Figure 1.

Generation of MG29-knockout mice. Homologous recombination at the MG29 locus (A). Restriction enzyme maps of the wild-type allele, targeting vector, and mutant allele (MG29^m1) are illustrated. The exons in the gene (E1–E3) are indicated by filled boxes, and the neomycin resistance gene (neo) and diphtheria toxin gene (DTA) are indicated by open boxes. The directions of transcription are indicated by arrows. The genomic DNA probe and PCR primers for detection of the mutant gene are indicated by a hatched box and open arrows, respectively. The predicted sizes of the DNA fragments in Southern blot analysis and PCR are also shown. Southern blot analysis of DNAs from mice bearing the MG29^m1 mutation (B). Genomic DNAs digested with BamHI were analyzed using the hybridization probe indicated in A. Size markers are indicated in kb pairs. Blot hybridization analysis of skeletal muscle RNAs from mice bearing the MG29^m1 mutation (C). Mouse MG29 cDNA was used as the hybridization probe, and size markers are indicated in kb. Immunoblot analysis of skeletal muscle microsomal proteins from mice bearing the MG29^m1 mutation (D). Microsomal proteins from hind limb muscles were analyzed with antibody against mouse MG29, and size markers are indicated in kD.