尼罗罗非鱼CD80基因的分子克隆与mRNA表达分析

发布时间：2018-03-06 04:18

本文选题：尼罗罗非鱼　切入点：CD　出处：《基因组学与应用生物学》2017年01期 　论文类型：期刊论文

【摘要】：CD80是T细胞活化和增殖过程中不可缺少的膜抗原,为T细胞的增殖分化提供必要的共刺激信号,在特异性细胞免疫过程中具有重要作用。本研究从罗非鱼脾脏组织中克隆获得CD80基因的编码区序列(Gen Bank登录号:KU884324;命名为On-CD80),该基因全长810 bp,可编码269个氨基酸,理论分子量为29.83 k D,等电点为5.97。氨基酸序列分析显示On-CD80具有2个保守的免疫球蛋白样结构域(Ig-like domain),5'端有一个典型的信号肽序列,3'端具有1个跨膜结构域。系统进化树中罗非鱼与伯氏朴丽鱼CD80蛋白聚为一支,显示了较高的同源性。荧光定量PCR分析显示,On-CD80在健康尼罗罗非鱼组织中均有表达,且在脾脏组织中表达量最高。经灭活的无乳链球菌免疫后,On-CD80在肾脏、脾脏、胸腺、肠道、鳃、脑组织中的表达量均显著上调,且表现出较为明显的时间依赖性表达模式,肠道、鳃和脑分别在8 h、24 h出现表达高峰,并于48 h、96 h再次出现表达高峰。脾脏和肾脏、胸腺的On-CD80表达则分别是在48 h、72 h达到表达高峰。综上所述,On-CD80表达能被无乳链球菌所诱导,提示该分子可能参与罗非鱼的抗菌免疫应答,该结果为进一步研究On-CD80在罗非鱼感染无乳链球菌过程中的作用奠定了一定的基础。
[Abstract]:CD80 is an indispensable membrane antigen in the process of T cell activation and proliferation, which provides necessary costimulatory signals for T cell proliferation and differentiation. In this study, the coding region sequence of CD80 gene was cloned from spleen tissue of tilapia, Genin Bank accession number: KU884324, named On-CD80, which is 810 BP in length and encodes 269 amino acids. The theoretical molecular weight is 29.83 KD and the isoelectric point is 5.97.The amino acid sequence analysis shows that On-CD80 has two conserved immunoglobulin-like domains (Ig-like domain) and a typical signal peptide sequence (3'terminal) with a transmembrane domain. The phylogenetic analysis shows that On-CD80 has two conserved immunoglobulin-like domains. The CD80 protein of Tilapia and Puccinia berryi are clustered into one branch in the tree. Fluorescence quantitative PCR analysis showed that On-CD80 was expressed in the tissues of healthy Nile tilapia and had the highest expression in spleen. On-CD80 was detected in kidney, spleen, thymus, intestine and Gill after inactivated streptococcus acus immunization. The expression levels in brain tissues were significantly up-regulated and showed a time-dependent expression pattern. The expression peaks in intestine, Gill and brain were observed at 8 h, 24 h, and 48 h, 96 h, respectively, and in spleen and kidney. The expression of On-CD80 in thymus reached its peak at 48 h and 72 h, respectively. In conclusion, the expression of On-CD80 could be induced by Streptococcus acuminata, suggesting that this molecule may be involved in the antimicrobial immune response of tilapia. The results laid a foundation for the further study of the role of On-CD80 in the process of infection of Streptococcus actinoides in tilapia.
【作者单位】：广东海洋大学水产学院;广东省水产经济动物病原生物学及流行病学重点实验室;广东省水产经济动物病害控制重点实验室;
【基金】：广东海洋大学创新强校项目(GDOU2013050108,GDOU2013050202) 湛江市非资助性项目(2010C3107001)共同资助
【分类号】：S943

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