| 马红悦，李玲，乌亚汗，田羽，李艳艳，庞保平,2021,沙葱萤叶甲核糖体蛋白基因GdRpS3a的克隆、分子特性及表达分析[J].环境昆虫学报,(5):1220-1228 |
| 沙葱萤叶甲核糖体蛋白基因GdRpS3a的克隆、分子特性及表达分析 |
| Molecular cloning, characterization and expression analysis of ribosomal protein S3a gene GdRpS3a in Galeruca daurica (Coleoptera: Chrysomelidae: Galerucinae) |
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| DOI： |
| 中文关键词: 沙葱萤叶甲 核糖体蛋白 基因克隆 表达分析 夏滞育 |
| 英文关键词:Galeruca daurica ribosomal protein gene cloning expression analysis summer diapause |
| 基金项目:国家自然科学基金（31760517） |
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| 中文摘要: |
| 核糖体蛋白（ribosomal protein, Rp）是一类参与蛋白质合成、细胞代谢、机体免疫及信号传导等重要功能的蛋白质。本研究旨在克隆沙葱萤叶甲Galeruca daurica核糖体蛋白S3a基因cDNA全长序列，分析其分子特性和表达模式，以期为进一步研究其在沙葱萤叶甲生长发育及滞育中的作用提供必要的基础。根据现有的沙葱萤叶甲转录组测序数据，应用cDNA末端快速扩增（RACE）技术克隆获得了沙葱萤叶甲RpS3a基因的全长cDNA序列，命名为GdRpS3a（GenBank登录号：MN660144）。该基因全长为800 bp，开放阅读框全长为687 bp，编码228个氨基酸，具有核糖体蛋白S3a家族的保守功能域；预测分子量为25.63 kDa，等电点pI为10.53，无信号肽和跨膜结构。同源序列比对和系统发育分析表明，GdRpS3a与玉米根萤叶甲Diabrotica virgifera virgifera RpS3a的亲缘关系最近，其氨基酸序列一致性最高为54.10%。采用qPCR技术检测该基因在沙葱萤叶甲不同发育阶段（卵、1~3龄幼虫、预蛹、蛹及成虫羽化3、7、10、15、25、40、60、80和100 d）和3日龄成虫在不同温度（0、5、10、15、20、25、30、35和40℃）处理1 h后的表达谱。结果表明：GdRpS3a在不同发育阶段的沙葱萤叶甲体内均有表达，在成虫滞育结束后表达量最高，在卵期和成虫滞育期间低表达。温度对GdRpS3a的表达有显著的影响，30℃下最高，0℃下最低。GdRps3a的表达与沙葱萤叶甲生长发育阶段及环境温度有关，可能在沙葱萤叶甲成虫夏滞育中起着重要作用。 |
| 英文摘要: |
| Ribosomal proteins are a kind of important proteins which are involved in the functions of protein synthesis, cellular metabolism, organism immunity, signal transduction and etc. This study aims to clone a ribosomal protein S3a (RpS3a) gene from Galeruca daurica, and to analyze its molecular characteristics and expression patterns so as to build a foundation for investigation on its functions in the development and diapause of G. daurica. The full-length cDNA of RpS3a was successfully cloned from G. daurica by RACE according to the sequence information available in the transcriptome data, and named GdRpS3a (GenBank accession number: MN660114), whose length was 800 bp with an open reading frame (ORF) of 687 bp, encoding a protein of 228 amino acids with the predicted molecular weight of 25.63 kDa and pI of 10.53. The encoded protein contains a conserved domain of RpS3a family without signal peptide and transmembrane domain. Homologous sequence alignment and phylogenetic analysis showed that GdRpS3a has the closest affinity with Diabrotica virgifera virgifera RpS3a with the highest amino acid sequence identity (54.10%）. Its expression profiling at different development stages (egg, 1~ 3 instar larva, prepupa, pupa, and adult of 3, 7, 10, 15, 25, 40, 60, 80, and 100 days after eclosion) and in the three-day old adults exposed to different temperatures (0, 5, 10, 15, 20, 25, 30, 35, and 40℃) for 1 h was performed by qPCR. The results showed that GdRpS3a was expressed at different development stages with the highest expression levels after termination of adult diapause and the lowest during the egg stage and adult diapause. Temperature had significant effects on the expression levels of GdRpS3a with the highest level at 30℃ and the lowest at 0℃. In conclusion, the expression of GdRpS3a was related to the developmental stages d of G. daurica and enviromental temperature, and may played an important role in summer diapause of G. daurica adults. |
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