舞毒蛾LdGSTe1基因的克隆及表达

Cloning and functional analysis of GSTe1 gene from Lymantria dispar

从舞毒蛾(Lymantria dispar)3龄幼虫转录组文库中鉴定获得舞毒蛾谷胱甘肽S–转移酶基因全长c DNA,命名为Ld GSTe1。该基因全长651 bp,编码216个氨基酸。序列分析显示,Ld GSTe1蛋白氨基酸序列含有GST_C_Delta_Epsilon与GST_N_Delta_Epsilon 2个保守结构域,属于类硫氧还蛋白和谷胱甘肽S–转移酶2个超家族。系统进化树分析表明,舞毒蛾Ld GST蛋白属于GST Epsilon家族。蛋白结构显示,Ld GSTe1蛋白包含一个N端和一个C端结构,α–螺旋与β–折叠为主要结构元件。实时荧光定量PCR结果表明,在4.0、10.0 mg/L鱼藤酮药剂处理下,舞毒蛾3龄幼虫Ld GSTe1基因表达先下降后上升,推测该基因参与舞毒蛾解毒应答。

Accroding to the transcriptome of the 3rd instar larva of Lymantria dispar(L. dispar), a GlutathioneS–transferase gene was determined and obtained, which named LdGSTe1. The open reading frame (ORF) of LdGSTe1was 651 bp encoding a protein of 216 amino acid residues. Sequence analysis showed that the amino acid sequences ofLdGSTe1 protein contained two conserved domains, namely GST_C_Delta_Epsilon and GST_N_Delta_Epsilon,belonging to the thioredoxin-like superfamily and glutathione S–transferase superfamily. Phylogenetic tree analysisindicated that the LdGST belonged to GST Epsilon family. Protein structure showed LdGSTe1 contains an N-terminaland C-terminal, and organized mainly into α–helix and β–sheet. The expression of LdGSTe1 in 3rd instar larvae L. Disparunder 4.0, 10.0 mg/L rotenone treatment was investigated using real-time fluorescence quantitative PCR. The resultsshowed that the expression of LdGSTe1 in L. dispar was first down-regulated and then up-regulated. Therefore, LdGSTe1gene was speculated to participate in the detoxification response of L. dispar.