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Characterization of a small constitutive promoter from Arabidopsis translationally controlled tumor protein (AtTCTP) gene for plant transformation

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Abstract

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A plant-derived 0.3 kb constitutive promoter was obtained from AtTCTP expression analysis, and successfully applied to the expression of a selectable marker gene for production of transgenic creeping bentgrass plants.

Abstract

The isolation and use of an efficient promoter is essential to develop a vector system for efficient genetic transformation of plants, and constitutive promoters are particularly useful for the expression of selectable marker genes. In this study, we characterized a small size of the constitutive promoter from the expression analysis of Arabidopsis thaliana translationally controlled tumor protein (AtTCTP) gene. Histochemical and fluorometric GUS analyses revealed that a 303 bp upstream region from the start codon of the AtTCTP gene showed strong GUS expression throughout all plant tissues, which is approximately 55 % GUS activity compared with the cauliflower mosaic virus 35S promoter (35Spro). To examine the possible application of this promoter for the development of genetically engineered crops, we introduced pCAMBIA3301 vector harboring the 0.3 kb promoter of AtTCTP (0.3kbpro) that was fused to the herbicide resistance BAR gene (0.3kb pro ::BAR) into creeping bentgrass. Our transformation results demonstrate that transgenic creeping bentgrass plants with herbicide resistance were successfully produced using 0.3kb pro ::BAR as a selectable marker. Northern blot analysis revealed that the transgenic plants with 0.3kb pro ::BAR showed reduced but comparable expression levels of BAR to those with 35S pro ::BAR. Moreover, the transcription activity of the 0.3 kb promoter could be increased by the fusion of an enhancer sequence. These results indicate that the 0.3 kb AtTCTP promoter can be used as a plant-derived constitutive promoter for the expression of selectable marker genes, which facilitates its use as an alternative to the 35S promoter for developing genetically engineered crops.

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Acknowledgments

We thank the Kumho Life Science Laboratory in Chonnam National University for providing plant growth facilities. This work was supported by Next-Generation BioGreen 21 Program, Rural Development Administration, Republic of Korea (to J.-I. Kim, grant no. PJ00797804), and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (to Y.-J. Han, grant no. 2011-0012362).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biotechnology and Kumho Life Science Laboratory, Chonnam National University, Gwangju, 500-757, Korea

    Yun-Jeong Han, Yong-Min Kim, Ok-Jin Hwang & Jeong-Il Kim

  2. Korea Bioinformation Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, Korea

    Yong-Min Kim

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  1. Yun-Jeong Han
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  2. Yong-Min Kim
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Correspondence to Jeong-Il Kim.

Additional information

Communicated by Youn-Il Park.

Y.-J. Han, Y.-M. Kim contributed equally to this work.

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Han, YJ., Kim, YM., Hwang, OJ. et al. Characterization of a small constitutive promoter from Arabidopsis translationally controlled tumor protein (AtTCTP) gene for plant transformation. Plant Cell Rep 34, 265–275 (2015). https://doi.org/10.1007/s00299-014-1705-5

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  • DOI: https://doi.org/10.1007/s00299-014-1705-5

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