DataSheet1.docx

Toll receptors are involved in innate immunity of invertebrates. In this study, we identify and characterize two Toll genes (named HcToll4 and HcToll5) from triangle sail mussel Hyriopsis cumingii. HcToll4 has complete cDNA sequence of 3,162 bp and encodes a protein of 909 amino acids. HcToll5 cDNA is 4,501 bp in length and encodes a protein of 924 amino acids. Both deduced HcToll4 and HcToll5 protein contain signal peptide, extracellular leucine rich repeats (LRRs), and intracellular Toll/interleukin-1 (IL-1) receptor domains. Quantitative real-time PCR analysis revealed that HcToll4 and HcToll5 were largely distributed in the hepatopancreas and could be detected in the gills and mantle. HcToll4 and HcToll5 expression could respond to Staphylococcus aureus, Vibrio parahaemolyticus, White spot syndrome virus (WSSV) or Poly I:C challenge. RNA interference by siRNA results showed that HcToll4 and HcToll5 were involved in the regulation of theromacin (HcThe) and whey acidic protein (HcWAP) expression. Based on these results, HcToll4 and HcToll5 might play pivotal function in H. cumingii innate immune response.Toll受体（Toll receptors）参与无脊椎动物的先天免疫应答。本研究从三角帆蚌（Hyriopsis cumingii）中鉴定并表征了两个Toll基因，分别命名为HcToll4与HcToll5。其中HcToll4的完整cDNA序列长度为3162 bp，编码含909个氨基酸残基的蛋白质；HcToll5的cDNA序列全长为4501 bp，编码含924个氨基酸残基的蛋白质。推导获得的HcToll4与HcToll5蛋白均包含信号肽、胞外亮氨酸重复序列（LRRs）以及胞内Toll/白细胞介素-1（IL-1）受体结构域。实时荧光定量PCR分析结果显示，HcToll4与HcToll5主要在肝胰腺中表达，同时在鳃与外套膜中也可检测到其转录产物。HcToll4与HcToll5的表达可在金黄色葡萄球菌（Staphylococcus aureus）、副溶血性弧菌（Vibrio parahaemolyticus）、白斑综合征病毒（WSSV）以及聚肌胞苷酸（Poly I:C）刺激下发生应答。小干扰RNA（siRNA）介导的RNA干扰实验结果表明，HcToll4与HcToll5参与调控theromacin（HcThe）与乳清酸性蛋白（HcWAP）的基因表达。基于上述实验结果，HcToll4与HcToll5可能在三角帆蚌的先天免疫应答中发挥关键作用。