Transgene structures in T-DNA-inserted rice plants
Transgene structures in T-DNA-inserted rice plants
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DOI：
10.1023/a:1025093101021
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发表时间：
2003-07-01
影响因子：
5.1
通讯作者：
An, G
中科院分区：
文献类型：
--
作者：
Kim, SR;Lee, J;An, G
DNA通常用于递送外源基因和作为插入诱变剂。虽然双子叶植物中存在大量关于T-DNA组织的信息，但对单子叶水稻却知之甚少。在此，我们研究了大量转基因水稻植株中T-DNA的结构。T-DNA边界的分析显示，超过一半的右端位于切割位点，而左端不保守，并且从左边界（LB）切割位点缺失多达180 bp。T-DNA与基因组DNA之间存在三种类型的连接。在第一个中，存在多达七个核苷酸重叠。这种类型的频率在LB区域比在右边缘（RB）高得多。在第二种类型中，这在RB中更常见，链接是直接的，没有任何重叠或填充DNA。第三类是T-DNA和植物序列之间的填充DNA。在检查的171个样品中，77个携带载体骨架序列，其中大多数是由于在LB处T链终止失败引起的。然而，显著部分也是由T-DNA和载体骨架共整合到单个基因座产生的。T-DNA与载体骨架之间的连接大多数形成于转移DNA的两个3'端或两个5'端之间。3'端主要通过3-6 bp的互补序列连接，而5'端通过与填充DNA的精确连接或不精确连接连接。
DNA is commonly used for delivery of foreign genes and as an insertional mutagen. Although ample information exists regarding T-DNA organization in dicotyledonous plants, little is known about the monocot rice. Here, we investigated the structure of T-DNA in a large number of transgenic rice plants. Analysis of the T-DNA borders revealed that more than half of the right ends were at the cleavage site, whereas the left ends were not conserved and were deleted up to 180 bp from the left border (LB) cleavage site. Three types of junctions were found between T-DNA and genomic DNA. In the first, up to seven nucleotide overlaps were present. The frequency of this type was much higher in the LB region than at the right border (RB). In the second type, which was more frequent in RB, the link was direct, without any overlaps or filler DNA. Finally, the third type showed filler DNA between T-DNA and the plant sequences. Out of 171 samples examined, 77 carried the vector backbone sequence, with the majority caused by the failure of T-strand termination at LB. However, a significant portion also resulted from co-integration of T-DNA and the vector backbone to a single locus. Most linkages between T-DNA and the vector backbone were formed between two 3' ends or two 5' ends of the transferred DNAs. The 3' ends were mostly linked through 3-6 bp of the complementing sequence, whereas the 5' ends were linked through either precise junctions or imprecise junctions with filler DNA.