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ZFIN ID: ZDB-PUB-150211-14
Isolation and expression of grass carp toll-like receptor 5a (CiTLR5a) and 5b (CiTLR5b) gene involved in the response to flagellin stimulation and grass carp reovirus infection
Jiang, Y., He, L., Ju, C., Pei, Y., Ji, M., Li, Y., Liao, L., Jang, S., Zhu, Z., Wang, Y.
Date: 2015
Source: Fish & shellfish immunology   44(1): 88-99 (Journal)
Registered Authors: Zhu, Zuoyan
Keywords: GCRV, MyD88, NF-κB, flagellin, grass carp, toll-like receptor 5
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Carps*
  • Cloning, Molecular
  • DNA, Complementary/genetics
  • DNA, Complementary/metabolism
  • Fish Diseases/genetics*
  • Fish Diseases/immunology*
  • Fish Diseases/virology
  • Fish Proteins/chemistry
  • Fish Proteins/genetics*
  • Fish Proteins/metabolism
  • Flagellin/pharmacology*
  • Gene Expression Regulation
  • Molecular Sequence Data
  • Organ Specificity
  • Phylogeny
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • Reoviridae/physiology
  • Reoviridae Infections/genetics
  • Reoviridae Infections/immunology
  • Reoviridae Infections/veterinary*
  • Reoviridae Infections/virology
  • Salmonella typhimurium/physiology
  • Toll-Like Receptor 5/chemistry
  • Toll-Like Receptor 5/genetics*
  • Toll-Like Receptor 5/metabolism
PubMed: 25665802 Full text @ Fish Shellfish Immunol.
Toll-like receptor 5 (TLR5), a member of Toll-like receptors (TLRs) family and is responsible for the bacterial flagellin recognition in vertebrates, play an important role in innate immunity. In the study, two TLR5 genes of grass carp (Ctenopharyngodon idellus), named CiTLR5a and CiTLR5b, were cloned and analyzed. Both CiTLR5a and CiTLR5b are typical TLR proteins, including LRR motif, transmembrane region and TIR domain. The full-length cDNA of CiTLR5a is 3054bp long, with a 2646bp open reading frame (ORF), 78bp 5' untranslated regions (UTR), and 330bp 3' UTR. The full-length cDNA of CiTLR5b is 3326bp, with a 2627bp ORF, 95bp 5' UTR, and 594bp 3' UTR. Phylogenetic analysis showed that CiTLR5a and CiTLR5b were closed to the TLR5 of cirrhinus mrigala, cyprinus_carpio, and danio rerio. Subcellular localization indicated that CiTLR5a and CiTLR5b shared similar localization pattern and may locate in the plasma membrane of transfected cells. Real-time quantitative PCR revealed CiTLR5a and CiTLR5b were constitutively expressed in all examined tissues, whereas the highest expressed tissue differed. Following exposure to flagellin and GCRV, CiTLR5a and CiTLR5b were up-regulated significantly. Moreover, the downstream genes of TLR5 signal pathway such as MyD88, NF-κB, IRF7, IL-1β, and TNF-α also up-regulated significantly, whereas the IκB gene was down-regulated, suggesting that CiTLR5a and CiTLR5b involved in response to flagellin stimulation and GCRV infection. The results obtained in the study would provide a new insight for further understand the function of TLR5 in teleost fish.