文章摘要
环朋朋,吕建建,孙东方,高保全,刘 萍.三疣梭子蟹PtDNMT1基因的克隆及其 在低盐适应中的表达分析.渔业科学进展,2019,40(1):92-100
三疣梭子蟹PtDNMT1基因的克隆及其 在低盐适应中的表达分析
The Cloning of the PtDNMT1 Gene of Portunus trituberculatus and Its Expression Analysis in Low Salinity Adaptation
投稿时间:2017-12-01  修订日期:2017-12-29
DOI:
中文关键词: 三疣梭子蟹  PtDNMT1  盐度胁迫  基因克隆  基因表达
英文关键词: Portunus trituberculatus  Methyltransferase 1  Salinity stress  Gene cloning  Gene expression
基金项目:
作者单位
环朋朋 上海海洋大学水产与生命学院 上海 201306农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071 
吕建建 农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071 
孙东方 上海海洋大学水产与生命学院 上海 201306农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071 
高保全 农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071 
刘 萍 农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071 
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中文摘要:
      甲基转移酶是维持基因组甲基化状态的重要基因,本研究利用SMART-RACE技术克隆了三疣梭子蟹(Portunus trituberculatus)甲基转移酶基因(PtDNMT1)。PtDNMT1基因cDNA序列全长5919 bp,包括4832 bp的开放阅读框,编码1610个氨基酸,预测分子量为148.15 kDa,理论等电点为4.68。结构预测发现,PtDNMT1有2个特殊的结构域,分别是锌指结构域(zf-CXXC)和甲基转移酶家族特有的Dcm结构域。进化树分析显示,PtDNMT1基因与昆虫类的DNMT基因聚为一支。组织表达分析发现,PtDNMT1基因在肝胰腺、鳃、卵巢、肌肉、胃、心脏、血液中均有表达,其中在肝胰腺中表达最高,卵巢和鳃次之。进一步研究了低盐胁迫后PtDNMT1基因在鳃、肝胰腺和肌肉组织中的表达变化规律为胁迫6 h时鳃组织中PtDNMT1基因的表达即达到峰值(5.3倍),并一直持续到12 h (4倍),随后逐步下降,在72 h时仍显著高于对照组(2.3倍);PtDNMT1基因在肝胰腺中的表达规律类似于鳃,然而其达到峰值的时间稍晚于鳃(24 h),且上调倍数高于鳃(8倍);低盐胁迫后PtDNMT1基因在肌肉中的表达最初呈现下调趋势,之后(24 h)上调表达至峰值(2.2倍),且一直上调表达至72 h。本研究首次克隆了PtDNMT1基因,根据其在各组织中的表达分布特征以及盐度胁迫后的表达变化情况,推测DNA甲基化在三疣梭子蟹低盐适应中发挥了重要作用。
英文摘要:
      Methyltransferase (DNMT1) is an important gene that maintains the methylation state of the genome. This study uses the SMART-RACE technique to clone the DNMT1 gene of Portunus trituberculatus (PtDNMT1). The cDNA sequence of PtDNMT1 gene is 5919 bp, including the open reading frame of 4832 bp, and 1610 amino acids are encoded. The predicted molecular weight is 148.15 kDa, and the theoretical isoelectric point is 4.68. The structure prediction found that there are two special PtDNMT1 domain structures: the zf-CXXC zinc finger domain and the methyltransferase familial Dcm structure domain. The analysis of the evolutionary tree showed that PtDNMT1 gene and insects DNMT gather into one. PtDNMT1 was expressed in the hepatopancreas, gill, ovary, muscle, stomach, heart, and blood. The highest expression was found in the hepatopancreas, followed by the ovaries and gills. Then we get a further research about the PtDNMT1 gene expression during low salinity stress in gill, l hepatopancreas and muscle tissue. In gill tissue, we found that PtDNMT1 gene expression peaked (5 folds) after 6 h, and continued to 12 h (6 folds), then declined gradually, but was still significantly higher than the control group (3 folds). The expression of PtDNMT1 in the hepatopancreas was similar to the gills, however the peak time was later than in gill tissue (24 h), and the peak was higher than in the gills (8 folds). After low salinity stress, the expression of the PtDNMT1 gene in muscle was firstly reduced, then (24 h) increased to a peak (2.2 times), and the expression was raised to 72 h. This study is the first to clone the PtDNMT1 gene. According to its distribution characteristics and occurrence in various organizations, expression changes under salinity stress. We speculate that low salinity adaptations in DNA methylation in P. trituberculatus played an important role.
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