Takahashi M, Nishihara M, Yamamura S, Nishizawa S, Irifune K, Morikawa H
Graduate Department of Gene Science, Faculty of Science, Hiroshima University, Kagamiyama, Higashi-Hiroshima 739-8526, Japan, , , , , , JP.
Iwate Biotechnology Research Center, Kitakami, Iwate 024-0003, Japan, , , , , , JP.
Plant Cell Rep. 1998 Apr;17(6-7):504-507. doi: 10.1007/s002990050432.
Explants (7.5±2.5 mm) cut from stems and roots of 3-week-old Eustoma grandiflorum Grise, (lisianthus) cv. Glory White seedlings were bombarded with plasmid pBI221, which harbors the uidA gene encoding β-glucuronidase (GUS) driven by the cauliflower mosaic virus (CaMV) 35S promoter. More than 800 blue spots of GUS-expressing cells were observed per 90 explants. Explants bombarded with pARK22 harboring the bar gene encoding phosphinothricin acetyltransferase driven by the CaMV 35S promoter were selected for bialaphos resistance. Putative transgenic plants were obtained about 3 months after bombardment. Southern blot analysis of putative transgenic plants revealed the presence of the bar gene in their genome.
从3周龄的大花龙胆（洋桔梗）品种Glory White幼苗的茎和根上切下外植体（7.5±2.5毫米），用质粒pBI221进行轰击，该质粒含有由花椰菜花叶病毒（CaMV）35S启动子驱动的编码β-葡萄糖醛酸酶（GUS）的uidA基因。每90个外植体中观察到800多个表达GUS的蓝色细胞斑点。用含有由CaMV 35S启动子驱动的编码膦丝菌素乙酰转移酶的bar基因的pARK22轰击外植体，筛选对双丙氨膦具有抗性的植株。轰击后约3个月获得了推定的转基因植株。对推定的转基因植株进行Southern印迹分析，结果显示其基因组中存在bar基因。
