邓思平,李嘉颖,张婉玲,宋春杰,李广丽,朱春华,吴天利

• 1:广东海洋大学水产学院
• 2:广东海洋大学南海水产经济动物增养殖广东普通高校重点实验室

摘要(Abstract)：

以勒氏笛鲷(Lutjanus russellii)基因组DNA为模板,采用同源克隆的方法,获得2 887 bp的肌肉生长抑制素(myostatin,MSTN)基因组序列,该MSTN序列具有3个外显子和2个内含子,包括101 bp的5′-UTR、385bp的外显子1、354 bp的内含子1、370 bp的外显子2、761 bp的内含子2、381 bp的外显子3和1 932 bp的3′-UTR。整个开放阅读框编码了378个氨基酸,前面的22个氨基酸为信号肽,具有9个保守的半胱氨基酸及一个RVRR的蛋白酶解加工位点。氨基酸序列分析发现,该基因编码的蛋白质与其他鱼类的I型同源性较高,与其他鱼类的2型MSTN同源性较低,且与鲈形目的同源性最高,与鲤形目的同源性较低,与人、鼠和鸡的同源性最低。采用邻接法(Neighbor-Joining)构建的MSTN的系统发育树表明,勒氏笛鲷MSTN与鲈形目鱼类的狼鲈属亲缘关系较近,且与鱼类的MSTN-1聚为1支。这表明该基因属于Ⅰ型MSTN基因。

关键词(KeyWords)： 勒氏笛鲷;肌肉生长抑制素(MSTN);基因;克隆;序列分析

Abstract:

Keywords:

基金项目(Foundation): 广东海洋大学博士启动项目(0712103);; 广西科学研究与技术开发计划重点项目(桂科合1140009-4)

作者(Author): 邓思平,李嘉颖,张婉玲,宋春杰,李广丽,朱春华,吴天利

参考文献(References)：

• [1]McPherron A C,Lawler A M,Lee S J.Regulation of skeletal musclemass in mice by a new TGF-beta superfamily member[J].Nature,1997,387(6628):83–90.
• [2]Whittemore L A,Song K,Li X,et al.Inhibition of myostatin inadultmice increases skeletal muscle mass and strength[J].Biochemical andBiophysical Research Communications,2003,300(4):965-971.
• [3]Bogdanovich S,Krag T O,Barton E R,et al.Functional improvementof dystrophic muscle by myostatin blockade[J].Nature,2002,420(6914):418-421.
• [4]McPherron A C,Lee S J.Double muscling in cattle due to mutations inthe myostatin gene[J].Proc Natl Acad Sci,1997,94(23):12457–12461.
• [5]Kambadur R,Sharma M,Smith T P,et al.Mutations in myostatin(GDF8)in double-muscled Belgian Blue and Piedmontese cattle[J].Genome Res,1997,7(9),910-916.
• [6]Grobet L,Martin L J,Poncelet D,et al.A deletion in the bovinemyostatin gene causes the double-muscled phenotype in cattle[J].NatGenet,1997,17(1):71-74.
• [7]Acosta J,Carpio Y,Borroto L,et al.Myostatin gene silenced byRNAi show a zebrafish giant phenotype[J].J Biotchenol,2005,119(4):324-331.
• [8]Amali A A,Lin C J F,Chen Y H,et al.Up-regulation of musclespecific transcription factors during embryonic somitogenesis ofzebrafish(Danio rerio)by knock-down of myostatin-1[J].Dev Dynam,2004,229(4):847-856.
• [9]Lee S B,Kim Y S,Oh M Y,et al.Improving rainbow trout(Oncorhynchus mykiss)growth by treatment with a fish(Paralichthysolivaceus)myostatin prodomain expressed insoluble forms in E.coli[J].Aquaculture,2010,302(3-4):270-278.
• [10]李绍华,熊远著,郑嵘,等.猪MSTN基因多态性及其SNPs的研究[J].遗传学报,2002,29(4):326-331.
• [11]杨威,王海霞.鸡生长分化因子GDF-8CDNA的克隆表达及蛋白质纯化[J].生物工程学报,2001,17(4):460-462.
• [12]徐建勇,陈松林.牙鲆肌肉生长抑制素(MSTN)基因克隆[J],水产学报,2008,32(4):497-506.
• [13]郁建锋,张营,王星果,等.鳡肌肉生长抑制素(MSTN)基因的克隆及其组织特异性表达分析[J].水产学报,2010,34(10):1486-1494.
• [14]Maccatrozzo L,Bargelloni L,Cardazzo B,et al.A novel secondmyostatin gene is present in teeost fish[J].FEBS Lett,2001,509(1):36-40.
• [15]Garikipati1D K,Gahr S A,Rodgers B D.Identification,characterization,and quantitative expression analysis of rainbow troutmyostatin-1a and myostatin-1b genes[J].J Endocrinol,2006,190(3):879-888.
• [16]Garikipati D K,Gahr S A,Roa lson E H,et al.Characterization ofrainbow trout myostatin-2 genes(rtMSTN-2a and-2b):genomicorganization,differential expression,and pseudogenization[J].Endocrinol,2007,148(5):2106-2115.
• [17]Helterline D L,Garikipati D,Stenkamp D L,et al.Embryonic andtissue-specific regulation of myostatin-1 and-2 gene expression inzebrafish[J].Gen Comp Endocrinol,2007,151(1):90-97.
• [18]Thomas M,Langley B,Berry C,et al.Myostatin,a negative regulatorof muscle growth,functions by inhibiting myoblast proliferation[J].JBiol Chem,2000,275(51):40235-40243.
• [19]Xu C,Wu G,Zohar Y,et al.Analysis of myostatin gene structure,expression and function in zebrafish[J].J Exp Biol,2003,206(Pt22):4067-4079.
• [20]Bogdanovich S,Perkins K J,Krag T O,et al.Myostatinpropeptide-mediated amelioration of dystrophic pathophysiology[J].The FASEB J,2005,19(6):543-549.