Figure 1.

Targeted disruption of the mouse Gα_o gene and its molecular analysis. (A) Scheme of targeting vector, genomic XbaI fragment spanning the Gα_o targeting site, and predicted structure of the disrupted gene. β-geo, β-galactosidase and neomycin phosphotransferase gene; TK, herpes virus thymidine kinase gene. (B) Southern blot analysis of XbaI-digested genomic DNA from wild-type, Gα_o+/−, and Gα_o−/− mice. As predicted from A, the wild type displays a single 19.9-kb band when probed with the 3′ and 5′ flanking probe, and the Gα_o−/− lacks the 19.9-kb band but has new bands at 15.2 and 9.4 kb when hybridized to the 3′ and 5′ flanking probes, respectively. Gα_o+/− displays the bands found in both wild-type and the Gα_o−/− animal. (C) Immunoblot of brain membrane and cytosol from wild-type and Gα_o−/− mice probed with R4 antibody. The size of molecular weight markers are indicated at the right.