眼斑拟石首鱼肌肉生长抑制素基因克隆及组织表达分析
Cloning and tissue expression analysis of the myostatin gene in the red drum,Sciaenops ocellatus
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摘要: 肌肉生长抑制素(MSTN)基因对肌肉生长起负调控作用,在动物育种上具有潜在的应用前景.报道了眼斑拟石首鱼(Sciaenops ocellatus)的MSTN基因的序列结构特征及其组织特异性表达.MSTN基因具有3个外显子和2个内含子,编码376个氨基酸.在内含子Ⅰ和3'-UTR区内,分别发现了(TC)4CC(TC)4CT(TC)4和(AC)7微卫星序列.推断的眼斑拟石首鱼的MSTN氨基酸序列具很强的保守性,与石鼓鱼、金鲷、条纹石鮨、白鲈、金眼石鮨之间的相似性在90%以上.在结构上,眼斑拟石首鱼的MSTN包含N端信号序列(1~22氨基酸残基),TGF-β前肽域(41~256氨基酸残基),RXXR蛋白水解位点(264~267位的RARR)和TGF-β区域(282~376氨基酸残基).在检测的10种组织中,MSTN只在眼斑拟石首鱼的肌肉、脑和眼组织中表达,其中在肌肉表达最强.Abstract: Myostatin is a negative regulator of skeletal muscle growth and has a potential application in aquaculture.The characterization of the myostatin gene and its expression in the red drum fish,Sciaenops ocellatus,were reported.The myostat in gene possessed three exons encoding 376 amino acids.The cDNA was 1750 bp long with a 108 bp 5'-UT R and a 511 bp 3'-UTR.Two micr osat ellite sequences,(TC)4CC(TC)4CT(TC)4 and(AC)7,existed in intron Ⅰand 3'-UTR,respectively.The deduced amino acid sequence was highly conserved,and had more than 90% identical to Umbrinacirrosa,Sparus aurata,Morone saxatilis,Morone amer icana,and Morone chrysops proteins.The myostatin of the red drum had a putative amino terminal signal sequence(residues 1-22),a TGF-β propeptide domain(residues 41~256),a RXXR proteolytic processing site(RARR,residues 264~267,matching the RXXR consensus site),and a TGF-B domain(residues 282~376).There were 13 conserved cysteine residues in the red drum myostatin,nine of which are common to all TGF-β superfamily members.The most conserved region of vertebrate myostatins is the TGF-β domain,which was the mature bioactive domain of the myostat in protein.In the examined tentissues,the myostatingene was expressed only in three,the skeletal muscle,brain and eye.The strongest expression was in the skeletal muscle.
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Key words:
- Sciaenop socellatus /
- myostatin /
- cloning /
- tissue expression
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MCPHENON A C,LAWLER A M,LEE S J.Regulation of skeletal muscle mass by a new TGF——8 superfamily member[J].Nature,1997,387(6628):83-90. RODGERS B D,WEBER G M.Sequence conservation among fish myostatin orthologuea and the characterization of two additional cDNA clones from Morone saxatilis and Morone americana[J].Comp Biochem Physiol B Biochem Moi Biol,2001,129(2-3):597-603. LIN J,ARNOLD H B,DELLA-FERA M A,et al.Myostatin knockout in mice increases myogenesis and decreases adipogenesis[J].Biochem Biophys Rea Commun,2002,291(3):701-706. MCPHERRON A C,LEE S J.Double muscling in cattle due to mutations in the myostatin gene[J].Proc Natl Acad Sci,1997,94(23):12457-12461. KAMBADUR R,SHARMA M,SIMTH T P,et al.Mutations in myostatin (GDF8) in double——muscled Belgian Blue and Piedmontese cattle[J].Genome Rea,1997,7(9):910-916. XU C,WU G,ZOHAR Y,et al.Analysis of myostatin gene structure,expression and function in zebrafish[J].J Experi Biol,2003,206(22):4 067-4 079. ACOSTA J,CARPIO Y,BORROTO I,et al.Myostatin gene silenced by RNAi show a zebrafish giant phenotype[J].J Biotechnolo,2005,119(4):324-331. 姜运良,李宁.吴常信,等.不同品种猪myostatin基因的单核苷酸多态性分析[J].遗传学报,2001,28(9):804-805. 顾志良,张海峰,朱大海,等.鸡myostatin基因单核苷酸多态性的群体遗传学分析[J].