紫大麦草中DREB类转录因子基因Hvi917DREB2的克隆、原核表达及序列分析

Cloning,prokaryotic expression and sequence analysis of DREB-like gene Hvi917DREB2 in Hordeum violaceum

DREB类转录因子是一类与植物逆境胁迫响应相关的重要蛋白,本研究利用同源克隆方法和RACE技术从紫大麦草(Hordeum violaceum)中分离到1个DREB类转录因子基因,命名为Hvi917 DREB2。序列分析表明Hvi917 DREB2基因含有1个792bp的开放阅读框,编码264个氨基酸残基,含有1个保守的AP2结构域,属于AP2大家族。该基因编码的氨基酸序列与Genebank中短芒大麦草AP2蛋白HbDREB2、DREB1(登录号分别为:AAU29412.1和AER42620.1)具有99%的氨基酸序列一致性。将该基因构建成原核表达载体,经IPTG诱导表达了1个分子量为34.5kD的蛋白。

DREB-like transcription factor is a kind of important protein for plant to respond abiotic stresses. Using a combination of RACE and homology cloning, a DREB-like gene from Hordeurn violaceurn, named as Hvi917DREB2, was isolated. Sequence analysis indicates that it contains an open reading frame of 792 bp encoding a putative protein with 264 amino acids, and a highly conserved AP2-super family domain. Compared with HbDREB2 （accession no. AAU29412.1 ） and DREBl（accession no AER42620.1） from Hordeum brevisubulaturn, sequence of Hvi917DREB2 showed 99% identity. In addition, cprokaryotic expression vector fused with Hvi917DREB2 sequence was constructed and a 34.5 kD DREB-like protein was expressed. The research prepared the foundation for researching the interaction between the DREB transcription factor and DRE cis-element, also provided a candidate gene for wheat molecular breeding.