文献类型：期刊文献

中文题名：尼罗罗非鱼CD63基因克隆及表达特征分析

英文题名：Cloning and expression characteristics of CD63 gene in Nile tilapia(Oreochromis niloticus)

作者：李梓昕[1];陈杨慧[1];卢春兰[1];黄瑜[1];蔡佳[1];王蓓[1]

机构：[1]广东海洋大学水产学院/广东省水产动物病害防控与健康养殖重点实验室/水产经济动物病害控制广东省普通高校重点实验室,广东湛江524088

年份：2024

卷号：55

期号：12

起止页码：3779

中文期刊名：南方农业学报

外文期刊名：Journal of Southern Agriculture

收录：北大核心2023、、CSCD_E2023_2024、北大核心、CSCD

基金：国家自然科学基金项目(32073006)。

语种：中文

中文关键词：尼罗罗非鱼;CD63基因;组织表达;无乳链球菌

外文关键词：Nile tilapia(Oreochromis niloticus);CD63 gene;tissue expression;Streptococcus agalactiae

中文摘要：【目的】克隆尼罗罗非鱼(Oreochromis niloticus)抗原分化簇63基因(CD63)并分析其在病原菌刺激后的表达情况,探究尼罗罗非鱼CD63基因在免疫应答中发挥的作用,为进一步研究CD63的免疫功能提供理论依据。【方法】对尼罗罗非鱼CD63基因进行克隆,利用生物信息学软件对尼罗罗非鱼CD63蛋白进行理化性质分析、跨膜结构域预测和系统进化分析等,采用实时荧光定量PCR检测尼罗罗非鱼CD63基因的组织表达特征及在无乳链球菌感染后的表达变化。【结果】尼罗罗非鱼CD63基因开放阅读框(ORF)长度为723 bp,编码240个氨基酸残基。尼罗罗非鱼CD63蛋白分子量为25.88 kD,理论等电点(pI)为8.38,蛋白平均亲水性系数(GRAVY)为0.745,为疏水性蛋白,不含信号肽,含4个跨膜结构域。进行比对分析发现,尼罗罗非鱼与奥利亚罗非鱼等其他鱼类的CD63氨基酸序列相似性较高;系统进化分析结果表明,尼罗罗非鱼与奥利亚罗非鱼亲缘关系最近。CD63基因在尼罗罗非鱼的脑、脾脏、肝脏、皮肤、头肾、鳃、心脏、后肠、血液和肌肉10种组织中均有表达,在肌肉中相对表达量最高,其次为血液和后肠,在脑中相对表达量最低。在无乳链球菌感染3~96 h,CD63基因在尼罗罗非鱼的鳃、心脏、头肾、脾脏、后肠、肌肉和皮肤中相对表达量最高值均较未感染时(0 h)显著上调(P<0.05,下同);在感染96 h时,CD63基因在尼罗罗非鱼的脑、鳃、头肾、肝脏、脾脏、后肠、肌肉和皮肤中的相对表达量较最高值均显著下调。【结论】在无乳链球菌感染后,尼罗罗非鱼CD63基因在各组织中的表达均发生明显变化,表明CD63基因参与尼罗罗非鱼对病原菌刺激的免疫应答过程。

外文摘要：【Objective】To clone the cluster of differentiation 63 gene(CD63)of Nile tilapia(Oreochromis niloticus)and analyze its expression after stimulation by pathogenic bacteria,so as to explore the role of CD63 gene in immune response of Nile tilapia,which could provide theoretical basis for study of the immune function of CD63.【Method】CD63 gene of Nile tilapia was cloned,and the CD63 protein of Nile tilapia was analyzed by bioinformatics softwares,including physical and chemical properties,transmem brane domain prediction and phylogenetic analysis.The tissue expression characteristics of CD63 gene in Nile tilapia and its expression changes after infection with Streptococcus agalactiae were detected by real-time fluorescence quantitative PCR.【Result】The CD63 gene of Nile tilapia had an open reading frame(ORF)length of 723 bp and encoded 240 amino acid residues.The molecular weight of CD63 protein of Nile tilapia was 25.88 kD,the theoretical isoelectric point(pI)was 8.38,and the mean hydrophilic coefficient(GRAVY)was 0.745.It was a hydrophobic protein without signal peptide,containing 4 transmembrane domains.The similarity of CD63 amino acid sequence between Nile tilapia and other species such as Oreochromis aureus was high through comparative analysis.Phylogenetic analysis shows that Nile tilapia was closely related to Oreochromis aureus.CD63 gene of Nile tilapia was expressed in 10 tissues(brain,spleen,liver,skin,head kidney,gills,heart,intestines,blood and muscle),with the highest relative expression in muscle,followed by blood and intestines,and the lowest relative expression in brain.The highest relative expression levels of CD63 gene in gills,heart,spleen,intestines,muscle and skin of Nile tilapia were significantly up-regulated at 3-96 h after infection with Streptococcus agalactiae compared with without infection(0 h)(P<0.05,the same below).At 96 h after infection,the relative expression of CD63 gene in brain,gills,head kidney,spleen,intestines,muscle and skin was significantly down-regulated compared with the highest value.【Conclusion】The expression of CD63 gene in tissues of Nile tilapia after infection with Streptococcus agalactiae is greatly changed,indicating that CD63 gene is involved in the immune response of Nile tilapia to pathogens.