扁穗冰草PLD基因cDNA序列克隆及生物信息学分析

cDNA clonging of phospholipase D gene from Agropyron cristatum(L) Gaertn and its bioinformatic analysis

利用RT-PCR技术和RACE方法克隆得到扁穗冰草(Agropyron cristatum(L.)Gaertn)PLD基因cDNA的全长,并对该cDNA序列、氨基酸序列的相似性、组成成分、理化性质、疏水性/亲水性、进化树、二级结构、三级结构、无序化特性进行了预测和分析.结果显示:扁穗冰草PLD基因具有2967个核苷酸,含有PLD所特有的HKD基序和C2结构域,属亲水性蛋白,二级结构以无规卷曲为主,蛋白质的三级结构显示两个HKD基序靠近形成具功能的活性位点,蛋白质无序化分析发现无序化区域较少;进化树分析表明与蒙古冰草亲缘关系最近,与草莓亲缘关系最远.为研究扁穗冰草PLD在干旱胁迫信号转导及应答过程的机理奠定基础.

Full-length cDNA sequence of PLD gene from Agropyron cristatum（L.） Gaertn was acquired by RT-PCR and RACE methods.Then,analysis and prediction on the acquired sequences and its amino similarity,composing component,physicochemical properties,hydrophobicity/hydrophilicity,sequence alignment,phylogenetic tree,secondary structure,tertiary structure and disordering characteristics of amino acid were conducted comprehensivly by bioinformatic.The results showed that PLD gene cDNA was 2967 bp in length from A.cristatum and it contain HKD motifs and C2 domain;PLD is hydrophilic protein,and secondary structure of PLD was mainly composed of andom coil.In addition,tertiary structure of PLD were formed by two HKD motifs which are close to each other to form a functional active site,however,less disordering domains were found.The molecular evolution trees show that PLD of A.cristatum had high genetic relationship with A.mongolicum keng and had low genetic relationship with Fragaria.The aim of this study is that the further research of the A.cristatum PLD especially lays a foundation for signal transduction and responses process in drought stress.