| 王片片，康桦华，刘振兴，王 芳，刘 春.剑尾鱼 IgD 基因的克隆及其在嗜水气单胞菌疫苗免疫后的表达变化[J].广东农业科学,2022,49(2):143-150 |
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剑尾鱼 IgD 基因的克隆及其在嗜水气单胞菌疫苗免疫后的表达变化 |
| Cloning of IgD Gene in Swordtail Fish and Changes of Its Responses to Aeromonas hydrophila Vaccine Challenges |
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| DOI：10.16768/j.issn.1004-874X.2022.02.017 |
| 中文关键词: 剑尾鱼 免疫球蛋白 D 基因克隆 实时荧光定量 PCR 免疫应答 |
| 英文关键词: Xiphophorus helleri immunoglobulin D gene cloning real-time PCR immune response |
| 基金项目:广东省农业科学院创新基金（202128）；广东省自然科学基金（2021A1515010498）；广州市科技计划项目（201906040005） |
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| 中文摘要: |
| 【目的】以剑尾鱼（Xiphophorus helleri）为试验对象，克隆获得其免疫球蛋白 D（Immunoglobulin D，
IgD）的 cDNA 序列，初步探讨该基因在剑尾鱼中的组织分布及在疫苗免疫后的表达规律。【方法】根据已知的
EST 库设计引物，利用 PCR 等方法获得剑尾鱼 IgD 基因 cDNA 序列并进行分析。运用实时荧光定量 PCR（RT-PCR）
方法检测 IgD 的组织分布及嗜水气单胞菌（Aeromonas hydrophila）疫苗免疫后，在头肾、脾脏和肠中的相对表
达变化。【结果】 剑尾鱼 IgD 完整开放阅读框（Open Reading Frame, ORF）为 3 879 bp，编码 1 298 个氨基酸，
预测其分子量大小为 145.5 ku；由可变区和恒定区组成，恒定区包含 CH1-CH2-CH3-CH4-CH5-CH6-CH7 和
1 个跨膜区（Transmembrane region，TM）。系统进化分析显示，剑尾鱼与鳜（Siniperca chuatsi）IgD 的基因序列
一致性最高、为 70%。IgD 在剑尾鱼头肾、脾和肠中均有分布，经嗜水气单胞菌疫苗腹腔注射免疫后，IgD 在头
肾、脾和肠中发生差异性表达，肾和脾脏中 IgD 表达量显著下调，肠中 IgD 表达量显著上调。【结论】IgD 可能
在剑尾鱼机体免疫应答与肠道粘膜免疫中发挥重要作用。 |
| 英文摘要: |
| 【Objective】The swordtail fish (Xiphophorus helleri) was employed to clone the gene sequence of
immunoglobulin D (IgD), and the tissue distribution and expression laws after vaccine immunization of the IgD in swordtail
fish were explored.【Method】With the known EST libraries, the cDNA sequences of IgD gene from swordtail fish were cloned and analyzed, and the tissue distribution and the relative expression changes in head kidney, spleen and intestine
after vaccination were detected by real-time PCR.【Result】The complete open reading frame (ORF) of swordtail fish was
3 879 bp, which encoded 1 298 amino acids, predicting that the molecular weight was 145.5 ku. The IgD of swordtail fish was
composed of variable region and constant region, and the constant region contained CH1-CH2-CH3-CH4-CH5-CH6-CH7
and a transmembrane region (TM). The systematic evolution analysis of gene sequences showed that swordtail fish IgD shared
similarities of 70% with mandarin fish (Siniperca chuatsi). IgD mRNA expression of swordtail fish was highly detected in head
kidney, spleen and intestine. After being immunized with Aeromonas hydrophila vaccine, the expression of IgD in the head
kidney and spleen was significantly down-regulated, and it was significantly up-regulated in the intestine.【Conclusion】IgD
may play an important role in immune response and intestinal mucosal immunity of swordtail fish. |
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