Capping protein (CP), a heterodimer of alpha and beta subunits, is found in all eukaryotes. CP binds to the barbed ends of actin filaments in vitro and controls actin assembly and cell motility in vivo. Vertebrates have three alpha isoforms (alpha 1, alpha 2, alpha 3) produced from different genes, whereas lower organisms have only one gene and one isoform. We isolated genomic clones corresponding to the alpha subunits of mouse CP and found three alpha 1 genes, two of which are pseudogenes, and a single gene for both alpha 2 and alpha 3. Their chromosomal locations were identified by interspecies backcross mapping. The alpha 1 gene (Cappa1) mapped to chromosome 3 between D3Mit11 and D3Mit13. The alpha 1 pseudogenes (Cappa1-ps1 and Cappa1-ps2) mapped to Chromosomes 1 and 9, respectively. The alpha 2 gene (Cappa2) mapped to Chromosome 6 near Ptn. The alpha 3 gene (Cappa3) also mapped to Chromosome 6, approximately 68 cM distal from Cappa2 near Kras2. One mouse mutation, de, maps in the vicinity of the alpha 1 gene. No known mouse mutations map to regions near the alpha 2 or alpha 3 genes.