Ramanathan V, Veluthambi K
Department of Plant Molecular Biology, School of Biotechnology, Madurai Kamaraj University, India.
Plant Mol Biol. 1995 Sep;28(6):1149-54. doi: 10.1007/BF00032676.
We introduced a plant selection marker, nptII, to the left of border A in the Agrobacterium Ti plasmid pTiA6. Infection of tobacco leaf discs with the modified Agrobacterium strain gave rise to kanamycin-resistant calli which grew in a hormone-dependent manner. Southern hybridization analysis of DNA isolated from four transformants indicated initiation of DNA transfer at or near border A and absence of T-DNA sequences. These results demonstrate that DNA transfer events starting at a left border on a native Ti plasmid and moving away from the T-DNA region occur and that they can be detected by designing a suitable selection strategy.
我们将植物选择标记基因nptII导入根癌土壤杆菌Ti质粒pTiA6中边界A的左侧。用改良的根癌土壤杆菌菌株感染烟草叶盘,产生了对卡那霉素具有抗性的愈伤组织,这些愈伤组织以激素依赖的方式生长。对从四个转化体中分离的DNA进行的Southern杂交分析表明,DNA转移在边界A处或其附近起始,并且不存在T-DNA序列。这些结果表明,从天然Ti质粒上的左边界起始并远离T-DNA区域的DNA转移事件会发生,并且可以通过设计合适的选择策略来检测。