尼罗罗非鱼CD247基因克隆及其组织表达分析Cloning and Tissue Expression Analysis of Gene CD247 in Nile Tilapia(Oreochromis niloticus)
樊博琳,黎源,汪志文,鲁义善,王蓓,简纪常
摘要(Abstract)：
【目的】克隆尼罗罗非鱼(Oreochromis niloticus)CD247基因(Gen Bank登录号:MF975639;On-CD247),分析On-CD247的生物信息学特征及其在正常和经灭活无乳链球菌注射刺激的尼罗罗非鱼组织中的表达。【方法】设计CD247基因引物并对CD247基因片段进行扩增,运用荧光定量PCR技术分析尼罗罗非鱼CD247组织分布。【结果与结论】CD247基因编码区为453 bp,编码150个氨基酸,理论分子质量为16.9 ku,等电点为6.33,在跨膜区域有保守的ASP27、Asp35残基,胞内具有三段保守的免疫受体酪氨酸激活基序(ITAM)。On-CD247序列与红笛鲷(Lutjanus sanguineus)相似性最高,为74.0%,与其他鱼类的相似性介于35.8%～73.0%之间。On-CD247在健康尼罗罗非鱼各组织中均有表达,其中在胸腺中表达量最高,其次是皮肤、肌肉、鳃,在头肾中表达量最低。经灭活无乳链球菌刺激后,On-CD247的表达量在胸腺、头肾和肠道中均显著上调,而在脾脏中却显著下调,且有明显的时序性。胸腺、头肾和肠道均在3 h表达量最高,脾脏在12 h表达量最低,头肾中的表达量也在12 h回落,表明On-CD247表达可被无乳链球菌所诱导。
关键词(KeyWords)： 尼罗罗非鱼;CD247;基因;克隆;组织表达;无乳链球菌
基金项目(Foundation): 国家自然科学基金青年基金项目(31702386);; 广东省科技厅国际科技合作领域项目(2017A050501037)
作者(Author): 樊博琳,黎源,汪志文,鲁义善,王蓓,简纪常
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