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. 1998 Jun;8(6):666–672. doi: 10.1101/gr.8.6.666

Figure 2.

Figure 2

Figure 2

Targeting of the D11S12 locus. (A; Top) The genomic 5.5-kb EcoRI fragment includes a 5.0-kb BamHI-targeted region. (Middle) Scheme of the pVT-3 targeting vector linearized with EcoRI and NotI. Yeast sequences, CEN6, ARSH4, and TRP1, and the mammalian ZeoR gene are shown. The vector can replace the D11S12 region by double crossover. (Bottom) Structure of the targeted locus. As a result of the targeting replacement, the size of the EcoRI fragment becomes 8.5 kb. (B) Southern blots of EcoRI-digested DNA from the parental DT40 11-3 line (lane 1) and five drug-resistant clones (lanes 2–6). DNA was probed with a 5′ external probe (a 5′ external 0.5-kb fragment of the D11S12 locus), showing a 5.5-kb parental fragment and the expected 8.5-kb targeted fragment in three of the five clones (lanes 4–6). The same filter was probed with the Zeo probe (a 1.3-kb NaeI-ZeoR gene-containing fragment), showing an 8.5-kb ZeoR-containing fragment in the same three clones (lanes 10–12). (E) EcoRI; (Ba) BamHI; (B) BclI.