团头鲂肌腱发育相关基因tnmd/xirp2a的克隆和表达
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国家自然科学基金项目(31872559);国家大宗淡水鱼类产业技术体系建设项目(CARS4608);中央高校基本科研业务费专项(2662018PY035);武汉市生态渔业产业技术体系品种改良岗位


Cloning and expression analysis of tnmd/xirp2a genes relating to tendon development in Megalobrama amblycephala
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    摘要:

    为分析肌腱发育相关基因tnmd/xirp2a与团头鲂肌间骨发育之间的相关性,通过克隆获得团头鲂tnmd/xirp2a的cDNA序列,其中tnmd的cDNA全长为1 420 bp,开放阅读框为906 bp,共编码301个氨基酸;xirp2a的cDNA全长为11 715 bp,包括开放阅读框9 522 bp,编码3 174个氨基酸。系统进化研究表明,tnmd/xirp2a基因在不同脊椎动物中保守性很低,且相比哺乳类和家禽类,鱼类tnmd基因存在一段氨基酸的缺失。采用荧光定量方法比较分析tnmd/xirp2a在团头鲂成鱼不同组织及肌间骨发生发育4个关键时期的表达情况,结果显示,tnmd/xirp2a在肌肉和肌间骨中的表达量均显著高于其他组织,且肌肉中tnmd基因表达高于肌间骨(P<0.05),而xirp2a在肌间骨中的表达高于肌肉组织(P<0.05)。在肌间骨发育4个关键时期(Ⅰ:尾部尚未出现肌间骨;Ⅱ:尾部出现少量肌间骨;Ⅲ:尾部肌间骨大量出现且长度增加;Ⅳ:肌间骨从尾部到背部全部出现且形态成熟)中,tnmd在第Ⅲ时期的表达水平显著高于另外3个时期;在第Ⅱ、Ⅲ、Ⅳ时期之间,xirp2a的表达水平没有显著性差异。此结果表明,tnmd基因在肌间骨的发育过程中存在潜在作用,而xirp2a基因与肌间骨的发育之间的相关性还需进一步研究。

    Abstract:

    In order to investigate the relationship between the tendon development related genes (tnmd/xirp2a) and the intermuscular bone in blunt snout bream (Megalobrama amblycephala),we firstly obtained the cDNA sequence of tnmd/xirp2a by cloning. The fulllength cDNA of tnmd is 1 420 bp,including an open reading frame (ORF) of 906 bp encoding 301 amino acids. The cDNA sequences of xirp2a is 11 715 bp in length,including an ORF of 9 522 bp encoding 3 174 amino acids. Phylogenetic analysis showed that the tnmd gene was lowly conserved among different vertebrates. The tnmd protein had a sequence deletion compared to mammals and poultry. The tnmd/xirp2a expression in the muscle and intermuscular bone was significantly higher than that in other tissues (P<0.05). The tnmd expression in the muscle was significantly higher than that in the intermuscular bone (P<0.05),while the result of xirp2a was the exact opposite. During intermuscular bone development,the tnmd mRNA was significantly higher at stage Ⅲ than that at the other three stages. However,there was no significant differences of the xirp2a expression among stages Ⅱ,Ⅲ and Ⅳ (P>0.05). Our results indicate that the tnmd gene may play possible regulation roles during the intermuscular bone development,while the relationship between the xirp2a gene and the intermuscular bone development needs to be further studied. The results of this study provide theoretical support for the derivation of intermuscular bone from the tendon.

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陈宇龙,张丽红,周佳佳,孟佑廉,高泽霞.团头鲂肌腱发育相关基因tnmd/xirp2a的克隆和表达[J].华中农业大学学报,2019,38(2):1-8

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  • 收稿日期:2018-10-14
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  • 在线发布日期: 2019-01-30
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