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细胞内 ,经G4 18/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。
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 -/- .
基金
国家自然科学基金重点项目 (3 983 0 3 60 )~~
