Department of Plant Molecular Biology, University of Leiden, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands.
Plant Mol Biol. 1985 Mar;5(2):85-9. doi: 10.1007/BF00020090.
Agrobacterium strains harbouring the T-region and the virulence-region of the Ti plasmid on separate replicons still display efficient T-DNA transfer to plants. Based on this binary vector strategy we have constructed T-region derived gene vectors for the introduction of foreign DNA into plants. The vectors constructed can replicate in E. coli, thus the genetic manipulations with them can be performed with E. coli as a host. They can be transferred to Agrobacterium as a cointegrate with the wide host range plasmid R772. Their T-regions are transferred to plant cells from Agrobacterium strains conferring virulence functions.The plasmid pRAL 3940 reported here is 11.5 kb large, contains a marker to identify transformed plant cells and unique restriction sites for direct cloning of passenger DNA, flanked by the left- and right-hand border fragments of the T-region (including the 25 bp border repeats). The plasmid is free of onc-genes. Therefore, is does not confer tumorigenic traits on the transformed plant cells and mature, fertile plants can thus be regenerated from them.
含有 Ti 质粒 T 区和毒性区的农杆菌菌株在独立的复制子上仍能有效地将 T-DNA 转移到植物中。基于这种二元载体策略，我们构建了 T 区衍生的基因载体，用于将外源 DNA 导入植物。构建的载体可以在大肠杆菌中复制，因此可以用大肠杆菌作为宿主进行遗传操作。它们可以与广谱质粒 R772 一起作为共整合体转移到农杆菌中。它们的 T 区从赋予毒性功能的农杆菌菌株转移到植物细胞中。本文报道的质粒 pRAL 3940 大小为 11.5 kb，含有一个用于鉴定转化植物细胞的标记物和用于直接克隆载体 DNA 的独特限制位点，侧翼是 T 区的左边界和右边界片段（包括 25 bp 的边界重复序列）。该质粒不含致癌基因。因此，它不会赋予转化植物细胞致癌特性，因此可以从它们中再生出成熟、可育的植物。
