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来自墨吉对虾(Fenneropenaeus merguiensis)的一种与蜕皮相关的肌肉生长抑制素基因(FmMstn)的特征分析

Characterization of a molt-related myostatin gene (FmMstn) from the banana shrimp Fenneropenaeus merguiensis.

作者信息

Zhuo Rui Qun, Zhou Ting Ting, Yang Shi Ping, Chan Siuming Francis

机构信息

Fisheries College, Guangdong Ocean University, Zhanjiang, PR China.

Fisheries College, Guangdong Ocean University, Zhanjiang, PR China; South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, PR China.

出版信息

Gen Comp Endocrinol. 2017 Jul 1;248:55-68. doi: 10.1016/j.ygcen.2017.03.010. Epub 2017 Mar 18.

Abstract

Myostatin is an important member of the transforming growth factor (TGF) family that functions to regulate muscle growth in animals. In this study, the myostatin gene (FmMstn) and two slightly different (short and long forms) cDNAs of the banana shrimp Fenneropenaeus merguiensis were cloned and characterized. Similar to Mstn gene of the scallop, fish and mammal, FmMstn gene consists of 3 exons and 2 introns. The 2kb upstream promoter region of the FmMstn gene consists of putative response elements for myocyte enhancing factor (MEF2) and E-box factors. The longest open reading frame of the short Mstn consists of 1260bp encoding for a protein with 420 amino acid residues. The long FmMstn is almost identical to the short FmMstn with the exception of 8 amino acid insertions. FmMstn is most similar to the Mstn of Litopenaeus vannamei and Penaeus monodon sharing >92-98% amino acid sequence identity. Multiple sequence alignment results revealed high degree of amino acid conservation of the cysteine residues and mature peptide of the FmMstn with Mstn from other animals. FmMstn transcript was detected in the heart, muscle, optic nerve and thoracic ganglion. FmMstn transcript level in muscle is higher in early postmolt, decreases in intermolt and increases again towards ecdysis. Higher expression level of FmMstn is also observed in smaller shrimp of the same age. Knock-down of FmMstn gene by RNAi can cause a significant increase in molt cycle duration and failure of some shrimp to undergo ecdysis. Direct DNA sequencing results revealed that FmMstn gene is highly polymorphic and several potential SNPs have been identified. Some SNPs are associated with the size difference of the shrimp. In summary, the result of this study indicates that shrimp FmMstn gene is molt/growth-related and the presence of SNP suggests that it could be a candidate gene for shrimp genetic improvement research.

摘要

肌肉生长抑制素是转化生长因子(TGF)家族的重要成员,其作用是调节动物的肌肉生长。在本研究中,克隆并鉴定了墨吉对虾(Fenneropenaeus merguiensis)的肌肉生长抑制素基因(FmMstn)以及两个略有不同(短型和长型)的cDNA。与扇贝、鱼类和哺乳动物的Mstn基因相似,FmMstn基因由3个外显子和2个内含子组成。FmMstn基因2kb的上游启动子区域包含心肌增强因子(MEF2)和E-box因子的假定反应元件。短型Mstn的最长开放阅读框由1260bp组成,编码一个含有420个氨基酸残基的蛋白质。长型FmMstn与短型FmMstn几乎相同,只是有8个氨基酸的插入。FmMstn与凡纳滨对虾(Litopenaeus vannamei)和斑节对虾(Penaeus monodon)的Mstn最为相似,氨基酸序列同一性大于92%-98%。多序列比对结果显示,FmMstn的半胱氨酸残基和成熟肽与其他动物的Mstn具有高度的氨基酸保守性。在心脏、肌肉、视神经和胸神经节中检测到FmMstn转录本。肌肉中FmMstn转录本水平在蜕皮后早期较高,在蜕皮间期降低,并在蜕皮期再次升高。在相同年龄的较小对虾中也观察到FmMstn的较高表达水平。通过RNAi敲低FmMstn基因可导致蜕皮周期持续时间显著增加,并且一些对虾无法蜕皮。直接DNA测序结果显示FmMstn基因具有高度多态性,已鉴定出几个潜在的单核苷酸多态性(SNP)。一些SNP与对虾的大小差异有关。总之,本研究结果表明对虾FmMstn基因与蜕皮/生长相关,SNP的存在表明它可能是对虾遗传改良研究的候选基因。

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