Institute of Molecular Biology, Austrian Academy of Sciences, Billrothstrasse 11, A-5020, Salzburg, Austria.
Plant Mol Biol. 1986 Sep;7(5):357-65. doi: 10.1007/BF00032565.
We describe in this paper the construction and use of a set of novel Ti plasmid-derived vectors that can be used to produce transgenic plants. These vectors are based on one of two strategies: 1) double recombination into the wild-type Ti plasmid of genetic information flanked by two T-DNA fragments on a wide-host range plasmid; 2) the binary vector strategy. The vector based on the double recombination principle contains a kanamycin resistance gene for use as a plant selectable marker, a polylinker for the insertion of foreign genes, and a nopaline synthase gene. The vector was constructed such that a disarmed T-DNA results from the double recombination event. The binary vector combines several advantageous features including an origin of replication that is stable in Agrobacterium in the absence of selection, six unique sites for insertion of foreign genes, an intact nopaline synthase gene, and a kanamycin resistance marker for selection of transformed plant cells. All of these vectors have been used to produce tobacco plants transformed with a variety of foreign genes.
我们在本文中描述了一组新型 Ti 质粒衍生载体的构建和使用,这些载体可用于生产转基因植物。这些载体基于两种策略之一:1)将侧翼带有两个 T-DNA 片段的遗传信息双重组到野生型 Ti 质粒上,位于广泛宿主范围质粒上;2)二元载体策略。基于双重组原理的载体包含一个卡那霉素抗性基因,用作植物选择标记,一个多克隆位点,用于插入外源基因,和一个胭脂碱合成酶基因。该载体的构建使得双重组事件产生一个无功能的 T-DNA。二元载体结合了几个有利的特点,包括在没有选择的情况下在农杆菌中稳定的复制起点、用于插入外源基因的六个独特位点、完整的胭脂碱合成酶基因,以及用于转化植物细胞选择的卡那霉素抗性标记。所有这些载体都已被用于生产转化了多种外源基因的烟草植物。