| 牛肌肉生成抑制素基因真核表达载体的构建及其在COS-7细胞中的表达 |
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| 引用本文： | 吕文发,赵静,姚淑珍,王军.牛肌肉生成抑制素基因真核表达载体的构建及其在COS-7细胞中的表达[J].中国兽医科技,2005,35(12):997-999. |
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| 作者姓名： | 吕文发 赵静 姚淑珍 王军 |
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| 基金项目： | 国家自然科学基金项目（30500366） |
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| 摘 要： | 将构建的牛pMD 18-T-MSTN克隆载体与真核表达载体pef-dhfr1a酶切,回收牛MSTN目的片段及pef-dhfr1a载体,构建了牛MSTN基因的真核表达质粒pef-dhfr1a-MSTN,然后转染COS-7细胞,将牛MSTN成熟蛋白编码序列在COS-7细胞中进行了表达.提取转染细胞的总RNA,采用RT-PCR和Western-blotting方法,分别从mRNA水平和蛋白质水平上检测到了牛MSTN基因在COS-7细胞中的表达产物,证明已经成功构建出该基因的真核表达载体.
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| 关 键 词： | 牛 肌肉生成抑制素基因 真核表达 COS-7细胞 |
| 文章编号： | 1000-6419（2005）12-0997-03 |
| 收稿时间： | 2005-10-08 |
Construction of eukaryotic expression vector with cattle myostatin gene and its expression in COS-7 cells |
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| Abstract: | In order to express encoding sequence of the cattle myostatin gene in eukaryotic system, a cloning plasmid and a pef-dhfrla vector was digested and then retrieved, respectively. The constructed eukaryotic expression plasmid pef-dhfrla-MSTN was induced by liposome. The cattle myostatin gene encoding a protein of 375 amino acids was expressed in COS-7 cells. The RT-PCR in mRNA level and Westernblotting analysis in protein level confirmed that the eukaryotic expression plasmid pef-dhfrla-MSTN was constructed successfully. |
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| Keywords: | cattle myostatin gene eukaryotic expression COS-7 cell |
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