黄子威,汪志文,黎源,夏洪丽,简纪常,鲁义善

• 1:广东海洋大学水产学院
• 2:广东省水产经济动物病原生物学及流行病学重点实验室
• 3:广东省水产经济动物病害控制重点实验室
• 4:广东海洋大学深圳研究院

摘要(Abstract)：

【目的】研究尼罗罗非鱼(Oreochromis niloticus)清道夫受体基因(On-SRA4)的重组表达、亚细胞定位及组织分布,为深入研究On-SRA4基因功能奠定基础。【方法】根据NCBI上公布的On-SRA4基因序列设计克隆引物,构建原核表达载体并诱导重组蛋白rOn-SRA4的表达。同时,采用生物信息学、亚细胞定位手段分析On-SRA4的结构特征,用实时定量PCR分析On-SRA4在头肾、胸腺、脾脏、肝、肠道、鳃、脑、肌肉、皮肤、血液的表达水平,用灭活无乳链球菌(Streptococcus agalactiae)、病毒类似物PolyI:C刺激健康罗非鱼,分析On-SRA4在头肾、胸腺、脾脏、肠道的表达水平。【结果与结论】成功构建原核表达载体pET28a-OnSRA4,诱导获得目的蛋白rOn-SRA4,rOn-SRA4分子质量约80 ku。亚细胞定位显示,rOn-SRA4定位于细胞核与细胞质。组织定量分析表明,On-SRA4在所有组织中均有表达,且在血液中表达量最高,在灭活无乳链球菌与Poly I:C刺激后,健康罗非鱼胸腺、肠道、脾脏和头肾中On-SRA4表达量均显著升高,表明On-SRA4参与了病原引起的免疫反应,是罗非鱼抵抗病原的相关基因。

关键词(KeyWords)： 尼罗罗非鱼;On-SRA4;重组表达;RT-qPCR;亚细胞定位

Abstract:

Keywords:

基金项目(Foundation): 广东省现代农业产业体系项目(2019KJ141);; 深圳市科技计划项目（JCYJ20180306173023022）;; 深圳市大鹏新区科技创新和产业发展专项资金项目（PT202101-23）

作者(Author): 黄子威,汪志文,黎源,夏洪丽,简纪常,鲁义善

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