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刘林, 何丽, 吴富盛, 阮记明, 周颖, 隗黎丽. 草鱼IRE1-like基因的克隆、组织表达及其对微囊藻毒素-LR的响应[J]. 水生生物学报, 2020, 44(6): 1168-1173. DOI: 10.7541/2020.135
引用本文: 刘林, 何丽, 吴富盛, 阮记明, 周颖, 隗黎丽. 草鱼IRE1-like基因的克隆、组织表达及其对微囊藻毒素-LR的响应[J]. 水生生物学报, 2020, 44(6): 1168-1173. DOI: 10.7541/2020.135
LIU Lin, HE Li, WU Fu-Sheng, RUAN Ji-Ming, ZHOU Ying, WEI Li-Li. CLONING AND TISSUES EXPRESSION OF INOSITOL-REQUIRING ENZYME 1-LIKE (IRE1-LIKE) GENE IN GRASS CARP AND ITS RESPONSE TO MICROCYSTIN-LR[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(6): 1168-1173. DOI: 10.7541/2020.135
Citation: LIU Lin, HE Li, WU Fu-Sheng, RUAN Ji-Ming, ZHOU Ying, WEI Li-Li. CLONING AND TISSUES EXPRESSION OF INOSITOL-REQUIRING ENZYME 1-LIKE (IRE1-LIKE) GENE IN GRASS CARP AND ITS RESPONSE TO MICROCYSTIN-LR[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(6): 1168-1173. DOI: 10.7541/2020.135

草鱼IRE1-like基因的克隆、组织表达及其对微囊藻毒素-LR的响应

CLONING AND TISSUES EXPRESSION OF INOSITOL-REQUIRING ENZYME 1-LIKE (IRE1-LIKE) GENE IN GRASS CARP AND ITS RESPONSE TO MICROCYSTIN-LR

  • 摘要: 为了研究内质网应激相关基因需肌醇酶1(Inositol-requiring enzyme 1, IRE1-like)的结构和生物学功能及其在草鱼(Ctenopharyngodon idella)响应微囊藻毒素-LR(MC-LR)中的作用, 研究根据草鱼转录组测序结果得到该基因家族成员IRE1-like的EST序列, 采用RACE技术获得了草鱼IRE1-like基因的cDNA全长序列(登录号: MG797683)。该基因序列全长3595 bp, 包括3093 bp开放阅读框, 编码1030个氨基酸, 分子量为116.24 kD, 理论等电点为6.26, 具有跨膜结构和信号肽。草鱼IRE1-like基因包含Luminal (39—307 aa)、STKc (569—834 aa)和RNase (837—915 aa)三个结构域。同源性和系统进化树结果表明草鱼IRE1-like基因与斑马鱼IRE1-α基因亲缘关系最近。荧光定量PCR(qRT-PCR)检测结果表明, 草鱼IRE1-like基因在肝脏组织中的表达量最高, 在肌肉、脾脏、鳃和心脏等其他8种不同组织中也均有表达。不同剂量MC-LR诱导草鱼24h和96h后, 25 μg MC-LR/kg BW和100 μg MC-LR/kg BW剂量组中草鱼肝脏IRE1-like基因表达水平分别在24h和96h显著上升(P<0.05)。以上结果初步说明了草鱼IRE1-like基因可能在MC-LR诱导草鱼内质网应激中起着重要的作用, 为进一步研究草鱼IRE1-like基因的功能奠定了基础。

     

    Abstract: To investigate the structure and biological function of inositol-requiring enzyme 1(IRE1), an endoplasmic reticulum stress-related gene, and its role in response to MC-LR, the full-length cDNA sequence of grass carp IRE1-like gene was cloned basing on the EST sequence of IRE1-like members from transcriptome sequencing results of grass carp by using RACE technology (GenBank: MG797683). The total length of IRE1-like gene was 3595 bp, including 3093 bp open reading frame encoding 1030 amino acids. Its molecular weight was 116.2 kD with a theoretical isoelectric point of 6.26, and it has transmembrane structure and signal peptide with three structural domains, including Luminal domain (39—307 aa), STKc domain (569—834 aa) and RNase domain (837—915 aa). The results of homology and phylogenetic tree indicated that grass carp IRE1-like was most related to Danio rerio IRE1-α. The results of quantitative RT-PCR (qRT-PCR) showed that grass carp IRE1-like gene highly express in liver, and also modestly express in other tissues, such as muscle, spleen, gill and heart. MC-LR significantly induced the expression of grass carp IRE1-like gene in 25 μg Mc-LR /kg BW and 100 μg MC-LR /kg BW group at 24h and 96h, respectively. Grass carp IRE1-like gene may play an important role in MC-LR-induced endoplasmic reticulum stress, laying a foundation for further study on the function of grass carp IRE1-like gene.

     

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