何亮银, 史晓丽, 林佳阳, 黄淑怡, 黄伟卿, 韩坤煌. 大黄鱼adsl基因的克隆及其表达量与肌苷酸含量的关联性分析[J]. 南方水产科学. DOI: 10.12131/20230178
引用本文: 何亮银, 史晓丽, 林佳阳, 黄淑怡, 黄伟卿, 韩坤煌. 大黄鱼adsl基因的克隆及其表达量与肌苷酸含量的关联性分析[J]. 南方水产科学. DOI: 10.12131/20230178
HE Liangyin, SHI Xiaoli, LIN Jiayang, HUANG Shuyi, HUANG Weiqing, HAN Kunhuang. Cloning of adsl gene in Larimichthys crocea and analysis of its correlation between expression level and inosine content[J]. South China Fisheries Science. DOI: 10.12131/20230178
Citation: HE Liangyin, SHI Xiaoli, LIN Jiayang, HUANG Shuyi, HUANG Weiqing, HAN Kunhuang. Cloning of adsl gene in Larimichthys crocea and analysis of its correlation between expression level and inosine content[J]. South China Fisheries Science. DOI: 10.12131/20230178

大黄鱼adsl基因的克隆及其表达量与肌苷酸含量的关联性分析

Cloning of adsl gene in Larimichthys crocea and analysis of its correlation between expression level and inosine content

  • 摘要: 大黄鱼 (Larimichthys crocea) 风味品质劣化严重制约着其产业的健康发展。为探究大黄鱼腺苷琥珀酸裂解酶 (Adenylosuccinate lyase, ADSL) 在风味关键物质肌苷酸 (Inosine monophosphate, IMP) 合成中的作用,克隆了大黄鱼adsl基因的开放阅读框 (ORF) 序列,分析了其基因结构和进化特征;在明确其组织分布特性的基础上,比较了不同养殖规格大黄鱼肌肉组织中的IMP含量和adsl基因的表达差异。结果显示:大黄鱼adsl基因ORF全长1 446 bp,编码481个氨基酸。对应的蛋白质相对分子质量为54.6 kD,等电点为6.19,包含N末端的裂解酶1和C末端的ADSL_C两个结构域。大黄鱼与棘头梅童鱼 (Collichthys lucidus) 的亲缘关系最近,相似性达98.54%。大黄鱼adsl基因由12个外显子和11个内含子组成,其mRNA在肌肉组织中表达量最大,且显著高于其他组织 (P<0.05)。随着体质量的增加,养殖大黄鱼肌肉组织中的IMP含量显著增加 (P<0.01);adsl基因mRNA的相对表达量也呈类似规律。相关性分析显示,大黄鱼肌肉组织中adsl基因的相对表达量与对应的IMP含量呈显著正相关 (r=0.962)。研究结果为进一步探明adsl基因在大黄鱼IMP合成及风味形成中的作用机制提供了参考资料。

     

    Abstract: Declining flesh quality in farmed large yellow croaker (Larimichthys crocea) seriously restricts the healthy development of its industry. To investigate the role of adenylsuccinate lyase (ADSL) in the synthesis of the key flavor substance inosine monophosphate (IMP) in L. crocea, we cloned the open reading frame (ORF) sequence of its adsl gene and analyzed the gene structure and evolutionary characteristics. After clarifying its tissue distribution characteristics, we compared the differences in IMP content and adsl gene expression in muscle tissues of cultured L. crocea with different sizes. The results show that the ORF cDNA of adsl was 1 446 bp and encoded a polypeptide of 481 amino acids (aa). The corresponding protein had a relative molecular weight of 54.6 kD and an isoelectric point of 6.19, including Pfam Lyase_1 (Ala24−Leu309) and ADSL_C (Leu374−Leu458) structural domains. Phylogenetic analysis of the deduced AA sequence reveals that the genetic relationship between L. crocea and Collechthys lucidus was the closest, with a similarity of 98.54%. The genomic DNA sequence of adsl gene was subdivided into 12 exons and 11 introns. Its mRNA had the highest expression level in muscle tissue and was significantly higher than other tissues (P<0.05). With the increase of body mass, the IMP content in L. crocea muscle increased significantly (P<0.01). The relative expression level of adsl gene mRNA also followed a similar pattern. Correlation analysis shows that the relative expression level of adsl gene in L. crocea muscle was significantly positively correlated with the corresponding IMP content (r=0.962). The results provide references for further sudy on the mechanism of action of adsl gene in IMP synthesis and flavor formation in L. crocea.

     

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