The Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Fish Shellfish Immunol. 2010 Feb;28(2):326-32. doi: 10.1016/j.fsi.2009.11.016. Epub 2009 Nov 24.
Galectins are a family of beta-galactoside-binding lectins which play crucial roles in innate immunity of vertebrates and invertebrates. In the present study, the cDNA of a galectin with multiple carbohydrate-recognition domains (CRDs) was cloned from bay scallop Argopectens irradians (designated AiGal1) by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of AiGal1 was of 2235 nucleotides, encoding a polypeptide of 549 amino acids. SMART program analysis revealed that AiGal1 contained four galectin CRDs, and all the CRDs contained the two consensus motifs essential for ligand-binding. Quantitative real-time PCR was employed to investigate the tissue distribution of AiGal1 mRNA and temporal expression in haemocytes of scallops challenged with Vibrio anguillarum, Micrococcus luteus and Pichia pastoris. The AiGal1 mRNA could be detected in all tested tissues with the highest expression level in hepatopancreas. After challenged by V. anguillarum and M. luteus, the expression level of AiGal1 mRNA was both up-regulated and reached the maximum level at 9 h (1.52 fold, P < 0.05) and 18 h (2.89 fold, P < 0.01) post challenge, respectively. However, there was no significant difference in the mRNA expression of AiGal1 in haemocytes after P. pastoris challenge (P > 0.05). These results collectively indicated that AiGal1 was a new member of the galectin family and involved in the immune responses against bacterial infection.
半乳凝集素是一类β-半乳糖苷结合凝集素,在脊椎动物和无脊椎动物的固有免疫中发挥着关键作用。本研究采用表达序列标签(EST)和快速扩增 cDNA 末端(RACE)技术,从海湾扇贝 Argopectens irradians 克隆到一个具有多个糖识别结构域(CRD)的半乳凝集素 cDNA(命名为 AiGal1)。AiGal1 的全长 cDNA 为 2235 个核苷酸,编码 549 个氨基酸的多肽。SMART 程序分析表明,AiGal1 包含四个半乳凝集素 CRD,所有 CRD 都包含配体结合所必需的两个保守基序。采用定量实时 PCR 技术研究了 AiGal1 mRNA 在扇贝组织中的分布及其在血细胞中对鳗弧菌、藤黄微球菌和毕赤酵母刺激的时间表达。AiGal1 mRNA 可在所有检测组织中检测到,在肝胰腺中的表达水平最高。在受到鳗弧菌和藤黄微球菌刺激后,AiGal1 mRNA 的表达水平均上调,并在刺激后 9 h(1.52 倍,P < 0.05)和 18 h(2.89 倍,P < 0.01)达到最高水平。然而,毕赤酵母刺激后血细胞中 AiGal1 mRNA 的表达没有显著差异(P > 0.05)。这些结果共同表明,AiGal1 是半乳凝集素家族的一个新成员,参与了对细菌感染的免疫反应。