首页 >  广东海洋大学学报 >  尼罗罗非鱼marco基因克隆、亚细胞定位及表达分析

2022, 42(5): 27-37. doi: 10.3969/j.issn.1673-9159.2022.05.004

尼罗罗非鱼marco基因克隆、亚细胞定位及表达分析

1. 广东海洋大学水产学院;

2. 广东省水产动物病害防控与健康养殖重点实验室, 广东 湛江 524088;

2. 广东海洋大学深圳研究院;

4. 广东省水生动物健康评估工程技术研究中心;

5. 深圳市海水经济动物种苗健康评价公共技术服务平台, 广东 深圳 518120

收稿日期:2022-03-16
修回日期:2022-03-16

基金项目:   广东省现代农业产业体系项目(2019KJ141);广东省基础与应用基础研究基金项目(2021A1515110481) 

关键词: 尼罗罗非鱼 , marco , 无乳链球菌 , 清道夫受体 , 亚细胞定位 , 表达分析

Molecular Cloning, Subcellular Localization and Expression Analysis of marco Gene in Nile Tilapia(Oreochromis niloticus)

1. Fisheries College of Guangdong Ocean University;

2. Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy culture, Zhanjiang 524088, China;

2. Shenzhen Institute of Guangdong Ocean University;

4. Guangdong Provincial Engineering Research Center for Aquatic Animal Health Assessment;

5. Shenzhen Public Service Platform for Evaluation of Marine Economic Animal Seedings, Shenzhen 518120, China

Received Date:2022-03-16
Accepted Date:2022-03-16

Keywords: Oreochromis niloticus , marco , Streptococcus agalactiae , scavenger receptor , subcellular localization , expression analysis

摘要

【目的】研究尼罗罗非鱼(Oreochromis niloticus)胶原样结构巨噬细胞受体(macrophage receptor with collagenous structure,Marco)的结构特征和免疫功能,为揭示尼罗罗非鱼免疫应答机制提供理论基础。【方法】采用聚合酶链式反应从尼罗罗非鱼脾脏克隆marco基因序列,利用生物信息学和亚细胞定位等手段分析其结构特征,用实时荧光定量PCR(qRT-PCR)技术检测其表达情况。【结果】尼罗罗非鱼marco 基因(Onmarco)的开放阅读框(open reading frame,ORF)长1 071 bp,共编码356 个氨基酸,具有跨膜结构域、典型的胶原样结构域和C 端富半胱氨酸受体(scavenger receptor cystein-rich,SRCR)结构域;多序列比对和系统进化分析表明,Marco 在硬骨鱼类中具有较高的保守性;亚细胞定位显示,尼罗罗非鱼Marco(OnMarco)在HEK-293T细胞中为全细胞分布,主要在细胞膜上表达;qRT-PCR结果显示,Onmarco在健康尼罗罗非鱼各组织中均有表达,但在血液中的表达量最高;经灭活的无乳链球菌和聚肌胞苷酸(Poly I:C)刺激后,Onmarco 在罗非鱼肠道、头肾和脾脏中的表达量均呈时序性变化;用脂多糖(LPS)、Poly I:C 和灭活后的无乳链球菌刺激罗非鱼头肾白细胞,Onmarco 表达量均显著升高。【结论】Onmarco基因可能在鱼体抵御细菌和病毒入侵的免疫反应中发挥重要作用。

【目的】研究尼罗罗非鱼(Oreochromis niloticus)胶原样结构巨噬细胞受体(macrophage receptor with collagenous structure,Marco)的结构特征和免疫功能,为揭示尼罗罗非鱼免疫应答机制提供理论基础。【方法】采用聚合酶链式反应从尼罗罗非鱼脾脏克隆marco基因序列,利用生物信息学和亚细胞定位等手段分析其结构特征,用实时荧光定量PCR(qRT-PCR)技术检测其表达情况。【结果】尼罗罗非鱼marco 基因(Onmarco)的开放阅读框(open reading frame,ORF)长1 071 bp,共编码356 个氨基酸,具有跨膜结构域、典型的胶原样结构域和C 端富半胱氨酸受体(scavenger receptor cystein-rich,SRCR)结构域;多序列比对和系统进化分析表明,Marco 在硬骨鱼类中具有较高的保守性;亚细胞定位显示,尼罗罗非鱼Marco(OnMarco)在HEK-293T细胞中为全细胞分布,主要在细胞膜上表达;qRT-PCR结果显示,Onmarco在健康尼罗罗非鱼各组织中均有表达,但在血液中的表达量最高;经灭活的无乳链球菌和聚肌胞苷酸(Poly I:C)刺激后,Onmarco 在罗非鱼肠道、头肾和脾脏中的表达量均呈时序性变化;用脂多糖(LPS)、Poly I:C 和灭活后的无乳链球菌刺激罗非鱼头肾白细胞,Onmarco 表达量均显著升高。【结论】Onmarco基因可能在鱼体抵御细菌和病毒入侵的免疫反应中发挥重要作用。

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目录

尼罗罗非鱼marco基因克隆、亚细胞定位及表达分析