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| Cloning and Functional Analysis of Sequestosome 1 (sqstm1) Gene in Nile Tilapia (Oreochromis niloticus) |
| HUANG Mei-Ling, JIANG Bai-Jian, FENG Jia-Min, LI Xing, HUANG Yu*, JIAN Ji-Chang |
| 1 College of Fisheries/Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy culture/Key Laboratory of Control for Disease of Aquatic Economic Animals of Guangdong Higher Education Institutes, Guangdong Ocean University, Zhanjiang 524088, China |
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Abstract Sequestosome 1 receptor protein (Sqstm1) is a selective autophagic adaptor protein in Nile tilapia (Oreochromis niloticus), which selectively participates in the removal of intracellular damaged organelles and proteins. To analyse the function of sqstm1 gene, and expression distribution of this gene in healthy Nile tilapia tissues and its expression pattern in the process of resisting bacterium and virus. In this study, the ORF of the gene On-sqstm1 (GenBank No. XP_005463852) was cloned, and the characteristics of bioinformatics and the subcellular localization were analysed, its expression patterns in healthy Nile tilapia tissues and after stimulation with different stimulant were analyzed by qPCR. The effect on nuclear factor kappa-B (NF-κB）signal path of On-sqstm1 was examined by using the dual luciferase reporting system. The results revealed that the ORF of the gene On-sqstm1 was successfully cloned and identified. which was 1 287 bp in length and encoded 428 amino acids. The results of subcellular localization showed that On-Sqstm1 was localized in the cytoplasm, the qPCR results revealed that On-sqstm1 was expressed in all tissues of tilapia and the highest in the liver. After challenged with the inactivated Streptococcus agalctiae and Poly I: C virus analogues, the expressions of brain, head kidney, liver, intestine and spleen significantly increased (P<0.05). The results of dual luciferase reporting system revealed that On-Sqstm1 significantly activated the NF-κB signal pathway activity.The above results indicated that On-Sqstm1 protein may participate in the immune response to pathogen infection by mediating NF-κB signaling pathway activity. This study provides a reference for further research on the mechanism of On-Sqstm1.
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Received: 25 May 2022
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Corresponding Authors:
* huangyu@gdou.edu.cn
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