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5 Inactivating the Function of Specific Genes in Eukaryotes 389 (a) Formation of ES cells carrying a knockout mutation neo r tk HSV Homologous recombination Gene X ES cells ES-cell DNA ES forex tehlikesi Nonhomologous recombination ES-cell DNA Other genes Random insertion No mutation in gene X Cells are resistant to G-418 but sensitive to ganciclovir Gene X replacement construct Gene-targeted insertion Mutation in gene X Cells are resistant to G-418 and ganciclovir (b) Positive and negative selection of recombinant ES cells Nonrecombinant cells Recombinants with random insertion Recombinants with gene-targeted insertion Treat with G-418 (positive selection) Treat with ganciclovir (negative selection) ES cells with targeted disruption in gene X EXPERIMENTAL FIGURE 9-38 Isolation of mouse ES cells with a gene-targeted disruption is the first stage in production of knockout mice.
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