毛尖紫萼藓抗旱相关基因Gp-LEA的克隆与表达分析

以毛尖紫萼藓干旱cDNA文库中获得的一段与LEA基因同源性较高的EST序列为基础,采用RACE技术分离该基因cDNA全长序列,命名为Gp-LEA。Gp-LEA基因的cDNA全长814bp,开放阅读框456bp,编码含151个氨基酸蛋白质。生物信息学分析结果显示,Gp-LEA蛋白为稳定蛋白,分子质量为16.612kD,理论等电点（pI）为5.06,含有LEA₂功能结构域,不属于跨膜蛋白且不存在信号肽。系统发生分析表明,Gp-LEA基因编码蛋白与花旗松LEA蛋白亲缘关系最近。荧光定量PCR分析显示,Gp-LEA基因在复水和快速干旱模式下均能表达。推测Gp-LEA基因在毛尖紫萼藓的复水和干旱过程中起着重要作用。

Cloning and Expression Analysis of Drought-tolerance Related Gene Gp-LEA in Grimmia pilifera

A full-length cDNA encoding late embriogenesis abundant protein (LEA) named Gp-LEA,was cloned from Grimmia pilifera through the method of rapid amplification of cDNA ends (RACE).The full length of this gene was 814 bp,and contained a 456 bp open reading frame which encoded a protein containing 151 amino acids.The result of bioinformatics showed that this protein was a stable protein with a function domain-LEA_2,which molecular weight was 16.612 kD and theoretical pI was 5.06,and it was not a transmembrane protein and had no signal peptide.Phylogenetic analysis indicated that this gene had the closest genetic relationship with that of Pseudotsuga menziesii.The quantitative RT-PCR results suggested that the expression of Gp-LEA gene was induced in both rehydration and dehydration,and this gene might play an important role in the two processes.