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引用本文:史宝,孙冉冉,柳学周,徐永江,姜燕,王滨,张正荣.黄条鰤(Seriola aureovittata)MyHC基因克隆及其在早期发育阶段表达研究.海洋与湖沼,2020,51(2):422-432.
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黄条鰤(Seriola aureovittata)MyHC基因克隆及其在早期发育阶段表达研究
史宝1, 孙冉冉1,2, 柳学周1,2, 徐永江1, 姜燕1, 王滨1, 张正荣1
1.青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.大连海洋大学水产与生命学院 大连 116023
摘要:
为解析肌球蛋白重链(MyHC)基因在黄条鰤(Seriola aureovittata)生长发育过程中作用机制,本研究采用RACE技术,克隆了黄条鰤MyHC基因全长cDNA序列,采用荧光定量PCR(qRT-PCR)方法对MyHC在黄条鰤不同组织、胚胎发育过程以及仔稚幼鱼发育过程中的表达特征进行了研究。结果表明:黄条鰤MyHC基因全长为6143bp,开放阅读框为5811bp,编码了1936个氨基酸,由3个保守结构域组成即MYSc-class II、Myosin tail1和SH3;系统进化树分析显示黄条鰤MyHC与高体鰤进化关系最近。qRT-PCR分析发现黄条鰤MyHC在各组织中均有表达,但在肌肉中表达量最高(P<0.05);随着黄条鰤胚胎发育的进行,MyHC在16细胞期之前表达量较高,在胚体下包2/3时期表达量显著升高,且在孵化期达到峰值(P<0.05);在仔稚幼鱼发育阶段,MyHC在孵化后20d后表达量显著升高,30d表达水平达到峰值(P<0.05),随后的35d到40d表达水平略有下降但仍保持较高表达趋势,黄条鰤MyHC表达具有发育阶段表达的特异性。MyHC表达特征揭示其参与了调控黄条鰤的早期生长发育。
关键词:  黄条鰤  MyHC  胚胎发育  仔稚幼鱼发育  基因克隆  表达分析
DOI:10.11693/hyhz20191100205
分类号:S917.4
基金项目:青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室开放课题,2017-3A01号;国家重点研发计划项目,2018YFD0901204号,2019YFD0900503号;国家自然科学基金项目,31772829号;中国水产科学研究院院级基本科研业务费-农业部海洋渔业可持续发展重点实验室开放课题资助,2019HY-XKQ01号;国家海水鱼产业技术体系项目,CARS-47号。
MOLECULAR CLONING AND mRNA EXPRESSION OF MyHC GENE IN THE EARLY DEVELOPMENT OF YELLOWTAIL KINGFISH SERIOLA AUREOVITTATA
SHI Bao1, SUN Ran-Ran1,2, LIU Xue-Zhou1,2, XU Yong-Jiang1, JIANG Yan1, WANG Bin1, ZHANG Zheng-Rong1
1.Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and Technology, Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;2.College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China
Abstract:
The myosin heavy chain (MyHC) is one of the major structural and contractile proteins of muscle. Study on the early growth and development and gene resources is relatively lacking in yellowtail kingfish Seriola aureovittata. To understand the regulatory roles of MyHC in the early growth and development of S. aureovittata, the full-length cDNA sequences of MyHC was cloned by the RACE technology. The real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression patterns of MyHC in different tissues, embryonic development stages, and larval and juvenile stages. The full length of the MyHC cDNA sequence was 6143bp, and the open reading frame was 5811bp, encoding 1936 amino acids. Additionally, common features were found in the MyHC of S. aureovittata, including MYSc-class II, myosin tail l and SH3 domain. Molecular phylogenetic analysis confirmed that S. aureovittata was closely related to greater amberjack S. dumerili. MyHC was expressed in various tissues, but it had the highest expression in muscle. Developmentally, there was a gradual increase in MyHC expression before the 16-cell stage. And the expression of MyHC increased significantly in the embryo encircling 2/3 of yolk sac, and reached the peak in the hatching larva (P<0.05). In the larval and juvenile stages, MyHC was significantly up-regulated on 20 days post hatching (dph), with peak expression occurring on 30 dph and decreased slightly between 35 and 45 dph (P<0.05). The MyHC expression of S. aureovittata showed a development-stage-specific characteristic. Therefore, MyHC plays a regulatory role in the embryo and early developmental stage of S. aureovittata.
Key words:  Seriola aureovittata  MyHC  embryonic development  larval and juvenile development  gene cloning  expression analysis
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