Komari Toshihiko, Takakura Yoshimitsu, Ueki Jun, Kato Norio, Ishida Yuji, Hiei Yukoh
Plant Innovation Center, Japan Tobacco Inc., Iwata, Shizuoka, Japan.
Methods Mol Biol. 2006;343:15-41. doi: 10.1385/1-59745-130-4:15.
A binary vector is a standard tool in the transformation of higher plants mediated by Agrobacterium tumefaciens. It is composed of the borders of T-DNA, multiple cloning sites, replication functions for Escherichia coli and A. tumefaciens, selectable marker genes, reporter genes, and other accessory elements that can improve the efficiency of and/or give further capability to the system. A super-binary vector carries additional virulence genes from a Ti plasmid, and exhibits very high frequency of transformation, which is valuable for recalcitrant plants such as cereals. A number of useful vectors are widely circulated. Whereas vectors with compatible selectable markers and convenient cloning sites are usually the top criteria when inserting gene fragments shorter than 15 kb, the capability of maintaining a large DNA piece is more important for consideration when introducing DNA fragments larger than 15 kb. Because no vector is perfect for every project, it is recommended that modification or construction of vectors should be made according to the objective of the experiments. Existing vectors serve as good sources of components.
双元载体是根癌农杆菌介导高等植物转化的标准工具。它由T-DNA边界、多克隆位点、大肠杆菌和根癌农杆菌的复制功能、选择标记基因、报告基因以及其他能提高系统效率和/或赋予系统更多功能的辅助元件组成。超级双元载体携带来自Ti质粒的额外毒力基因,表现出非常高的转化频率,这对于像谷物这样难转化的植物很有价值。许多有用的载体广泛流通。当插入长度小于15 kb的基因片段时,具有兼容选择标记和方便克隆位点的载体通常是首要标准,而当引入大于15 kb的DNA片段时,维持大片段DNA的能力则更重要。由于没有一种载体对每个项目都是完美的,建议根据实验目的对载体进行修饰或构建。现有的载体是很好的元件来源。