamycin and 4.5 mg/L 6 BA for shoot differentiantion. Shoots with kanamycin resistance were subcultured on selection medium with 30～50 mg/L kanamycin for eliminating escaped non transformants, and then rooted on MS medium containing 20mg/L kanamycin and 0.5 mg/L IBA. Transgenic plants were transplanted into soil and grew in the field. DNA So...