FIG. 1.
SDS-PAGE analysis of PV66/AMA-1 expression in P. pastoris. (A) N-glycosidase F digestion of secreted recombinant PV66/AMA-143–487. PV66/AMA-143–487, present in the culture supernatant after methanol induction for 72 h, was digested with N-glycosidase F. Samples representing 10 μl of culture supernatant were analyzed. Lanes: 1, culture supernatant; 2, mock N-glycosidase F digestion; 3, N-glycosidase F-digested material. (B) Expression of PV66Δglyc43–487. Ten microliters of culture supernatant, at 48 h postinduction, was analyzed. Lanes: 1, PV66Δglyc43–487; 2, wild-type PV66/AMA-143–487. (C) Purified mid-scale-produced PV66Δglyc43–487. PV66Δglyc43–487 was purified by ion-exchange and gel filtration chromatographies from a mid-scale culture and chemostat fermentor supernatant. Ten micrograms of purified material was loaded on a nonreducing SDS gel. All gels were stained with Coomassie brilliant blue.