Fig. 2.

Disruption of the pig SMN locus by gene targeting. a Diagram of the pig SMN locus, the corresponding pig SMN genomic sequence used to construct the 5′ and 3′ recombination arms for the pML8 targeting vector and the targeted locus following homologous recombination with the pML8 targeting sequence. The primers 360L and 6632R were used to screen for recombination of the 5′ arm. The primers 1610L and 460R were used to screen for recombination of the 3′ arm. The primers 360L and 460R were used for long range PCR. Primers 360L and 635neoR and 764 neoL and 460R were used to screen for the presence of neo. b PCR of pig fetal fibroblast cells from colony X8D2. Lanes 1, 8 and 15 are 1 kb DNA ladders. For each primer pair, the templates are no template control, JR1 wildtype and X8D2 pig cell lysates respectively. Lanes 2–4 are PCRs using primers 360L and 6632R. Lanes 5–7 are PCRs using primers 360L and 635neoR. Lanes 9–11 are PCRs using primers 1610L and 460R. Lanes 12–14 are PCRs using primers 764 neoL and 460R. Lanes 16–18 are PCRs using primer pairs 360L and 460R