Figure 1.

Genotyping Aag +/+, +/−, and −/− mice. (A) Schematic representation of a portion of the wild-type and targeted Aag alleles indicating the location of three primers and HindIII (H) sites. Note that Aag exon II is disrupted by insertion of a neo expression cassette to create an Aag null allele. Primers PI and PIII yield a 164-bp fragment in the presence of the wild-type allele. In the presence of the targeted allele, primers PII and PIII yield a 523-bp fragment. HindIII sites are loosely indicated to show that disruption of Aag by neo insertion increases the size of the HindIII fragment (details have been presented elsewhere) (12). (B) (Upper) PCR genotyping of MEFs (lanes 1–5). PCR genotyping of DNA from spleens of mice used in subsequent biochemical analysis (lanes 6–7). (Lower) Southern blot analysis of the DNA used for PCR analysis above. HindIII-digested DNA was probed with a fragment of the Aag gene that lies outside of the targeting construct (12). The Aag null allele (8.4 kb) and the wild-type Aag allele (6.3 kb) are indicated.