Development of a simple and efficient system for excising selectable markers in Arabidopsis using a minimal promoter::Cre fusion construct |
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Authors: | Hyun-Bi Kim Jung-Il Cho Nayeon Ryoo Shaohong Qu Guo-Liang Wang Jong-Seong Jeon |
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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: | The development of rapid and efficient strategies to generate selectable marker-free transgenic plants could help increase
the consumer acceptance of genetically modified (GM) plants. To produce marker-free transgenic plants without conditional
treatment or the genetic crossing of offspring, we have developed a rapid and convenient DNA excision method mediated by the
Cre/loxP recombination system under the control of a −46 minimal CaMV 35S promoter. The results of a transient expression assay showed
that −46 minimal promoter::Cre recombinase (−46::Cre) can cause the loxP-specific excision of a selectable marker, thereby connecting the 35S promoter and β-glucuronidase (GUS) reporter gene. Analysis of stable transgenic Arabidopsis plants indicated a positive correlation between loxP-specific DNA excision and GUS expression. PCR and DNA gel-blot analysis further revealed that nine of the 10 tested T1 transgenic lines carried both excised and nonexcised constructs in their genomes. In the subsequent T2 generation plants, over 30% of the individuals for each line were marker-free plants harboring the excised construct only.
These results demonstrate that the −46::Cre fusion construct can be efficiently and easily utilized for producing marker-free transgenic plants. |
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Keywords: | -46 promoter Arabidopsis Cre loxP marker-free plant |
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