| 小鼠锌指蛋白基因ZF-12基因剔除的ES细胞系的建立 |
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| 引用本文： | 厉建中,杨桦,傅继梁. 小鼠锌指蛋白基因ZF-12基因剔除的ES细胞系的建立[J]. 遗传学报, 2002, 29(10): 860-864 |
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| 作者姓名： | 厉建中 杨桦 傅继梁 |
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| 作者单位： | 1. 第二军医大学医学遗传学教研室,上海,200433
2. 第二军医大学医学遗传学教研室,上海,200433;上海南方模式生物研究中心,上海,201203 |
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| 基金项目： | 国家自然科学基金重点项目 (3 983 0 3 60 )~~ |
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| 摘 要： | 利用小鼠锌指蛋白ZF－12基因组DNA片段，构建了针对小鼠ZF－12基因座的替换型打靶载体pSSC-TV-10.5。经限制性核酸内切酶酶切及部分测序鉴定其结构正确后，通过电穿孔将线性化打靶载体导入ES人，经G418／GANC双药筛选和分子鉴定，获得4个ZF－12^ /-基因的ES细胞杂合子克隆，其生长状态良好，为进一步建立ZF－12基因剔除的小鼠动物模型创造了条件。
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| 关 键 词： | 小鼠 锌指蛋白基因 ZF－12 基因剔除 ES细胞系 基因打靶 胚胎干细胞 |
| 文章编号： | 0379-4172(2002)10-0860-05 |
| 修稿时间： | 2002-03-25 |
Establishment of Mouse ZF-12+/- Embryonic Stem Cells |
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| Abstract: | By using the mouse zinc finger protein gene ZF 12 genomic DNA fragment, pSSC TV 10.5 was designed and constructed as a replacement vector. Structure of pSSC TV 10.5 was identified by restrictive digestion analysis and partly sequencing. Then linearized vector was electroporated into ES cells, and transfected cells were screened by G418 and GANC selection. Among 508 G418 r/GANC r colonies, 4 were proved to have taken place the homologous recombination of ZF 12 by PCR and southern blotting analysis. This study lays the foundations of preparing mouse models of ZF 12 +/- or ZF 12 -/- . |
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| Keywords: | gene targeting ES cell ZF 12 ZNF191 |
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