Figure 1.
Introduction of point mutations at the PDGF-β receptor locus. (A) Changes introduced by in vitro mutagenesis in the amino acid sequence encompassing the PI3K-binding sites in the PDGF-β receptor are marked in bold letters. (B) Targeting of the locus. (Top) Targeting vector. (Middle) Wild-type locus. (Bottom) Targeted locus. Shaded bar, wild-type exon; solid bar, mutated exon. Oligonucleotide primers (arrowheads) used for PCR detection and probes used for Southern analysis are indicated. The position and expected fragment sizes of wild-type and targeted HindIII (H) fragments detected with the 5′ Southern are shown. E.RV, EcoRV; S, SalI; E.RI, EcoRI; H, HindIII; X, XhoI; K, KpnI; [S], site destroyed; HSV-Tk, herpes virus simplex thymidine kinase; neo-lox2, neomycin phosphotransferase expression cassette flanked by lox sites.