中华鳖MHCⅡα基因cDNA的克隆及组织表达分析  

Cloning and Expression Analysis of MHCⅡαcDNA in Pelodiscus sinensis

作　　者：童正飞 葛玲瑞 胡亚洲[1,3] 杨涛[1] 涂开发 王佩[1] 周先文 王晓清[1,3] 熊钢[2,3] TONG Zhengfei;GE Lingrui;HU Yazhou;YANG Tao;TU Kaifa;WANG Pei;ZHOU Xianwen;WANG Xiaoqing;①;XIONG Gang(College of Animal Science and Technology,Hunan Agricultural University,Changsha,Hunan410128,China;Hunan Biological and Electromechanical Polytechnic,Changsha,Hunan410127,China;Technology System of Aquatic Industry in Hunan Province,Changsha,Hunan410128,China)

机构地区：[1]湖南农业大学动物科学技术学院,湖南长沙410128 [2]湖南生物机电职业技术学院,湖南长沙410127 [3]湖南省水产产业技术体系,湖南长沙410128

出　　处：《渔业科学进展》2021年第6期102-108,共7页Progress in Fishery Sciences

基　　金：国家重点研究开发项目-蓝色粮仓(2018YFD0900200);湖南省自然科学基金青年基金项目(2018JJ3265);国家自然科学基金(31672640);湖南省教育厅资助科研项目(18C0170)共同资助。

摘　　要：为了探究中华鳖(Pelodiscus sinensis)的主要组织相容性复合体(major histocompatibility complex,MHC)的结构和功能,本研究利用RACE技术成功克隆获得中华鳖MHCⅡα基因的cDNA全长序列,其长度为1296 bp,开放阅读框(ORF)为807 bp,共编码268个氨基酸,分为信号肽序列、Ⅱ类组织相容性抗原α结构域、IGc1结构域及跨膜结构域4个功能结构域。通过NJ法构建的系统进化树分析显示,中华鳖与西部锦龟(Chrysemys picta bellii)亲缘较近,而与哺乳类、鸟类及鱼类亲缘关系较远。荧光定量PCR结果显示,MHCⅡαmRNA在中华鳖8个组织中均有表达,在脾、心、肝及肠组织中表达水平较高,在肌肉组织中表达量最低。感染嗜水气单胞菌(Aeromonas hydrophila)12 h后,中华鳖MHCⅡαmRNA在肝和肠组织中的表达显著上调;感染1 d时,在脾组织中的表达显著上调;感染1 d及5 d时,在肾组织中的表达显著上调。研究表明,中华鳖MHCⅡα基因参与了中华鳖免疫反应。To study the structure and function of MHC(major histocompatibility complex)in Pelodiscus sinensis,we obtained the full-length cDNA of MHCⅡαusing RACE-PCR(rapid amplification of cDNA ends PCR)technology.The resulting sequence was 1296 bp in length,including an ORF(open reading frame)region of 807 bp.The peptide-encoded MHCⅡαgene could be divided into four parts,including the signal peptide,MHCⅡalpha domain,IGc1 domain,and transmembrane region.A neighbor-joining tree showed that the MHCⅡαgene from P.sinensis and Chrysemys picta bellii cluster into one cluster.The P.sinensis gene had a close genetic-relationship with C.picta bellii and a farther genetic-relationship with mammals,aves,and fish.Using qRT-PCR in all 8 tissues,the highest expression of the MHCⅡαgene was observed in the spleen,heart,liver,and intestine,while the lowest level was found in muscle.Meanwhile,MHCⅡαmRNA expression levels were significantly up-regulated in the liver and intestine(after 12 h),the spleen(on the 1st day),and the kidney(on 1st and 5th day)after being infected with Aeromonas hydrophila.The liver,spleen,intestine,and kidney are closely related to immunity,which indicates that this gene has important effects on the immune response.

关 键 词：中华鳖 MHCⅡα 序列分析 基因表达 

分 类 号：S917.4[农业科学—水产科学]