Tagging of PF3D7_1019700) with GFP and pulldown of associated proteins.

(A) The diagram shows GFP tagging of the PF3D7_1019700 at its C-terminus by single-crossover homologous recombination. Purple blocks show the fragment used for homologous recombination. (B) Integration-specific PCR using primers P1 and P2. WT, Wildtype 3D7; C1 and C2 are two transgenic clones. (C) Live cell imaging shows the localization of PF3D7_1019700::GFP in the nuclei by fluorescence microscopy. R: ring; ET: early trophozoite; LT: late trophozoite; ES: early schizont; LS: late schizont. (D) Proteins identified from parasite nuclear extracts by IP and LC-MS/MS. Single-step IPs with anti-GFP beads were done using the PF3D7_1019700::GFP (R1 and R2 indicate two replicates). The wild-type 3D7 was used as the IP control. The proteomic data were analyzed by SAINT using a threshold of probability >94% and 1% FDR. Gene ID and annotations are shown on the right. (PDF)