Serotype 4 rotavirus strains have been classified into two antigenic "subtypes" by a solid phase immune electron microscopy technique in which cross-absorbed animal polyclonal immune sera are used as the source of antibodies. The sequences of the gene encoding the outer capsid glycoprotein VP7 from a single serotype 4 rotavirus field strain identified as subtype A ("ST3-like") and from three field strains identified as subtype B ("VA70-like") were determined. A comparison of the deduced amino acid sequences indicated that 15 amino acid residues were divergent between subtypes but were conserved within a subtype. Three of these 15 amino acid residues (at positions 96, 212, and 217) were located in regions of the VP7 that have been defined as serotype-specific antigenic sites. These data suggest that VP7 subtype differences may result from critical amino acid substitutions within an immunodominant serotype 4-specific antigenic site. Whether these differences are an important mechanism in the epidemiology of rotaviruses requires further investigation.