斜带石斑鱼 BAFF 基因的克隆、分子特征 及溶藻弧菌感染后表达特征相关分析

Cloning, molecular characteristics and expression characteristics of BAFF gene in Grouper(Epinephelus coioides) after infection with Vibrio alginolyticus

摘要

【目的】B细胞激活因子（B cell activating factor，BAFF）是肿瘤坏死因子超家族的关键成员，在B细胞的存活、增殖与分化中起主导作用。为深入探究溶藻弧菌感染下鱼体的免疫调控机制，本研究聚焦于B细胞激活因子，探究其动态表达与免疫应答的关联。【方法】克隆斜带石斑鱼（Epinephelus coioides）BAFF基因并进行生物信息学分析，建立溶藻弧菌（Vibrio alginolyticus，10⁵、10⁷和10⁹ CFU·mL^-1）感染模型，分析BAFF在感染过程中的表达变化，为鱼类抗病机制与病害防控提供参考。【结果】斜带石斑鱼BAFF基因的开放阅读框全长为792 bp，编码264个氨基酸；预测蛋白分子质量为29.23 kDa，理论等电点（pI）为9.23，不稳定系数为57.37，平均亲水性为−0.296；该蛋白无信号肽，但包含一个TNF家族结构域。溶藻弧菌感染后，3个处理组的BAFF表达在0~6 h均显著上升，6~72 h下调且趋于稳定；其中10⁹CFU·mL^-1组在72 h时出现回升趋势且表达量高于其余两组。【结论】本研究初步揭示了斜带石斑鱼BAFF基因的结构特性及其在病原感染下的表达调控模式，为深入探究石斑鱼的免疫应答机制提供了数据支持。

Abstract

[Objective] B-cell activating factor (BAFF), a key member of the tumor necrosis factor superfamily, plays a pivotal role in the survival, proliferation, and differentiation of B cells. To further investigate the immune regulatory mechanisms in fish upon pathogenic infection, this study focused on BAFF and examined the relationship between its dynamic expression and immune responses. [Method] The BAFF gene of Epinephelus coioides was cloned and analyzed by bioinformatics. Infection models with Vibrio alginolyticus (10⁵, 10⁷, and 10⁹ CFU·mL^-1) were established to analyze the expression changes of BAFF during infection, providing a reference for disease resistance mechanisms and disease prevention and control in fish. [Result] The open reading frame of the BAFF gene in the slant-band grouper spans 792 bp, encoding 264 amino acids. The predicted protein molecular weight is 29.23 kDa, with a theoretical isoelectric point (pI) of 9.23, a stability coefficient of 57.37, and an average hydrophilicity of -0.296. The protein lacks a signal peptide but contains one TNF family domain. Following Vibrio alginolyticus infection, BAFF expression significantly increased in all three treatment groups between 0-6 h, then decreased and stabilized between 6-72 h. Notably, the 10⁹ CFU·mL^-1 group exhibited a rebound trend at 72 h with higher expression levels than the other two groups. [Conclusion] This study elucidates the structural features of the BAFF gene in oblique-banded grouper and its expression profile in response to bacterial infection, laying a molecular foundation for further understanding of immune response mechanisms in grouper.

关键词

斜带石斑鱼 / BAFF / 浸泡感染 / 免疫应答

Key words

Epinephelus coioides / BAFF / immersion infection / immune response

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基金资助

广东省普通高校特色创新类项目(2024KTSCX201)