Fig. 1

C-terminally tagged PfAlba1-Ty1 was successfully expressed from an episome in P. falciparum asexual blood stages. a The transfection and maintenance of the pPfAlba1-Ty1C plasmid was confirmed by PCR of genomic DNA. Primer pairs targeted endogenous ALBA1 (p1+p2; see Additional file 11 for primer sequences), the ALBA1-TY1 locus on the episome (p3+p4 and p5+p6), and the unrelated ALBA4 genomic locus (A4F+R), in either untransfected 3D7 parasites or parasites harboring the empty vector or pPfAlba1-Ty1C. The scheme (top panel) is not drawn to scale; UTR = untranslated region. The leftmost lane (bottom panel) represents the size marker, GeneRuler 1 kb Plus DNA Ladder (Fermentas, Life Technologies). b PfAlba1-Ty1 was detected in protein lysates prepared from empty vector or PfAlba1-Ty1 parasites by denaturing gel electrophoresis and western blotting with mouse anti-Ty1 antibodies or anti-PfAlba1 antibodies. PfAldolase served as a loading control. c Immunofluorescence assays were used to determine the localization of PfAlba1-Ty1 in trophozoite (T) and schizont (S) stages of PfAlba1-Ty1 parasites. Empty vector transfectants served as a negative control. Antibodies used included mouse anti-Ty1 (green) or anti-PfAlba1 (red). Nuclei were labeled with DAPI (blue). Scale bar represents 1 μM