Huang Bo, Liu Jin-Yuan
Laboratory of Molecular Biology and MOE Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Biochim Biophys Acta. 2006 Jun;1759(6):263-9. doi: 10.1016/j.bbaexp.2006.04.006. Epub 2006 May 20.
A novel cDNA encoding DRE-binding transcription factor, designated GhDBP3, was cloned from Gossypium hirsutum. This protein was classified into A-4 group of DREB subfamily based on multiple sequence alignment and phylogenetic characterization. Semiquantitative RT-PCR showed that GhDBP3 was expressed in the leaves, cotyledons, roots and stems of 2-week-old cotton seedlings under non-stress conditions and was greatly induced in the cotton cotyledons by drought, NaCl, low temperature and ABA treatment. EMSA revealed that GhDBP3 was able to bind to the DRE cis-element in vitro. Transient assay using the particle bombardment method showed that GhDBP3 was a transcriptional activator, capable of activating expression of a reporter gene driven by the LEA D113 promoter containing a DRE like sequence in tobacco cells. Our results indicate that GhDBP3 could be a new member of DRE-binding transcription factor family and may play an important role in response to ABA and environmental stresses.
从陆地棉中克隆出一个编码DRE结合转录因子的新cDNA，命名为GhDBP3。基于多序列比对和系统发育特征分析，该蛋白被归类于DREB亚家族的A-4组。半定量RT-PCR结果显示，在非胁迫条件下，GhDBP3在2周龄棉花幼苗的叶片、子叶、根和茎中均有表达，并且在干旱、NaCl、低温和ABA处理下，棉花子叶中的表达量显著上调。凝胶迁移实验（EMSA）表明，GhDBP3在体外能够与DRE顺式作用元件结合。利用粒子轰击法进行的瞬时表达分析表明，GhDBP3是一个转录激活因子，能够激活烟草细胞中由含有类DRE序列的LEA D113启动子驱动的报告基因的表达。我们的结果表明，GhDBP3可能是DRE结合转录因子家族的一个新成员，并且可能在响应ABA和环境胁迫中发挥重要作用。
