柞蚕铜锌超氧化物歧化酶基因的克隆与表达分析

Cloning and Expression Analysis of Copper/Zinc Superoxide Dismutase Gene From Antheraea pernyi

超氧化物歧化酶(SOD)是生物体内的重要保护性酶类。采用RT-PCR方法克隆了柞蚕(Antheraea pernyi)铜锌超氧化物歧化酶(Cu/Zn-SOD)基因的ORF序列。生物信息学分析表明,该序列共编码154个氨基酸,具有Cu/Zn-SOD的保守性结构特征,与家蚕(Bombyxmori)、美国白蛾(Hyphantria cunea)、野桑蚕(Bombyx mandarina)以及果蝇(Drosophila melanogaster)Cu/Zn-SOD基因的同源性分别是81.5%、81.7%、81.5%、66.7%。柞蚕蛹经紫外线照射处理后用半定量PCR方法检测,发现照射不同时间脂肪体内Cu/Zn-SOD基因的转录水平存在差异,在照射5 min后开始增加,至10 min时为转录高峰,15 min时又呈下降趋势。

Superoxide dismutase（SOD） is an important antioxygenic enzyme in living organisms.The open reading frame（ORF） sequence of Antheraea pernyi copper/zinc superoxide dismutase gene was obtained through RT-PCR method.Sequence analysis indicated that this partial cDNA sequence encodes a 154 amino acid polypeptide containing the conserved domain of Cu/Zn-SODs.The evolutionary analysis showed that its sequence identities with Bombyx mori,Hyphantria cunea,Bombyx mandarina and Drosophila melanogaster are 81.5%,81.7%,81.5% and 66.7%,respectively.The transcription activity of Cu/Zn-SOD gene in fat body,which was determined by RT-PCR,increased after 5 min of UV radiation,reaching the peak at 10 min and decreased at 15 min after UV treatment.