大口黑鲈肌肉生长抑制素基因单核苷酸多态性位点的筛选及其与生长性状关联性分析
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农业部公益性行业科研专项(200903045);国家自然科学基金项目(3090112);广东省科技计划项目(2007B020708008);广东省自然科学基金(9151038001000005);广东省海洋渔业科技推广专项(A200899F01);广东省科技兴渔项目(B200701A06,B200601G09)


SNPs detection in largemouth bass myostatin gene and its association with growth traits
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    摘要:

    采用PCR-SSCP和PCR-RFLP技术对大口黑鲈肌肉生长抑制素(myostatin,MSTN)基因全序列进行了SNPs位点筛选和分型,共筛选到2个单核苷酸多态性(SNPs)位点(C-1453T和T+33C),其中C-1453T位于启动子E8 box和Octamer(+)调控元件之间的区域,T+33C的突变位于第一外显子区域,属于同义突变,氨基酸没有发生变化;利用一般线性模型分析单标记位点与大口黑鲈生长性状(体重、体长、体高、体宽和眼间距)相关性,均未达到显著水平(P>0.05)。将2个SNPs位点不同基因型组合成6种双倍型(去掉频率小于3%的组合),关联分析表明,双倍型D2在体重、体长、体高、体宽和眼间距的均值均高于其它双倍型,而双倍型D5在体重、体长、体高、体宽和眼间距的均值均低于其它双倍型,双倍型D2与D5之间在5个主要生长性状均存在差异显著(P<0.05),推测双倍型D2对生长性状起正相关,而双倍型D5与大口黑鲈的生长性状呈负相关,因此推断MSTN基因突变位点双倍型D2与D5可作为大口黑鲈生长性状的两个标记位点,用其分子标记位点来辅助大口黑鲈育种工作以期加快育种进程。

    Abstract:

    Myostatin or GDF-8,a member of the transforming growth factorβ(TGF-β)surperfamily,has been demonstrated to be a negative regulator of myogenesis and inhibited myoblast proliferation during cell cycle and myogenic differentiation in animals.In present research,two SNPs sites(C-1453T and T+33C)were identified from total MSTN gene in larghmouth bass by using PCRSSCP technique and sequencing; The mutational site of C-1453 was between the regions of E8 box and Octamer(+).the mutation of T+33C was situated in the first extron of the MSTN gene,however the mutation of the first extron has not changed the encoded amino which belonged to the synonymy mutation.The three kinds of genotypes per mutation site were obtained by PCRRFLP method and the genotype frequency of the mutation was caculated in the population of 128 individuals.A general linear model was used to statistically analyze associations of MSTN gene polymorphisms with major growth traits respectively,and the results indicated that there was no significant difference(P>0.05)by all genotypes of single mutation site.Six diplotypes,with the minor allelic frequencies of above 3%,were constructed based on two SNPs in the experiment population.The means of diplotype D2 were over other diplotypes and the means of diplotype D5 were lower other diplotypes in terms of body width,body height,body length,body weight,and snout length therfore associations analysis indicated that there were significant associations between diplotype D2 and D5 in the five growth traits of largemouth bass.The study implies that diplotypes D2 and D5 of MSTN gene could be used as the moleculer genetic marker for selection breeding.

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于凌云,白俊杰,樊佳佳,李小慧,叶星.大口黑鲈肌肉生长抑制素基因单核苷酸多态性位点的筛选及其与生长性状关联性分析[J].水产学报,2010,34(6):665~671

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历史
  • 收稿日期:2010-02-05
  • 最后修改日期:2010-03-20
  • 录用日期:2010-03-30
  • 在线发布日期: 2010-06-10
  • 出版日期: 2010-06-15