赤眼鳟Mx1基因的cDNA克隆及表达特性  

MOLECULAR CLONING AND EXPRESSION CHARACTERISTICS OF MX1GENE IN SQUALIOBARBUS CURRICULUS

作　　者：王静安[1] 李耀国[1,2] 赵鑫 金生振[1] 陈开健 肖调义[1,2] WANG Jing-An;LI Yao-Guo;ZHAO Xin;JIN Sheng-Zhen;CHEN Kai-Jian;XIAO Tiao-Yi(Hunan Engineering Research Center for Utilization of Characteristics of Aquatic Resources,Hunan Agricultural University,Changsha 410128,China;Collaborative Innovation Center for Efficient and Health Production of Fisheries in Hunan Province,Changde 415000,China)

机构地区：[1]湖南农业大学湖南省特色水产资源利用工程技术研究中心,长沙410128 [2]水产高效健康生产湖南省协同创新中心,常德415000

出　　处：《水生生物学报》2020年第4期756-763,共8页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金面上项目(31572615和31272652);中国博士后科学基金特别资助项目(2018T110833);中国博士后科学基金资助项目(2017M612560)资助。

摘　　要：为探究赤眼鳟(Squaliobarbus curriculus)是否存在Mx1(Myxovirus resistance)基因及参与抗病毒免疫反应,研究利用RACE技术获得了赤眼鳟Mx1基因(ScMx1)的cDNA全长序列,并对其进行了生物信息学分析;采用荧光定量PCR技术,检测了ScMx1在赤眼鳟健康组织中的表达情况以及感染GCRV后ScMx1和ScIFN-Ⅰ的表达特征。结果表明,Sc Mx1的c DNA全长为3000 bp,包含5′非编码区124 bp,开放阅读框1893 bp,3′非编码区983 bp,共编码630个氨基酸。预测的ScMx1蛋白包含GTP酶结合区域、中央核心结构域和GTP酶效应结构域。ScMx1与青鱼Mx1的相似性最高(97%),与ScMx的相似性仅为50%。ScMx1在所检测的10种组织中均有表达,其中在脾脏中表达量最高。经GCRV感染开始至168h,ScMx1和ScIFN-Ⅰ在肝脏和体肾中的表达量持续上调;在脾脏和头肾中于感染后72h达到峰值。相关性分析显示脾脏中ScMx1和ScIFN-Ⅰ的表达水平呈显著相关(r=0.94,P=0.018)。研究发现赤眼鳟存在Mx1基因,且可能参与了抗GCRV免疫应答反应。In order to investigate whether the Myxovirus resistance isoform(Mx1) exists in Squaliobarbus curriculus and gets involved in antiviral immune response, the full-length cDNA of ScMx1 was cloned and characterized by using RACE-PCRs technology. The expression levels of ScMx1 and ScIFN-I were detected after injection with grass carp reovirus(GCRV) with qPCR. The full-length cDNA of ScMx1 was 3000 bp including a 5′-terminal untranslated region of124 bp, a 3′-terminal untranslated region of 893 bp and an open reading frame of 1893 bp encoding a polypeptide of630 amino acid residues. The ScMx1 protein consisted of Dynamin GTPase domain, Dynamin central domain and Dynamin GTPase effector domain. The BLAST analysis showed that ScMx1 shared high identity(97%) with Mylopharyngodon piceus Mx1, and low identity(50%) with ScMx. The expression of ScMx1 could be detected in all tested tissues,with the highest expression in spleen. After injection with GCRV, the ScMx1 and ScIFN-I expression levels were upregulated to 168 h in liver and trunk kidney, and peaked at 72 h in spleen and head kidney. There was a significant correlation between ScMx1 and ScIFN-I expressions in spleen(r=0.94, P=0.018). These results indicated that there were ScMx1 gene existed in Squaliobarbus curriculus, and it may play an important role in the immune response of against GCRV.

关 键 词：赤眼鳟 Mx1 基因克隆 IFN-I GCRV 组织表达 

分 类 号：Q344.1[生物学—遗传学]