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  

Construction of eukaryotic expression vector with cattle myostatin gene and its expression in COS-7 cells

作　　者：吕文发[1] 赵静[1] 姚淑珍[1] 王军[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118

出　　处：《中国兽医科技》2005年第12期997-999,共3页Chinese Journal of Veterinary Science and Technology

基　　金：国家自然科学基金项目(30500366)

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。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.

关 键 词：牛 肌肉生成抑制素基因 真核表达 COS-7细胞 

分 类 号：S859.79[农业科学—临床兽医学] Q786[农业科学—兽医学]