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. 2016 Feb 25;17(3):233. doi: 10.3390/ijms17030233

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© 2016 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

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Identification of a regulator of the CBF1 gene. (a) A rice complementary DNA (cDNA) library was generated using a pGAD424 vector in which the coding sequences of the protein of interest were C-terminally fused to the activation domain (AD). A 2.0-kb section of the CBF1 gene promoter was cloned into the pHISi vector in which His was a reporter gene; (b) A yeast one-hybrid assay was performed to analyze the regulator of the CBF1 gene (ROC1) activation of the CBF1 promoter. Yeast cells harboring either AD-ROC1 and pCBF1-His or AD and pCBF1-His were grown on SD media lacking Leu or His and containing the indicated concentrations of 3-amino-1,2,4-triazole (3AT), a competitive inhibitor of HIS3.