PfMAP2 is essential for male gametogenesis in the malaria parasite Plasmodium falciparum

Transcriptional profiling of transgenic P. falciparum stage V gametocytes of the strain NF54/MAP-2GFPDD before and after activation for gametogenesis. The DD (FKBP destabilisation domain) allows for the conditional expression of fusion proteins: DD fusion proteins are rapidly degraded or stably expressed in absence or presence of the stabilising ligand Shield-1, respectively (Banaszynski LA, Chen LC, Maynard-Smith LA, Ooi AG, Wandless TJ. A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules.Cell. 2006 Sep 8;126(5):995-1004). The goal of this experiment was to identify genes differentially expressed in PFMAP-2GFPDD-expressing compared to PfAMP-2GFPDD-depleted stage V gametocytes. Four condition experiment using RNA extracted from mature stage V gametocytes (day 13) of the strain NF54/MAP-2GFPDD expressing PfMAP-2GFPDD from the endogenous locus. Gametocytes were cultured in presence of Shield-1 (ON) (stable PfMAP-2GFPDD expression) or had Shield-1 removed from the culture medium one day prior to sample harvesting (OFF) (PfMAP2-GFPDD protein degraded). Each population was split and one half was activated for gametogenesis by treatment with xanthurenic acid for 10 minutes (ON act. and OFF act.) while the other half was not (ON and OFF). Two biological replicates were performed (eight samples in total).