| 何恺轩，温 彬，高建忠，陈再忠.七彩神仙鱼 BTN1A1 基因克隆及其在嗜水气单胞菌胁迫下的表达分析[J].广东农业科学,2024,51(4):45-53 |
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七彩神仙鱼 BTN1A1 基因克隆及其在嗜水气单胞菌胁迫下的表达分析 |
| Cloning of BTN1A1 Genes from Symphysodon aequifasciatus and Analysis of Its Expression under Aeromonas hydrophila Stress |
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| DOI：10.16768/j.issn.1004-874X.2024.04.004 |
| 中文关键词: 七彩神仙鱼 嗜乳脂蛋白 嗜水气单胞菌 免疫调节 基因表达 |
| 英文关键词: Symphysodon aequifasciatus butyrophilin Aeromonas hydrophila immunomodulation gene expression |
| 基金项目:上海市自然科学基金（20ZR1423600）；上海海洋大学科技发展专项基金（A2-2006-22-2002）；国家自然科学基金（31902376） |
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| 中文摘要: |
| 【目的】克隆七彩神仙鱼（Symphysodon aequifasciatus）嗜乳脂蛋白 1A1（Butyrophilin 1A1, BTN1A1）基因，分析其在病原刺激下的表达模式，为了解七彩神仙鱼 BTN1A1 基因功能提供依据。【方法】利用生物信息学对七彩神仙鱼 2 个 BTN1A1 基因（BTN1A1-1 和 BTN1A1-2）进行结构和进化分析。根据前期获得的七彩神仙鱼皮肤转录组数据，选取 BTN1A1-1 和 BTN1A1-2 基因的 CDS 区设计引物进行克隆。采用 qRT-PCR 分析 BTN1A1 在各组织中的表达及嗜水气单胞菌刺激下的表达模式。【结果】BTN1A1-1 和 BTN1A1-2 的 ORF 序列长度为 894、1 275 bp，分别编码 298、424 个氨基酸。BTN1A1-1 和 BTN1A1-2 蛋白均具有 1 个信号肽、1 个 Ig 结构域、1 个 lg_like 结构域和 1个跨膜结构域的经典结构，BTN1A1-2 还具有 1 个胞质结构域。七彩神仙鱼 BTN1A1-1 与慈鲷科其他鱼类相似性较高，与尼加拉瓜湖始丽鱼（Archocentrus centrarchus）BTNIAI 对应氨基酸序列同源性最高、为 94.14%。但 BTN1A1-2 与其他鱼类发生分离，形成独特分支。BTN1A1-1 和 BTN1A1-2 在七彩神仙鱼各组织中均有表达，但在鳃、肠道、皮肤等与免疫相关的组织中表达量相对较高。七彩神仙鱼 BTN1A1-1 在嗜水气单胞菌胁迫后表达量先显著下降后上升，而 BTN1A1-2 表达量则先显著上升后下降。【结论】七彩神仙鱼 BTN1A1-1 基因相对保守，而 BTN1A1-2 基因在进化上较为特殊。二者在免疫刺激下的差异表达暗示其可能在七彩神仙鱼免疫防御中发生功能分化。 |
| 英文摘要: |
| 【Objective】Butyrophilin 1A1 (BTN1A1) gene from Symphysodon aequifasciatus was cloned and its express pattern under pathological infection was analyzed, to provide basis for better understanding of the function of BTN1A1 gene in S. aequifasciatu.【Method】The structure and evolutionary characteristics of two BTN1A1 genes (BTN1A1-1 and BTN1A1-2) were analyzed by bioinformatics analysis. Based on the transcriptome data of the skin of the S. aequifasciatu obtained at early stage, the CDS regions of BTN1A1-1 and BTN1A1-2 genes were used to design the primers for gene cloning. The expressions of BTN1A1-1 gene in various tissues and its expression pattern under the stimulus of Aeromonas hydrophila were analyzed by qRT-PCR.【Result】The ORF regions of BTN1A1-1 and BTN1A1-2 were 894 and 1 275 bp, encoding 298 and 424 amino acids, respectively. Both BTN1A1-1 and BTN1A1-2 had a classical structure of BTN, which contained a signal peptide, an Ig domain, an lg_like domain and a transmembrane domain. BTN1A1-2 also had a cytoplasmic domain. BTN1A1-1 showed a closer relationship with the genes found in cichlid fish. It shared the highest homology of 94.14% with the corresponding amino acid sequences of Archocentrus centrarchus; while BTN1A1-2 showed lower similarity with other types of fish, forming a special branch. The expressions of BTN1A1-1 and BTN1A1-2 were highly in gill, intestine and skin of S. aequifasciatu. The expression of BTN1A1-1 decreased significantly and then increased after the infection of A. hydrophila, while the expression of BTN1A1-2 increased significantly and then decreased.【Conclusion】The BTN1A1-1 gene was more evolutionarily conservative than BTN1A1-2. The differential expressions of BTN1A1-1 and BTN1A1-2 in response to immune stimulation suggests that they may be functionally differentiated in the immune defence of S. aequifasciatus. |
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