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Mol. Cells 2012; 33(1): 61-69

Published online January 31, 2012

https://doi.org/10.1007/s10059-012-2212-6

© The Korean Society for Molecular and Cellular Biology

Development of a Simple and Efficient System for Excising Selectable Markers in Arabidopsis Using a Minimal Promoter::Cre Fusion Construct

Hyun-Bi Kim1,4, Jung-Il Cho1,4, Nayeon Ryoo1, Shaohong Qu2, Guo-Liang Wang3, and Jong-Seong Jeon1,*

1Graduate School of Biotechnology and Crop Biotech Institute, Kyung Hee University, Yongin 446-701, Korea, 2Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang 310021, China, 3Department of Plant Pathology, Ohio State University, Columbus, OH 43210, USA, 4 These authors contributed equally to this work.

Correspondence to : *Correspondence: jjeon@khu.ac.kr

Received: September 30, 2011; Revised: November 9, 2011; Accepted: November 11, 2011

Abstract

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.

Keywords -46 promoter, Arabidopsis, Cre, loxP, marker-free plant

Article

Research Article

Mol. Cells 2012; 33(1): 61-69

Published online January 31, 2012 https://doi.org/10.1007/s10059-012-2212-6

Copyright © The Korean Society for Molecular and Cellular Biology.

Development of a Simple and Efficient System for Excising Selectable Markers in Arabidopsis Using a Minimal Promoter::Cre Fusion Construct

Hyun-Bi Kim1,4, Jung-Il Cho1,4, Nayeon Ryoo1, Shaohong Qu2, Guo-Liang Wang3, and Jong-Seong Jeon1,*

1Graduate School of Biotechnology and Crop Biotech Institute, Kyung Hee University, Yongin 446-701, Korea, 2Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang 310021, China, 3Department of Plant Pathology, Ohio State University, Columbus, OH 43210, USA, 4 These authors contributed equally to this work.

Correspondence to:*Correspondence: jjeon@khu.ac.kr

Received: September 30, 2011; Revised: November 9, 2011; Accepted: November 11, 2011

Abstract

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.

Keywords: -46 promoter, Arabidopsis, Cre, loxP, marker-free plant

Mol. Cells
Jun 30, 2022 Vol.45 No.6, pp. 353~434
COVER PICTURE
ERα is modified by UFM1 and this modification (ufmylation) plays a crucial role in promoting the stability of ERα and breast cancer development. However, when ERα is deufmylated and then ubiquitinated, it disappears by proteasome-mediated degradation (Yoo et al., pp. 425-434).

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