The calpains comprise a family of heterodimeric (80+28 kDa) Ca2+-dependent cysteine proteases, probably having roles in signal transduction and cytoskeletal remodelling. We describe cloning and sequencing of the 28 kDa calpain subunit cDNA from mouse (coding for 268 amino acids), and characterization of its gene. The gene spans 7 kb and contains 11 exons. The promoter region, like those of other calpain genes, lacks an obvious TATA box, but contains several Sp1 binding sites.