2022, 42(5): 27-37. doi: 10.3969/j.issn.1673-9159.2022.05.004
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
Keywords： Oreochromis niloticus , marco , Streptococcus agalactiae , scavenger receptor , subcellular localization , expression analysis
【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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