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| Cloning, Expression of MsIL-21 Gene in Largemouth Bass (Micropterus salmoides) and Its Response to Bacterial Infection |
| ZHU Xue-Qing1,2, ZHAO Fei2,*, DENG Yu-Ting2, TAN Ai-Ping2, LAI Ying-Tiao2, GONG Hua2, HUANG Zhi-Bin2, JING Peng-Hua2 |
1 National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;
2 Pearl River Fisheries Research Institute/Key Laboratory of Fishery Drug Development of Ministry of Agriculture and Rural Affairs/Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Chinese Academy of Fishery Sciences, Guangzhou 510380, China |
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Abstract Interleukin-21 (IL-21) is a member of the common gamma chain (γc) cytokine family and plays an important role in defense against a variety of inflammatory and immune diseases. In order to explore the immune response function of IL-21 against pathogen infection, in this study, the ORF of largemouth bass (Micropterus salmoides) MsIL-21 (GenBank No. XP_038555572.1) was cloned, and bioinformatics and expression analysis were performed. The results showed that the full length of MsIL-21 ORF was 441 bp and encoding 146 amino acids. Sequence analysis revealed that MsIL-21 had the typical structure of IL-21 protein, including 4 groups of α-helices and 4 cysteine residues. Homology analysis revealed that MsIL-21 was most closely related to IL-21 of European sea bass (Dicentrarchus labrax). qPCR analysis showed that MsIL-21 was commonly expressed in all the healthy largemouth bass tissues tested, with the highest expression in the liver. After infection with Aeromonas veronii or Nocardia seriolae, the up-regulation of expression was detected in the liver, head kidney, and spleen. Similarly, after stimulation with pathogenic analogs lipopolysaccharide (LPS), lipoteichoic acid (LTA), and polyinosinic-polycytidylic acid [Poly(I:C)], the expression of MsIL-21 could be induced in head kidney leukocytes. The recombinant MsIL-21 protein was expressed and purified by prokaryotic expression system, which induced the expression of IL-10, IL-22, IL-1β, and interferon-γ (IFN-γ) in head kidney leukocytes. Taken together, the present study initially suggests that MsIL-21 may be involved in defense against pathogenic bacterial invasion and infection. This study further enriches the functional studies of fish IL-21.
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Received: 25 February 2022
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Corresponding Authors:
*zhaofei@prfri.ac.cn
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