文章摘要
中华鳖MHCⅡα基因的克隆及组织表达分析
Cloning and expression analysis of MHCⅡα gene inPelodiscus Sinensis
投稿时间:2020-08-02  修订日期:2020-08-16
DOI:
中文关键词: 中华鳖  MHCⅡα  序列分析  基因表达
英文关键词: Pelodiscus sinensis  MHCⅡα  sequence analysis  gene expression
基金项目:国家自然科学基金项目、国家重点研究开发项目-蓝色粮仓、湖南省自然科学基金青年基金项目
作者单位E-mail
童正飞 湖南农业大学动物科学技术学院 长沙 1101142532@qq.com 
胡亚洲 湖南农业大学动物科学技术学院 长沙  
杨涛 湖南农业大学动物科学技术学院 长沙  
涂开发 湖南农业大学动物科学技术学院 长沙  
王佩 湖南农业大学动物科学技术学院 长沙  
王晓清 湖南农业大学动物科学技术学院 长沙 wangxiao8258@126.com 
葛玲瑞 湖南生物机电职业技术学院  
熊钢 湖南生物机电职业技术学院  
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中文摘要:
      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基因的全长序列,其长度为1296 bp,开放阅读框(ORF)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、鸟类及两栖类亲缘关系较远。荧光定量PCR结果分析表明MHCⅡα mRNA在中华鳖肝脏、脾脏、肠道及肾组织中均有表达。感染嗜水气单胞菌(Aeromonas hydrophila)后,实验组肝脏在1、3和7天时的MHCⅡα mRNA上调表达,分别为对照组的2.04、2.28和4.02倍;实验组脾脏在7天时上调表达,为对照组的1.55倍;实验组肠道在3天和7天时上调表达,分别为对照组的1.40和1.79倍,而肝脏、脾脏及肠道都是与免疫密切相关的组织器官,以上研究表明中华鳖MHCⅡα基因参与了中华鳖免疫反应。
英文摘要:
      To study the structure and function of MHC (major histocompatibility complex) in Pelodiscus sinensis, we obtained the fullength cDNA of MHCⅡα by RACE-PCR (rapid amplification of cDNA ends polymerase chain reaction) technology. This sequence was 1296 bp, including an ORF (open reading frame) region of 807 bp. The peptide encoded MHCⅡα gene could be divided into four parts, including signal peptide, MHC II alpha domain, IGc1 domain and transmembrane region. A neighbor-joining tree shows that the MHCⅡα gene from P. sinensis and Chrysemys picta bellii clustered into one cluster. It had close genetic-relationship with C. picta bellii and father genetic-relationship with mammals, aves and amphibians. Quantitative real-time polymerase chain reaction analysis showed that MHCⅡα mRNA was expressed in liver, spleen, intestine and kidney of P. sinensis. Meanwhile, MHCⅡα mRNA expression levels were up-regulated in liver (on 1st, 3rd and 7th day), spleen (on 7th day) and intestine (on 3rd and 7th day ) after being infected with Aeromonas hydrophila, and the liver, spleen and intestine are closely related to immunity, which indicates that this gene has importanteffects on immune response。
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