Molecular cloning and expression analysis of the myostatin gene in sea perch (Lateolabrax japonicus).
Myostatin (MSTN) is a member of the transforming growth factor-beta (TGF-beta) superfamily that functions as a negative regulator of skeletal muscle development and growth in mammals. However, few reports are available about the structure and function of MSTN in teleost. Here, the MSTN gene was cloned from sea perch (Lateolabrax japonicus) by homology cloning and genomic walking. In the 4873-bp genomic sequence, three exons, two introns, and 5' and 3' flanking sequences were identified. The sea perch MSTN gene encodes a 374-amino acid protein, including a signal peptide, conserved cysteine residues, and a RXXR proteolytic cleavage domain. Expression analysis of MSTN revealed that MSTN was highly expressed in eyes, brain, and muscle; intermediately in intestine; and weakly in gill, spleen, liver, and heart. It was demonstrated that MSTN mRNA was highly expressed in embryonic stem cell line (LJES1), but it was undetectable in several types of somatic cell lines from sea perch, including fibroblast-like cell, epithelioid cell, and lymphocyte-like cell. Further, it was demonstrated that the 5' flanking region of the MSTN gene can drive the expression of green fluorescent protein (GFP) reporter gene in LJES1 cells and transgenic zebrafish (Danio rerio). This is the first report on the expression profile of MSTN gene in various types of cell cultures.
Ye
Han-Qing
HQ
Key Lab For Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China.
Chen
Song-Lin
SL
Sha
Zhen-Xia
ZX
Liu
Yang
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression / Q000502:physiology*; D020869:Gene Expression Profiling / Q000662:veterinary*; D023061:Gene Order; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000502:physiology; D055435:Myostatin; D010471:Perches / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000235:genetics*; D029961:Zebrafish Proteins
D017931:DNA Primers; C525914:Mstnb protein, zebrafish; D055435:Myostatin; D016212:Transforming Growth Factor beta; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins; D004247:DNA