水产科学  2021, Vol. 40 Issue (1): 1-9    DOI: 10.16378/j.cnki.1003-1111.20020
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仿刺参SARM基因的克隆、表达和功能研究
孙红娟1, 陈仲1, 贺迪2, 高杉1, 王摆1, 关晓燕1, 蒋经伟1, 董颖1, 周遵春1
1.辽宁省海洋水产科学研究院,辽宁省海洋水产分子生物学重点试验室,辽宁 大连 116023;
2.凌海市达莲海珍品养殖有限责任公司,辽宁 锦州 121209
Characterization, Expression and Function of SARM in Sea Cucumber Apostichopus japonicus
SUN Hongjuan1, CHEN Zhong1, HE Di2, GAO Shan1, WANG Bai1, GUAN Xiaoyan1, JIANG Jingwei1, DONG Ying1, ZHOU Zunchun1
1.Liaoning Key Laboratory of Marine Fishery Molecular Biology, Liaoning Ocean and Fisheries Science Research Institute, Dalian 116023, China;
2.Linghai Dalian Marine Treasures Breeding Co., Ltd., Jingzhou 121209, China
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摘要 采用RACE方法克隆了仿刺参TLR信号通路MyD88非依赖途径接头分子SARM,命名为AjSARM。AjSARM基因的序列全长3874 bp,开放阅读框为2412 bp,编码803个氨基酸。通过SMART软件对AjSARM蛋白进行蛋白结构预测,发现AjSARM蛋白由2个SAM结构域和1个C端TIR结构域组成。采用实时荧光定量PCR的方法检测AjSARM基因在仿刺参发育不同时期的表达量,发现在小耳幼虫期开始高表达,神经发育也是从这一时期开始的,表明AjSARM蛋白在仿刺参的发育过程中发挥了重要作用。仿刺参体腔细胞经过4种病原菌(灿烂弧菌、假交替单胞菌、希瓦氏菌和蜡样芽孢杆菌)刺激后,在灿烂弧菌、假交替单胞菌和希瓦氏菌刺激组,AjSARM基因在4 h和24 h的表达均受到抑制,然而在蜡样芽孢杆菌组,24 h表达上调。除了希瓦氏菌组外,其他3个组的AjSARM基因在96 h表达量达到最高。AjSARM在应对不同病原菌刺激后不同时间点所表现出的动态表达模式表明,它参与了仿刺参体腔细胞免疫应答的过程。通过对AjSARM的结构以及在不同发育阶段和不同病原菌刺激后的表达模式进行分析,探索仿刺参MyD88非依赖途径接头分子SARM的功能,可以为棘皮动物的发育和免疫系统进化研究提供参考。
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孙红娟
陈仲
贺迪
高杉
王摆
关晓燕
蒋经伟
董颖
周遵春
关键词 仿刺参SARM结构分析发育阶段免疫刺激基因表达    
Abstract:SARM, an adaptor of MyD88 independent signal pathway, was cloned in sea cucumber Apostichopus japonicus, named AjSARM by RACE method. The full length of AjSARM was 3874 bp with an ORF of 2412 bp encoding 803 amino acids. Structure prediction by SMART indicated that AjSARM contained two sterile motifs (SAM) and a C-terminal TIR domain. Quantitative real-time PCR was selected to examine AjSARM expression at different developmental stages of sea cucumber. AjSARM was rapidly up-regulated from auricularia at nervous development stage. It was indicated that AjSARM played an important role in the process of development. After four pathogens stimulus, the expression of AjSARM was inhibited at 4 h and 24 h in the groups of Vibrio splendidus, Pseudoalteromonas nigrifacien and Shewanella baltica chanlenged, but upregulated at 24 h in Bacillus cereus group. Except in S. baltica group, AjSARM reached top expression level all in other three groups at 96 h. The dynamic expression patterns of AjSARM at different times in response to different pathogens proved that it participated in the immune reaction in sea cucumber coelomocytes. The structure and expression analyses of AjSARM help to explore its functions as an adaptor and will lay foundations for the evolution study of immune system in echinoderm and disease prevention in sea cucumber.
Key wordssea cucumber Apostichopus japonicus    SARM    structure analysis    developmental stage    immune challenge    gene expression
收稿日期: 2020-02-10     
PACS:  S968.9  
基金资助:国家自然科学基金资助项目(31672688);辽宁省兴辽英才计划项目(XLYC1802091);大连市顶尖及领军人才项目(2018RD10);辽宁省中央引导地方科技创新项目(2019416034);辽宁省农业科学院院长基金资助项目(2020QN2423).
通讯作者: 周遵春(1967—),男,研究员,博士;研究方向:海洋生物技术. E-mail:zunchunz@hotmail.com.   
作者简介: 孙红娟(1987—),女,助理研究员,硕士;研究方向:生物化学与分子生物学. E-mail:826146350@qq.com.
引用本文:   
孙红娟, 陈仲, 贺迪, 高杉, 王摆, 关晓燕, 蒋经伟, 董颖, 周遵春. 仿刺参SARM基因的克隆、表达和功能研究[J]. 水产科学, 2021, 40(1): 1-9.
SUN Hongjuan, CHEN Zhong, HE Di, GAO Shan, WANG Bai, GUAN Xiaoyan, JIANG Jingwei, DONG Ying, ZHOU Zunchun. Characterization, Expression and Function of SARM in Sea Cucumber Apostichopus japonicus. 水产科学, 2021, 40(1): 1-9.
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