Characterization and expression analysis of Toll-like receptor 2 gene in large yellow croaker, Larimichthys crocea
Characterization and expression analysis of Toll-like receptor 2 gene in large yellow croaker, Larimichthys crocea
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DOI：
10.1016/j.fsi.2015.01.037
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发表时间：
2015-05-01
影响因子：
4.7
通讯作者：
Yao, Cui-Luan
中科院分区：
文献类型：
--
作者：
Fan, Ze-Jun;Jia, Qiao-Jing;Yao, Cui-Luan
Toll样受体2（TLR 2）在天然免疫应答中起重要作用。本文报道了大黄鱼Toll样受体2（LcTLR 2）全长cDNA序列的分离和鉴定。LcTLR 2 cDNA的5 '端非翻译区（5'-UTR）长135 bp，开放阅读框（ORF）长2478 bp，编码825个氨基酸，3 '端非翻译区（5'-UTR）长50 bp。亚细胞定位分析表明LcTLR 2-pEGFP主要表达于细胞浆。实时定量逆转录PCR（qRT-PCR）分析显示LcTLR 2在大多数检查的组织中广泛表达，其中在血液中的表达最多，其次是脾脏，并且在胃中的表达最弱。研究了注射副溶血性弧菌、脂多糖（LPS）和多聚肌苷酸：胞苷酸（polyI：C）后脾脏、头肾和肝脏中LcTLR 2的表达水平。我们的结果显示，在所有三次免疫攻击后，LcTLR 2转录物显著增加（p < 0.05）。然而，与polyI：C和LPS相比，LcTLR 2在副溶血性链球菌刺激后的所有检测组织中诱导了更高的表达水平。此外，我们还研究了鞭毛蛋白、多聚I：C、肽聚糖（PGN）和LPS刺激对LCK中LcTLR 2表达的影响，结果表明，鞭毛蛋白和PGN刺激后LcTLR 2的表达量显著增加，提示LcTLR 2可能在鱼类防御细菌感染中发挥重要作用。此外，与LPS相比，鞭毛蛋白和肽聚糖可能在大黄鱼LcTLR 2的诱导中起重要作用。(C)2015爱思唯尔有限公司版权所有。
Toll-like receptor 2 (TLR2) plays an important role in innate immune responses. Here we describe the isolation and characterization of the full-length cDNA sequence of toll-like receptor 2 in large yellow croaker Larimichthys crocea (LcTLR2). The LcTLR2 cDNA contains a 5'-terminal untranslated region (5'-UTR) of 135 bp, an open reading frame (ORF) of 2478 bp encoding a polypeptide of 825 amino acid residues and a 3'-UTR of 50 bp. Subcellular localization analysis suggested that the LcTLR2-pEGFP was mainly expressed in cytoplasm. Quantitative real-time reverse transcription PCR (qRT-PCR) analysis revealed a broad expression of LcTLR2 in most examined tissues, with the most predominant expression in blood, followed by spleen, and the weakest expression in stomach. The expression levels of LcTLR2 after injection with Vibrio parahaemolyticus, Lipopolysaccharides (LPS) and poly inosinic:cytidylic (polyI:C) were investigated in spleen, head-kidney and liver. Our results showed that LcTLR2 transcripts increased significantly after all the three immune challenges (p < 0.05). However, compared with polyI:C and LPS, higher expression levels of LcTLR2 were induced in all examined tissues after parahaemolyticus stimulation. In addition, the expression levels of LcTLR2 after flagellin, polyl:C, peptidoglycan (PGN) and LPS challenge in LCK were investigated, our findings showed that high LcTLR2 transcripts were induced after flagellin and PGN stimulation, suggesting that LcTLR2 might play a vital role in fish defense against bacterial infection. Furthermore, compared with LPS, flagellin and peptidoglycan might play an important role in LcTLR2 induction in large yellow croaker. (C) 2015 Elsevier Ltd. All rights reserved.