猪Myostatin同源重组载体的构建  

Construction of Myostatin gene of pig homologous recombination vector

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

机构地区：[1]中国农业大学动物科技学院,北京100094 [2]北京锦绣大地农业股份有限公司,北京100049 [3]东北农业大学,黑龙江哈尔滨150030

出　　处：《黑龙江畜牧兽医》2006年第10期10-13,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然基金项目(30371036);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方法扩增了猪Myostatin基因的3′臂、5′臂,其长度分别为1.4 kb、4.4 kb。将扩增的片段与T载体连接后进行部分序列测定,与已发表的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,构建了猪的Myostatin表达Neor、Tk基因的双标记转基因载体。In this study, mammal double marker transgenic expression vector containing Neo^r 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 gent of the pig which length were about 1.4 kb and 4.4 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

关 键 词：Neo^r基因 TK基因 ploxP质粒 聚合酶链式反应 

分 类 号：S828[农业科学—畜牧学]