Within warm-blooded animals, Toxoplasma gondii switches from an actively replicating form called a tachyzoite into a slow growing encysted form called a bradyzoite. To uncover the genes involved in bradyzoite development, we screened over 8000 T. gondii insertional mutants by immunofluorescence microscopy. We identified nine bradyzoite development mutants that were defective in both cyst wall formation and expression of a bradyzoite specific heat shock protein. One of these mutants, named 42F5, contained an insertion into the predicted gene TGME49_097520. The disrupted protein is serine/proline-rich with homology to proteophosphoglycans from Leishmania. T. gondii proteophosphoglycan (GU182879) expressed from the native promoter was undetectable in tachyzoites, but bradyzoites show punctate spots within the parasite and staining around the parasitophorous vacuole. Complementation of the 42F5 mutant with GU182879 expressed from either the alpha-tubulin or native promoter restores cyst wall formation. Overall, GU182879 is upregulated in bradyzoites and enhances cyst wall component expression and assembly.