p53-/-细胞是什么意思 ?这个细胞的式这么得到的 The targeting vectors used in this experiment were constructed so that the first codons of the drug resistance markers replaced the first codon of the p53 gene within exon 2. Transfection was done with LipofectAmine (Life Technologies) and subconfluent monolayers of cells. The LipofectAmine-DNA mixture was removed 3 hours after transfection and cells were grown in complete medium for 18 hours before they were replated in selective medium. After transfection with the neomycin-resistance vector, resistant colonies were grown in the presence of geneticin (0.4 mg/ml) (Gibco). One geneticin-resistant clone of 600 assayed was found to harbor a homologous recombinant, and this clone was expanded and transfected with the hygromycin B–resistance vector. Colonies were selected in hygromycin B (0.1 mg/ml). One clone of 940 assayed had a homologous recombinant of the second p53 allele.这种细胞是完全敲除的，目前经常使用的干扰方法能实现目的基因的下调，而不能实现完全敲除。 这种完全敲除的方法一般都是用同源重组的方法替换基因组上的exon区序列。 查看更多
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