Genomic sequence comparison, promoter activity, SNP detection of RIG-I gene and association with resistance/susceptibility to grass carp reovirus in grass carp (Ctenopharyngodon idella)

Dev Comp Immunol. 2013 Apr;39(4):333-42. doi: 10.1016/j.dci.2012.12.004. Epub 2012 Dec 28.

Abstract

As an intracellular pattern recognition receptor (PRR), retinoic acid-inducible gene-I (RIG-I) is responsible for detection of nucleic acids from pathogens in infected cells and activation of type I interferon (IFN). In the present study, the 5'-flanking region, introns and single nucleotide polymorphisms (SNPs) of CiRIG-I (Ctenopharyngodon idella RIG-I) were identified and characterized. The genomic CiRIG-I was 12810 bp in length, consisted of an 1864 bp 5'-flank region whose promoter activity was confirmed, 15 exons and 14 introns. By pooled DNA sequencing, two SNPs were detected in the 5'-flanking region; 10 SNPs were discovered in introns; and one SNP was found in exons. After a challenge experiment, these SNPs were selected to analyze their association with the resistance/susceptibility of C. idella to grass carp reovirus (GCRV), using case-control study. Chi-square test was employed to assess the association. The result showed that -780 C/T, 4731 C/T, 4945 A/G, 8461 C/T, and haplotype 3428A-3432G were significantly associated with the phenotype (P<0.05). To confirm the correlation, another independent challenge experiment was performed, in which the cumulative mortality of -780 genotype CC, 4731 genotype CC and 4945 genotype AA were significantly lower than that of -780 genotype TT, 4731 genotype TT and 4945 genotype GG, respectively (P<0.05). In addition, the SNP-SNP interaction analysis revealed that there was no significant interaction among those SNPs (P>0.05). These significant SNPs and the haplotype might be potential genetic markers for the molecular selection of C. idella strains that are resistant to GCRV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Biomarkers
  • Carps / genetics*
  • Carps / immunology
  • Carps / virology
  • Case-Control Studies
  • Cells, Cultured
  • Disease Susceptibility
  • Fish Diseases / immunology*
  • Fish Diseases / virology
  • Genetic Predisposition to Disease
  • Genotype
  • Interferon Type I / immunology
  • Polymorphism, Single Nucleotide
  • Promoter Regions, Genetic
  • Receptors, Pattern Recognition / genetics*
  • Receptors, Pattern Recognition / immunology
  • Receptors, Retinoic Acid / genetics
  • Receptors, Retinoic Acid / metabolism*
  • Reoviridae / immunology
  • Reoviridae Infections / immunology
  • Reoviridae Infections / veterinary*
  • Reoviridae Infections / virology
  • Sequence Analysis, DNA / veterinary

Substances

  • Biomarkers
  • Interferon Type I
  • Receptors, Pattern Recognition
  • Receptors, Retinoic Acid