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Excision of a selective marker in transgenic rice using a novel Cre/loxP system controlled by a floral specific promoter

机译:使用由花特异性启动子控制的新型Cre / loxP系统切除转基因水稻中的选择标记

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摘要

Based on the Cre/loxP system, we have developed a novel marker-free system mediating a direct auto-excision of loxP-flanked marker genes from T(1 )transgenic rice without any treatment or further offspring crossing. To achieve this, the floral-specific promoter OsMADS45 was isolated from rice and the expression patterns of OsMADS45 promoter was characterised by using the pOs45:GUS transgenic plants. Furthermore, the binary vector with Cre recombinase under the control of OsMADS45 promoter was constructed and introduced into rice by Agrobacterium-mediated transformation and transgenic rice plants were generated. Southern blot analysis showed that auto-excision of the selective markers occurred in some T(1) progeny of the transgenic plants, suggesting that a high auto-excision frequency can be achieved with our Cre/loxP system. This auto-excision strategy provides an efficient way of removing the selectable marker gene from transgenic rice.
机译:基于Cre / loxP系统,我们开发了一种新颖的无标记系统,该系统介导了T(1)转基因水稻中loxP侧翼标记基因的直接自动切除,无需任何处理或进一步的后代杂交。为此,从水稻中分离出了花特异性启动子OsMADS45,并使用pOs45:GUS转基因植物对OsMADS45启动子的表达模式进行了表征。此外,构建了在OsMADS45启动子控制下具有Cre重组酶的二元载体,并通过农杆菌介导的转化将其引入水稻,并产生了转基因水稻植物。 Southern印迹分析表明,选择性标记的自动切除发生在转基因植物的某些T(1)后代中,这表明我们的Cre / loxP系统可以实现较高的自动切除频率。这种自动切除策略提供了从转基因水稻中去除选择标记基因的有效方法。

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