| 赵琰明1, 朱家颖1, 泽桑梓2, 赵 宁1, 杨 斌1*.云南切梢小蠹超氧化物歧化酶基因的克隆与序列分析[J].植物保护,2015,41(5):32-38. |
| 云南切梢小蠹超氧化物歧化酶基因的克隆与序列分析 |
| Molecular cloning and sequence analysis of superoxide dismutase gene from Tomicus yunnanensis |
| 投稿时间：2014-08-27 修订日期：2015-02-14 |
| DOI： |
| 中文关键词: 云南切梢小蠹 超氧化物歧化酶 基因克隆 序列分析 实时荧光定量PCR |
| 英文关键词:Tomicus yunnanensis superoxide dismutase gene clone sequence analysis real time quantitative PCR |
| 基金项目:国家自然科学基金项目 (31260179); 国家林业局林业公益性行业科研专项 (201004067) |
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| 中文摘要: |
| 超氧化物歧化酶(superoxide dismutase, SOD)是一种广泛存在于生物体各组织, 在机体抗氧化过程中起着重要作用的抗氧化酶。采用RACE技术, 克隆获得了云南切梢小蠹超氧化物歧化酶基因。该基因cDNA序列全长768 bp, 开放阅读框为462 bp, 编码153个氨基酸。预测蛋白质分子量为15.8 ku, 等电点为5.68。同源性比对分析发现, 云南切梢小蠹SOD与中欧山松大小蠹SOD在氨基酸水平上相似性高达93%, 与赤拟谷盗、点蜂缘蝽、侵扰锥猎蝽、丽蝇蛹集金小蜂、佛罗里达弓背蚁、柑橘凤蝶、棉铃虫、黑果蝇、家蝇的相似性也都在67%~71%之间。其推导的氨基酸序列中含有2个Cu/Zn SOD的特异序列(GFHIHEFGDNT42-52和GNAGGRLACAVI136-147), Cu原子分别与His44、His46、His61和His118配位, Zn原子分别与His61、His69、His78和Asp81配位, 半胱氨酸Cys55和Cys144基之间形成Cu/Zn SOD链内二硫键。系统进化树分析表明, 克隆获得的云南切梢小蠹SOD隶属于铜锌SOD家族(icCu/Zn SOD)。实时荧光定量PCR表明, icCu/Zn SOD基因在云南切梢小蠹各发育阶段和不同部位均有表达, 但表达量各不相同。 |
| 英文摘要: |
| Superoxide dismutase (SOD) is a kind of antioxidant enzymes, existing widely in biological groups. It plays a key role in insects’ antioxidant protection system. A cDNA encoding Cu/Zn SOD was first isolated from Tomicus yunnanensis using rapid amplification of cDNA ends (RACE) strategy. This gene was 768 bp in full length with an open reading frame (ORF) of 411 bp, which encoded 153 amino acid residues. Its predicted molecular weight and isoelectric point were 15.8 ku and 5.68, respectively. The deduced amino acid sequence of T.yunnanensis SOD showed the highest similarity (93%) with that of Dendroctonus ponderosae, and had 67%-71% similarity with other insects, including Tribolium castaneum, Riptortus pedestris, Triatoma infestans, Nasonia vitripennis, Camponotus floridanus, Papilio xuthus, Helicoverpa armigera, Drosophila virilis and Musca domestica. Two typical structure and functional domains of Cu/Zn SOD (GFHIHEFGDNT42-52and GNAGGRLACAVI136-147) were located in the deduced amino acid sequence. Several highly conserved motifs including Cu, Zn binding sites H(44), H(46), H(61), H(118) for Cu binding, and H(61), H(69), H(78), D(81) for Zn binding, and Cys55 and Cys144 for the only intrachain disulfide bond were also identified in T.yunnanensis Cu/Zn SOD. Phylogenetic tree indicated that T.yunnanensis Cu/Zn SOD belonged to icCu/Zn SOD family. The real time quantitative PCR showed that icCu/Zn SOD gene expressed at different developmental stages and in different tissues of T.yunnanensis, but the expression profiles were different. |
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