Fig. 1. Knock-in of IRES-lacZ into the VEGFA locus. (A) Vegf IRES-lacZ knock-in alleles with insertion of the reporter cassette at different positions of the Vegf 3'UTR. LoxP sites remaining after excision of the selectable marker are indicated by arrowheads. Sequences are not drawn to scale. (B) Schematic representation of the wild-type, targeted (Vegflo+Neo) and Neo-excised (Vegflo) allele. Exons are depicted as boxes. Probes used in Southern blot analysis and expected fragment lengths are indicated. (C) Southern blot analysis with Probes 1 and 2 on EcoRV digested genomic DNA showed proper homologous recombination both at the 5' and 3' homology arms of the target vector, respectively. The Southern blot analysis with Probe 1 on BglII digested genomic DNA showed successful removal of the loxP flanked neo selectable marker.
