牡丹PsDREB转录因子基因的克隆及亚细胞定位

Clonging and Subcellular Location of a DREB Transcription Gene of Paeonia suffruticosa

DREB转录因子是一个干旱应答元件的结合蛋白,它在植物对干旱、高盐及低温胁迫的分子反应中起着非常重要的调控作用。本研究以芍药科芍药属牡丹‘洛阳红’品种叶片提取的总DNA为模板,通过PCR扩增获得一个1 661 bp的DREB(dehydration responsive element binding protein)基因的cDNA序列,包含1 089 bp的开放阅读框、448 bp的5'非编码序列和124 bp的3'非编码序列,命名为PsDREB。该基因的cDNA编码363个氨基酸,与葡萄、梧桐、杨树和咖啡的同源性分别为61%、60%、56%和55%。为进一步验证其功能,本研究通过基因枪法对该基因编码的蛋白进行了亚细胞定位检测,结果发现该蛋白定位于细胞核,符合转录因子细胞核定位的特征。研究结果为进一步研究该基因编码蛋白的结构和功能奠定了基础。

DREB transcription factor is a dehydration responsive element binding protein,which plays a very important role in regulating its molecular response in plants to drought,high salt and cold stress.In this research,a full-length cDNA of DREB gene from Paeonia suffruticosa‘Luo Yanghong'leaves was obtained and named Ps DREB.The cDNA sequence was 1 661 bp,which contained 1 089 bp open reading frame for 363 amino acid residues,448 bp 5'-terminal and 124 bp 3'-terminal sequence.Homology analysis indicated that the Ps DREB protein showed 61%,60%,56% and 55% similarity to the DREB homologs from Vitis vinifera,Theobroma cacao,Populus trichocarpa and Coffea Arabica.In order to further verify the function of Ps DREB,subcellular localization analysis was carried out by gene gun method.The results showed that the PsDREB protein was localized in the nucleus,indicating that the protein was consistent with the characteristics of nuclear localization of transcription factors.These results formed a basis to further study the structure and function of the gene encoded protein.