van der Graaff E, den Dulk-Ras A, Hooykaas P J
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Netherlands.
Plant Mol Biol. 1996 Jun;31(3):677-81. doi: 10.1007/BF00042239.
We analyzed 29 T-DNA inserts in transgenic Arabidopsis thaliana plants for the junction of the right border sequences and the flanking plant DNA. DNA sequencing showed that in most lines the right border sequences transferred had been preserved during integration, corroborating literature data. Surprisingly, in four independent transgenic lines a complete right border repeat was present followed by binary vector sequences. Cloning of two of these T-DNA inserts by plasmid rescue showed that in these lines the transferred DNA consisted of the complete binary vector sequences in addition to the T-region. On the basis of the structure of the transferred DNA we propose that in these lines T-DNA transfer started at the left-border repeat, continued through the vector part, passed the right border repeat, and ended only after reaching again this left-border repeat.
我们分析了转基因拟南芥植株中29个T-DNA插入片段与右侧边界序列及侧翼植物DNA的连接情况。DNA测序表明，在大多数株系中，转移的右侧边界序列在整合过程中得以保留，这与文献数据相符。令人惊讶的是，在4个独立的转基因株系中，存在完整的右侧边界重复序列，其后跟着二元载体序列。通过质粒拯救对其中两个T-DNA插入片段进行克隆表明，在这些株系中，转移的DNA除了T-DNA区域外还包括完整的二元载体序列。基于转移DNA的结构，我们推测在这些株系中，T-DNA转移始于左侧边界重复序列，接着穿过载体部分，经过右侧边界重复序列，直到再次到达左侧边界重复序列才结束。
