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齐口裂腹鱼肌肉生长抑制素基因的cDNA克隆与表达分析

[cDNA cloning and expression analysis of MSTN gene from Schizopygopsis pylzovi].

作者信息

Chao Yan, Zhao Lan-Ying, Li Chang-Zhong, Xie Bao-Sheng, Shen Zhi-Xin, Wang Guo-Jie, Wang Zhen-Gang, Li Chao, Bai Bin-Qiang, Zhang Hui, Qi De-Lin

机构信息

Animal Science Department of Agriculture and Animal Husbandry College, Qinghai University, Xining 810016, China.

出版信息

Dongwuxue Yanjiu. 2012 Oct;33(5):473-80. doi: 10.3724/SP.J.1141.2012.05473.

Abstract

Myostatin (MSTN) is a member of the TGF-β superfamily that acts as a negative regulator of skeletal muscle growth. A full-length, 2 180 bp, cDNA sequence of the myostatin gene from Schizopygopisis pylzovi was cloned with RT-PCR,5'-RACE and 3'-RACE and the cDNA clone included a 1 128 bp ORF, encoding a 375 amino acid peptide. Using PCR, we obtained the sequences of two introns of the MSTN gene and found that its structure in Schizopygopsis pylzovi was similar to that of other vertebrates, including three exons and two introns. Likewise, the putative MSTN peptide of Schizopygopsis pylzovi contains a conserved RXXR proteolytic cleavage domain, and 8 conserved cysteine residues in the C terminal of the protein, similar to other vertebrates. Phylogenetic analysis showed that the MSTN of Schizopygopsis pylzovi has high homology with other cyprinid fishes, but a low homology with mammals and birds. In the 9 examined tissues, the MSTN gene was highly expressed in heart, kidney, intestine and spermary, while weakly expressed in muscle, brain, fat, gill and hepatopancreas. Quantitative real-time PCR analysis showed that the expression of MSTN gene was different during embryo development, suggesting that the fish MSTN may not only play roles in muscle development but also contribute to other biological functions.

摘要

肌肉生长抑制素(MSTN)是转化生长因子-β超家族的成员,作为骨骼肌生长的负调节因子。采用逆转录聚合酶链反应(RT-PCR)、5'-cDNA末端快速扩增(5'-RACE)和3'-cDNA末端快速扩增(3'-RACE)技术,克隆了齐口裂腹鱼肌肉生长抑制素基因全长2180bp的cDNA序列,该cDNA克隆包含一个1128bp的开放阅读框(ORF),编码一个375个氨基酸的肽段。通过聚合酶链反应(PCR),获得了肌肉生长抑制素基因两个内含子的序列,发现其在齐口裂腹鱼中的结构与其他脊椎动物相似,包括3个外显子和2个内含子。同样,齐口裂腹鱼推定的肌肉生长抑制素肽段含有一个保守的RXXR蛋白水解切割结构域,且在该蛋白的C末端有8个保守的半胱氨酸残基,这与其他脊椎动物相似。系统发育分析表明,齐口裂腹鱼的肌肉生长抑制素与其他鲤科鱼类具有高度同源性,但与哺乳动物和鸟类的同源性较低。在所检测的9个组织中,肌肉生长抑制素基因在心脏、肾脏、肠道和精巢中高表达,而在肌肉、脑、脂肪、鳃和肝胰腺中低表达。实时定量聚合酶链反应(qRT-PCR)分析表明,肌肉生长抑制素基因在胚胎发育过程中的表达存在差异,提示鱼类肌肉生长抑制素可能不仅在肌肉发育中发挥作用,还参与其他生物学功能。

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