摘要
【目的】克隆强抗寒性牧草——短芒大麦DREB1(dehydration responsive element binding protein 1)转录因子,分析其生理生化特性,为理想抗逆工程基因的筛选和利用奠定理论基础。【方法】利用RACE-PCR(Rapidamplification of cDNAends-polymerase chain reaction)技术分离短芒大麦DREB1转录因子全长cDNA序列,North-ern杂交和凝胶滞留试验分析其在逆境条件下的表达情况,及其与DRE(dehydration responsive element)元件的结合活性。【结果】从强抗寒性短芒大麦中成功分离了1个新的DREB1类转录因子HbDREB1,该基因全长899 bp,其蛋白序列中含有1个典型的AP2/EREBP DNA结构域及"PKK/RPAGRxKFxETRHP"和"DSAWR"、"LWSY"3个DREB1特征标签序列;序列比对分析表明,HbDREB1与其他植物的DREB1类转录因子的同源性较高。HbDREB1在转录水平上明显受冷胁迫诱导表达,具有结合DRE-顺式作用元件的功能及作为转录因子必备的核定位特性。【结论】HbDREB1基因参与了非生物胁迫信号转导,具有提高植物抗寒性的潜能。
[Objective] Cloning and physiological and biochemical characteristics analysis of DREB1 gene in Hordeurn brevisubulaturn provided a theoretical basis for the screening and utilization of ideal resistant gene. [Method] The cDNA sequence of DREB1 gene in Hordeurn brevisubulaturn was isolated by RACE-PCR;and its expression in adversity condition and binding activity with DRE element were analyzed by northern hybridization and electrophoretic mobility shift assay respectively. [Result] A new gene, HbDREB1 ,was isolated from DREB1 successfully. A full length of 899 bp, HbDREB1 contains a typical AP2/ EREBP DNA structure zone in its protein sequence and PKK/RPAGRxKFxETRHP and DSAWR sequences. Sequence comparison showed HbDREB1 has high homology with DREB1 genes of other plants. Expression of HbDREB1 at transcription level is induced obviously by cold stress, and, HbDREB1 can bind with DRE cisacting element and has nucleic localization characteristics. [Conclusion] HbDREB1 gene participated in signal transduction in abiotic stress condition and has a potential for enhancing cold resistance of plants.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2009年第7期59-67,共9页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家自然科学基金项目(30471229)
