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首页> 外文期刊>Fish & Shellfish Immunology >A galectin from roughskin sculpin, Trachiderrnus fasciatus: Molecular cloning and characterization
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A galectin from roughskin sculpin, Trachiderrnus fasciatus: Molecular cloning and characterization

机译：来自粗皮杜鹃的半乳糖凝集素：分子克隆和表征

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Galectins are a family of beta-galactoside-binding lectins, which have been proved to be involved in host-pathogen interactions by recognizing pathogen associated molecular patterns (PAMPs) on the surface of virus, bacteria, fungi and protozoa. In this study, a galactoside-binding lectin homolog was identified from roughskin sculpin Trachidermus fasciatus, named TfGal. The full-length of TfGal cDNA was 1016 bp with a 5' untranslated region (UTR) of 134 bp and a 3' UTR of 474 bp, and the open reading frame (ORF) is 408 bp. The deduced protein was composed of 135 amino acids, including a carbohydrate-recognition domain and a galactoside-type carbohydrate-binding motif H-NPR/W-E-R. The deduced amino acid sequence shared 58.52%-87.4% similarities with the galectins of the other fishes. Quantitative real-time PCR (qRT-PCR) analysis demonstrated that TfGal mRNA was abundantly expressed in the ovary, heart, stomach, skin, moderately expressed in the liver, brain, gills, spleen, and rarely expressed in the hemocytes, meat and intestine. The expression of TfGal mRNA in the hemocytes and the skin was dramatically up-regulated after challenged with LPS, reaching the highest level at 2 h post-challenge, and then dropped abruptly, while the expression of TfGal mRNA in the liver was upregulated at 2-6 h post-challenge, and then returned to the normal level, with an increase at 96 h post-challenge again. However, no obvious change of the expression of TfGal mRNA was detected in the stomach. Recombinant TfGal purified from Escherichia coil (BL21) could agglutinate and/or bind microorganisms in Ca2+-independent manner. These results suggested that TfGal might be involved in the innate immune response of roughskin sculpin. (C) 2013 Elsevier Ltd. All rights reserved.
机译：半乳凝素是β-半乳糖苷结合凝集素的一个家族，已通过识别病毒，细菌，真菌和原生动物表面的病原体相关分子模式（PAMP）被证明参与宿主-病原体相互作用。在这项研究中，半乳糖苷结合的凝集素同系物是从粗皮雕刻的Trachidermus fasciatus中鉴定出来的，名为TfGal。 TfGal cDNA的全长为1016 bp，5'非翻译区（UTR）为134 bp，3'UTR为474 bp，开放阅读框（ORF）为408 bp。推导的蛋白质由135个氨基酸组成，包括一个碳水化合物识别结构域和一个半乳糖苷型碳水化合物结合基序H-NPR / W-E-R。推导的氨基酸序列与其他鱼类的半乳凝素共有58.52％-87.4％的相似性。实时定量PCR（qRT-PCR）分析表明TfGal mRNA在卵巢，心脏，胃，皮肤中大量表达，在肝，脑，腮，脾中中等表达，而在血细胞，肉和肠中很少表达。 LPS攻击后，血细胞和皮肤中TfGal mRNA的表达显着上调，在攻击后2小时达到最高水平，然后突然下降，而肝脏中TfGal mRNA的表达在2时上调。攻击后-6小时，然后恢复到正常水平，并在攻击后96小时再次增加。然而，在胃中未检测到TfGal mRNA表达的明显变化。从大肠杆菌（BL21）中纯化的重组TfGal可以以不依赖Ca2 +的方式凝集和/或结合微生物。这些结果表明，TfGal可能参与了粗皮杜鹃的天然免疫反应。 （C）2013 Elsevier Ltd.保留所有权利。

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