Analysis of TBP gene sequences from a variety of species for clustering of short sequence motifs and for over- and underrepresentation of short sequence motifs suggests involvement of slippage in the recent evolution of the TBP N-terminal domains in metazoans, Acanthamoeba and wheat. AGC, GCA and CAG are overrepresented in TBP genes of other species, suggesting that opa arrays were amplified from motifs overrepresented in ancestral species. The phylogenetic distribution of recently slippage-derived sequences in TBP is similar to that observed in the large subunit ribosomal RNAs, suggesting a propensity for certain evolutionary lineages to incorporate slippage-generated motifs into protein-coding as well as ribosomal RNA genes. Because length increase appears to have taken place independently in lineages leading to vertebrates, insects and nematodes, TBP N-terminal domains in these lineages are not homologous. All gene duplications in the TBP gene family appear to have been recent events despite strong protein sequence similarity between TRF and P. falciparum TBP. The enlargement of the TBP N-terminal domain may have coincided with acquisition of new functions and may have accompanied molecular coevolution with domains of other proteins, resulting in the acquisition of new or more complex mechanisms of transcription regulation.