沙冬青AmDREB3基因的克隆及植物表达载体构建

CLONING AND PLANT EXPRESSION VECTOR CONSTRUCTION OF AMDREB3 FROM Ammopiptanthus mongolicus

脱水应答元件结合蛋白(Dehydration-responsive element/DRE binding proteins,DREBs)属于植物AP2/EREBP(APETALA2/Ethylene-responsive element binding protein)转录因子大家族中的一个亚族,在植物对低温、干旱和盐碱等非生物胁迫的适应中起重要作用。本论文从强耐寒耐旱植物蒙古沙冬青(Ammopiptanthus mongolicus)中克隆到AmDREB3基因的编码区cDNA,对其编码蛋白的结构域和系统进化关系进行了分析,并将其成功构建到分别由CaMV35S和拟南芥rd29A启动子驱动的植物表达载体pCAMBIA3301上,为进一步通过转基因植物等方法验证该基因在耐逆性中的功能奠定了基础。

DREBs(Dehydration-responsive element/DRE binding proteins) are a subfamily in AP2/EREBP transcription factor family in plant and play important roles in plant tolerance to multiple abiotic stresses such as low temperature,drought and salinity.In this paper,the coding region cDNA of a DREB-like protein,namely AmDREB3,was cloned from Ammopiptanthus mongolicus,a plant with very strong tolerance to both freezing and drought stresses.The conserved domains and phylogenetic relationship of AmDREB3 with other known DREBs were analyzed.Moreover,the cDNA fragment of AmDREB3 was successfully ligased to the downstream of CaMV35S and Arabidopsis rd29A promoters in plant expression vector pCAMBIA3301.The study paved way for elucidating the gene`s function by transgenic plants and other methods subsequently.