Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou, Guangdong 515063, China.
Fish Shellfish Immunol. 2013 Jan;34(1):129-35. doi: 10.1016/j.fsi.2012.10.018. Epub 2012 Oct 26.
Infectious diseases have seriously inhibited the aquaculture of mud crab Scylla paramamosain in southeastern China. Identification of the immune molecules and characterization of the defense mechanisms will be pivotal to the reduction of these diseases. Available data show that leucine-rich repeat (LRR) proteins play a crucial role in protein-protein interactions, recognition processes or immune reactions in both invertebrates and vertebrates. In the present study, we cloned and characterized a LRR cDNA from the mud crab Scylla paramamosain (SpLRR) by using the RACE strategy. Bioinformatics analysis predicted that SpLRR contains one open reading frame of 1893 bp and encodes a LRR protein of 630 amino acids with 17 LRR domains and 5 potential N-glycosylation sites. Further, SpLRR and other arthropod LRR proteins could be clustered into one branch in a phylogenetic tree. SpLRR transcripts were detected using RT-PCR from examined tissues including heart, gill, stomach, intestine, muscle and hemocytes, whereas not from hepatopancreas. More importantly, the SpLRR mRNA expression was up-regulated after infection with Vibrio alginolyticus, Beta streptococcus or Poly I: C for 12-48 h, suggesting a novel LRR homolog in crab might be associated with the resistance to pathogens. The result could be important for future investigation of crab immune defense mechanisms.
传染病严重抑制了中国东南地区泥蟹锯缘青蟹的水产养殖。鉴定免疫分子和描述防御机制对于减少这些疾病至关重要。现有数据表明,富含亮氨酸重复(LRR)蛋白在无脊椎动物和脊椎动物的蛋白质-蛋白质相互作用、识别过程或免疫反应中起着关键作用。本研究采用 RACE 策略,从泥蟹锯缘青蟹中克隆并鉴定了一个 LRR cDNA(SpLRR)。生物信息学分析预测 SpLRR 包含一个 1893 bp 的开放阅读框,编码一个 630 个氨基酸的 LRR 蛋白,具有 17 个 LRR 结构域和 5 个潜在的 N-糖基化位点。此外,SpLRR 和其他节肢动物 LRR 蛋白可以在系统发育树中聚类到一个分支中。使用 RT-PCR 从心、鳃、胃、肠、肌肉和血细胞等检测组织中检测到 SpLRR 转录物,而在肝胰腺中未检测到。更重要的是,在感染溶藻弧菌、β链球菌或聚肌胞后 12-48 小时,SpLRR mRNA 的表达上调,这表明蟹中一种新的 LRR 同源物可能与对病原体的抗性有关。该结果对未来研究蟹的免疫防御机制具有重要意义。