Fig. 3. Targeted disruption of the RIM-BP3 gene. (A) The strategy for the generation of a targeted RIM-BP3 allele. Black bars represent coding regions of the single-exon gene. BamHI restriction sites (B) and the probe used for Southern analysis (R), and PCR primers (arrows) used for genotyping are indicated. (B) Genotype confirmation of the knockout mouse by PCR (upper panel) and Southern analysis (lower panel). The DNA fragments derived from the wild-type (WT) and mutant (MT) alleles are indicated on the right. +/+, wild type; +/-, heterozygote; -/-, homozygote. (C) Genotype confirmation of the knockout mouse by western analysis. The blot containing testis protein extracts from wild-type (+/+), heterozygous (+/-) and homozygous (-/-) adult mice was probed with antibodies specific for N- and C-terminal regions (Fig. 1A). The detection of β-actin serves as a loading control.