title: Characterization and Inductive Expression Analysis of Interferon Gamma-Related Gene in the Indian Major Carp, Rohu (Labeo rohita).
abstract: Interferon gamma (IFN-γ) is one of the key cytokines that plays a major role against viral and intracellular bacterial infection. In addition to the IFN-γ gene, teleost fish possess a second copy known as IFN-γ-related (IFN-γrel) gene. This report describes structural and functional properties of IFN-γrel gene in the Indian major carp, rohu (Labeo rohita), a commercially important freshwater fish species in the Indian subcontinent. The rohu IFN-γrel gene consisted of four exons with three intervening introns and phylogenetically closely related to grass carp. The full-length IFN-γrel cDNA comprised 927 bp nucleotides with a single open reading frame of 504 bp, encoding 167 amino acids (aa) polypeptide with a signal peptide of 24 aa. The mature rohu IFN-γrel protein was 143 aa with a predicted molecular weight of 16.85 kDa. Basal expression analysis of IFN-γrel showed its wide range of expression in all examined tissues: The highest was in the skin and the lowest was in the liver. In response to LPS, poly I:C, iE-DAP, muramyl dipeptide stimulations, and bacterial infections, IFN-γrel gene expression was significantly (p<0.05) induced in treated fish tissues as compared with their control. The IFN-γrel was expressed as recombinant protein (rIFN-γrel) and confirmed through western blot. Stimulation of peripheral blood leukocytes with rIFN-γrel protein resulted in the activation of IFN-γ receptor and marked induction of inducible nitric oxide synthase gene expression. These results together may suggest the important role of IFN-γrel as an antimicrobial cytokine in fish.
authors.affiliation: 1 Fish Health Management Division, Central Institute of Freshwater Aquaculture (CIFA) , Bhubaneswar, Odisha, India .
authors.forename: Banikalyan
authors.initials: B
authors.lastname: Swain
authors.forename: Madhubanti
authors.initials: M
authors.lastname: Basu
authors.forename: Saswati S
authors.initials: SS
authors.lastname: Lenka
authors.forename: Surajit
authors.initials: S
authors.lastname: Das
authors.forename: Pallipuram
authors.initials: P
authors.lastname: Jayasankar
authors.forename: Mrinal
authors.initials: M
authors.lastname: Samanta
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000254:growth & development / Q000378:metabolism; D001424:Bacterial Infections / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D001483:Base Sequence; D015153:Blotting, Western; D002347:Carps / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005393:Fish Diseases / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D007371:Interferon-gamma / Q000235:genetics* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D017471:Receptors, Interferon / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D000096743:Interferon gamma Receptor
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D007371:Interferon-gamma; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D017471:Receptors, Interferon; D000096743:Interferon gamma Receptor