Cloning and expression analysis of the myostatin gene in Elopichthys bambusa

In order to clone full-length myostatin gene and characterize its tissue expression profile in E. bambusa,RT-PCR and rapid-amplification of cDNA ends(RACE)were used to acquire full-length cDNA.A full-length of 2 177 bp myostatin cDNA sequence in E. bambusa was cloned,which included a 1 125 bp complete ORF encoding a 375 amino acids peptide,a 89 bp long 5′-untranslated region,and a 1 052 bp long 3′-untranslated region.The 375 amino acids long putative peptide contains a 22 amino acids long signal peptide,a conserved RIRR proteolytic processing site,and 9 conserved cysteine residues in the C terminal of the protein.The E. bambusa MSTN had high similarity to the Megalobrama pellegrini,Ctenopharyngodon idella,Cyprinus carpio myostatin while had low similarity to the M. perciformfishes myostatin and lower similarity to the mammalian and bird myostatin.The phylogenetic analyses showed that the E. bambusa myostatin had closest relationship with the M. pellegrini myostatin.Furthermore,myostatin mRNA tissue expression profile was assessed by semi-quantitative RT-PCR.In the seven examined tissues,the myostatin gene was expressed strongly in muscle and brain,weakly in heart but not expressed in kidney,intestine,gill,heart and liver.These results suggested that the fish myostatin gene might not only play roles in muscle development but also contribute to other biological functions.