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牦牛MSTN基因分子克隆及序列分析

DOI: 10.3969/j.issn.1000-7091.2013.01.015, PP. 76-81

Keywords: 牦牛,MSTN基因,克隆,序列分析

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Abstract:

设计特定引物对牦牛MSTN基因PCR分段扩增并克隆和测序,利用分子生物学软件进行序列拼接,获得牦牛MSTN基因序列(GenBank登录号EU926670)。该基因由3个外显子和2个内含子组成,CDS序列全长为1128bp(GenBank登录号EU926671),由375个氨基酸组成。外显子大小分别为373,374,381bp,内含子大小分别为1843,2028bp。牦牛与普通牛的MSTN基因编码区中,在417位发生一次碱基转换(C→T),但未造成氨基酸改变。不同物种间在该基因编码区核苷酸序列和氨基酸序列上有较高的相似性,牦牛与普通牛、绵羊、猪、人、狗、小鼠、马、兔子、鸡、猩猩各物种间核苷酸相似性大小分别为99.9%,96.5%,94.3%,89.1%,91.9%,91.3%,93.6%,91.7%,82.0%,92.0%。牦牛与普通牛MSTN氨基酸序列相似性最高,为100%;而与绵羊、猪、小鼠、人、狗、马、兔子、鸡、猩猩各物种间相似性大小分别为93.3%,95.5%,92.5%,94.1%,93.3%,94.9%,94.4%,88.0%,94.4%。生物信息学软件分析发现:牦牛MSTN基因编码蛋白的理论分子量约为42.6kDa,PI值为6.14,Leu的含量最高(9.9%),其次是Lys(7.2%)。牦牛MSTN基因编码蛋白二级结构以β折叠为主,属于跨膜蛋白,跨膜区位于AA6-AA23之间;具有一个分泌信号肽结构,其氨基酸序列MQKLQICVYIYLFMLIVA具有TGF-β家族的特征。

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