| 摘要: |
| 目的制备brevican基因敲除小鼠。 方法采用ET克隆方法,构建brevican打靶载体。线性化打靶载体,电转化胚胎干细胞(embryonic stem cell, ES),将正确同源重组的ES细胞注射入囊胚腔,生育嵌合体,再交配繁育杂合子及纯合子。 PCR法鉴定小鼠的基因型。 结果将PGK启动子指导的NEO表达框敲进Bcan第3外显子,敲除第3~8外显子序列,得到打靶载体,上游臂为2.4 kb,下游臂为4.8 kb。基因打靶后,得到双臂均发生正确同源重组的克隆数14个。利用阳性ES细胞克隆注射入囊胚, |
| 关键词: brevican 基因敲除小鼠 胚胎干细胞 同源重组 |
| DOI：10.3724/SP.J.1008.2011.0818 |
| 投稿时间：2011-05-13修订日期：2011-06-15 |
| 基金项目:国家自然科学基金(30571912). |
|
| Generation of brevican knockout mice |
| HAN Xi1,DONG Yan2* |
| (1. Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, China;2. Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200433, China) |
| Abstract: |
| ObjectiveTo generate brevican knockout mutant mice. MethodsThe targeting construct to inactivate the brevican gene was made by using the ET cloning technology. The construct was linearized and electroporated into embryonic stem (ES) cells. Homologous reco |
| Key words: brevican knockout mice embryonic stem cells homologous recombination |
