Figure 1.

RIZ gene targeting. (A) Schematics of murine RIZ gene (top), targeting vector (middle), and targeted allele (bottom) are shown with horizontal bars representing genomic DNA and horizontal lines representing plasmid DNA. The positive and negative selection cassettes are boxes labeled Neo and TK, respectively. Restriction sites listed include: EcoRI (E), Xba I (X), BglII (B), HindIII (H), NsiI (N), KpnI (K), Kpn2I (K2), SacII (S), NotI (No), and XhoI (Xh). (B) Southern blot analysis of genomic DNA of RIZ-targeted mice. Genomic DNA was digested with XbaI and was hybridized with the 5′ probe. (C) Northern blot analysis. Total RNA isolated from adult mouse brain was analyzed by Northern blotting with the RIZ1-specific probe or the common (exon 8) probe, as indicated. A similar amount of loaded RNA was shown by ethidium bromide staining of rRNA. (D) Immunoblot analysis. Protein extracts were prepared from the testis of adult animals and immunoprecipitated with anti-KG7.1S serum against the N terminus of RIZ2. The immunoprecipitated products were resolved on 5% SDS gel followed by immunoblot by use of 1715 serum against the C terminus of RIZ1/2 proteins. (E) RT–PCR analysis. Size in base pairs (bp) of molecular weight standards in lane M are indicated.