ppat.1011831.s001.tif (2.93 MB)
Epitope tagging of TgPP6 holoenzyme in Toxoplasma gondii.
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posted on 2023-12-13, 18:35 authored by Qin-Li Liang, Lan-Bi Nie, Hany M. Elsheikha, Ting-Ting Li, Li-Xiu Sun, Zhi-Wei Zhang, Meng Wang, Bao-Quan Fu, Xing-Quan Zhu, Jin-Lei Wang(A) Schematic showing endogenous tagging of the gene of interest (GOI) at the C-terminus, and the mAID system used to conditionally inactivate GOI, dihydrofolate reductase (DHFR) resistance cassette for selection using pyrimethamine. (B) PCR1 and PCR2 showed the correct integration of tags in the indicated strains. (C) Western blotting detected the expression of the hemagglutinin (HA)-tagged TgPP6 in RH tachyzoites. The bands of 6×HA tagged TgPP6C, TgPP6R and TgPP6S were detected at approximately 97, 121 and 45 kDa, respectively.
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six conserved motifsphosphoproteomic analysis revealedfunctional studies showeddiv >< pcell cycle progressiontgpp6c significantly reducedtgpp6c significantly attenuatedtgpp6c predominately alteredparasite lytic cycleparasite cell cycletoxoplasma gondii cell divisiontoxoplasma gondii tgpp6c contributesparasite ’parasite growthtranslational regulatorstaken togethersynchronous divisionresults showpys tachyzoitespru cystsproteins involvedprotein phosphatasesphosphorylation statuslow homologylethal dosedisordered rosettesdeficient typedaughter cellscomplex whereasbradyzoite differentiation
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