水稻转录因子DREB1的克隆与原核表达  

Cloning and prokaryotic expression of transcription factor RDREB1 in rice (Oryza satire L.)

作　　者：高峰[1] 姚泉洪[2] 秦秋琳[1] 刘金戈[2] 熊爱生[2] 彭日荷[2] 洪义欢[1] 陈建民[1] 

机构地区：[1]扬州大学生物科学与技术学院,扬州225009 [2]上海市农业科学院生物技术研究所,上海201106

出　　处：《上海农业学报》2007年第2期1-6,共6页Acta Agriculturae Shanghai

基　　金：江苏省自然科学基因项目(BK2003042);国家转基因研究专项(J00-A-007);上海市科委国际合作项目(055407068);上海市优秀学科带头人计划(06XD14017);上海市重大基础研究项目(03DJ14016;05DJ14008);上海市青年科技启明星项目A类(0SQMX1445);上海市科委重点项目(05dz223266;03dz19311)

摘　　要：DREB类转录因子特异地与DRE顺式作用元件结合,在非生物逆境胁迫(干旱、低温、高盐)中激活胁迫诱导基因的表达,使植物能够耐受水分胁迫的影响。以顺式元件rd29A3 cis构建诱饵质粒.通过酵母单杂交方法从水稻cDNA文库中筛选获得7个阳性质粒,序列分析显示阳性质粒中插入的cDNA序列一致,根据核苷酸序列推导出cDNA编码一219个氨基酸组成的多肽,该多肽有保守的EREBP/AP2结构域,N端为核定位信号(NLS),而C端则为转录激活区域,属于EREBP/AP2类转录因子,命名为RdreB1。重新合成的RdreB1 I基因降低了GC比含量,诱导处理产生了约45 kD的特异表达的融合蛋白。The DREB （Dehydration responsive element binding） transcription factors, specifically binding the dehydration responsive element （DRE） , play important roles in the expression of many stress inducible genes under drought, low-temperature and high-salt conditions. In this study, the bait plasmid was constructed with the cis element rd29A3 and seven positive plasmids were selected from rice cDNA library by using the yeast one-hybrid system. The sequence analysis showed that the cDNA sequence inserted into the positive plasmids was identical. According to the nucleotide sequence a cDNA-coded polypeptide consisting of 219 amino acids was deduced. This polypeptide had a conserved AP2/EREBP domain in which the N-terminal was a nuclear localization signal and the C-terminal was a transcriptional activation domain and was named RdreB1. The re-synthesized RdreBlI decreased GC contents and the induction treatment produced a 45 kD specifically expressed fusion protein.

关 键 词：DREB 转录因子 酵母单杂交 表达 水稻 

分 类 号：S511[农业科学—作物学] Q943.2[生物学—植物学]