Troponin I (TnI) is a constituent protein of the troponin complex located on the thin filament of striated muscle that provides a calcium-sensitive switch for striated muscle contraction. Unlike other contractile proteins, the cardiac isoform of troponin I (TnIc) is expressed only in cardiac muscle and therefore offers a model for cardiac-specific expression. It is also subject to developmental regulation with increased expression occurring at the time of birth. Here we describe the isolation and characterization of the human TnIc gene (HGMW-approved symbol TNNI3) and its promoter. The gene comprises eight exons contained within 6.2 kb of genomic DNA. The proximal promoter and 1.1-kb 5'-flanking region were sequenced, and several putative cis-acting elements that are conserved between the human and the mouse TnIc genes were identified. In addition, multiple copies of a 37-bp chromosome 19-specific mini-satellite sequence were identified within this region. Following transfection, 2300 bp of 5' sequence is active in both cardiac myocytes and skeletal muscle cells but is inactive in fibroblasts, indicating that it can drive expression but is insufficient to confer cardiac specificity.