Elimination of the nptII marker gene in transgenic apple and pear with a chemically inducible R/Rs recombinase
Elimination of the nptII marker gene in transgenic apple and pear with a chemically inducible R/Rs recombinase
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DOI：
10.1007/s11240-014-0443-2
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发表时间：
2014-02
期刊：
影响因子：
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通讯作者：
L. Righetti;L. Righetti;L. Righetti;S. Djennane;P. Berthelot;P. Berthelot;P. Berthelot;R. Cournol;R. Cournol;R. Cournol;N. Wilmot;N. Wilmot;N. Wilmot;K. Loridon;K. Loridon;K. Loridon;E. Vergne;E. Vergne;E. Vergne;E. Chevreau;E. Chevreau;E. Chevreau
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文献类型：
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作者：
L. Righetti;L. Righetti;L. Righetti;S. Djennane;P. Berthelot;P. Berthelot;P. Berthelot;R. Cournol;R. Cournol;R. Cournol;N. Wilmot;N. Wilmot;N. Wilmot;K. Loridon;K. Loridon;K. Loridon;E. Vergne;E. Vergne;E. Vergne;E. Chevreau;E. Chevreau;E. Chevreau
无标记转基因植物的高效生产对于大多数水果物种来说仍然是一个挑战，尽管此类植物是许多“新育种技术”的必要组成部分，特别是顺式和内生技术。无标记植物生产对于优良水果转基因品系中基因的连续堆积也是必要的。在这里，我们使用了一种 R/Rs 位点特异性重组酶，该重组酶通过与该激素的配体结合域融合而受到地塞米松的翻译后调节，以及编码胞嘧啶脱氨酶/新霉素转移酶 (codA-nptII) 蛋白的双功能选择标记基因；这实现了卡那霉素阳性选择的第一步，然后是 5-氟胞嘧啶阴性选择的第二步。我们研究的目的是在苹果简历上优化这个系统。银河和梨CV。会议对地塞米松和 5-氟胞嘧啶治疗的效果进行了详细研究，并比较了早期与延迟选择策略。我们首次能够生产出无标记转基因梨植物，并证实了使用化学诱导重组酶系统生产无标记转基因苹果植物的可行性。我们建议对梨品种使用早期选择策略。苹果简历的会议和延迟选择策略。星系。
The efficient production of marker-free transgenic plants is still a challenge in most fruit species even though such plants are a necessary component of many “new breeding technologies”, particularly cis- and intragenesis. Marker-free plant production is also necessary for the successive stacking of genes in an elite fruit transgenic line. Here, we used a R/Rs site-specific recombinase that is post-translationally regulated by dexamethasone through fusion with a ligand-binding domain for this hormone, and a bi-functional selectable marker gene coding for a cytosine deaminase/neomycin transferase (codA–nptII)protein; this enabled a first step of positive kanamycin selection, followed by a second step of negative 5-fluorocytosine selection. The aim of our study was to optimize this system on the apple cv. Galaxy and on the pear cv. Conference by conducting a detailed study of the effects of dexamethasone and 5-fluorocytosine treatments, and by comparing an early versus a delayed selection strategy. We were able to produce marker-free transgenic pear plants for the first time, and confirm the feasibility of producing marker-free transgenic apple plants using a chemically inducible recombinase system. We recommend the use of an early selection strategy for the pear cv. Conference and a delayed selection strategy for the apple cv. Galaxy.