Molecular characterization, tissue distribution and functional analysis of galectin 1-like 2 in grass carp (Ctenopharyngodon idella).
Molecular characterization, tissue distribution and functional analysis of galectin 1-like 2 in grass carp (Ctenopharyngodon idella).
复制标题
DOI：
10.1016/j.fsi.2019.09.041
复制
发表时间：
2019-11
影响因子：
4.7
通讯作者：
Denghui Zhu;P. Fu;Rong Huang;Lv Xiong;Yumeng Wang;Libo He;L. Liao;Yongming Li;Zuoyan Zhu;Yaping Wang
中科院分区：
文献类型：
--
作者：
Denghui Zhu;P. Fu;Rong Huang;Lv Xiong;Yumeng Wang;Libo He;L. Liao;Yongming Li;Zuoyan Zhu;Yaping Wang
半乳糖凝集素是凝集素超家族中进化保守的一类，具有识别病原菌、抗细菌和抗病毒等功能。本研究从草鱼（Ctenopharyngodon idella）中获得了一段405 bp的半乳凝素1-like 2（CiGal 1-L2）cDNA序列，编码134个氨基酸，预测分子量为15.143 kDa，等电点为5.33。在糖结合结构域（CRD）中检测到糖结合基序（H-N-R、V-N和W-E-R）。氨基酸序列同源性分析表明，CiGal 1-L2与哺乳动物和鱼类的同源性分别为40.30-42.54%和66.42-81.20%。系统进化树分析表明，CiGal 1-L2与鱼类galectin-1 s聚在一起，与鲤鱼（Cyprinus carpio）亲缘关系较近。实时荧光定量PCR（RT-qPCR）分析表明，CiGal 1-L2在所有受试组织中广泛表达。此外，在草鱼呼肠孤病毒（GCRV）、脂多糖（LPS）和聚肌胞苷酸（poly I：C）的诱导下，CiGal 1-L2的表达量显著上调。在LPS和poly I：C处理后，CiGal 1-L2-GFP的荧光分布于HEK 293 T细胞的胞浆和胞核，并有核转位的趋势。重组CiGal 1-L2（rCiGal 1-L2）蛋白对LPS具有较强的结合能力。总之，这些结果为深入了解galectin-1在硬骨鱼类中的免疫作用提供了依据。
Galectins, as an evolutionary conserved group of lectin superfamily, has the functions of pathogen recognition, anti-bacteria and anti-virus. In this study, a 405 bp cDNA sequence of galectin 1-like 2 (CiGal1-L2) was obtained from grass carp (Ctenopharyngodon idella), which encoded 134 amino acids with a predicted molecular mass of 15.143 kDa and an isoelectric point of 5.33. The sugar binding motifs (H–N-R, V–N and W--E-R) were detected in carbohydrate-binding domain (CRD). The amino acid sequence similarity showed that CiGal1-L2 was 40.30–42.54% and 66.42–81.20% similarity to mammalian and fish counterparts, respectively. The phylogenetic tree showed that CiGal1-L2 was clustered with fish galectin-1s and closely related toCyprinus carpio. Real-time quantitative PCR (RT-qPCR) analysis revealed thatCiGal1-L2was widely expressed in all tested tissues. In addition, the expression ofCiGal1-L2was differentially up-regulated challenged with grass carp reovirus (GCRV), lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C). The fluorescence of CiGal1-L2-GFP was distributed in the cytoplasm and nucleus of HEK 293T cells and showed a trend of nuclear translocation after LPS and poly I:C treatment. Finally, the recombinant CiGal1-L2 (rCiGal1-L2) protein showed strong binding ability to LPS. In conclusion, the results provided further insight into the immune roles of galectin-1 in teleost.