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首页> 外文期刊>Fish & Shellfish Immunology >Molecular characterization and expression analysis of TLR 7 and TLR 8 homologs in large yellow croaker (Pseudosciaena crocea)
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Molecular characterization and expression analysis of TLR 7 and TLR 8 homologs in large yellow croaker (Pseudosciaena crocea)

机译：大黄鱼中TLR 7和TLR 8同源物的分子表征和表达分析

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摘要

The two toll-like receptor (TLR) genes, LycTLR7 and LycTLR8, were cloned from large yellow croaker (Pseudosciaena crocea), an economically important marine fish in China. The full-length cDNAs of LycTLR7 and LycTLR8 are 3544 and 3593 bp, with an open reading frame (ORF) of 3165 and 3093 bp, encoding 1053 and 1030 amino acids, respectively. The TLR family motifs, such as leucine rich repeat (LRR) and Toll/interleukin (IL)-1 receptor (TIR) domain, are conserved in the LycTLR7 and LycTLR8, with 17 and 14 LRRs, and with a TIR domain, respectively. It is also noted that an LRR N-terminal domain (LRR-NT, residues 24-60) is present in the LycTLR7 but not in the LycTLR8. Both LycTLR7 and LycTLR8 contain a conserved extracellular CxRCxxxxxPCxxC motif, which was found in TLR7/TLR8 of other species and required for stimulus-induced signal transduction. Homology comparison shows that LycTLR7 has 79%, 71.9%, 65.9% and 65.8% identity to fugu, rainbow trout, carp and catfish TLR7, while LycTLR8 has 67.1%, 60.7%, 60.6%, 52.4%, and 51.5% identity to fugu TLR8, rainbow trout TLR8a1, rainbow trout TLR8a2, catfish TLR8-2, and catfish TLR8-1, respectively. Subcellular localization analysis revealed that both LycTLR7 and LycTLR8 are located in the endoplasmic reticulum of epithelioma papulosum cyprini (EPC) cells, which is similar to TLR7/TLR8 in mammals. The two TLRs were constitutively expressed in all tissues tested, especially in immune-related tissues such as spleen, head kidney and gills. An increased expression of LycTLR7 and LycTLR8 was observed in head kidney and spleen of large yellow croakers stimulated by poly (I:C), a viral mimic. In head kidney, their mRNA expression was up-regulated more than 10 times compared to the controls at 12 h after poly (I:C) stimulation. These results suggested that LycTLR7 and LycTLR8 may play a role in the defense against viral infection like their mammalian homologs. (C) 2013 Elsevier Ltd. All rights reserved.
机译：这两个通行费样受体（TLR）基因LycTLR7和LycTLR8是从大型黄鱼（Pseudosciaena crocea）（中国经济上重要的海鱼）中克隆而来的。 LycTLR7和LycTLR8的全长cDNA为3544和3593 bp，开放阅读框（ORF）为3165和3093 bp，分别编码1053和1030个氨基酸。 TLR家族基序，例如富含亮氨酸的重复序列（LRR）和Toll /白介素（IL）-1受体（TIR）域，在LycTLR7和LycTLR8中保守，分别具有17和14个LRR和一个TIR域。还应注意，LyrcTLR7中存在LRR N末端结构域（LRR-NT，残基24-60），而LycTLR8中不存在。 LycTLR7和LycTLR8均包含保守的细胞外CxRCxxxxxPCxxC基序，该基序在其他物种的TLR7 / TLR8中发现，并且是刺激诱导的信号转导所必需的。同源性比较显示LycTLR7与河豚，虹鳟鱼，鲤鱼和cat鱼TLR7的同一性分别为79％，71.9％，65.9％和65.8％，而LycTLR8与河豚的同一性为67.1％，60.7％，60.6％，52.4％和51.5％ TLR8，虹鳟鱼TLR8a1，虹鳟鱼TLR8a2，cat鱼TLR8-2和cat鱼TLR8-1。亚细胞定位分析表明，LycTLR7和LycTLR8都位于乳头丘疹上皮细胞（EPC）细胞的内质网中，与哺乳动物中的TLR7 / TLR8类似。在所有测试的组织中，尤其是在免疫相关的组织（如脾脏，头部肾脏和腮）中，两个TLR均组成性表达。在病毒模拟的聚（I：C）刺激下，在大的黄花鱼的头部肾脏和脾脏中观察到LycTLR7和LycTLR8的表达增加。在头肾中，在多（I：C）刺激后12小时，与对照组相比，其mRNA表达上调了10倍以上。这些结果表明LycTLR7和LycTLR8可能像它们的哺乳动物同系物一样在防御病毒感染中发挥作用。 （C）2013 Elsevier Ltd.保留所有权利。

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