Characterization of a myostatin gene (MSTN1) from spotted halibut (Verasper variegatus) and association between its promoter polymorphism and individual growth performance.
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.
National Marine Environmental Monitoring Center, Dalian 116023, China. hjli@nmemc.gov.cn
Hongjun
H
Li
Jingfeng
J
Fan
Shuxi
S
Liu
Qing
Q
Yang
Guiqiang
G
Mu
Chongbo
C
He
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001947:Breeding / Q000379:methods; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D005432:Flounder / Q000235:genetics* / Q000254:growth & development*; D040461:Gene Components; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D011110:Polymorphism, Genetic / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D044366:Transition Temperature
D017931:DNA Primers; D055435:Myostatin