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. 2006 Jun 5;103(24):9333–9338. doi: 10.1073/pnas.0600905103

Fig. 1.

Fig. 1.

Generation of DAT mutant knockin mice. (A) Targeting strategy. Thin lines, mouse genomic DNA; thick lines, sequences included in the targeting construct; K, KpnI sites; open box, mDAT exon 3; open box with a line, exon 3 with the triple mutation L104V/F105C/A109V and a KpnI site; open arrow, Neo cassette; triangles, LoxP sites; shaded arrow, thymidine kinase gene; shaded box, probe for Southern blot analysis; arrows, PCR primers. (B) PCR using primer f1 external of the short arm and mutant-specific primer r1. (C) PCR using specific primer f2 and primer r2 external of the long arm. (D) PCR using primers f3 and r3, which amplify a 564-bp fragment from the WT allele and a 667-bp fragment from the mutant allele. (E) Southern blot analysis of mouse genomic DNA digested with KpnI; the additional KpnI in the mutant allele reduces the probed fragment from 4.2 to 3.4 kb. HET, heterozygous; DAT-CI, homozygous mutant mice with cocaine-insensitive DAT; M, DNA marker.