| 摘要: |
| 【目的】研究不同类型农药胁迫下松墨天牛核糖体蛋白基因表达的变化，为靶标防治松树重要害虫松墨天牛提供理论依据和参考。【方法】从构建的松墨天牛 cDNA 文库中获得表达序列标签（EST），用 Blast 程序从GenBank数据库中查找同源序列，克隆得到松墨天牛的一个核糖体蛋白基因；用实时荧光定量 PCR (RT-qPCR) 检测 11 种农药胁迫下该基因的表达。【结果】从松墨天牛 cDNA 文库中获得1条 EST 序列与昆虫核糖体蛋白 S15A (RPS15A)同源的基因，命名为MaRPS15A。MaRPS15A cDNA长504 bp，含有 5′、 3′ 非编码区和长 393 bp 的开放阅读框，编码 130个氨基酸，预测的蛋白质分子质量为 14.75 ku，理论等电点为 9.95。MaRPS15A与赤拟谷盗（Tribolium castaneum）核糖体蛋白RPS15A的相似性为98%，与其他昆虫核糖体蛋白RPS15A的相似性大于88%。除溴氰菊酯外，其他农药胁迫导致MaRPS15A相对表达量下降。【结论】 成功克隆了松墨天牛核糖体蛋白基因MaRPS15A (GenBank登录号:KJ930032)。大多数农药抑制了MaRPS15A的表达，表明松墨天牛受到了不利影响。 |
| 关键词: 松墨天牛 核糖体蛋白 RPS15A 实时荧光定量 PCR (RT-qPCR) 农药胁迫 |
| DOI： |
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| 基金项目:国家自然科学基金项目（31170612） |
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| Cloning cDNA of ribosomal protein (RPS15A） from Monochamus alternatus and effects of insecticides on its expression |
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LUO Lin-lin,CAI Zi-ling,LIN Tong
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| Abstract: |
| 【Objective】This study explored the expression of ribosomal protein gene under different insecticide stresses to provide theoretical information for control of Monochamus alternatus as an important of pines.【Method】 Expressed sequence tags (ESTs) screened from the formerly constructed cDNA library of M.alternatus were selected randomly for sequencing.The homogenous sequences were obtained from GenBank by Blast.Real time quantitative RT-PCR (RT-qPCR) was used to determine transcript abundance patterns after treatment by 11 insecticides.【Result】A EST from GenBank relating to insect ribosomal protein S15A (RPS15A) was obtained and named MaRPS15A.The isolated cDNA of MaRPS15A containing both 5′ and 3′ untranslated regions was 504 nucleotide long with an open reading frame of 393 bp and 130 amino acids.The calculated molecular weight was 14.75 ku and the pI was 9.95.The deduced protein sequence of MaRPS15A showed 98% amino acid identity to Triboliumc astaneum RPS15A and >88% identity to RPS15A ribosomal proteins of other insects.All insecticides resulted in decreased transcript abundances except deltamethrin.【Conclusion】The ribosomal protein gene MaRPS15A was successfully cloned from M.alternatus (GenBank No.KJ930032).Most of the tested insecticides suppressed MaRPS15A expression,indicating that adverse impact on M.alternatus. |
| Key words: Monochamus alternatus ribosomal protein RPS15A RT-qPCR treatment with insecticides |
