赤眼鳟Toll样受体9基因cDNA全长克隆及表达分析
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- 英文论文题名：Molecular cloning and expression analysis of Toll-like receptor 9 gene in Squaliobarbus curriculus
- 论文作者：金生振 ; 王荣华 ; 周智愚 ; 刘巧林 ; 徐莹 ; 肖调义
- 英文论文作者：JIN Shengzhen ; WANG Ronghua ; ZHOU Zhiyu ; LIU Qiaolin ; XU Ying ; XIAO Tiaoyi ; Hunan Engineering Research Center for Utilization of Characteristics of Aquatic resources ; Hunan agricultural university ; Collaborative Innovation Center for Efficient and Health Production of Fisheries in Hunan Province
- 年：2016
- 作者机构：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;水产高效健康生产湖南省协同创新中心;
- 论文关键词：赤眼鳟 ; Toll样受体9 ; 分子克隆 ; 表达特征
- 英文论文关键词：Squaliobarbus curriculus ; Toll-like Receptor 9 ; Molecular cloning ; Expression characterizations
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S943
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：561k
- 原文格式：D
- 会议级别：全国
摘要
为研究赤眼鳟(Squaliobarbus curriculus)Toll样受体9(Toll-like receptor 9,ScTLR9)基因是否参与抗病毒免疫反应,通过RACE技术,首次克隆和分析了ScTLR9基因cDNA全长序列;通过Real-Time qPCR技术,检测了ScTLR9 mRNA在健康赤眼鳟10个组织中的分布以及感染草鱼呼肠孤病毒(GCRV)后在赤眼鳟和草鱼(Ctenopharyngodon idellus)头肾中的表达特征。结果表明:ScTLR9基因cDNA序列全长3687 bp,编码1059个氨基酸残基,推导的蛋白质分子量为121.61 k D,等电点为8.91;SMART结构域预测显示,ScTLR9由N端的信号肽(SP)、富亮氨酸结构域(LRRs)、跨膜结构域(TM)和C端的Toll/白介素-1受体结构域(TIR)组成。实时荧光定量结果显示,ScTLR9 mRNA在检测的各组织中均有表达,其在头肾中的相对表达量极显著高于其它组织;感染GCRV后,ScTLR9和Ci TLR9表达水平表现出明显的差异。研究显示,ScTLR9具有TLRs家族基因的典型结构特征,并能被GCRV诱导,推测其在赤眼鳟抗GCRV入侵免疫反应中发挥了重要作用。
In order to investigate whether the toll-like receptor 9(ScTLR9) gene involved in antiviral immune response, The S.curriculus TLR9 cDNA of full sequence was cloned and analyzed for the first time with RACE technique. Using real-time qPCR to analysis relative expression of ScTLR9 mRNA in 10 different tissues and the relative expression in head kidney after infected by GCRV at different times in Barbel chub and Grass carp.The results showed that the complete sequence of ScTLR9 was 3687 bp, encoding 1059 amino acid residues. The putative p I and Mw of ScTLR9 was 8.91 and 121.61 k D. The ScTLR9 motifs was consisted of SP, LRRs,TM, and TIR. The relative expression in head kidney significantly higher than other groups; After GCRV infection, ScTLR9 and Ci TLR9 expression in head kidney showed obvious difference.These results suggest that ScTLR9 had typical structure characteristics of TLRs family, and play an important role in the immune response of barbel chub against GCRV infection.
In order to investigate whether the toll-like receptor 9(ScTLR9) gene involved in antiviral immune response, The S.curriculus TLR9 cDNA of full sequence was cloned and analyzed for the first time with RACE technique. Using real-time qPCR to analysis relative expression of ScTLR9 mRNA in 10 different tissues and the relative expression in head kidney after infected by GCRV at different times in Barbel chub and Grass carp.The results showed that the complete sequence of ScTLR9 was 3687 bp, encoding 1059 amino acid residues. The putative p I and Mw of ScTLR9 was 8.91 and 121.61 k D. The ScTLR9 motifs was consisted of SP, LRRs,TM, and TIR. The relative expression in head kidney significantly higher than other groups; After GCRV infection, ScTLR9 and Ci TLR9 expression in head kidney showed obvious difference.These results suggest that ScTLR9 had typical structure characteristics of TLRs family, and play an important role in the immune response of barbel chub against GCRV infection.
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