半滑舌鳎骨形态发生蛋白4基因的克隆和表达分析
作者:
作者单位:

山东省渔业资源与生态环境重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071

作者简介:

马骞(1983-), 女, 博士, 研究方向为渔业资源分子生态学. E-mail: maqian@ysfri.ac.cn

中图分类号:

S917

基金项目:

国家自然科学基金项目(31201981);中国水产科学研究院黄海水产研究所级基本科研业务费资助项目(20603022012032).


Molecular cloning and expression analysis of BMP4 gene in tongue sole (Cynoglossus semilaevis)
Author:
Affiliation:

Key Laboratory for Fishery Resources and Eco-environment, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China

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    摘要:

    本研究克隆了半滑舌鳎()骨形态发生蛋白4(BMP4)基因的cDNA序列,并分析其在半滑舌鳎成鱼的组织表达分布及其在早期发育阶段的定位和表达变化趋势。结果表明,半滑舌鳎基因cDNA全长为1680 bp,包括了419 bp的5'非翻译区(5'UTR)、编码403个氨基酸的1212 bp的开放阅读框(ORF)以及49 bp的3'非翻译区(3'UTR)。同源性分析和分子进化聚类分析结果显示,半滑舌鳎与已报道慈鲷科(Cichlidae)各鱼种的同源性最高,并与之共同聚为鱼类BMP4分支。BMP4基因在早期胚胎发育阶段(卵裂期和原肠期)具有极高水平的表达,此后其表达水平逐渐降低;但在后期仔鱼期(孵化后3日龄和4日龄),其表达水平再次升高。整体原位杂交检测结果表明, 基因主要在冠状幼鳍、上下颌及胸鳍处表达;进入稚鱼期后在鳃盖骨及各鳍均有表达。上述研究结果揭示了BMP4在半滑舌鳎早期骨骼发育中的重要作用,为揭示海水硬骨鱼类骨骼发生、发育的调控机制奠定了理论基础。

    Abstract:

    The growth and development of the vertebrate skeleton is regulated by the bone morphogenetic proteins (BMPs), among which BMP4 is one of the most active osteoinduction factors. In this study, we cloned the ) and described its expression profiling. The cDNA sequence was 1680 bp, and contained a 5'-untranslated region (5'-UTR) of 419 bp, an open reading frame (ORF) of 1212 bp encoding a protein of 403 amino acids, and a 3'-UTR of 49 bp. Homology and phylogenetic analyses revealed that the deduced amino acid sequence of the BMP4 protein shares highest identity with those of Cichlidae species, and fish BMP4 forms a single lineage distinct from the BMP4 proteins of other vertebrates. The tissue expression distribution of the mRNA was predominantly expressed in the gill, and less in the dorsal fin, cartilage, etc. In the early developmental stages, mRNA was also detected at the egg, larval, juvenile, and fingerling stages, with the highest expression in the egg stage (cleavage and gastrula periods), and the second highest in the larval stage (3-4 days old). Whole-mount in situ hybridization showed that mRNA was firstly detected in head and anterior region in the early larval stage, and then appeared in the crown-like larval fin, jaw, and pectoral fin. mRNA was then predominantly expressed in all the fins and the operculum during the early juvenile stage. These results demonstrate the important role of BMP4 in the fin development of , and provide a theoretical foundation for investigating the mechanisms regulating fish bone development.

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马骞,冯文荣,柳淑芳,庄志猛,马慧.半滑舌鳎骨形态发生蛋白4基因的克隆和表达分析[J].中国水产科学,2016,23(3):500-512
MA Qian, FENG Wenrong, LIU Shufang, ZHUANG Zhimeng, MA Hui. Molecular cloning and expression analysis of BMP4 gene in tongue sole (Cynoglossus semilaevis)[J]. Journal of Fishery Sciences of China,2016,23(3):500-512

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  • 在线发布日期: 2016-05-18
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