Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites

Purpose: Toxoplasma must import inorganic phosphate via a transporter at the plasma membrane. Methods: WT and TgPiT-deficient parasites were grown in human foreskin fibroblasts for 2 days. The parasites were syringed, filter-purified, and resuspended n ice-cold PBS. A trizol-based method was used to extract total RNA from both strains. Toral RNA was subjected to cDNA library construction using Ilumina. Results: The parasite expresses a unique TgPiT as selective Pi transporter, located at the plasma membrane and on VAC internal membranes of endosomal organelles. Conclusion: TgPiT is essential for parasite survival, virulence, ionic homeostasis and osmoregulation Total RNAs of WT and ∆TgPiT parasites were converted to cDNA libraries. The assay was replicated in duplicate.