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| 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) |
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| DOI： |
| 中文关键词: 桔小实蝇 SOD3 基因克隆 半定量PCR |
| 英文关键词:Bactrocera dorsalis SOD3 gene cloning semi-quantitative PCR |
| 基金项目:广东省科技计划项目（2012B050500009）;2014年大学生创新创业训练项目（201411347001） |
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| 中文摘要: |
| 采用反转录聚合酶链式反应 (RT-PCR) 和快速扩增 cDNA 末端 (RACE) 技术克隆桔小实蝇 SOD3 基因，并命名为 BdorSOD3。BdorSOD3 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 BdorSOD3 基因大肠杆菌诱导后的表达情况，结果表明，BdorSOD3 在处理与对照的24 h、48 h都有表达，但 BdorSOD3 在处理后48 h表达量明显升高，结果暗示 BdorSOD3 与桔小实蝇蛹对大肠杆菌的免疫机制有关。 |
| 英文摘要: |
| 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%）withAGE89778.1, another SOD protein formBactrocera 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 SOD3mRNA 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 hpupae treated with E. coli. Higher expression level in 48 h pupae treated with E. coli impliedBdorSOD3 involved in immune metabolism of B.dorsalis pupa. |
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