山羊Myostatin基因打靶载体的构建  

Construction of Myostatin Gene of Goat Gene Targeting Vector

作　　者：潘求真[1] 孙书锋[1] 陆泉枝[1] 王海[2] 连正兴[1] 杨宁[1] 李宁[1] 谭景和[3] 

机构地区：[1]中国农业大学动物科技学院 [2]北京锦绣大地农业股份有限公司 [3]东北农业大学

出　　处：《中国草食动物》2006年第3期10-13,共4页China Herbivores

基　　金：国家"863"计划项目(2001AA213031)

摘　　要：根据绵羊、猪、小鼠和牛的Myostatin基因序列,设计引物,通过PCR方法扩增了山羊的Myostatin基因的3′臂、5′臂,其长度分别为1·4kb、4·5kb。将扩增的片段与T载体连接后进行部分序列测定,与已知发表的Myostatin序列进行同源性比较,同源性为100%。对目的片段与载体进行酶切后定向连接形成转基因结构,经序列测定后,证明其插入方向和位置完全正确,构建表达Neo、Tk基因的哺乳动物双标记转基因载体,并命名为ploxp-M。In the study, mammal double marker transgenic expression vector containing Neo gene and Tk gene was constructed. According to Myostatin gene sequence of the sheep, pig, mouse and cattle,we designed primers, and had gotten by PCR 3′-arm and 5′-arm of Myostatin gene of the goat which length were about 1.4kb and 4.5kb respectively. Amplified segments were connected with T-vector, and had portion sequencing.Then the sequence were compared with that had already published of Myostatin, and the homology was 100%. We had enzymed the objective segments on the vector, and then directional connected come into being transgenic structure, After sequencing, the inset direction and position was proved to be correct completely, and the vector constructed was named ploxp-M.

关 键 词：Neo基因 TK基因 ploxP质粒 聚合酶链式反应 

分 类 号：S827-2[农业科学—畜牧学]