We have identified at the molecular level two murine myosin heavy chain (MHC) genes which encode adult skeletal isoforms. As is the case for the murine cardiac MHC genes, the genes are closely linked, head to tail. To identify the isoform encoded by each gene, we located and sequenced the 3′ terminai and assessed the genes' temporal and tissue-specific expression patterns using probes specific for the 3′ untranslated regions. These analyses indicate that the myosin encoded by the gene located 5′ in the linked pair is the murine 2A isoform. The isoform encoded by the 3′-most gene of the linked pair appears to encode an additional isoform that is expressed in a number of adult skeletal muscles. The pattern of expression of the 3′-most gene indicates that it is tightly controlled developmentally. Transcripts from this gene, when compared to those from the MHC2A and β-MHC genes, are the predominant MHC transcripts found in the diaphragm, tongue, soleus, and masseter, indicating that it encodes a major skeletal MHC isoform. © 1992.