瘤背石磺肌肉生长抑制素基因的克隆及组织表达分析

发布时间：2018-11-20 05:11

【摘要】：为探究石磺由海洋向陆地进化过程中肌肉生长和发育的分子机制,实验以石磺科贝类转录组数据为基础,采用RACE方法从瘤背石磺肌肉中首次克隆到MSTN cDNA的全长并做了相应的组织表达分析。结果显示,瘤背石磺MSTN基因cDNA全长2667 bp,包括1650 bp的开放阅读框,374 bp的5′端非翻译区,643 bp的3'端非翻译区,共编码549个氨基酸。预测该基因编码的蛋白质原子总量为8776,分子式为C2774H4331N783O862S26,分子质量约为63.27 ku,理论等电点为6.02,信号肽预测结果显示,N端具有21个氨基酸长度的信号肽。瘤背石磺MSTN具有MSTN的共同特征,包括蛋白酶水解位点RSRR和C端多肽生物活性区以及9个保守的半胱氨酸残基。通过进化树分析,瘤背石磺MSTN与加州海兔MSTN的亲缘关系最近。RT-PCR结果显示,MSTN基因在各个组织中均有表达;含肌纤维组织中的表达量低于内脏器官的表达量,在肝胰腺中的表达量最高,腹足表达量最低。MSTN基因一级结构具有很高的保守性,说明该基因在进化上的限制性和功能的重要性;同时该基因在石磺非肌肉组织中表达,表明该基因不仅有抑制肌肉生长的作用,还参与其他生命活动的调节。
[Abstract]:In order to explore the molecular mechanism of muscle growth and development during the evolution of stone sulfonate from ocean to land, the experiment was based on the transcriptome data of shellfish. RACE method was used to clone the full length of MSTN cDNA from the sarcomatous muscle for the first time and the corresponding tissue expression was analyzed. The results showed that the total length of cDNA of MSTN gene was 2667 bp, including open reading frame of 1650 bp, 5 'untranslated region of 374 bp and 3' terminal untranslated region of 643 bp, encoding 549 amino acids. The total amount of protein atoms encoded by the predicted gene is 8776, the molecular formula is C2774H4331N783O862S26, and the molecular mass is about 63.27 ku, the theoretical isoelectric point is 6.02. The signal peptide prediction results show that the N-terminal has 21 amino acid length signal peptides. MSTN has the common characteristics of MSTN, including protease hydrolysis site (RSRR), C-terminal polypeptide bioactive region and 9 conserved cysteine residues. According to phylogenetic tree analysis, the relationship between MSTN and MSTN was close. RT-PCR results showed that MSTN gene was expressed in all tissues. The expression of MSTN gene in the primary structure of the MSTN gene was highly conserved, but the highest in the hepatopancreas and the lowest in the gastropods, as compared with that in the visceral organs, and the expression level in the muscle-containing fibers was lower than that in the visceral organs. It shows the importance of the restriction and function of the gene in evolution. At the same time, the gene was expressed in non-muscle tissue, indicating that the gene not only inhibits muscle growth, but also participates in the regulation of other life activities.
【作者单位】：上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室;
【基金】：国家自然科学基金(41276157) 上海高校水产学一流学科建设项目~~
【分类号】：S917.4

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