遗传学报,2002,29(7):565-570. MACCATROZZO L,BARGELLONI L,PATARNELLO P,et al.Characterization of the myostatin gene and a linked microsatellite marker in shi drum (Umbrina cirrosa,Sciaenidae)[J].Aquaculture,2002,205(1/2):49-60. BIGA P R,CAIN K D,HARDY R W,et al.The isolation,characterization,and expression of a novel GDFII gene and a second myostatin form in zebrafish,Danio redo[J].Gener Comp Endocrinol,2004,138(1):32-41. KOCABAS A M,KUCUKTAS H,DUNHAM R A,et al.Molecular characterization and differential expression of the myostatin genein channel catfish (Ictalurus punctatus)[J].Biochem Biophysiol Acta——Gene Struc Express,2002,1575(1/3):99-107. XUE L,QIAN K,QIAN H,et al.Molecular cloning and characterization of the myostatin gene in croceine croaker,Pseudosciaena crocea[J].Mol Biol Rep,2006,33(2):129-136. BARZIZA D E,BUENTELLO J A,GATLIN D M.Dietary arginine requirement of juvenile red drum (Sciaenops cellatus) based on weight gain and feed efficiency[J].J Nutr,2000,130(7):1798-1799. TURNER T F,RICHARDSON L R,GOLD J R.Polymorphic microsatellite DNA markers in red drum (Sciaenops cellatus).Mol Ecol,1998,7(12):1771-1773. MOHAMED J S,KHAM l A.Molecular cloning and differential expression of three GnRH mRNAs in discrete brain areas and lymphocytes in red drum[J].J Endocrinol,2006,188(3):407-416. SAMBROOK J,FRITSCH E F,MANIATIS T.Molecular Cloning:A Laboratary Manual[M].2ed.New York:Cold Spring Harbor Laboratory Press,Cold Spring Harbor,1989:55-56. ALTSCHUL S F,MADDEN T L,SCHAFFER A A,et al.Gapped BLAST and PSI——BLAST:a new generation of protein database search programs[J].Nucl Acids Res,1997,25(17):3389-3402. THOMPSON J D,HIGGINS D G,GIBSON T J.Clustal W:improving the sensitivity of progressive multiple sequence alignment through sequence weighting,position——specific gap penalities and weight matrix choice[J].Nuel Acids Res,1994,22(22):4673-4680. KUMAR S,TAMURA K,NEI M.MEGA3:Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment[J].Briefings in Bioinformatics,2004,5(2):150-163. MACCATROZZO L,BARGELLONI L,RADAELLI G,et al.Characterization of the myostatin gene in the gihhead seabream (Sparus aurata):sequence,genomic structure,and expression pattern[J].Mar Biotechnol,2001,3(3):224-230. 姜运良,李宁,赵兴波,等.猪肌肉生长抑制素基因侧翼区新徼卫星标记的鉴定及分析[J].遗传学报,2004-31(5):480-484. THIES R S,CHEN T,DAVIES M V,et al.GDF-8 propeptide binds to GDF-8 and antagonizes biological activity by inhibiting GDF-8 receptor binding[J].Growth Factors,2001,18(4):251-259. LEE SJ,MCPHERRON A C.Regulation of myostatin activity and muscle growth[J].Proc Natl Acad Sci USA,2001,98(16):9306-9311. GROBET L,MARTIN L J,PONCELET D,et al.A deletion in the bovine myostatin gene causes the double——muscled phenotype in cattle[J].Nat Genet,1997,17(1):71-74. WIENER P,SMITH JA,LEWIS A M,et al.Muscle——related traits in cattle:the role of the myostatin gene in the South Devon breed[J].Genet Sel Evol,2002,34(2):221-232. SHARMA M,KAMBADUR R,MATTHEWS K G,et al.Myostatin,a transforming growth factor——beta superfamily member,is expressed in heart muscle and is upregulated in cardiomyocytes after infarct[J].J Cell Physiol,1999,180(1):1-9. JI S,LOSINSKI R L,CCRNELIUS R L,et al.Myostatin expression in porcine tissues:tissue specificity and developmental and postnatal regulation[J].Am J Physiol,1998,275(4):1265-1273. RODGERS B D,WEBER G M,SULLIVAN C V,et al.Isolation and characterization of myostatincomplementary deoxyribonucleic acid clones from two commercially important fish:Oreochromis mossambicus and Morone chrysop[J].Endocrinol,2001,142(4):1412-1418. ROBERTS S B,GOETZ F W.Differential skeletal muscle expression of myostatin across teleost species,and the isolation of multiple myostatin isoform[J].FEBS Lett,2001,491 (3):212-216. MACCATROZZO L,BARGELLONI L,CARDAZZO B,et al.A novel second myostatin gene is present in teleost fish[J].FEBS Lett,2001,509(1):36-40.
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