七彩神仙鱼β-胸腺素基因克隆及表达分析  

Cloning and expression analysis ofβ-thymosin gene in Symphysodon aquifasciatus

作　　者：彭啸 温彬 陈再忠[1,2,3] 高建忠 PENG Xiao;WEN Bin;CHEN Zaizhong;GAO Jianzhong(Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,Ministry of Education,Shanghai Ocean University,Shanghai 201306,China;National Demonstration Centre for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]上海海洋大学,农业农村部淡水水产种质资源重点实验室,上海201306 [2]上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海201306 [3]上海海洋大学,水产科学国家级实验教学示范中心,上海201306

出　　处：《淡水渔业》2024年第3期78-84,共7页Freshwater Fisheries

基　　金：上海市自然科学基金(20ZR1423600)。

摘　　要：为了探究胸腺素基因在七彩神仙鱼(Symphysodon aquifasciatus)中的特征、进化和功能,基于转录组序列比对及生物信息学的方法对七彩神仙鱼的胸腺素进行了鉴定、基因与蛋白结构分析、系统发生分析,通过RACE技术克隆获得了七彩神仙鱼胸腺素基因全长,利用qRT-PCR技术研究其在各组织中的表达规律,通过病原感染实验,研究了七彩神仙鱼β-胸腺素基因在病原胁迫下的表达规律。结果显示,七彩神仙鱼的胸腺素基因鉴定为胸腺素β1型,命名为dfTβ。克隆结果显示,dfTβ的全长为779 bp,ORF区序列长度为303 bp,编码100个氨基酸,其5′-UTR为43 bp,3′-UTR为433 bp,含有22个poly-A。系统发育进化分析结果显示,七彩神仙鱼β-胸腺素同尼加拉瓜湖始丽鱼β-胸腺素聚为一簇。qPCR结果显示,β-胸腺素在七彩神仙鱼各组织中均有表达,其中在肠、皮肤、鳃中表达最高,在头肾和脾中表达最低。嗜水气单胞菌(Aeromonas hydrophila)感染后,七彩神仙鱼皮肤、头肾、脾和肠中dfTβ均在短时间内显著上调,并随着时间出现先上升后下降的趋势,但各组织中上调倍数最高峰的时间不同。研究结果表明,dfTβ基因参与了七彩神仙鱼受病原刺激后的免疫应答,且表达模式不同,表明其可能在不同组织、免疫的不同阶段发挥不同的作用。β-thymosins are a family of highly conserved polar 5-kDa polypeptides present in high concentra-tions in almost every cell.It has multiple and diverse intracellular and extracellular functions in vertebrates.It has a potentially strong immune function and has not been studied in Symphysodon aquifasciatus.The experiment used comparative genomics and bioinformatics methods to identify the thymosin of Symphysodon aquifasciatus,gene and protein structure analysis,and phylogenetic analysis to explore the characteristics,evolution and function of the thymosin gene in S.aquifasciatus.Through comparing the genome of S.aquifasciatus with NCBI database to construct the phylogenetic tree.The full length of S.aquifasciatus gene was obtained by RACE-PCR,and its ex-pression in various tissues was studied by quantitative real-time PCR.Then,we examined the expression of the thymosin gene under pathogen stress.The results showed that the thymosin gene of the fish was identified as thymosinβType 1,named dfTβ.The results of RACE-PCR showed that the full-length of dfTβgene was 779 bp,which contained 303 bp open reading frame,encoding 100 amino acids,and 43 bp 5′non-coding re-gion,433 bp 3′non-coding region.The results of phylogenetic evolution analysis showed that S.aquifasciatusβ-Thymosin gathered with Archocentrus centrarchus.The qPCR results showed that thymosin was expressed in all fish tissues,with the highest expression in the intestine,skin and gills and the lowest expression in the head,head kidney and spleen.After the infection of Aeromonas hydrophila,dfTβin the skin,head kidney and spleen of the colorful angelfishβ1 were significantly up-regulated in a short period,and showed a trend of first rising and then falling with time,but the time of the highest up-regulation multiple in each tissue was different.The re-sults show that the dfTβgene is involved in the immune response of the fish after being stimulated by the patho-gen,and its expression patterns are different,indicating that it may play different roles i

关 键 词：七彩神仙鱼(Symphysodon aquifasciatus) β-胸腺素 基因克隆 表达分析 

分 类 号：S965.8[农业科学—水产养殖]