Study of PfI2-PfPP1 interaction and mapping of binding motifs in yeast two hybrid system. pGBKT7, pGBKT7-lam, pGBKT7-PfPP1 constructs were inserted into mat α (Y187) yeast and pGADT7, pGADT7-PfI2, pGADT7-PfI2(1–94), pGADT7-PfI2(19–144), pGADT7-PfI2W16A and pGADT7-PfI2Y103A constructs into mat a (AH109) yeast. These transformations were followed by mating (A). Yeast diploids were checked on SD-LW plates (B) and interactions were identified by growth on SD-LWH (C) or SD-LWHA (D). Western-blot analysis of extracts prepared from yeast diploids revealed with mAb anti-Gal4-AD (E) or mAb anti-Gal4-BD (F) showing the expected expression of the Gal4-AD tagged PfI2 proteins (PfI2WT: lanes 4,5,6; PfI2(1–94): lanes 7,8,9; PfI2(19–144): lanes 10,11,12; PfI2W16A: lanes 13,14,15 and PfI2Y103A: lanes 16,17,18) and Gal4-BD tagged laminin : lanes 2,5,8,11,14,17 or Gal4-BD tagged PfPP1: lanes 3,6,9,12,15,18.
Fréville et al. BMC Biology 2013 11:80 doi:10.1186/1741-7007-11-80