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【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】 The objective of this study was to clone the transcription factor DREB1 (Dehydrated responsive element binding protein 1), a key gene of drought-resistance herb, to analyze the physiological and biochemical characteristics of DREB1 and lay the theoretical foundation for the screening and utilization of ideal resistance genes. 【Method】 The full-length cDNA sequence of DREB1 was isolated by RACE-PCR (rapid amplification of cDNA ends-polymerase chain reaction) technique. North-ern hybridization and gel retention assays were used to analyze the expression of DREB1 under stress conditions. And DRE (dehydration responsive element) element binding activity. 【Result】 A novel DREB1-type transcription factor HbDREB1 was successfully isolated from the strong-cold-resistant B. barley. The full-length HbDREB1 gene was 899 bp in length and contained a typical AP2 / EREBP DNA domain and a “ Three DREB1 signature sequences of ”PKK / RPAGRxKFxETRHP “ and ”DSAWR “ and ”LWSY " were obtained. Sequence alignment analysis showed that HbDREB1 shared high homology with other DREB1 transcription factors. HbDREB1 is significantly induced by cold stress at the transcriptional level, has the function of binding to the DRE-cis-acting element and is an essential nuclear localization characteristic as a transcription factor. 【Conclusion】 The HbDREB1 gene is involved in abiotic stress signal transduction and has the potential to improve plant cold tolerance.