Abstract:The coding sequence of interleukin-21 (IL-21) gene was cloned and expressed in bastard halibut Paralichthys olivaceus by using transcriptome sequencing data and PCR technology, and analyzed by bioinformatics analysis. The results showed that the open reading frame of IL-21 gene was 429 bp, encoding 142 amino acids with a molecular weight of 16.0 ku, and had a 19 bp signal peptide sequence at the N-terminal. Molecular phylogenetic tree indicated that IL-21 protein of bastard halibut had the closest relationship with IL-21 of perch Lates calcarifer and Stegastes partitus. Real-time fluorescent PCR revealed that IL-21 gene was expressed in all seven healthy tissues of bastard halibut, with the maximal expression level in liver. The expression of IL-21 was found to be up-regulated in head kidney and spleen of bastard halibut immunized with the extracted lipopolysaccharide from Edwardsiella tarda, indicating that IL-21 was involved in the immune response induced by lipopolysaccharide. When MyD88 gene was silenced by RNA interference, the expression of IL-21 was down-regulated, and the expression of MyD88 and IL-21 genes began to increase gradually in the bastard halibut immunized with the extracted lipopolysaccharide from E. tarda, indicating that that IL-21 is related to MyD88 pathway.
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