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Identification and characterization of IKKε gene from grass carp Ctenopharyngodon idella

Li, Jun, Zhou, Man, Peng, Lingzhi, Sun, Wenzheng, Yang, Peilin, Yan, Jun, Feng, Hao
Fish & shellfish immunology 2015 v.47 no.1 pp. 255-263
Ctenopharyngodon idella, Danio rerio, Pimephales promelas, Western blotting, amino acids, complementary DNA, fish, fluorescent antibody technique, gene overexpression, genes, immune response, messenger RNA, nucleotides, polypeptides, staining, tissues
IKKε is an IKK-related kinase implicated in antiviral immune response in higher vertebrates. To elucidate the function of IKKε in teleost fish, grass carp IKKε (gcIKKε) has been cloned and characterized in this paper. The full-length cDNA of gcIKKε is composed of 2529 nucleotides and encodes a polypeptide of 723 amino acids. The mRNA transcription of gcIKKε was constitutively detected in all the selected tissues and the gcIKKε mRNA level increased at 36 h after GCRV infection. Western blot data of both HEK293T cells and EPC cells demonstrated that gcIKKε was around 80 KDa; and immunofluorescence staining data of both NIH3T3 cells and EPC cells determined gcIKKε was a cytosolic protein. The mRNA level of gcIKKε in CIK cells was increased more than 150 times right after poly(I:C) treatment and PMA treatment triggered gcIKKε mRNA transcription in CIK cells more than 100 times. Over-expression of gcIKKε in EPC cells activated the promoter activity of both zebrafish IFN and fathead minnow IFN. gcIKKε mRNA transcription level in CIK cells was increased from 48 h post GCRV infection with different MOIs. All the data support the idea that gcIKKε is a novel teleost IκB kinase recruited in the IFN-mediated antiviral immunity of grass carp.