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. 2022 Feb 9;298(3):101713. doi: 10.1016/j.jbc.2022.101713

Figure 1.

Figure 1

PfPGM1 is expressed throughout the parasite developmental stages.A, the integration strategy schematic for endogenous tagging of PfPGM1 at the genomic locus by single crossover homologous recombination. The primers used in cloning (1, 2) and the diagnostic PCR (3, 4) are indicated by arrows and numbered accordingly. GFP and glmS tag are represented by green and red boxes, respectively. B, diagnostic PCR with the indicated primer sets to confirm integration at the PfPGM1 genomic locus in the obtained transgenic parasites (denoted PGM-Kd). 3D7 parasites, having the unmodified genomic locus, were used as control. C, immunoblotting using antibodies against GFP confirms the expression of GFP-fusion protein in the PGM-Kd parasite line. PfActin served as a loading control. D, live cell microscopy images demonstrating the abundance and subcellular localization of the GFP-tagged PfPGM1 protein (green) in the PGM-Kd parasites. 4′,6-diamidino-2-phenylindole (DAPI) was used to stain the nuclei. The scale bar represents 2 μm. The sequences of the primers (1–4) are listed in the Table S3 (S.No. 11–14). Kd, knockdown; PGM, phosphoglycerate mutase.