Figure 2. Recombinant protein expression, purification, and analysis of PvGAMA fragments.

(a) PvGAMA-Ecto and PvGAMA-Tr1 were synthesized using the wheat germ cell-free protein expression system and purified on Ni-Sepharose columns. The purified fragments of PvGAMA were found in the soluble elution fractions. The arrowheads indicate specific bands for each recombinant protein. T, total translation mix; S, supernatant; P, pellet; Ft, flow through; E, elution. (b) The purified PvGAMA-Ecto and PvGAMA-Tr1 were resolved by SDS-PAGE, transferred to a PVDF membrane, and probed with an anti-His-tag antibody (His), vivax-infected human sera (I), or rabbit PvGAMA-Ecto immunized sera (R). Healthy human sera (H) and non-immunized rabbit sera were used as negative controls (N). (c) Schizont lysates under reducing condition probed with PvGAMA-Ecto immune sera (R). A pre-immunized rabbit serum was used as a negative control (U). The arrowheads indicate specific bands for each recombinant protein.