FIG. 1.

Targeted disruption of the mouse Rap250 gene. (A) Structures of the wild-type Rap250 allele, targeting vector, and targeted allele are shown with the BamHI (B), HindIII (H), KpnI (K), and XbaI (X) restriction sites and primers for PCR screening. The locations of probes used in Southern blot analysis are indicated. Black boxes indicate exons. The exon numbering is adapted from the work of Zhu et al. (42). (B) Homologous recombination in ES cells. Southern blot analysis of DNA from two positive ES clones, clone 1B8 and 2D2, digested with BamHI is shown. The 5-kb band for the wild-type allele (WT) and the 11-kb band for the targeted allele (KO) are indicated. The probes used are indicated in Fig. 1A. (C) Southern blot analysis of mouse yolk sac DNA digested with BamHI. (D) PCR genotyping of mouse embryo DNA. The bottom band, using the U6 and L8 primers, represents the wild-type allele and the top band, using the KOF1 and L3 primers, represents the mutated allele. M, molecular marker (1-kb ladder). (E) Photo of wild-type and Rap250^−/− embryos at E13.5. No obvious abnormalities or significant size differences were detected between wild-type and Rap250^−/− littermates.