Figure 3 | Scientific Reports

Figure 3

From: Construction of stable mouse artificial chromosome from native mouse chromosome 10 for generation of transchromosomic mice

Figure 3

Strategy for the construction of 10MAC2 and 10MAC3 vectors. (A) Strategy for the targeted integration of NeoR and 5’HPRT-loxP into the 10MAC to generate 10MAC2 vector by the targeting vector, p10MAC2. Arrowheads indicate genomic PCR primers. (B) FISH analysis of the 10MAC2 vector in the DT40 (10MAC2) cells using the digoxigenin-labeled mouse Cot-1 DNA (red) and the biotin-labeled 5’HPRT-loxP gene (green). The arrow indicates the 10MAC2 vector, and the inset shows an enlarged image of the 10MAC2 vector. (C) Strategy for the targeted integration of EGFP, NeoR and 5’HPRT-loxP into the 10MAC to generate 10MAC3 vector by the targeting vector, p10MAC3. Arrowheads indicate genomic PCR primers. (D) Electroporation of the p10MAC3 plasmid yielded GFP-expressing, G418-resistant transfectants from the DT40 (10MAC3) cells. Phase-contrast (top panel) and fluorescence (bottom panel) micrographs are shown. Scale bar: 100 μm. (E) FISH analysis of the 10MAC3 vector in the DT40 (10MAC3) cells using the digoxigenin-labeled mouse Cot-1 DNA (red) and the biotin-labeled 5’HPRT-loxP gene (green). The arrow indicates the 10MAC3 vector, and the inset shows an enlarged image of the 10MAC3 vector.

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