Orotate phosphoribosyltransferase (OPRTase; EC 2.4.2.10) catalyzes phosphoribosyl group transfer between alpha-d-5-phosphoribosyl-1-pyrophosphate and orotate to form orotidine-5′-monophosphate and pyrophosphate, the nucleotide-forming step in pyrimidine biosynthesis. It is one of ten PRTases that perform viral roles in de novo and salvage pathways for purine, pyrimidine and pyridine nucleotides. Although the PRTases are important drug targets, they are poorly understood mechanistically, and no three-dimensional structures exist. Here, we report the complete sequence of the Salmonella typhimurium pyrE gene and the deduced sequence of the OPRTase gene product. OPRTase forms tetragonal crystals from polyethylene glycol solutions; these crystals diffract to better than 2 Å resolution, and are stable to radiation damage. The space group is P41212 (or P43212) with unit cell dimensions of a = b = 48·5 Å, c = 210·5 Å, and α = β = γ = 90°. A crystalline form of the selenomethionine derivative of the protein is also reported.