文章摘要
罗红,李耀国,李东放,邹钧,肖调义.赤眼鳟信号转导与转录激活因子1基因(ScSTAT1)序列结构及免疫功能[J].水产学报,2021,45(3):381~395
赤眼鳟信号转导与转录激活因子1基因(ScSTAT1)序列结构及免疫功能
Sequence structure and immune function of signal transduction and transcriptional activator STAT1 of barbel chub (Squaliobarbus curriculus)
投稿时间:2019-11-26  修订日期:2020-06-05
DOI:10.11964/jfc.20191112082
中文关键词: 赤眼鳟  STAT1  干扰素通路  草鱼出血病  结构与功能
英文关键词: Squaliobarbus curriculus  signal transduction and activator of transcription 1 gene (STAT1)  interferon pathway  grass carp hemorrhage  structure and function
基金项目:国家自然科学基金(31802288,31572615);中国博士后科学基金(2018T110833,2017M612560);湖南省自然科学基金(2019JJ50231)
作者单位E-mail
罗红 湖南农业大学湖南省特色水产资源利用工程技术研究中心湖南 长沙 410128  
李耀国 湖南农业大学湖南省特色水产资源利用工程技术研究中心湖南 长沙 410128 yaoguolijkl@163.com 
李东放 湖南农业大学湖南省特色水产资源利用工程技术研究中心湖南 长沙 410128  
邹钧 湖南农业大学湖南省特色水产资源利用工程技术研究中心湖南 长沙 410128  
肖调义 湖南农业大学湖南省特色水产资源利用工程技术研究中心湖南 长沙 410128 tyxiao1128@163.com 
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中文摘要:
      为探索赤眼鳟信号转导与转录激活因子1(signal transduction and activator of transcription 1, STAT1)的免疫功能,实验利用cDNA末端快速扩增(RACE)技术获得了赤眼鳟STAT1(命名为ScSTAT1),ScSTAT1的cDNA全长为2 922 bp,共编码718个氨基酸。ScSTAT1含STAT_int、STAT_alpha、STAT_bind和SH2 4个保守功能域。系统进化分析显示,ScSTAT1先与草鱼STAT1聚为一支,再与青鱼、鲫以及黄颡鱼等聚为一支。相对于哺乳类和爬行类物种而言,ScSTAT1缺少含丝氨酸磷酸化位点的C-末端转录激活功能结构域(transcriptional activation domain,TAD)。基因表达分析显示,脾脏ScSTAT1表达量极显著高于其他组织,而肝脏中表达量最低。经细菌类似物脂多糖(LPS)、双链 RNA类似物聚肌胞苷酸(Poly I: C)及草鱼呼肠孤病毒(GCRV)刺激后,赤眼鳟脾脏和体肾组织中ScSTAT1表达水平总体上调。GCRV感染后12 h脾脏组织ScSTAT1表达先显著下降,随后极显著上升;GCRV刺激后12和72 h,体肾组织ScSTAT1表达量显著高于对照组;赤眼鳟鳍条细胞系(ScF) ScSTAT1经RNA干扰后基因表达水平极显著下调,且IRF3、IRF9以及Mx均在GCRV刺激48 h时极显著下调。本研究为进一步深入探索ScSTAT1的抗病功能奠定了基础。
英文摘要:
      The grass carp hemorrhage disease poses a serious threat to the long-term expansion of the aquaculture industry. The barbel chub (Squaliobarbus curriculus) could hybrid with the grass carp to produce progeny possessing resistance to GCRV infection and, therefore, are considered valuable genetic resources for studying the molecular mechanisms of grass carp hemorrhage. To investigate the immune function of barbel chub STAT1 (ScSTAT1) against GCRV infection, the RACE (rapid amplification of cDNA ends), qPCR (quantitative polymerase chain reaction) and RNAi (RNA interference) techniques were applied to obtain the full-length cDNA sequence of ScSTAT1, to detect its expression profile in healthy and GCRV-infected tissues, and to explore its basic immune function. The ScSTAT1 was 2 922 bp in length and encoded a protein of 718 amino acid residues. The ScSTAT1 contained conserved domains for STAT_int, STAT_alpha, STAT_binding and SH2. Phylogenetic analysis revealed that the ScSTAT1 was closely clustered with the homologue from Ctenopharyngodon idella, forming an extended clade with those from Mylopharyngodon piceus, Carassius auratus and Tachysurus fulvidraco. Compared to the homologues from mammals and reptiles, ScSTAT1 lacked a C-terminal TAD domain where a serine phosphorylation site is present. The expression level of ScSTAT1 was shown to be the highest in the spleen among the tissues analyzed, with the lowest level detected in the liver. Treatment of fish with LPS, Poly I:C or GCRV resulted in upregulation of expressions of ScSTAT1 in the spleen and kidney. At 12 h post GCRV infection, the ScSTAT1 expression was down-regulated in the spleen, followed by increases. At 12 h and 72 h post GCRV infection, in trunk kidney the expression levels of ScSTAT1 were significantly higher than those in the control group. In the S. curriculus fin cell (ScF) line, knockdown 60% of ScSTAT1 expression by RNA interference led to decreased expression of IRF3, IRF9 and Mx at 48 h post GCRV infection. The results of the present study proved that ScSTAT1 participated in the signal transduction of the IFN system and played a key role in immune reaction against GCRV infection. The results also provide research basis for further study on the functions of ScSTAT1 in disease resistance in fish.
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