Perez P, Tiraby G, Kallerhoff J, Perret J
BIOSEM, Laboratoire de biologie moléculaire et cellulaire, Aubiere, France.
Plant Mol Biol. 1989 Oct;13(4):365-73. doi: 10.1007/BF00015548.
Tobacco cells are sensitive to bleomycin and phleomycin. The Tn5 and the Streptoalloteichus hindustanus (Sh) bleomycin resistance ('Ble') genes conferring resistance to these antibiotics have each been inserted into two plant expression vectors. They are flanked by the nopaline synthase (nos) or the cauliflower mosaic virus (CaMV) 35S promoters on one side, and by the nos polyadenylation signal on the other. These four chimaeric genes were introduced into the binary transformation vector pGA 492, which were thereafter mobilized into Agrobacterium tumefaciens strain LBA 4404. The resulting strains were used to transform Nicotiana tabacum cv. Xanthi nc using the leaf disc transformation procedure. In all cases, phleomycin- and bleomycin-resistant tobacco plants were regenerated from transformed cells under selective conditions; however, the highest frequency of rooted plants was obtained when transformation was carried out with the 'Sh Ble' gene under the control of the 35S promoter. Phleomycin resistance was stably transmitted to sexual offspring as a dominant nuclear trait as confirmed by Southern blotting.
烟草细胞对博来霉素和腐草霉素敏感。赋予对这些抗生素抗性的Tn5和印度斯坦链霉菌（Sh）博来霉素抗性（“Ble”）基因已分别插入两种植物表达载体中。它们一侧由胭脂碱合成酶（nos）或花椰菜花叶病毒（CaMV）35S启动子侧翼，另一侧由nos多聚腺苷酸化信号侧翼。这四个嵌合基因被引入二元转化载体pGA 492，随后将其转入根癌农杆菌菌株LBA 4404中。所得菌株用于通过叶盘转化程序转化烟草品种Xanthi nc。在所有情况下，在选择条件下从转化细胞中再生出对腐草霉素和博来霉素抗性的烟草植株；然而，当用35S启动子控制下的“Sh Ble”基因进行转化时，获得了最高频率的生根植株。如Southern印迹法所证实，腐草霉素抗性作为显性核性状稳定地传递给有性后代。
