二穗短柄草钙依赖型蛋白激酶BdCDPK4基因的克隆及其编码蛋白质的生物信息学分析

Cloning and encoding protein bioinformatics analysis of BdCDPK4 from Brachypodium distachyon

二穗短柄草(Brachypodium distachyon L.)因与小麦、大麦等有很高的同源性,作为新型禾本科模式生物吸引了越来越多的研究者。试验采用基因克隆技术分离了二穗短柄草Bd CDPK4基因的编码区序列,并采用生物信息学方法分析了该基因及其编码的蛋白质的序列特征、结构和功能。结果表明,分离了BdCDPK4基因1 851 bp的cDNA序列(Gen Bank登录号为XM_003559516),编码区长度为1 752 bp,编码583个氨基酸,分子量为64.78 ku,等电点为9.12,含有N端可变区、蛋白激酶区、自抑制区、EF手型结构、类似钙调素等结构域。亚细胞定位预测位于叶绿体和液泡上。进化树分析结果显示,该蛋白与小麦Ta CDPK19蛋白的同源性最近。根据各物种间CDPKs蛋白的高同源性,结合小麦TaCDPK19基因被高盐和小麦白粉病菌Blumeria graministritici(Bgt)诱导,推测Bd CDPK4基因也可能参与逆境胁迫下的信号传导途径,在抗逆等生物学过程中发挥重要功能。

Brachypodium distachyon,the genome of which shares high homology with wheat and barley,has been used as the new model species of Gramineae. In order to clarify the structure and function of Brachypodium distachyon CDPKs gene,the coding region sequence（ CDS） of BdCDPK4 gene of Brachypodium distachyon was isolated,and its character,structure and function were analyzed by bioinformatics methods. The results showed that1 851 bp c DNA（ Gen Bank ID： XM_003559516） of coding sequence of BdCDPK4 gene included the complete 1 752 bp CDS,coded 583 amino acids,and located on chloroplast and vacuole. The molecular weight and isoelectric pointof BdCDPK4 protein were 64. 78 ku and 9. 12,respectively. BdCDPK4 protein contained several domains including N-variable domain,kinase domain,autoinhibitory domain,EF-hands and Ca M-like domain. Phylogenetic analysis results suggested that BdCDPK4 protein had high homology with wheat Ta CDPK19. Ta CDPK19 were induced by high salt and white powder Blumeria graministritici（ Bgt）. BdCDPK4 protein may participate in the regulation of signal transduction pathways under stresses according to the homologies of protein domain among all species. Taken together,our data could establish a good foundation for studying the biological functions of BdCDPK4 gene.