Molecular characterization, tissue expression and sequence variability of the barramundi (Lates calcarifer) myostatin gene.
BACKGROUND
Myostatin (MSTN) is a member of the transforming growth factor-beta superfamily that negatively regulates growth of skeletal muscle tissue. The gene encoding for the MSTN peptide is a consolidate candidate for the enhancement of productivity in terrestrial livestock. This gene potentially represents an important target for growth improvement of cultured finfish.


RESULTS
Here we report molecular characterization, tissue expression and sequence variability of the barramundi (Lates calcarifer) MSTN-1 gene. The barramundi MSTN-1 was encoded by three exons 379, 371 and 381 bp in length and translated into a 376-amino acid peptide. Intron 1 and 2 were 412 and 819 bp in length and presented typical GT...AG splicing sites. The upstream region contained cis-regulatory elements such as TATA-box and E-boxes. A first assessment of sequence variability suggested that higher mutation rates are found in the 5' flanking region with several SNP's present in this species. A putative micro RNA target site has also been observed in the 3'UTR (untranslated region) and is highly conserved across teleost fish. The deduced amino acid sequence was conserved across vertebrates and exhibited characteristic conserved putative functional residues including a cleavage motif of proteolysis (RXXR), nine cysteines and two glycosilation sites. A qualitative analysis of the barramundi MSTN-1 expression pattern revealed that, in adult fish, transcripts are differentially expressed in various tissues other than skeletal muscles including gill, heart, kidney, intestine, liver, spleen, eye, gonad and brain.


CONCLUSIONS
Our findings provide valuable insights such as sequence variation and genomic information which will aid the further investigation of the barramundi MSTN-1 gene in association with growth. The finding for the first time in finfish MSTN of a miRNA target site in the 3'UTR provides an opportunity for the identification of regulatory mutations on the expression of this gene.
De Santis
Christian
C
Aquaculture Genetics Research Program, School of Marine and Tropical Biology, James Cook University, Townsville, Queensland, 4811, Australia. christian.desantis@jcu.edu.au
Evans
Brad S
BS
Smith-Keune
Carolyn
C
Jerry
Dean R
DR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D029941:Fish Proteins / Q000145:classification / Q000235:genetics*; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D055435:Myostatin; D010473:Perciformes / Q000235:genetics*; D010802:Phylogeny; D020641:Polymorphism, Single Nucleotide; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D016212:Transforming Growth Factor beta / Q000145:classification / Q000235:genetics*
D029941:Fish Proteins; D055435:Myostatin; D016212:Transforming Growth Factor beta