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Properties of the maize transposable element Activator in transgenic tobacco plants: a versatile inter-species genetic tool.

机译:玉米转座因子激活子在转基因烟草植物中的特性:一种通用的种间遗传工具。

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摘要

The maize controlling element Activator (Ac) transposes autonomously from an integrated T-DNA vector to new sites in the genomes of tobacco and other heterologous plant species. Here we demonstrate that critical functions required for transposition of Ac in maize are conserved in tobacco and that Ac transposes at high frequency for at least five generations. Ac structure and terminal sequences are conserved upon transposition and a characteristic 8-bp duplication of target sequences is generated upon integration. Ac remains unmethylated, transcriptionally active, and capable to trans-activate transposition of the nonautonomous Dissociation (Ds) element throughout several generations. In tobacco, as in maize, Ac transposes adjacent to low copy or unique DNA, and transcriptional analysis of unique target DNA provides evidence that an Ac element transposed into a gene. In maize, increasing copies of Ac delay the timing and reduce the frequency of early transposition of Ac and Ds. In tobacco, increasing copies of Ac correlate with an increased frequency of Ds trans-activation. These data firmly establish that the Ac and Ds transposable elements are versatile genetic tools well suited for use as insertional mutagens and demonstrate that thorough investigation of mechanism and regulation of transposition is facilitated in heterologous settings.
机译:玉米控制元件激活剂(Ac)从整合的T-DNA载体自动转座到烟草和其他异源植物物种基因组中的新位点。在这里,我们证明了玉米中Ac的转座所需的关键功能在烟草中得以保留,并且Ac至少在五代内以高频转座。易位时,Ac结构和末端序列会保守,整合后会生成目标序列的特征性8 bp复制。 Ac保持未甲基化,具有转录活性,并能够在整个几代中转位激活非自主解离(Ds)元素。在烟草中,就像在玉米中一样,Ac邻近低拷贝或独特的DNA转座,独特目标DNA的转录分析提供了一个证据,表明Ac元素已转座到基因中。在玉米中,增加Ac的拷贝会延迟时间,并降低Ac和Ds早期转座的频率。在烟草中,Ac拷贝数的增加与Ds反式激活频率的增加有关。这些数据坚定地证明,Ac和Ds转座因子是通用的遗传工具,非常适合用作插入诱变剂,并证明在异源环境中,彻底研究了转座的机制和调控。

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