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Cis-regulatory element-based genome-wide identification of dreb1/cbf targets in Arabidopsis

Progress in Natural Science:Materials International 2008年第5期18卷 579-583页

作者： Huan Zhao a, Weizhong Liu a,b, Yong Hu a, Xianglin Liu a, Yikun He a, a College of Life Science, Capital Normal University, Beijing 100037, China b College of Life Sciences, Shanxi Normal University, Linfen 041004, China College of Life Science Capital Normal University Beijing 100037 China College of Life Sciences Shanxi Normal University Linfen 041004 China

Microarray analysis is used to identify transcriptional targets. However, the direct targets of transcription factors cannot be distin- guished from indirect ones; further, genes with low-expression levels cannot be identi ed by this method. In the present study, we exploit the cis-element dehydration-responsive element (DRE) that is known to be responsible for the transcription of DRE binding factor 1 (dreb1) targets in the promoter region of all Arabidopsis genes. Putative targets whose promoters contain the elements were veri ed by both microarray and reverse transcription-polymerase chain reaction (RT-PCR) analysis. Five new dreb1/cbf direct targets were identi ed. Compared with traditional microarray analysis, our method is convenient and cost-e ective for identifying the downstream targets of transcription factors.

关键词： Arabidopsis Cis-element Transcription factor dreb1/cbf Stress

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Molecular cloning and functional characterization of a dreb1/cbf-like gene (Ghdreb1L) from cotton

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Science China(Life Sciences) 2007年第1期50卷 7-14页

作者： HUANG Bo, JIN LongGuo & LIU JinYuan Laboratory of Molecular Biology and Protein Science Laboratory of the Ministry of Education, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China Laboratory of Molecular Biology and Protein Science Laboratory of the Ministry of Education Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China

The transcription factors dreb1s/cbfs play important roles in the regulation of plant resistance to environmental stresses and are quite useful for generating transgenic plants tolerant to these stresses. In the present work, a cDNA encoding dreb1/cbf-like protein (Ghdreb1L) from cotton was isolated, and its sequence features, DNA binding preference, and expression patterns of the transcripts were also characterized. Ghdreb1L contained one conserved AP2/ERF domain and its amino acid sequence was similar to the dreb1/cbf group of the dreb family from other plants. The DNA-binding domain of Ghdreb1L was successfully expressed as a fusion protein in Escherichia coli BL21 (DE3) and purified by Ni-NTA affinity chromatography. Electrophoretic mobility shift assay revealed that the purified Ghdreb1L fusion protein had a specific binding activity with the previously characterized DRE ele-ment (core sequence, ACCGAC) and also with the DRE-like sequence (core sequence, GCCGAC) in the promoter of the dehydration-responsive late embryogenesis-abundant gene LEA D113. Semi-quantita- tive RT-PCR showed that Ghdreb1L was induced in the cotton cotyledons by low temperature, as well as drought and NaCl treatments. These results suggested that the novel cotton Ghdreb1L might play an important role in response to low temperature as well as drought and high salinity through binding to the DRE cis-element.

关键词： cotton dreb1/cbf cold dehydration LEA D113

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