The expression in normal rat cells of modified polyoma virus genomes, separately encoding large T, middle T or small T antigens, has allowed the investigation of the roles of these proteins in oncogenic transformation. Middle T is sufficient to transform cells of established lines but the transformants are serum dependent. Large T lacks intrinsic oncogenic potential but can relieve the serum dependence of normal and transformed cells. Middle T alone cannot transform primary rat embryo fibroblasts.