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徐泳娴,邹子鸿,汤菊芬,鲁义善,简纪常,蔡佳.斜带石斑鱼ERP44基因的克隆与表达分析[J].广西科学院学报,2021,37(2):109-116. [点击复制]
- XU Yongxian,ZOU Zihong,TANG Jufen,LU Yishan,JIAN Jichang,CAI Jia.Cloning and Expression Analysis of ERP44 Gene from Orange-Spotted Grouper, Epinephelu coioides[J].Journal of Guangxi Academy of Sciences,2021,37(2):109-116. [点击复制]
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| 斜带石斑鱼ERP44基因的克隆与表达分析 |
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徐泳娴1, 邹子鸿1, 汤菊芬1, 鲁义善1, 简纪常1, 蔡佳1,2
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| (1.广东海洋大学水产学院, 广东省水产经济动物病原生物学及流行病学重点实验室, 广东省水产经济动物病害控制重点实验室, 广东湛江 524088;2.广西科学院, 广西北部湾海洋研究中心, 广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007) |
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| 摘要: |
| 为进一步了解内质网蛋白44基因（ERP44）在石斑鱼免疫反应中的作用，本研究根据实验室石斑鱼转录组数据中ERP44的表达序列标签（EST）设计引物，克隆了斜带石斑鱼（Epinephelus coioides）Ec-ERP44基因的开放阅读框序列（ORF），利用生物信息学手段分析Ec-ERP44基因及其编码蛋白的序列结构和特征，并通过实时荧光定量PCR检测该基因的组织分布特征，以及脂多糖（Lipopolysaccharide，LPS）、聚肌胞苷酸（Polyinosinic-polycytidylic acid，Poly I：C）刺激后该基因在石斑鱼脾脏中的表达变化。研究结果表明，Ec-ERP44基因ORF全长1 233 bp，编码 410个氨基酸；该蛋白具有蛋白质二硫键异构酶（PDI）家族保守的硫氧还蛋白结构域。同时，Ec-ERP44基因在健康石斑鱼体内的多种组织均有表达，其中在脑组织中的表达量最高；在LPS与Poly I：C刺激后，石斑鱼脾脏中Ec-ERP44基因的表达显著上调。本研究结果表明，Ec-ERP44基因参与了石斑鱼抗病原感染的免疫反应。 |
| 关键词: ERP44 斜带石斑鱼 基因克隆 抗感染免疫 表达分析 |
| DOI：10.13657/j.cnki.gxkxyxb.20210806.004 |
| 投稿时间：2021-05-31 |
| 基金项目:国家自然科学基金项目（U20A2065，32002426，32073006），广东省大学生创新创业训练计划项目（CXXL2018053），广西自然科学基金项目（2020GXNSFAA297243），南方海洋科学与工程广东省实验室（湛江）（湛江湾实验室）自主立项项目（ZJW-2019-06），广东海洋大学“冲一流”专项（231419013）和广西红树林滨海湿地生态保护与可持续利用人才小高地人才项目（BGMRC202101）资助。 |
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| Cloning and Expression Analysis of ERP44 Gene from Orange-Spotted Grouper, Epinephelu coioides |
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XU Yongxian1, ZOU Zihong1, TANG Jufen1, LU Yishan1, JIAN Jichang1, CAI Jia1,2
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| (1.Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Fisheries College, Guangdong Ocean University, Zhanjiang, Guangdong, 524088, China;2.Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Beibu Gulf Marine Research Center, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China) |
| Abstract: |
| In order to further study the roles of ERP44 in the grouper immune response, the open reading frame (ORF) sequence of ERP44 gene (Ec-ERP44) was cloned from orange-spotted grouper (Epinephelus coioides) through the primers designed by the EST sequence from the transcriptome data of laboratory. The structural characters of Ec-ERP44 gene and its coding amino acid sequence were analyzed using bioinformatics methods. Also, the tissue distribution and expression changes of Ec-ERP44 gene after LPS and Poly I:C stimulation was examined by qRT-PCR. The results showed that the ORF sequence of Ec-ERP44 gene was 1 233 bp, encoding 410 amino acids. It was deduced that Ec-ERP44 protein had a conserved thioredoxin domain of the PDI family. At the same time, Ec-ERP44 gene was widely expressed in various tissues, among which the highest expression level was in brain tissue. After LPS and Poly I:C stimulation, the expressions of Ec-ERP44 gene in spleen were significantly up-regulated. The results of this study indicated that Ec-ERP44 gene was involved in the immune response of grouper against pathogen infection. |
| Key words: ERP44 orange-spotted grouper gene cloning anti-infections immunity expression analysis |
