| 向 梅1,2, 谢应强2, 张洪志2, 李玉艳2, 王孟卿2, 臧连生1*, 张礼生2*.七星瓢虫乙酰辅酶A羧化酶基因克隆及表达分析[J].植物保护,2021,47(3):14-21. |
| 七星瓢虫乙酰辅酶A羧化酶基因克隆及表达分析 |
| Cloning and expression analysis of acetyl-CoA carboxylase in Coccinella septempunctata |
| 投稿时间：2020-02-25 修订日期：2020-06-17 |
| DOI：DOI:10.16688/j.zwbh.2020093 |
| 中文关键词: 七星瓢虫 乙酰辅酶A羧化酶 脂代谢 基因克隆 表达分析 |
| 英文关键词:Coccinella septempunctata acetyl-CoA carboxylase lipid metabolism gene clone gene expression |
| 基金项目:国家自然科学基金(31972339);国家重点研发计划(2017YFD0201000);国家烟草总局重大专项（110201901038（LS-01）） |
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| 中文摘要: |
| 乙酰辅酶A羧化酶(acetyl-CoA carboxylase,ACC)是脂肪酸合成第一步反应的限速酶,参与长链脂肪酸的合成。本试验基于七星瓢虫Coccinella septempunctata L.转录组数据,探究ACC在滞育七星瓢虫脂代谢中的调控作用。利用RT-PCR和RACE技术从七星瓢虫中克隆得到ACC基因全长,命名为CsACC(GenBank登录号:MT012819),该基因cDNA序列全长7 217 bp,开放阅读框(ORF)长为6 792 bp,编码2 263个氨基酸,预测蛋白质分子量为255.9 kDa,理论等电点(pI)为5.90,无跨膜螺旋结构,无信号肽。通过氨基酸多序列比对分析,结果显示CsACC与鞘翅目的赤拟谷盗Tribolium castaneum乙酰辅酶A羧化酶同源性较高。利用实时荧光定量PCR技术检测CsACC基因在七星瓢虫正常发育及滞育诱导不同阶段的相对表达量,结果显示,滞育诱导30 d表达量达到最高,滞育诱导60 d、滞育解除期以及正常发育30 d表达量几乎持平。本研究为揭示CsACC基因在脂代谢通路中的调控作用提供理论依据。 |
| 英文摘要: |
| Acetyl-CoA carboxylase (ACC) is a rate-limiting enzyme involved in the first step of fatty acid synthesis. In order to explore the function of ACC in lipid metabolism of Coccinella septempunctata at diapause stage, the full-length cDNA of a ACC gene was cloned using RT-PCR and RACE technique based on the transcriptome database. The gene was named CsACC (GenBank accession no. MT012819). The full length of cDNA sequence of this gene was 7 217 bp and the length of open reading frame (ORF) was 6 792 bp, encoding 2 263 amino acid residues. The predicted molecular weight of the protein was 255.9 kDa with an isoelectric point (pI) of 5.90 without a transmembrane helical structure or signal peptide. The results showed that CsACC had a high homology with that of Tribolium castaneum. The relative expression levels of CsACC gene in different stages of normal and diapause development were detected by real-time fluorescence quantitative PCR. The results showed that the expression levels of CsACC gene reached the highest level on 30th day after diapause induction, and the expression levels were almost flat on 60th day after diapause inductions, 30th day after normal development and after diapause termination. This study provides a theoretical basis for revealing the regulatory role of CsACC gene in the lipid metabolism pathway. |
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