斜带石斑鱼核苷酸结合寡聚化蛋白NOD及NOD的表达分析.doc

斜带石斑鱼核苷酸结合寡聚化蛋白NOD1及NOD2的表达分析 Comparative analysis of nuclear oligomerization domainproteins NOD -1 and NOD2 in orange-spotted grouper5101520253035 Epinephelus coioides #Abstract: Nucleotide-binding oligomerization domain-containing proteins-1 and-2 NOD1 and NOD2 are members of the NOD-like receptors NLRs family. They are both cytoplasmic receptors, and sensemicrobial infections/danger molecules to induce host innate immune response. In this study, thefull-length ORF sequences of NOD1 and NOD2 were cloned, and the putative amino acid sequenceswere identi?ed in orange-spotted grouper Epinephelus coioides . The complete open reading frame ORF of grouper NOD1 contained 2823 bp encoding a 940 amino acid protein. Grouper NOD2 cDNAcontained a 2967 bp ORF, encoding a protein of 988 amino acid residues. Both grouper NOD1 andNOD2 had similar domains to human and fish counterparts. Phylogenetic tree analysis showed thatgrouper NOD1 clustered with grass carp, zebra?sh and channel cat?sh, while NOD2 was most closelyrelated to fugu. Expression patterns of grouper NOD1 and NOD2 were next studied. NOD1 had thehighest level of expression in skin while NOD2 in trunk kidney. Post Vibrio alginolyticus strainEcGS020401 , lipopolysaccharide LPS or PolyI:C challenges, gene expression of grouper NOD1 andNOD2 was stimulated to different extents. NOD1 showed a significant enhancement after LPSstimulation, but NOD2 increased more significantly after PolyI:C invasion, indicating that NOD1 andNOD2 may exert different effects on the eradication of bacteria and virus. The adaptor proteinRIP-like-interacting CLARP kinase RICK and downstream molecule interleukin-8 IL-8 were alsoinduced at different levels after stimulation, which indicated that NOD1 and NOD2 signal transductionwas involved in grouper innate immune protection against bacterial and viral infections.Key words: Orange-spotted grouper; NOD1; NOD2; RICK; IL-80 IntroductionNucleotide-binding oligome