Department of Biochemistry, University of Arizona, 85721, Tucson, AZ, USA.
Plant Cell Rep. 1989 Jun;8(6):354-7. doi: 10.1007/BF00716672.
Daucus carota hypocotyl sections were transformed withAgrobacterium tumefaciens LBA4404 containing CaMV 35S promoter, β-glucuronidase coding sequence and the nopaline synthase (Nos) poly adenylation sequences in Bin 19. Sliced sterile seedling hypocotyl segments were preincubated for 2 days, co-cultivated withAgrobacterium for an additional 2 days, and then transferred to medium containing 100ug/ml of kanamycin and 400ug/ml carbenicillin. In 6 weeks kanamycin resistant calli were obtained in 5.8% of the explants from one variety. Calli were subcultured on solid medium, and in 4 weeks introduced into suspension culture. NPTII and Southern blot analysis confirmed that three selected lines were transformed with 1-3 copies of the GUSII construction. GUS activity in transformants was 5 to 250 fold over background.
胡萝卜下胚轴切片用含有 CaMV 35S 启动子、β-葡萄糖醛酸酶编码序列和胭脂碱合成酶（Nos）多聚腺苷酸化序列的根癌农杆菌 LBA4404 转化。无菌幼苗下胚轴切段预培养 2 天，与农杆菌共培养 2 天，然后转移到含有 100μg/ml 卡那霉素和 400μg/ml 羧苄青霉素的培养基中。在 6 周内，从一个品种的外植体中获得了 5.8%的卡那霉素抗性愈伤组织。愈伤组织在固体培养基上继代培养，4 周后引入悬浮培养。NPTII 和 Southern blot 分析证实，三条选定的品系被 1-3 个拷贝的 GUSII 构建体转化。转化体中的 GUS 活性比背景高 5 到 250 倍。
