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KMID : 0578320120330010061
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Molecules and Cells
2012 Volume.33 No. 1 p.61 ~ p.69
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Development of a Simple and Efficient System for Excising Selectable Markers in Arabidopsis Using a Minimal Promoter::Cre Fusion Construct
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Kim Hyun-Bi
Cho Jung-Il
Ryoo Na-Yeon
Shaohong Qu
Guo-Liang Wang
Jeon Jong-Seong
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Abstract
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The development of rapid and efficient strategies to gen-erate 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 ?-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 non-excised 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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KEYWORD
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-46 promoter, Arabidopsis, Cre, loxP, marker-free plant
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