桔小实蝇超氧化物歧化酶基因SOD3的克隆及表达分析

Cloning,expression analysis of SOD3 in Bactrocera dorsalis(Hendel )(Diptera: Tephritidae)

采用反转录聚合酶链式反应(RT-PCR)和快速扩增c DNA末端(RACE)技术克隆桔小实蝇SOD3基因,并命名为Bdor SOD3。Bdor SOD3阅读框全长531 bp,编码176个氨基酸,第1-20位氨基酸为其信号肽区域;该蛋白序列与桔小实蝇的另外一种SOD蛋白AGE89778.1序列的一致性最高,达98.7%;采用Swiss-model在线软件模拟构建Bdor SOD3蛋白的三维结构;采用半定量PCR方法,研究Bdor SOD3基因大肠杆菌诱导后的表达情况,结果表明,Bdor SOD3在处理与对照的24 h、48 h都有表达,但Bdor SOD3在处理后48 h表达量明显升高,结果暗示Bdor SOD3与桔小实蝇蛹对大肠杆菌的免疫机制有关。

Rapid amplification cDNA ends （RACE） method and reverse transcription PCR （RT- PCR） method were used to clone SOD3 gene. The whole sequence of SOD3 was gained and named as BdorSOD3. The full - length of open reading frame in BdorSOD3 is 531 bp in length and the putative amino acids shared the highest similarity （98.7%） with AGE89778. 1, another SOD protein form Bactrocera dorsalis. 1 -20 amino acids of N-terminal reveals the signal peptide of the deduced protein. SWISS - MOLDLE program was used to build simulated 3D structure of SOD3. Expression levels of SOD3 mRNA were investigated by semi - quantitative PCR method. Semi - quantitative PCR results indicated that BdorSOD3 expressed in all treatments and controls while the highest expression level was detected in 48 h pupae treated with E. coli. Higher expression level in 48 h pupae treated with E. coli implied BdorSOD3 involved in immune metabolism of B. dorsalis pupa.