| 摘要: |
| 采用RACE 技术克隆三疣梭子蟹核受体fzt-f1 基因, 全长cDNA 序列1763bp.该基因3'和 5'非编码区分别为1034bp 和141bp, 开放阅读框为588bp, 推测编码195 个氨基酸, 预测分子量为 22.8kDa, 理论等电点为6.35, 命名为PTNHr 基因。同源性和系统进化分析表明, 三疣梭子蟹fzt-f1 基因与刀额新对虾同源性高达75.4%, 并且与刀额新对虾聚为一支。对核受体基因fzt-f1、EcR 和RXR 在蜕皮周期中的表达情况进行实时荧光定量PCR 分析, 结果表明, 三个核受体基因在不同蜕皮时期, 均出现表达差异, 说明这三个基因都参与蜕皮调控过程。其中, fzt-f1 和RXR 基因在不同蜕皮时期的 表达模式上出现相反的表达特征, 预测这两个基因存在相互抑制的调节关系。EcR 和RXR 基因在不 同蜕皮时期的表达模式上出现部分相似的表达特征。 |
| 关键词: 三疣梭子蟹 核受体 基因克隆 基因表达 去眼柄 |
| DOI：10.11693/hyhz20150300077 |
| 分类号: |
| 基金项目:国家自然科学基金项目, 41576147 号, 41306177 号; 青岛海洋科学与技术国家实验室鳌山科技创新计划项目, 2015ASKJ02 号。 |
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| CLONING OF PORTUNUS TRITUBERCULATUS ftz-f1 cDNA AND EXPRESSION ANALYSIS OF RELATED NUCLEAR RECEPTORS DURING MOLTING CYCLE |
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ZHANG Long-Tao1,2,3, LV Jian-Jian1,3, GAO Bao-Quan1,3, LIU Ping1,3, FU Ping1,3
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1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;2.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;3.Function Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266235, China
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| Abstract: |
| We successfully cloned fzt-f1 gene of Portunus trituberculatus for the first time using RACE (rapid-amplification of cDNA ends) method, and named it PTNHr. The full-length cDNA sequence of ftz-f1 is 1763bp, which contains a 1034bp 5'-UTR, 141bp 3'-UTR, and 588bp open reading frame (ORF) that encodes 195 amino-acid polypeptides. It has a molecular mass of 22.8 kDa and isoelectric point (pI) of 6.35. Homologous analysis showed that ftz-f1 has the highest homology to Metapenaeus ensis, and phylogenetic analysis showed that fzt-f1 of P. trituberculatus was also in the same branch with M. ensis. In addition, using RT-PCR (reverse transcription-polymerase chain reaction), we investigated the functions of fzt-f1, EcR, and RXR in different molting stages. The results reveal different expression patterns of the molting-regulation function of them, of which fzt-f1 and RXR show opposite features, indicating a mutual suppression relation of them, while those of EcR and RXR are similar somewhat. |
| Key words: Portunus trituberculatus fzt-f1 Gene cloning expression analysis nuclear receptor |
