Abstract
With the use of in vivo recombination theory, the screening time of yeast one-hybrid system was decreased in the present study. A basic helix-loop-helix (bHLH) protein PsGBF was successfully obtained from a glutathione (GSH)-induced pea cDNA library using the G-box cis-element of the PsCHS1 promoter as a bait. Electrophoretic mobility shift assay (EMSA) and β-galactosidase assay results suggested that PsGBF possesses both G-box-specific binding and transcription-activating activities. The specific interaction of PsGBF with G-box was further confirmed by in vivo transient expression assays in tobacco. The current study examined the combination effect of G-box with Box I elements in the interaction with PsGBF or OsMYC. The results indicated that PsGBF bound with the G-box, but not the Box I element. Moreover, this combination effect of G-box and Box I only associated with PsGBF but not with other bHLH-type proteins such as OsMYC.
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Acknowledgments
We thank Prof. Fan Chen of the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, for providing the plasmids pHISi, pLacZi and the yeast strain YM4271. This work was supported by grants from the National Key Basic Science ‘973’ Program (No. G2000016204), the Ministry of Education Doctor's Foundation Program (No. 20030001083) and the National Natural Science Foundation (Nos. 39980003 and 30500342) of China.
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Qian, W., Tan, G., Liu, H. et al. Identification of a bHLH-type G-box binding factor and its regulation activity with G-box and Box I elements of the PsCHS1 promoter. Plant Cell Rep 26, 85–93 (2007). https://doi.org/10.1007/s00299-006-0202-x
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DOI: https://doi.org/10.1007/s00299-006-0202-x