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大头鳕TNFSF6基因的结构分析及其在发育早期和病毒暴发时的表达水平 预览

Structural analysis of TNFSF6 gene and its expression profiles in early development stages and virus outbreak in Gadus macrocephalus
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摘要 为揭示大头鳕TNFSF6基因的基本结构与功能及其在大头鳕发育和神经坏死病毒(Pacific cod nervous necrosis virus, PCNNV)暴发时的响应机制,本研究通过基因克隆获得TNFSF6 cDNA开放阅读框序列,并对序列进行生物信息学分析,运用相对荧光定量PCR(qRT-PCR)方法对TNFSF6在不同组织、孵化后不同日龄仔鱼和PCNNV感染前后仔稚鱼中的表达水平进行检测。结果显示,大头鳕TNFSF6 cDNA长1 388 bp,5′UTR占315bp,3′UTR占500 bp,ORF全长573 bp,编码190个氨基酸。qRT-PCR结果显示,TNFSF6在各组织均有表达,但在脾脏和鳃组织中的表达量较高;大头鳕孵化后15、20、37和40 dTNFSF6基因的转录水平分别是其在5 d转录水平的0.28、0.15、0.12和0.13倍。在24 d和46 d PCNNV暴发时,病鱼TNFSF6基因的转录水平高于对照组;在77 d PCNNV暴发时,病鱼TNFSF6转录水平则显著低于对照组。研究表明,TNFSF6基因在大头鳕发育早期和仔鱼暴发PCNNV时发挥了重要的作用。 In order to understand the structure and expression profiles of TNFSF6 gene in Gadus macrocephalus, the full-length cDNA of TNFSF6 was cloned and analyzed using the bioinformatics tools, and quantitative realtime PCR (qRT-PCR) was used to examine the TNFSF6 mRNA expression levels in various tissues of adult G. macrocephalus, at different development stages of larvae, and during PCNNV outbreak. The results showed that TNFSF6 cDNA contains a 5′UTR of 315 bp, a 3′UTR of 500 bp, and an ORF of 573 bp encoding a protein of 190 amino acids. The results revealed that TNFSF6 was expressed in various tissues and it was higher in the spleen and gill. QRT-PCR indicated that the transcript level of TNFSF6 on 15, 20, 37, and 40 d post hatching was 0.28, 0.15, 0.12, 0.13 times of that on 5 d respectively. Compared to the control, the transcript level of TNFSF6 decreased significantly during virus outbreak on 77 d, however, there was an upward trend of TNFSF6 expression on 24 and 46 d respectively. The results indicated that TNFSF6 plays an important role in the early development stages and in responding to PCNNV outbreak.
作者 刘瑞婷 蒋洁兰 毛明光 姜志强 布尔特 矫志伟 陈明康 孙谦 LIU Ruiting;JIANG Jielan;MAO Mingguang;JIANG Zhiqiang;BU Erte;JIAO Zhiwei;CHEN Mingkang;SUN Qian(Key Laboratory of Mariculture and Stock Enhancement in North China’s Liaoning Province Sea,Ministry of Agriculture and Rural Affairs,Key Laboratory of Fish Applied Biology and Aquaculture in North China,Liaoning Province,Dalian Ocean University,Dalian 116023,China;State Key Laboratory of Freshwater Ecology and Biotechnology,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China)
出处 《水产学报》 CAS CSCD 北大核心 2019年第2期523-531,共9页 Journal of Fisheries of China
基金 国家自然科学基金(31302202) 中国博士后基金(2016M602391) 中国水产科学研究院基本科研和农业部海洋渔业可持续发展重点实验室开放课题(2017HY-XKQ01-08) 辽宁省教育厅专项(QL201702,LR2016009).
关键词 大头鳕 肿瘤坏死因子 TNFSF6 发育 神经坏死病毒 Gadus macrocephalus tumor necrosis factor TNFSF6 development viral nervous necrosis
作者简介 通信作者:毛明光,E-mail:mmg@dlou.edu.cn.
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