Excision of a selective marker in transgenic rice using a novel Cre/<Emphasis Type="Italic">loxP</Emphasis> system controlled by a floral specific promoter |
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Authors: | Xianquan Bai Qiuyun Wang Chengcai Chu |
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Institution: | (1) State Key Laboratory of Plant Genomics and National Centre for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China;(2) Sci-Tech Innovation Centre of Hainan State Farm, Haikou, Hainan, 570206, China;(3) Graduate University of the Chinese Academy of Sciences, Beijing, 100039, China |
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Abstract: | 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 T1 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 T1 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.
Xianquan Bai and Qiuyun Wang contributed equally to the work. |
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Keywords: | OsMADS45 Floral-specific promoter Cre/loxP Marker-free Rice |
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