FIG. 1.

Generation of ALR2-deficient mice. (a) The gene targeting construct, containing the herpes simplex virus tk gene (tk) and the neomycin resistance gene (neo). The restriction map of the wild-type and mutant ALR2 genes are shown. Relative positions of all exons are shown in filled boxes. The NotI site used to linearize the construct and the outside probe (filled bar) and the expected sizes of EcoRV fragments used for analysis of genomic DNA are indicated. E, EcoRV; R, EcoRI; S, SpeI; B, BglII. (b) Genotyping the ALR2 allele by Southern blot analysis. The expected bands of ∼9.8 kb for the wild-type ALR2 allele and ∼6.1 kb for the mutant ALR2 allele were observed. (c) Northern analysis of total RNA (10 μg/lane) from kidney, testis, and brain tissue of F₂ mice. The blot was analyzed with an ALR2 cDNA probe containing exons 5 to 8 and normalized with β-actin. An ∼1.3-kb band was detected representing the ALR2 transcript. (d) Western analysis of total protein isolated from brain, liver, bladder, and kidney extracts of F₂ mice. ALR2 was detected using a polyclonal rabbit antibody specific for it.