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DREB1/CBF类转录因子在植物抵抗外界环境胁迫上起到非常重要的作用,而且对于利用基因工程技术来提高植物的抗逆性也非常有用.从棉花中分离出一个编码DREB1/CBF类蛋白(GhDREB1L)的cDNA,并对该蛋白质的序列特性、DNA结合特性及转录本的表达特征进行了分析.GhDREB1L含有一个保守的AP2/ERF结构域,其氨基酸序列与其他植物中DREB家族的DREB1/CBF组成员高度相似.采用Ni-NTA亲和层析技术,成功纯化了在BL21(DE3)菌株中表达的GhDREB1L蛋白的DNA结合区.凝胶滞留实验揭示,纯化的GhDREB1L融合蛋白能够特异性地与已经得到鉴定的DRE元件(核心区,ACCGAC)以及胚胎发育晚期丰富蛋白基因LEAD113启动子区中的类DRE元件(核心区,GCCGAC)结合.半定量RT-PCR显示GhDREB1L基因在棉花子叶中受到低温、干旱以及NaCl处理的诱导.这些结果暗示了棉花GhDREB1L转录因子可能是通过与DRE元件的结合,在低温、干旱及高盐的应答途径中起重要作用.
DREB1 / CBF transcription factors play a very important role in plant resistance to external environmental stress and are also very useful for improving the stress resistance of plants by using genetic engineering techniques.A DREB1 / CBF-like protein (DREB1 / CBF) GhDREB1L) was cloned and its sequence characteristics, DNA binding properties and transcript expression characteristics were analyzed.GhDREB1L contains a conserved AP2 / ERF domain whose amino acid sequence is identical to that of DREB1 / CBF in other plants The members of the group are highly similar.The DNA binding domain of the GhDREB1L protein expressed in BL21 (DE3) strain was successfully purified by Ni-NTA affinity chromatography.The gel retention assay revealed that the purified GhDREB1L fusion protein can specifically bind to The DRE element (core region, ACCGAC) and the DRE element (core region, GCCGAC) in the LEAD113 promoter region have been identified by semi-quantitative RT-PCR analysis, and the GhDREB1L gene is under low temperature in cotton cotyledons , Drought and NaCl treatment.These results suggest that cotton GhDREB1L transcription factor may be through the combination of DRE elements in low temperature, drought and high salt Of the response pathways play an important role.