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

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Title
Characterization of a small constitutive promoter from Arabidopsis translationally controlled tumor protein (AtTCTP) gene for plant transformation
Author(s)
Y J Han; Yong Min Kim; O J Hwang; J I Kim
Bibliographic Citation
Plant Cell Reports, vol. 34, no. 2, pp. 265-275
Publication Year
2015
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 (0.3kbpro::BAR) into creeping bentgrass. Our transformation results demonstrate that transgenic creeping bentgrass plants with herbicide resistance were successfully produced using 0.3kbpro::BAR as a selectable marker. Northern blot analysis revealed that the transgenic plants with 0.3kbpro::BAR showed reduced but comparable expression levels of BAR to those with 35Spro::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.
Keyword
Arabidopsis translationally controlled tumor proteinConstitutive promoterPlant transformationPromoter analysisSelectable marker
ISSN
0721-7714
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s00299-014-1705-5
Type
Article
Appears in Collections:
Division of Biomedical Research > Genome Editing Research Center > 1. Journal Articles
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