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Correlation between grass carp (Ctenopharyngodon idella) resistance to grass carp reovirus and the genetic insert-deletion polymorphisms in promoter and intron of RIG-I gene
Authors:Quanyuan Wan  Jianguo SuLan Wang  Limin PengLijun Chen
Institution:College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
Abstract:RIG-I (retinoic acid-inducible gene I) is an essential cytosolic pathogen recognition receptor that binds to a variety of viral RNA or DNA to induce type I interferons. In the present study, insert–deletion polymorphisms in promoter and introns of CiRIG-I (Ctenopharyngodon idella RIG-I) were explored, their associations with resistance/susceptibility to grass carp reovirus (GCRV) were analyzed. To this end, genomic sequence of CiRIG-I gene was obtained, and twenty pairs of primers were prepared for the detection of insert–deletion polymorphisms. Five insert–deletion mutations were found, a 2-bp mutation and an 8-bp mutation existed in the promoter and other three sizes in 74 bp, 146 bp and 53 bp were sited in the intron 8. After a challenge experiment, only the genotype and allele of − 740 insert–deletion mutation in the promoter and allele of 6804 insert–deletion mutation were significantly associated with resistance/susceptibility to GCRV among the five mutations (P < 0.05). To further identify this correlation, another independent challenge test was carried out. The result revealed that the cumulative mortality in ins/ins genotype individuals (43.75%) at − 740 insert–deletion mutation was significantly lower than that in ins/del (72.09%) and del/del (74.19%) genotypes (P < 0.05). Linkage disequilibrium and haplotype analysis showed 6610 insert–deletion mutation and 6804 insert–deletion mutation were linkage disequilibrium. The haplotype ins–ins (6610ins–6804ins) was significantly susceptible to GCRV, and ins–del (6610ins–6804del) was significantly resistant to GCRV (P < 0.05). Those could be potential gene markers for the future molecular selection of strains that are resistant to GCRV.
Keywords:CARD  N-terminal caspase recruitment domain  CASP8  caspase 8 gene  CLR  C-type lectin receptor  dsRNA  double strand RNA  ecCuZnSOD  extracellular copper&ndash  zinc superoxide dismutase gene  GCRV  grass carp reovirus  IFN  interferon  LGP2  laboratory of genetics and physiology 2  IL  interleukin  IPS-1  interferon-β promoter stimulator 1  IRF5 gene  interferon regulatory factor 5 gene  MDA5  melanoma differentiation associated gene 5  MHC gene  major histocompatibility complex gene  NLR  nucleotide-oligomerization domain (NOD)-like receptor  PAMP  pathogens associated molecular pattern  PRR  pattern recognition receptor  RD  C-terminal repression domain  RFLP  restriction-fragment length polymorphism  RIG-I  retinoic acid-inducible gene I  RLR  retinoic acid-inducible gene-I (RIG-I)-like receptor  TLR  toll-like receptor
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