【作者】 余丽丽； 林鹏； 郭松林； 王艺磊； 张子平； 王婷婷； 冯建军；

【Author】 YU Lili;LIN Peng;GUO Songlin;WANG Yilei;ZHANG Ziping;WANG tingting;FENG Jianjun;Engineering Research Centre of Eel Modern Technical Industry, Ministry of Education;Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture;Fisheries College, Jimei University;

【机构】 集美大学水产学院鳗鲡现代产业技术教育部工程研究中心农业部东海海水健康养殖重点实验室； 福建农林大学动物科学学院；

【摘要】 Toll样受体3(TLR3)是用于识别双链RNA的一种重要模式识别受体。为探索TLR3的序列特征及其功能,利用RACE技术克隆日本鳗鲡TLR3基因cD NA全长序列,命名为AjT LR3,全长3383bp,开放阅读框为2766bp,编码921个氨基酸,AjT LR3具有16个LRR的胞外结构域、跨膜结构域以及TIR结构域。采用实时荧光定量PCR(qR T-PCR)检测AjT LR3的表达情况,发现其在日本鳗鲡各组织器官中均有表达;经poly I:C刺激后鳃与肾无明显变化,其余均显著提高;而LPS刺激后,仅在肝脏和肠中有显著提高。日本鳗鲡肝脏细胞体外实验结果显示:poly I:C处理后12h和24h表达水平显著增高,CpG-DNA和肽聚糖处理后24h表达量均有显著增加;而细菌浓度达到107cfu/m L和108cfu/mL后,AjT LR3基因表达水平分别在24h、12h显著增高并达到峰值。以上研究表明,AjT LR3在日本鳗鲡抵御病毒及细菌的免疫应答过程中具有重要作用。更多还原

【Abstract】 Toll-like receptor 3(TLR3) is one of the most significant members in TLRs family which is renowned as pattern recognition receptor(PRR) recognize double-stranded RNA(ds RNA). TLR3 was characterized and analyzed for the first time in Anguilla japonica which named AjT LR3. The full-length cD NA sequence of AjT LR3 is composed of 3383 bp with an open reading frame(ORF) of 2766 bp encoding a polypeptide of 921 amino acid residues. Analysis of the deduced amino acid sequence indicated that AjT LR3 protein has three main structural domains, extracellular domain which contains 16 leucine-rich repeats(LRRs) motifs, a transmembrane region and a Toll/interleukin-1 receptor(TIR) domain. Quantitative real-time polymerase chain reaction(q RT-PCR) analysis revealed a broad expression for AjT LR3 in all tissues; The AjT LR3 expressions in blood, intestine, liver, spleen, skin, heart, and muscle were significantly induced after injection with the viral mimic poly I:C, only in the liver and intestine significantly improved with the stimulation LPS. In vitro, the AjT LR3 transcripts of Japanese eel liver cells up-regulated with poly I:C treatment at 12 h and 24 h, whereas there were significantly enhanced by PGN and Cp G-DNA stimulation after 24 h; And the expression level of AjT LR3 gene was significantly increased at 24 h and 12 h after the bacterial concentration reached 107 cfu/mL and 108 cfu/mL, respectively. These results collectively suggested AjT LR3 transcripts expression possibly play an important role in fish defense against viral and bacterial infection.更多还原