| 引用本文: | 王 伟,吴旭干,楼 宝,徐 蕾,刘智俊,成永旭.三疣梭子蟹(Portunus trituberculatus)RXR基因克隆及其在蜕皮过程中的表达分析.海洋与湖沼,2014,45(5):1105-1114. |
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| 三疣梭子蟹(Portunus trituberculatus)RXR基因克隆及其在蜕皮过程中的表达分析 |
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王 伟,吴旭干,楼 宝,徐 蕾,刘智俊,成永旭
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上海海洋大学 水产种质资源发掘与利用教育部重点试验室,上海海洋大学 水产种质资源发掘与利用教育部重点试验室,浙江省海洋水产研究所,上海海洋大学 水产种质资源发掘与利用教育部重点试验室,上海海洋大学 水产种质资源发掘与利用教育部重点试验室,上海海洋大学 水产种质资源发掘与利用教育部重点试验室
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| 摘要: |
| 本研究通过转录组测序和RACE技术克隆了三疣梭子蟹类维甲酸X受体2(PtRXR2)基因cDNA全长(登录号: KF914662), 并通过荧光定量PCR(qRT-PCR)技术研究了该基因在三疣梭子蟹不同组织及不同蜕皮阶段的表达情况。结果表明, (1) PtRXR2基因cDNA全长1718bp, 包括5’非编码区(5’-UTR)141bp、3’-UTR 209bp和开放阅读框1368bp, 编码455个氨基酸, 预测分子量和等电点为49.75kDa和6.79。(2) PtRXR2推导氨基酸序列的Blastp 结果显示, PtRXR2与已知甲壳动物RXR的一致性为74%—99%, 系统进化树分析表明PtRXR2与其它甲壳动物RXR聚为一支。(3) qRT-PCR结果显示PtRXR2在C期三疣梭子蟹Y器官、鳃、眼柄和大颚器中表达水平较高, 肝胰腺中的表达水平最低。其它6个组织中的表达水平居中。(4) 不同蜕皮阶段, PtRXR2在Y器官、眼柄和胸神经中均是AB期表达水平最高, E期最低, 这可能与上述器官的细胞增殖主要发生在AB期有关; 肌肉中的PtRXR2表达水平在AB和C期较高, E和D期最低, 与肌肉的生长和营养物质积累主要发生在这两个阶段相对应; 肝胰腺中的PtRXR2表达水平在AB期最高, C期最低, 这暗示PtRXR2主要参与肝胰腺中的上皮细胞增殖和分化, 对于C期的营养代谢调控可能不起关键作用。 |
| 关键词: 三疣梭子蟹 类维甲酸X受体(RXR) 基因克隆 表达分析 蜕皮 |
| DOI：10.11693/hyhz20131200218 |
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| 基金项目:国家自然科学基金项目, 41276158 号; 上海高校水产学一流学科建设项目, 沪教科2012-62 号; 上海高校创新团队(第二期)项目, 沪教科2009-26 号; 浙江省重大科技专项计划项目, 2011C12011 号; 浙江省农业科技成果转化资金项目, 2013-171-72 号; 上海市科委工程技术中心能力提升项目, 13DZ2280500 号 |
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| CLONING OF RETINOID X RECEPTOR (RXR) AND ITS EXPRESSION ANALYSIS DURING MOLTING IN PORTUNUS TRITUBERCULATUS |
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Wang Wei,Wu Xugan,Lou Bao,Xu Lei,Liu Zhijun and Cheng Yongxu
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Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University,Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University,Marine Fisheries Research Institute of Zhejiang,Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University,Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University,Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University
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| Abstract: |
| We cloned full-length cDNA of a retinoid X receptor (GenBank accession number: KF914662) in Portunus trituberculatus by transcriptome sequencing and RACE (rapid amplification of cDNA ends). The relative gene expression levels of PtRXR2 (P. trituberculatus RXR2) gene in various tissues were detected by qRT-PCR (quantitative real-time PCR) during the molting cycle. The full-length of receptor PtRXR2 cDNA is 1718bp, including a 1368bp ORF (encoded 455 amino acid residues), a 141bp 5'-UTR and a 209bp 3'-UTR, whereas its calculated molecular weight and isoelectric point are 49.75 kDa and 6.79, respectively. The predicted amino acid sequence of PtRXR2 shared 74%—99% identity with those of other crustaceans as showed in homologous analysis, and PtRXR2 is clustered with crustaceans RXRs in phylogenetic tree. The receptor had highest expression in Y-organ, gill, eyestalk, and mandibular organ, lowest in hepatopancreas, and intermediate in the other six tissues. PtRXR2 was expressed high in stages AB and C of molting cycle in Y-organ, eyestalk and thoracic ganglia, and very low in stage E due probably to cell proliferation in stage AB in these tissues, whereas in the muscle, PtRXR2-mRNA expression was higher in stages AB and C than that in stages E and D, corresponding to the muscular growth and nutrition accumulation. In addition, the peak value of PtRXR2-mRNA in hepatopancreas occurred in stage AB and the lowest level existed in stage C. Therefore, PtRXR2 is involved mainly in the proliferation and differentiation of epithelial cell in hepatopancreas in stage AB, but the metabolism of nutrition in stage C. |
| Key words: Portunus trituberculatus retinoid X receptor (RXR) gene cloning expression analysis molting |
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