National Marine Environmental Monitoring Center, Dalian 116023, China.
Comp Biochem Physiol B Biochem Mol Biol. 2012 Apr;161(4):315-22. doi: 10.1016/j.cbpb.2011.12.008. Epub 2011 Dec 24.
Myostatin (MSTN) is a member of the transforming growth factor-β superfamily which could play an important role in negatively regulating skeletal muscle growth and development in mammal and non-mammal species. In the present study, a MSTN1 gene (designated as VvMSTN1) was cloned and characterized in one flatfish species, spotted halibut (Verasper variegatus). In the 3078 bp genomic sequence, three exons, two introns and a promoter sequence were identified. Sequence analysis of the promoter region revealed that it contained several cis-regulatory elements such as CAAT-box, TATA-box and E-boxes. The deduced protein sequence included a signal peptide, a TGF-β propeptide in the N-terminal region and the TGF-β active peptide in the C-terminal region. Phylogenetic analysis suggested that VvMSTN1 is an orthologue of teleost MSTN1 proteins which arose along with MSTN2 during a duplication event at the base of teleost evolution. Quantitative real-time PCR analysis revealed that VvMSTN1 mRNA was ubiquitously expressed in all nine tested tissues, with the most transcriptionally abundant in skeletal muscle. A primary assessment of sequence variability revealed five single nucleotide polymorphisms (SNPs) existed in the promoter region, among which three (G-653T, T-355C and G-253A) were genotyped with an advanced melting temperature (T(m))-shift method and tested for their association with growth traits (body length, body depth and total mass). Results indicated that genotype CC of locus T-355C had significantly higher growth traits than genotype TC and TT (P<0.05) in female spotted halibut. These results suggest that V. variegatus MSTN could be selected as a candidate gene for the future molecular breeding of stains with enhanced individual growth performance.
肌肉生长抑制素 (MSTN) 是转化生长因子-β超家族的成员，它在调节哺乳动物和非哺乳动物的骨骼肌生长和发育中起着重要作用。本研究在一种比目鱼类——星斑川鲽（Verasper variegatus）中克隆并鉴定了 MSTN1 基因（命名为 VvMSTN1）。在 3078bp 的基因组序列中，鉴定出三个外显子、两个内含子和一个启动子序列。启动子区序列分析表明，它含有 CAAT 盒、TATA 盒和 E 盒等几个顺式调控元件。推导的蛋白质序列包括一个信号肽、N 端的 TGF-β前肽和 C 端的 TGF-β活性肽。系统进化分析表明，VvMSTN1 是硬骨鱼类 MSTN1 蛋白的同源物，它是在硬骨鱼类进化基础上的一次复制事件中与 MSTN2 一起产生的。定量实时 PCR 分析显示，VvMSTN1mRNA 在所有 9 种检测组织中均广泛表达，在骨骼肌中转录丰度最高。对序列变异性的初步评估显示，启动子区存在五个单核苷酸多态性（SNP），其中三个（G-653T、T-355C 和 G-253A）通过高级熔解温度（T(m))-shift 方法进行了基因分型，并检测了它们与生长性状（体长、体深和总质量）的相关性。结果表明，T-355C 位点的 CC 基因型雌星斑川鲽的生长性状显著高于 TC 和 TT 基因型（P<0.05）。这些结果表明，V. variegatus MSTN 可以作为未来具有增强个体生长性能的品系分子育种的候选基因。
