title: Molecular characterization, expression patterns, and subcellular localization of RIG-I in the Jinding duck (Anas platyrhynchos domesticus).
abstract: Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) have recently been identified as cytoplasmic sensors for RNA virus. Recent research has shown that RIG-I, a member of this family, play an important role in innate immunity. In this study, we cloned the RIG-I gene from Jinding duck by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). We determined that the cDNA of duRIG-I contains a 14-bp 5' UTR, a 2802-bp open reading frame, and alternative 3' UTRs (295-bp and 927-bp) and encodes a polypeptide of 933 amino acids. Based on this sequence, the duRIG-I protein is predicted to have conserved domains typical of RLRs. In addition, duRIG-I was found to be distributed throughout DF1 cells by indirect immunofluorescence, as predicted. duRIG-I mRNA was scarcely detected in healthy tissues by semi-quantitative RT-PCR (sqRT-PCR). To study the role of RIG-I in innate immunity, we used synthetic double-stranded RNA to mimic viral infection in vivo and detected duRIG-I transcripts in spleen and liver by quantitative real-time PCR (qRT-PCR). The expression of duRIG-I mRNA was significantly elevated at 8h post-injection (P < 0.05) and was indistinguishable from control levels at other time points (P > 0.05). These results suggest that duRIG-I plays an important role in innate immune responses to double-stranded RNA viruses and warrant further studies to reveal the possible mechanism.
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009 Jiangsu, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Chen
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zhengyang
authors.initials: Z
authors.lastname: Huang
authors.forename: Qi
authors.initials: Q
authors.lastname: Xu
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhu
authors.forename: Yiyu
authors.initials: Y
authors.lastname: Tong
authors.forename: Qinmin
authors.initials: Q
authors.lastname: Yu
authors.forename: Jiatong
authors.initials: J
authors.lastname: Ding
authors.forename: Guohong
authors.initials: G
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D053487:DEAD-box RNA Helicases / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004372:Ducks / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D007113:Immunity, Innate / Q000235:genetics / Q000276:immunology; D008099:Liver / Q000276:immunology / Q000378:metabolism; D010802:Phylogeny; D012330:RNA, Double-Stranded / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000276:immunology / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D053487:DEAD-box RNA Helicases
keywords: Duck; Expression pattern; RIG-I; Subcellular localization