| 摘要: |
| 克隆了三疣梭子蟹TLR家族的一个新基因, 将其命名为PtToll6。该基因全长为4 034 bp, 预测分子量为109.078 kDa, 理论等电点为6.06,共编码977个氨基酸。SMART预测显示, PtToll6具有Toll样受体典型的结构域, 包括前端的序列区(LRR)、跨膜区(TM)和胞内(TIR)区, 进化树分析表明其与同属节肢动物门的果蝇Dm toll9聚为一支。组织表达分布结果显示, PtToll6在肝胰腺中特异性地高表达, 其次表达于肠道, 在鳃中的表达量最低。采用3种PAMPs进行注射刺激, 发现PtToll6对脂多糖的响应最为强烈, 在感染48 h后的表达量为对照组的上千倍, 推测其可能为PtToll6基因主要识别的病原相关分子模式。PtToll6在人工感染副溶血弧菌后的12 h开始上调表达, 至24 h达到峰值(上调9.02倍)。采用RNAi敲降PtToll6后, MyD88的表达量相应下调, 同时发现感染副溶血弧菌后的死亡率显著提高, 为阴性对照组的1.8倍。实验结果表明: PtToll6基因在抗副溶血弧菌中发挥重要功能。本实验对解析三疣梭子蟹抵御病原入侵的分子机制具有重要指导作用。 |
| 关键词: 三疣梭子蟹 Toll样受体 模式识别受体 病原相关分子模式 |
| DOI：10.11693/hyhz20221100285 |
| 分类号: |
| 基金项目:国家虾蟹产业技术体系, CARS-48号; 国家自然科学基金项目, 42076116号; 中国水产科学研究院基本科研业务费, 2020TD46号。 |
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| CLONING OF PTTOLL6 GENE IN PORTUNUS TRITURATUS AND ITS FUNCTION IN IMMUNITY |
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ZHANG Wei-Wei1,2, LYU Jian-Jian2, LI Yu-Kun1,2, CHU Fan-Zhi1,2, GAO Bao-Quan2, LIU Ping2
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1.National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;2.National Key Laboratory of Marine Breeding and Sustainable Production, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
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| Abstract: |
| we cloned a new gene from the Portunus trituberculatus TLRs family and named it PtToll6. The gene is 4 034 bp in length and encodes 977 amino acids. PtToll6 has a typical domain of Toll-like receptor, including LRR, transmembrane region, and intracellular TIR region. Phylogenetic tree analysis showed that PtToll6 is clustered with Dm toll9 of the same phylum Arthropoda. The tissue expression distribution show that PtToll6 was specifically over-expressed in hepatopancreas, followed by the intestinal tract, and then the heart. Using three types of PAMPs for injection stimulation, it was found that PtToll6 had the strongest response to lipopolysaccharides, and the expression amount after 48 h of infection was thousands of times that of the control group, and it was speculated that it may be the pathogen-related molecular pattern mainly recognized by PtToll6 gene. The expression of PtToll6 was up-regulated at 12 h after Vibrio parahaemolyticus infection and peaked at 24 h (up-regulated 9.02-fold). After RNAi knocked down PtToll6, the expression of MyD88 was lowered, and the mortality rate of V. parahaemolyticus was significantly increased by 1.8 times that of negative control. This study showed that PtToll6 gene could play an important role in resisting V. parahaemolyticus, and provided an important reference for the understanding the molecular mechanism of P. trituberculatus. |
| Key words: Portunus trituberculatus toll-like receptors (TLRs) pattern recognition receptors (PRR) pathogen associated molecular patterns (PAMPs) |
