Abstract
The interferon-related developmental regulator 1 (IFRD1) protein is expected to play a role in the regulation of inflammatory responses in adult mice, since it is known to repress transcription of NF-κB in myoblasts that regenerate skeletal muscle after traumatic injury Micheli et al., 2011. The IFRD2 gene is expressed in many tissues including skeletal muscle, kidney, heart, brain, lung, placenta and liver in adult humans and is highly expressed in adult human skeletal muscle and heart. In mice, interferon-related developmental regulator 2 (IFRD2) may be associated with early haematopoiesis after gastrulation and in the hepatic primordium Buanne et al., 1998. In this study, we analysed the molecular characteristics of the IFRD2 gene identified from Pagrus major (PmIFRD2) and performed multiple alignments and phylogenetic analyses of the protein sequence. In addition, we examined the expression pattern of IFRD2 in healthy red sea bream tissues and the temporal expression pattern after challenging with various pathogens [Edwardsiella piscicida (E. piscicida), Streptococcus iniae (S. iniae) and red sea bream iridovirus (RSIV)]. This study characterises the non-specific immune response of the red sea bream after viral and microbial infections.
Highlights
NF-kB in myoblasts that regenerate skeletal muscle after traumatic injury Micheli et al, 2011
Analysed and Real-time PCR Open reading frame (ORF) of PmIFRD2 cDNA was obtained from generation sequencing (NGS) analysis from liver of rea sea bream challenged with S. iniae
PmIFRD2 gene expression level profiles were compared between healthy fish and fish challenged with various pathogens
Summary
PmIFRD2 was acquired from the liver of a red sea bream and the ORF identified by NGS analysis. Sanger sequencing was performed to verify the cDNA sequence of PmIFRD2. The amino acid sequence of PmIFRD2 was predicted using the GENETYX ver. 7.0 program (SDC Software Development, Japan) and the NCBI BLAST program. The molecular weight and isoelectric point of PmIFRD2 were predicted using the ProtParam tool of the ExPASy Proteomics Server, and the location of the specific domains of PmIFRD2 was predicted using SMART web software. Multiple sequence alignments were analysed by ClustalW between the predicted amino acid sequence of PmIFRD2 and the IFRD2 amino acid sequences of other species registered in the NCBI peptide sequence database. The phylogenetic analysis of PmIFRD2 was performed using the neighbour-joining (NJ) method of the Mega 4 program. Support for each node was derived from 2000 bootstrap replicates
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