摘要
为研究红鳍东方鲀Takifugur ubripes免疫系统相关细胞因子信号传导抑制因子-3(Suppressor of cytokine signaling,SOCS3)的表达情况,采用相对实时荧光定量PCR方法比较了体质量为(290.34±5.13)g的健康红鳍东方鲀脑、脾脏、肠、鳃、肌肉和肝脏6个不同组织中socs3基因表达量,以及感染哈氏弧菌Vibrio harveyi后肝脏、脾脏和肠组织socs3基因表达量的变化。结果表明:socs3基因在健康红鳍东方鲀肝脏组织、肌肉组织中均有显著表达(P<0.05),在鳃、脑、脾脏、肠组织中表达不显著(P>0.05);感染哈氏弧菌的红鳍东方鲀,在感染后的24、48 h时,肝脏组织socs3的表达量显著高于0 h(P<0.05),分别约为初始时的10倍和8倍,其他组织变化不明显(P>0.05);利用NCBI的ORF Finder程序鉴定所得红鳍东方鲀SOCS3(NP_001072096.1)序列的潜在开放阅读框(ORF),通过模块化结构研究工具来鉴定保守结构域和信号肽,表明socs3的cDNA序列全长1533 bp,其中5′端非编码区为215 bp,开放阅读框(ORF)长度为606 bp,3′端非编码区为713 bp,共编码201个氨基酸;利用MEGA 6.0软件进行氨基酸序列比对并构建系统发育树,结果显示,硬骨鱼类单独聚为一支,与哺乳动物、两栖动物及鸟类分开,硬骨鱼类中的SOC3b单独聚为一支,红鳍东方鲀与其同科的黑青斑河豚Tetraodon nigroviridis的SOC3聚在一起,并与三刺鱼Gasterosteus aculeatus属于同一分支。本研究结果可为红鳍东方鲀免疫应答的研究提供数据支持。
The expression of suppressor of cytokine signaling 3 ( socs3 ) gene in six tissues was comparatively investigated in healthy redfin puffer Takifugu rubripes and in liver of the redfin puffer challenged with Vibrio harveyi using real-time quantitative fluorescence PCR in order to probe for immune function of redfin puffer. The gene socs3 was shown to be significantly expressed in liver and muscle of healthy redfin puffer( P <0.05), without significant expression in gill, brain, spleen and intestine ( P >0.05). There was significantly higher expression level of socs3 in liver of the infected redfin puffer for 24 h and 48 h than for 0 h ( P <0.05), as much about ten times as the initial concentration in 24 h and as much eight times as the initial concentration in 48 h, without significant differences in other tissues( P >0.05). The NCBI s ORF Finder procedure revealed the potential Open Reading Frame(ORF) of SOCS3 sequence(NP_001072096), and conserved domain and signal peptide were identified by modular structural research tools. MEGA 6.0 revealed that the total length of sequence of socs3 cDNA was 1533 bp, SOCS3 protein including the 5′ end non-coding region length of 215 bp, the length ORF of 606 bp, the 3′ end non-coding region length of of 713 bp and coding 201 amino acids. The phylogenetic analysis of SOCS3 indicated that the teleost fishes were grouped into one, in the same branch as Tetradon nigrovividis , and Gasterosteus aculeatus . separated from mammals, amphibians and birds. The findings indicate that the immune expression and cytokine of gene socs3 play an important role in immune regulation process in redfin puffer, promoting understanding of immune function in redfin puffer.
作者
华昕彤
刘鹰
陈冰
徐岩
傅松哲
姜晨
HUA Xin-tong;LIU Ying;CHEN Bing;XU Yan;FU Song-zhe;JIANG Chen(Aquacultural Engineering Research & Development Center, Dalian Ocean University, Dalian 116023, China;College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;Fisheries College, Ocean University of China, Qingdao 266000, China)
出处
《大连海洋大学学报》
CAS
CSCD
北大核心
2019年第1期33-40,共8页
Journal of Dalian Ocean University
基金
国家重点研发计划项目(2017YFD0701700)
国家自然科学基金资助项目(31472312
31672673)
江苏省现代农业-研究开发示范类项目(BE2015325)
