Characterization of root-specific alpha-Dioxygenase1 promoter in tomato

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Characterization of root-specific alpha-Dioxygenase1 promoter in tomato
H A Lee; D J Lee; Cheol-Goo Hur; D Choi
Bibliographic Citation
Horticulture Environment and Biotechnology, vol. 50, no. 3, pp. 240-246
Publication Year
The availability of tissue-specific promoters can dramatically increase the successful application of transgenic technology. For isolation of root-specific promoter, 11 tomato ESTs, predicted as expressed in a tissue-specific manner by in silico Audic’s test, were selected for further analysis. An EST encoding α-Dioxygenase1 (α-DOX1) gene, catalyzing the oxygenation of fatty acids to yield oxylipin, was selected to be root-specific based on RT-PCR analysis. Promoter region of α-DOX1 was isolated by PCR method. Putative cis-acting elements, such as root-specificity and phytohormones responsiveness, were predicted by Plant cis-acting regulatory DNA elements program (PLACE). Promoter deletion fragments were amplified based on putative cis-acting element analysis, then cloned into pCAMBIA1391z for driving the expression of β-glucuronidase (GUS) gene. These constructs were transformed into Arabiopsis thaliana by floral dip method. T1 transgenic seeds were harvested for further characterization. The GUS expression of transgenic line SlDOX1P10 and SlDOX1P05 was only observed in root tissues. Phytohormones and salt-responsiveness in these transgenic Arabidopsis were also shown by Histochemical GUS staining. The promoter could be used as a useful tool for expression of foreign gene in plant root.
GUS histochemical assaysin silicotissue-specific promotersArabidopsis transformation
Korea Soc-Assoc-Inst
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