基于转录组的海州香薷COPT铜转运蛋白家族鉴定及响应铜胁迫表达分析

Transcriptome-based identification of copper transporter family of Elsholtzia splendens and expression analysis under copper stress

为了解海州香薷(Elsholtzia splendens)COPT(copper transporter)铜转运蛋白对铜胁迫的响应规律,运用生物信息学方法对海州香薷转录组数据库中的COPT家族成员进行鉴定与分析,构建植物表达载体探究蛋白的亚细胞定位.结果显示:海州香薷转录组中有11个EsCOPT成员,它们编码的蛋白质由72~159个氨基酸组成,等电点介于4.72~9.92,平均相对分子质量为12.870×10^(3).11个成员均为膜蛋白,系统进化树分析显示EsCOPT蛋白分别属于3个亚组.正常条件下,EsCOPT2和EsCOPT9表达量较高,铜处理后,除EsCOPT5的表达无明显变化外,其他EsCOPTs的表达均呈下降趋势.全长EsCOPT9编码的蛋白具有3个跨膜结构域且N末端较短,共定位实验表明EsCOPT9-GFP定位于液泡膜上.

To explore the response of COPT to copper stress in Elsholtzia splendens,bioinformatics method was used to identi-fy and analyze COPT family members in E.splendens transcriptome database.Furthermore,plant expression vector was con-structed to explore the subcellular localization of the protein.The results showed that there were 11 EsCOPT members in E.splendens transcriptome,which encode proteins with 72-159 amino acids,isoelectric point of 4.72-9.92 and the average rela-tive molecular mass of 12.870×10^(3).All EsCOPTs are membrane proteins.Phylogenetic tree analysis showed that EsCOPTs belong to three subgroups.Under normal conditions,the expression of EsCOPT2 and EsCOPT9 are higher than other members.After Cu treatment,except for EsCOPT5 with unchanged expression level,all EsCOPTs showed a downregulated expression.The full-length ESCOPT9-encoded protein has three transmembrane domains and the N-terminal is short.The colocalization experi-ment showed that EsCOPT9-GFP was located on the vacuolar membrane.