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作 者:卜云萍[1] 王广科[1] 胡国武[1] 孙红妍[1] 任勇[1] 李航[1] 李明春[1] 邢来君[1]
机构地区:[1]南开大学微生物系,天津300071
出 处:《生物技术》2003年第3期6-8,共3页Biotechnology
基 金:国家自然科学基金项目资助(编号 :30 2 0 0 1 76) ;天津市自然科学基金重点项目 (编号 :0 1 380 2 51 1 ) ;高等学校骨干教师资助计划项目
摘 要:采用农杆菌介导的大豆子叶节转化系统成功地将深黄被孢霉△6 -脂肪酸脱氢酶基因导入大豆。从发芽 5 - 7d的大豆无菌苗切取子叶节外植体 ,经农杆菌浸染和共培养后 ,在含 5 0mg L卡那霉素的选择培养基上培养 2 - 4w后 ,从子叶节处诱导出抗性不定芽。将不定芽转移到伸长培养基上 ,4 - 6w后长至 2 - 3cm高的再生苗。再将再生苗切下转入生根培养基 ,2 - 6w生根。生根后的再生植株经逐步锻炼移入花盆中 ,部分移栽成活的T0 植株能正常开花结荚。从T0 植株上收获T1 种子 ,按株系种植。T0 和T1 代经PCR检测和DNA分子杂交分析 。The △ 6-fatty acid desaturase gene fron Mortitieralla isabellina was introduced into soybean successfully with Agrobacterium -cotylendon node transformation system Cotylendon node explants were prepared from 5-7 day old seedlings,infected and co-cultivated with Agrobacterium tumefaciens Kanamycin resistant (Kmr) adventitious buds emerged after 2-4 weeks on selective medium The buds were transferred onto elongation medium and regenerated shoots grew to 2-3cm high after 4-6 weeks The shoots were cut off and transferred onto rooting medium After about 2-6 weeks,the rooted plantlets were hardened off and transferred into pots A part of the plants could bear flower and set seeds normally PCR and Southern blot analysis from the T0 and T1 plants proved that foreign gene were transferred and integrated into soybean genome
关 键 词:深黄被孢霉 Δ6-脂肪酸脱氢酶基因 导入 大豆 农杆菌介导法
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