水产学报2018，Vol.42Issue(6)：817-827,11.DOI:10.11964/jfc.20171010990
九孔鲍MSTN基因cDNA克隆及表达
Molecular cloning and expression analysis of the MSTN cDNA in Haliotis diversicolor supertexta
摘要
Abstract
Myostatin is an important member of the transforming growth factor (TGF) family that functions to reg-ulate muscle development and growth in animals, The purpose of this study was to characterize and predict func-tion of the myostatin gene of Haliotis diversicolor supertexta which is an important aquaculture shellfish. In this study, the myostatin (Hs-MSTN) cDNA of H. diversicolor supertexta were cloned and characterized by rapid amp-lification cDNA ends (RACE) methods. The full length of Hs-MSTN cDNA sequence consists of 3 755 bp contain-ing a 5′ untranslated region (UTR) of 324 bp, a 3′ UTR of 1 985 bp, and an open reading frame of 1 446 bp encod-ing a protein with 481 amino acid residues, with a calculated molecular mass of 54.96 ku, and the theoretical iso-electric point of 9.41, The structure of Hd-MSTN included a putative signal peptide (1-17 aa), a TGF-β propeptide domain (157-367 aa) and a conserved TGF-β domain (379-481 aa). Multiple sequence alignment results revealed conservation of the RRPR proteolytic site and nine conserved cysteines of the Hs-MSTN with MSTN from other animals, and two propeptide proteolytic sites RQRR (120-124 aa) and RYRR (235-239 aa) were found. Phylogen-etic analysis showed that the Hs-MSTN gene was clustered in the same subgroup with the H. rufescens, Quantitat-ive real-time PCR detection results indicated that the Hs-MSTN genes were expressed widely in adductor muscle, mantle, gonad, liver, gill, foot and the highest expression level was observed in the adductor muscle, mantle, foot, and Hs-MSTN transcript was widely detected in early developmental stages: unfertilized egg, fertilized egg, 4-cell embryos, 8-cell embryos, gastrulae, larvae, juvenile stage, and the higher in fertilized egg, in 4-cell embryos and 8-cell embryos and the expression level increases in gastrulae, larvae and decreases in juvenile stage. Our results in-dicate that MSTN is involved in muscle growth regulation of H. diversicolor supertexta.关键词
九孔鲍/肌肉生长抑制素/基因克隆/基因表达Key words
Haliotis diversicolor supertexta/myostatin/gene cloning/gene expression分类
生物科学引用本文复制引用
艾加林,栗志民,刘建勇..九孔鲍MSTN基因cDNA克隆及表达[J].水产学报,2018,42(6):817-827,11.基金项目
深圳市大鹏新区产业发展专项(KY20170211) Development Special Fund of Shenzhen Dapeng New District (KY20170211) （KY20170211）
