【作者】 陈开健； 王静安； 刘巧玲； 王荣华； 徐莹； 赵鑫； 肖调义；

【Author】 Chen Kaijian;Wang Jing-an;Liu Qiaolin;Wang Ronghua;Xu Ying;Zhao Xin;Xiao Tiaoyi;Hunan Engineering Research Center for Utilization of Characteristics of Aquatic resources,Hunan agricultural university;Collaborative Innovation Center for Efficient and Health Production of Fisheries in Hunan Province;

【机构】 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； 水产高效健康生产湖南省协同创新中心；

【摘要】 本文采用RACE技术获得了赤眼鳟(Squaliobarbus curriculus)TRAF6(ScTRAF6)基因cDNA全长,共2621 bp,包含5’非编码区50 bp,开放阅读框1626 bp,3’非编码区942 bp。ScTRAF6共编码542个氨基酸,推导的蛋白质分子量为61.67k Da,等电点为6.01。SMART结构分析结果显示,ScTRAF6包含RING结构,两个锌指结构,螺旋卷曲结构和MATH结构域。系统进化树分析表明,ScTRAF6与团头鲂的亲缘关系最近。Real-time PCR结果显示:ScTRAF6在赤眼鳟的12种组织中均有表达,其中肝脏表达水平最高,中肠最低;经GCRV感染后,赤眼鳟脾脏和头肾中ScTRAF6均呈现3个周期波动上调表达,分别在6h、24h和96h达到峰值,表明TRAF6参与赤眼鳟抗GCRV免疫应答反应。更多还原

【Abstract】 The cDNA of tumor necrosis factor receptor-associated factor 6 in Squaliobarbus curriculus(ScTRAF6) was cloned by using rapid amplification of cDNA ends. The full-length cDNA of ScTRAF6 is 2621 bp including a 5’-terminal untranslated region of 50 bp, a 3’-terminal untranslated region of 942 bp and an open reading frame of 1626 bp encoding a polypeptide of542 amino acid residues. The putative isoelectric point(p I) and molecular weight(Mw) of ScTRAF6 was 6.01 and 61.67 k D, respectively. The ScTRAF6 protein includs one ring domain,two zinc fingers, one coiled-coil region, and one MATH domain. Phylogenetic analysis showed that the ScTRAF6 was evolutionarily closest to this in Megalobrama amblycephala. In normal barbel chub, expression of ScTRAF6 could be detected in almost all the 12 tested tissues by Real-time PCR, the highest expressed in the liver and the lowest expresed in midgut. After injection with GCRV, the ScTRAF6 expression level was up and down regulated three times in both spleen and head kidney, and reached the peak at 6 h, 24 h and 96 h. The results suggest that ScTRAF6 may play important roles in the immune response of barbel chub against GCRV.更多还原